Search results for “Cellular”

RBM45: Molecular, Cellular, and Evolutionary Biology

This article surveys RBM45 from molecular function and RNA binding to cellular roles and evolutionary conservation. It summarizes emerging links to neurobiology and disease, and highlights tools and models to probe RBM45 biology.

Changes in Cellular Morphology in Bronchoalveolar Lavage Fluid of Children with Mycoplasma Pneumoniae Pneumonia

Objective To study changes of cell morphology in BALF in children with Mycoplasma pneumoniae pneumonia (MPP). Methods From December 2021 to May 2022, a group of 32 children diagnosed with Acute MPP and admitted for treatment in the Pediatrics Department and PICU of the First Affiliated Hospital of Xinxiang Medical University were selected for our study. These patients underwent bronchoalveolar lavage as part of their clinical assessment. For comparison, we included a control group comprising 10 children who were not infected but had bronchial foreign bodies. We investigated the cellular composition in the bronchoalveolar lavage fluid (BALF) using Wright-Giemsa staining and microscopic evaluation, aiming to understand the relationship between shifts in cell proportions and extra-pulmonary symptoms associated with MPP. Results In this study, a total of 42 cases were enrolled, with 32 cases in the study group and 10 cases in the control group. There were no statistically significant differences in gender, age, height, weight, and BMI between the two groups (p > 0.05). The study group exhibited significantly higher levels of neutrophil percentage (GRA%), CRP, D-dimer, and LDH in blood routine tests compared to the control group (p < 0.05). Furthermore, the proportions of neutrophils (%) and macrophages (%) in BALF were significantly higher in the study group compared to the control group (p< 0.05), while the proportion of lymphocytes (%) in BALF showed no statistically significant difference between the two groups (p> 0.05). Conclusion In the acute phase of MPP in children, BALF is predominantly composed of neutrophils. A lower proportion of lymphocytes in BALF is associated with a higher incidence of extra-pulmonary manifestations and longer hospitalization duration.

First Analytical Solution of Intracellular life Cycle of SARS-CoV-2

Role of Tie2, CD14, Angiopoietin as Angiogenetic Markers in Hepatocellular Carcinoma Complicating Hepatitis C Virus Infection

Background Identifying biomarkers for early detection of hepatocellular carcinoma (HCC) remains quite challenging. In this study we aimed to estimate the number of TIE2-expressing monocytes (TEMs) cells, which display pro-tumoral activities and are defined as CD14+, TIE2+, and angiopoietin-2; and its potential use as a possible diagnostic marker in HCC patients complicating HCV induced cirrhosis. Methods Current study was conducted on 112 patients. They were divided into two groups: Group I (78 patients) with HCC complicating HCV induced cirrhosis; and group II chronic hepatitis C patients (34 patients). Both groups were compared to (age and sex-matched) healthy persons as group III (38 persons). Result The number of the circulating TEMs: CD14+ and TIE2+ monocytes were significantly higher in the peripheral blood of HCC patients than HCV LC patients and healthy controls, sensitivity and specificity for HCC diagnosis were respectively: CD14 (89.7%, 83.3%), TIE 2 (76.9%, 83.3%), and Ang-2 (76.9%, 66.7%). Moreover, analysis of the P-value and the odd’s ratio showed that CD14 has the highest predictive value for HCC. Conclusion Our results suggest that TEMs and Ang-2can be used as diagnostic markers for HCC, especially among the high-risk group of patients.

Developing Cellular & Molecular Biomarkers for Anti-Inflammatory Activities of Probiotic Bacteria in Fermented Foods

We present below a mechanistic cellular and molecular approaches for the development of Anti-Inflammatory biomarkersof Probiotic Bacteria in Fermented Foods. Probiotics are live microorganisms that promote human health by counteracting the noxious toxic gut microflora in human intestine, by modulating of the tight junctions, and by increasing mucin production, enforcing intestinal epithelial cell barrier function, modifying microbial community within the gut intestinal disorders, and improving immune responses associated with chronic inflammation in experimental animal models, collectively enhancing human health. Cytokine secretion by intestinal epithelial cells and macrophages are regulated by probiotics through key signaling pathways such as nuclear factor-κB and mitogen-activated kinases, resulting in alleviation of several disorders such as allergies, diabetes, obesity, heart diseases and cancer. MicroRNAs are small non-coding RNA molecules involved in transcriptional and post-translational regulation of gene expression by inhibiting gene translation. Using in vitro and in vivo approaches in cell lines and mice models to study effects of probiotic conditional media and heat-killed bacterial strains with anti-inflammatory effect to elucidate the mechanisms by which probiotics affect signaling pathways, and by using global cytokine and microRNA gene expression analyses approaches to develop biomarkers for studying different pro- and anti-inflammatory activities, and using statistical approaches to analyse the data, we show that cytokines and miRNAs have an essential role in regulation of cancerous and inflammatory pathways. This mechanistic approach will result in developing specific disease biomarkers for the early diagnosis of certain pathogenic states, as well as evaluating the effect of different dietary components on developed biomarkers in health states that will promote and enhance human health. Comparing the concordance of the in vitro to the in vivo research findings will confirm the correspondence of both approaches to each other. Moreover, this study will have a major public health relevance in elucidating the role of miRNAs and their targets in inflammation, paving the way to diagnosing and treating of pathogenic human disease stages.

The Biological Basis of Cellular Diabetes Mellitus

This letter presents a clinician's perspective on the biological basis of diabetes mellitus at the cellular level. Drawing on neurosurgical practice, the author argues that persistent dysglycemia hinders recovery and may reflect long-standing intracellular processes, calling for causal, interdisciplinary management beyond symptomatic care. The piece outlines testable hypotheses and invites further laboratory validation.

Cellular Mechanics and Therapeutic Resistance of the Cancer Relapse

The aims of this study are to investigate the variation in the mechanical behaviour of the primary cancer from cancer relapse, and measuring the therapeutic resistance acquired by cancer relapse. A431-cultured cells were irradiated for 7 months until 85 Gy. Then, a selected single cell was left to grow as stable A431-R cell line. 106 cells of A431 cells and 106 of A431-R cells suspended in 100 μL of medium were injected into subcutaneous tissues on the right thigh of athymic mice to generate tumor xenografts models of primary cancer (A431-P) and cancer relapse (A431-R). Radiotherapy of a low-dose of 30Gy was applied on xenoimplanted tumors after one week from inoculation. A mock process was performed on untreated groups of mice for controls. Tumor size was monitored starting from inoculation and tumor growth was measured along 42 days. Rates of mitosis and apoptosis and the histologic grade (HG) that characterize the tumor response were determined as described in earlier studies. Alterations induced on tumor HG in the treated models were 100% identical to the energy of the applied doses. The differences in response energy between cancer relapse and primary cancer irrespectively of the treatment (untreated vs. treated) or origin of the cells (A431-P vs. A431-R) in all phases of tumor responses (growth, shrinkage or regrowth) were 100% identical to the total differences in the administered regimens applied on those groups during those phases. Cancer relapse is characterized by a delay in growth before second line therapy for its relatively lower rate of mitosis compared by the primary cancer inducing a corresponding delay in the early detection. The therapeutic resistance of the cancer relapse is equivalent to the energy of the doses which have been delivered in the prior therapies, and requires increasing the administered dose by an amount equivalent to that resistance.

Tumor Growth Dynamics: Dietary Fish Oil Induced Inhibition of Human Breast Carcinoma Growth, A Phenomenon of Reduced Cellular DNA Synthesis or Increased Cell Loss?

Diets high in unsaturated fatty acids, especially those containing high levels of linoleic acid, e.g., corn oil, enhance mammary gland tumorigenesis in experimental animals. In contrast, diets high in long-chain polyunsaturated fatty acids such as eicosapentaenoic (EPA) and docosahexaenoic (DHA), e.g. menhaden oil, appear to have a suppressive effect on this tumorigenic process. Many mechanisms have been proposed to explain the tumor inhibitory action exerted by menhaden oil and other fish oils, e.g., differences in prostaglandin metabolism, energy efficiency, alterations of the immune system, changes in lipid peroxidation, etc. Fundamental to a mechanistic understanding of this phenomenon, however, is an understanding as to whether or not the tumor inhibitory activities of dietary fish oil is mediated via an inhibition of tumor cell proliferation or mediated via an enhancement of tumor cell loss. Whether the amount of dietary fat or the type of fat effects mammary tumorigenic processes, via an effect on tumor cell proliferation or tumor cell loss, has not been clearly established. In the studies described in this communication, three methods were utilized to study tumor cell proliferation, i.e., H3-thymidine autoradiographic analysis, 5-bromo 2'-deoxyuridine (Brdu) flow cytometric analysis, and proliferative cell nuclear antigen (PCNA) flow cytometric analysis. Two methods were used to study tumor cell loss, i.e., a determination of the I125Urd tumor emission rate and a determination of a cell loss factor from the formulas of Steel and Begg. The tumor examined was the human breast carcinoma cell line MDA- MB231 maintained in athymic nude mouse. No significant difference in cell proliferation between carcinomas of mice fed a high corn oil diet (20% w/w) and a diet high in fish oil (19% menhaden oil, 1% corn oil). In contrast, a significant (p<0.05) increase in the rate of I125Urd emission rate and cell loss factor from the carcinomas in the fish oil fed mice compared to the corn oil fed mice was observed. In summary, the decreased tumor volume in the human breast carcinomas maintained in athymic nude mice fed a fish oil diet as compared to those fed a corn oil diet, appears to be due, at least in part, to an increased rate of carcinoma cell loss rather than a decreased rate of carcinoma cell proliferation.

Central Retinal Vein Occlusion in Hepatocellular Carcinoma

A 66 year old Chinese male with a medical history of hypertension, diabetes mellitus and hepatitis B carrier was diagnosed with hepatocellular carcinoma in 2009. He underwent treatment with selective internal radiation spheres and sorafenib, and multiple cycles of chemotherapeutic agents such as bevacizumab, erlotinib, OXAFI ( intravenous oxaliplatin and doxorubicin given on days 1, 8 and 15 in a 28-day cycle, a daily continuous infusion of fluorouracil and subcutaneous interferon alfa-2b 5 million units administered thrice weekly), thalidomide, capecitabine, and rapamycin over the course of four years. Along the course of treatment, he developed pulmonary embolism and was initially started on anti-coagulation. Two months later, he developed hemoptysis and the anti-coagulants were stopped. During his routine ophthalmology visit for diabetic eye evaluation, he complained of blurring of vision of his left eye for the past four to five weeks. He was found to have central retinal vein occlusion (CRVO) of the left eye, associated with macular edema. Visual acuity was 6/15 for the right eye and 6/60 for the left eye. Eyelids, conjunctiva, cornea, anterior chamber, pupils, lens and ocular motility were normal. Humphrey visual field testing showed a superior arcuate and basal defect. This is the first reported case of CRVO in hepatocellular carcinoma. The etiology of CRVO is multifactorial, withhepatic malignancy, previous major surgery, multiple cycles of chemotherapy and cessation of anticoagulant therapyas possible aetiological factors. His background medical problems of diabetes and hypertension are further contributors.

High-Throughput Complex Disease Modeling for Ethical Drug Discovery: Clinical Relevance of a NAM Platform for Cancer Biomarker Development

The development of tumor biomarkers derived from blood, or its components, has become pivotal in advancing early cancer diagnosis. Malignant transformations induce cancer-specific alterations in the transcriptome, proteome, and secretome of tumor cells. Recent studies highlighted similar alterations in peripheral blood mononuclear cells (PBMCs) in cancer patients, which appear to mirror the state of transformation in tumor cells. These findings suggest an intercellular communication–driven mechanism rather than a systemic inflammatory response and, in addition to current ctDNA-based liquid biopsy biomarkers, point to a novel, simple, and highly robust approach for the early detection of cancer. Using this phenomenon to advance PBMC-based biomarker development, it will be essential to achieve 3D in vitro tumor models that reproduce a highly physiological tumor microenvironment (TME). Likewise, more enhanced 3D ex vivo models are required to enable the replication of cell-to-cell and organ-to-organ communication. These systems will guide the self-organization of mixed microenvironments derived from different tissues and enable them to accurately reproduce the molecular connections underlying these alterations. In this study, an innovative new modular 3D co-culturing approach was used to expose PBMCs to lung tumoroids, under physiologically relevant conditions. Changes in DNA fragmentation of PBMCs in the presence of lung cancer were quantified and used as a biomarker. To validate the predictiveness of this biomarker, our results were compared with clinical data from a clinical evaluation study. Similar to the clinical trial observations, PBMCs, when exposed to lung tumoroids, showed a significantly lower level of DNA fragmentation (37%). This modular 3D co-culturing model showed a predictiveness of the clinical data of > 90%, demonstrating its power to monitoring cell-to-cell communication effects and support the development of blood-based biomarkers.

Modulation of Summer Squash Growth and Productivity Via Spiritual Blessings (Biofield) Energy Treatment (SBET)

Background The increasing demand for sustainable and eco-friendly agricultural practices has led to the exploration of non-traditional methods to enhance crop yield and resilience. Spiritual Blessings (Biofield) Energy Treatment (SBET), a form of consciousness-driven energy healing, is increasingly being investigated for its potential to modulate biological systems at the cellular and molecular levels without use of chemical additives Objective This study aimed to evaluate the impact of SBET on the growth characteristics and overall productivity of summer squash (Cucurbita pepo L.). Methods The study was conducted using a controlled experimental design, where seeds and plots were divided into two groups: control and treated. The treated group received a remote SBET by a recognized practitioner, while the control group remained untreated. Both groups were maintained under identical environmental conditions (soil, water). Parameters such as germination rate, plant height, leaf area index, and total fruit yield were monitored over a full growth cycle. Results Results showed that plant height, number of branches, and total number of leaves per plant were significantly improved by 35.14% (p ≤ 0.001), 41.64% (p = 0.011), and 49.01% (p = 0.029), respectively, in the treatment group compared to the control group. Additionally, fruit length and total fruit yield (tons per hectare) were significantly increased by 39.68% (p = 0.002) and 15.92%, respectively, in the treatment group compared to the control group. Conclusion Exposure of SBET significantly improved both vegetative and reproductive development, yielding substantial increases in plant height, branching, and leaf production.

Diabetic Mouse Delayed Wound Model Following Treatment with the NerveStim™ Neuropathy System

Delayed wound healing in diabetes is characterized by impaired angiogenesis, persistent inflammation, extracellular matrix dysregulation, and peripheral neuropathy. A preclinical study was conducted using a diabetic mouse delayed wound model to evaluate the surrounding tissue of a wound, (its periwound) and its tissue responses following treatment with the NerveStim™ Neuropathy System, a combination topical gel and neuromuscular electrical stimulation platform. Periwound tissue was harvested at Day 14 and analyzed using NanoString gene expression profiling. Treated animals demonstrated visibly increased periwound tissue thickness compared to untreated controls. Differential expression analysis identified 76 significantly upregulated and 17 downregulated genes. Upregulated pathways included angiogenesis (Vegfa, Fgf2, Pdgfb, Nos3), neurotrophic signaling (Ngf, Bdnf, Scn9a, Trpv1), macrophage polarization (Arg1, Mrc1, Il10), and extracellular matrix remodeling (Col1a1, Col3a1, Mmp9, Timp1). Downregulation of select pro-inflammatory mediators (Nos2, Mif) was observed. These coordinated transcriptional changes are consistent with activation of reparative immune, neurovascular, and matrix remodeling pathways in diabetic periwound tissue.

Activation of Transcription Factor EB Protects Against Proinflammatory Insults Through NF-κB Inhibition in Keratinocytes

Keratinocytes are pivotal in mediating cutaneous inflammation. Identifying anti-inflammatory factors within these cells holds promise for developing novel therapeutic strategies to manage skin inflammation. Transcription factor EB (TFEB) has recently emerged as a key regulator linking cellular energy metabolism to inflammatory processes, primarily through its influence on autophagy and NF-κB signaling. However, whether TFEB activation exerts anti-inflammatory effects in keratinocytes remains unclear. In vitro inflammation model was established in HaCat cells by incubation with proinflammatory mediators LPS and IL-1β. Cell viability and TFEB expression and phosphorylation were measured. The effect of TFEB activation by C1 and adenoviral TFEB overexpression on the expression of proinflammatory genes including COX-2, MCP-1 and IL-6 were detected. Also, IκBα protein level were determined. TFEB phosphorylation is increased while TFEB total protein expression is inhibited by treatment with LPS and IL-1β. Pharmacological activation of TFEB by compound C1 and TFEB overexpression suppressed the expression of COX-2, MCP-1 and TNF-α induced by LPS and IL-1β. TFEB overexpression increased basal IκBα expression and restored IκBα level under LPS treatment. TFEB knockdown reduced TFEB expression and lowered basal expression level of COX-2, MCP-1 and TNF-α. Our findings indicate that TFEB activation can mitigate inflammatory gene expression in keratinocytes triggered by LPS and IL-1β. This implicates TFEB as a significant novel modulator of cutaneous inflammation, highlighting its potential as a therapeutic target. Targeting TFEB could thus be a viable strategy for developing new treatments for chronic inflammatory skin conditions.

The Dose Modulates the Body’s Physiology: Literature Review

The body interacts with endogenous and exogenous molecules through various receptor networks at the cellular and organismal levels by which the different physiological processes of the organ systems get activated. Life could no longer exist without the body’s interaction with these signaling molecules. The number of molecules interacted with a receptor type within the body determines the efficiency of biological processes that would determine the biochemistry and anatomy behind the basic body functions. The dose plays a complex role in multiple physiological processes by modulating the natural processes of the different biological systems. It is always connected to the physiological and anatomical aspects of the biological sciences in which it manifests the biological sensitivity that would determine the efficiency of biological responses of the body systems. There are always physiologic and non-physiologic doses for every compound administered into the biological systems. The does that facilitated the body to manifest that biological sensitivity which has maintained the physiological processes of the body systems was noted as physiologic dose, whereas the dose that has suppressed the biological sensitivity of the body that became inefficient in maintaining the physiological processes of the different biological systems was termed non-physiologic dose. It is important to adjust the dose or dosage based on physical and biological factors such as the efficiency of the different organ systems, body weight, and the timing of the dose triggering a biological response to maintain the natural processes of the body systems. This means that integrated biological data is required in order to be able to identify the physiologic and non physiologic doses for biological or pharmacological use.

The Effects of L Carnitine on in Vitro Maturation of Immature Bovine Oocytes

L-Carnitine (Lc) acts as an antioxidant that neutralizes free radicals, especially superoxide anions and protects cells against oxidative damage-induced apoptosis, as following ovulation, intracellular reactive oxygen species (ROS) accumulation increases in oocytes, Oocytes exhibit an intracellular defense mechanism against an oxidative attack. This outcome adversely affects fertilization and subsequent embryonic development, thereby increasing the risk of an early miscarriage and abnormal development of offspring. The purpose of this study was to see how adding LC to either maturation or fertilization medium affected the developmental competence of immature bovine oocytes. In this study, Ovaries from apparently normal reproductive organs of cattle were collected within 30 minutes from slaughter and evisceration of animals. Cumulus oocyte complexes (COCs) were collected by aspiration of medium sized ovarian follicles (4-8 mm). COCs of acceptable quality were selected, washed and incubated in tissue culture media 199 (TCM199) supplemented with 10% heat inactivated fetal calf serum, 5 μg/ml luteinizing hormone (LH), 0.5 μg/ml follicle stimulating hormone (FSH) and 1 μg/ml estradiol-17β for 20:22 hour at 38.5 C◦ under 5% CO2 in air with 90% humidity. different concentrations of LC (1.25,2.5 and 5mM) were used. The results were consistent for both maturation and fertilization and there is a significant increase in maturation, fertilization., cleavage and blastocyst rate. In conclusion, LC has important role in IVEP through addition of LC to maturation media or culture media it improved nuclear maturation and blastocyst formation rates in bovine oocytes.

Molecular Control of Human Embryonic Development: A Comprehensive Study

Human embryonic development is a highly coordinated and complex process that transforms a single fertilized cell into a fully formed human organism. This process is governed by intricate molecular mechanisms involving genetic regulation, signal transduction pathways, and intercellular communication. This study explores key molecular pathways controlling human embryonic development, focusing on the roles of morphogens, transcription factors, signaling molecules, and epigenetic modifications. By reviewing the most recent literature and experimental studies, we aim to highlight the molecular orchestration that directs cell fate decisions, tissue patterning, and organogenesis in humans.

Traumatic Maculopathy (Berlin´S Edema) Treated with Enhancement of Human Photosynthesis®

Commotio retinae (CR) is a condition frequently observed in clinical practice, particularly following closed globe trauma (CGT) due to sport, labor, or traffic accident injuries. It is the main cause of unilateral vision loss in male patients aged between. It is characterized by transient gray-white retinal coloration and reduction of visual acuity (VA). Symptoms depend mainly on the location and severity (deep) of the injury, with less complains when only the superficial or peripheral retina is affected. It may be confined to the posterior pole, when it is also called Berlin’s edema, after the first hypothesis of Berlin (1873). There is no specific treatment since the treatment depends on the region of the retina and choroid affected. In this work we report a case of Berlin edema, treated with ǪIAPI 1®, to restore the balance of oxygen, which is generated at the intracellular level.

REDOX ENZYMES: From Basics to Free Radical Scavenging

Redox enzymes are a type of enzyme that catalyzes redox reactions, that is, electron transfer reactions between two chemical species. Redox enzymes are essential for many biological processes, including cellular respiration, photosynthesis, energy production, and the elimination of free radicals. They are divided into two main types: oxidoreductases and electron transferases. Oxidoreductases catalyze the direct transfer of electrons between two chemical species, while electron transferases catalyze electron transfer by cofactors. Examples of redox enzymes include cytochrome c oxidase, NADH dehydrogenase, succinate dehydrogenase, and catalase. Each of these enzymes play an important role in cellular metabolism and organism homeostasis.

Malaria: A Driving Force to the Emergence and the Global Spread of Antibiotics Resistance

Malaria and bacteraemia are significant public health concerns and economic threats. In Africa, the intensity for simultaneous transmission and co-infection of Plasmodium spp and other bacteria pathogens are extremely high. It is believed that malaria suppress the immune system and enable the translocation of bacteria in the gastrointestinal tract to other cellular compartments in the body. Some of the factors that contributed to the co-emergence of these pathogens are poor access to clean water, sanitation and hygiene (WASH), poor infection control measures, inefficient health care systems. In addition, the similarities in the clinical signs and symptoms of these febrile diseases and the fact that the etiologic diagnostic testing can be complex, costly, and limited are the reasons why clinicians in resource-constrained setting often prescribe antibiotics empirically prior to or without laboratory testing to prevent severe outcomes in any patient hospitalized with malaria. However, this indiscriminate use of antibiotics has been identified as the driving force for antibiotic resistance, which is already at alarming rate in malaria endemic nations. In developed countries where malaria had been previously eradicated, there are increasing reports of imported malaria with concurrent bacteraemia. In this review, we emphasized the role of malaria in the indiscriminate use of antibiotics and the fact that eliminating malaria in Africa is one of the best strategies to address the emergence and the global spread of multi-drug resistance organisms.

Creation of Music-Induced Analgesia in Chronic Pain Patients through Endogenous Opioid Production: A Narrative Review

Chronic pain affects over 30% of the global population, and reliance on external drugs for treatment has led to major issues, including the present opioid epidemic. A healthier option is necessary, which is why music therapy’s analgesic effects have been extensively studied within the last 20 years. Not only is music relatively harmless but given that chronic pain patients require repeated treatment, musical intervention is far more accessible and economical. While the mechanisms underlying music-induced analgesia are relatively unclear, the production of endogenous opioids while listening to music through both the descending pain modulatory circuit and the limbic system, is postulated to play this role. This review describes the brain regions and pathways by which music may trigger the release of endogenous opioids such as enkephalins, endorphins, and dynorphins. More importantly, it discusses the cellular mechanisms through which these neuropeptides are thought to mediate pleasure-induced analgesia in chronic pain patients.

RETRACTED: Leap into Fetal Surgery; In Utero Placental Mesenchymal Stem Cell Therapy, A Contemporary Approach to Treating Myelomeningocele

This article has been retracted on April 10, 2025. VIEW THE RETRACTION NOTICE (https://doi.org/10.14302/issn.2997-2086.jfs-25-5857) Myelomeningocele (MMC), a class of spina bifida is a type of neural tube defect. According to the U.S. Centers for Disease Control and Prevention, each year approximately 1,400 babies born in the United States have spina bifida. The disease manifests with the lack of skin and bone covering the caudal part of the spinal cord. The patient developing such a condition often develops lifelong impaired lower limb mobility accompanied by hydrocephalus, and urinary and bowel incontinence. The available interventions include prenatal and postnatal surgery to fuse the dura. Prenatal surgery performed before 26 weeks of gestation reduces the risk of death or the need for ventriculoperitoneal shunting. It also enhanced results on a comprehensive index for mental and motor function. When compared to postnatal surgery, prenatal surgery reduces the manifestation of several secondary outcomes, including the degree of hindbrain herniation seen in the Chiari II malformation. Stem cell therapy for MMC on animal models of chick, ovine, and rodents with reported cases 15/63, 15, and 136, respectively, using human Embryonic Stem Cells (hESCs), Neural Stem Cells (NSCs), Mesenchymal Stem Cells (MSCs) showed significant coverage of MMC defect and slight neurogenesis was also observed. With an understanding of medical literature about in-utero regenerative capacity, it is to be appreciated that placental stem cells surgically seeded within a biocompatible scaffold of the cell patches can play a part in alleviating the spinal cord manifestation associated with MMC. Documented animal studies show that incorporating Placental Mesenchymal Stem Cells in prenatal surgery has reported improved neurogenesis and lower limb mobility. In an ovine myelomeningocele model, the development of in-utero myelomeningocele repair with human Placental Mesenchymal Stem Cells seeded onto an extracellular matrix (PMSC-ECM) enhances motor findings. The clinical trial for the first stem cell therapy on human subjects known as the “CuRe Trial: Cellular Therapy for In Utero Repair of Myelomeningocele.” is expected to be finished by 2030. So far, the cases undergoing treatment have shown significant leg movement and a greater degree of bowel and urinary control. This FDA-approved clinical trial is envisioned to be the future of treating MMC.

Rbm45 Phylogenetics, Protein Domain Conservation, and Gene Architecture in Clade Metazoa

Mammalian Rbm45 is predominately expressed in neuronal tissue and is integral in brain development and neuronal differentiation under physiological conditions. Dysregulation of Rbm45 has been strongly associated with neurodegenerative disorders in humans and can drive hepatocellular carcinoma through reprogramming lipid metabolism. Intriguingly, Rbm45 is an ancient protein, evolutionarily conserved throughout metazoans, including in sponges which lack a nervous system. Curiously, the evolution of Rbm45 gene structure and protein domain conservation across kingdom Animalia is largely unknown. We performed phylogenetic analysis of Rbm45 nucleotide and amino acid sequences from 36 species representing 9 phyla: Porifera, Cnidaria, Priapulida, Mollusca, Brachiopoda, Arthropoda, Echinodermata, Hemichordata, and Chordata. While the tree from Rbm45 nucleotide sequence data resulted in clades Protostomia and Deuterostomia showing paraphyly, the phylogeny derived from Rbm45 amino acid sequence largely recapitulated known monophyletic relationships among metazoans. Human RBM45 protein structure includes three RNA-binding domains (RBD), a homo-oligomerization association (HOA) domain, a nuclear localization sequence (NLS), and a nuclear export sequence (NES). Multiple sequence alignment across the same 36 taxa used for phylogenetic analysis revealed conservation of all three RBDs, the HOA, and NLS; in contrast the NES was only detected in clade Craniata and not in clades Ambulacraria and Protostomia. Rbm45 gene structure analysis revealed increasing gene complexity concomitant with increasing evolutionary complexity. Rbm45 from non-bilaterian taxa had from 2 to 4 large exons, while bilaterian taxa had between 6 to17 small exons. These findings demonstrate that Rbm45 is an ancient, highly conserved gene among metazoans suggesting a function in a breadth of neural/sensory systems.

The trajectory of immunoglobulins immune response against the different amounts of xenobiotics matches the trajectory of biological changes associated with ageing: A systematic review

Background Ageing is a life process in which progressive molecular, cellular, physiological and anatomical changes manifesting in humans and animals including other organisms lead to the decline of biological functions. Immunoglobulins (Igs) are glycoprotein molecules produced by white blood cells mainly B lymphocytes following signal transduction as a result of their interaction with pathogenic microbes or poisonous substances introduced into the body systems. They elicit responses against the side effects of pathogens and poisons in which their response efficiency usually declines as we are ageing. Objective Thus, the similarities between Igs’ immune response against the different amounts of xenobiotics and the biological changes associated with ageing have been systematically assessed using the reports of different study results on humans and animals. Methods First, a literature search was carried out in google, PubMed and google scholar using planned search terms related to the title of this study. Review and original articles were retrieved, downloaded and saved on a computer. And then the effects of different factors i.e. xenobiotics, age, sex and lifestyle-based practices on the levels of serum Igs (IgG, IgA and IgM) in animals and humans have been studied using a systematic review of different literature sources. Finally, the relationship between the findings of various studies has been assessed and judgment on the possible cause of ageing has been made. Results The findings of different research have demonstrated that the signaling efficiency of immunoglobulin M (IgM) has been limited by the amount of test compounds administered to study Balb c mice in the oral route. The response efficiency of IgM immune response against the lower doses of test compounds were high compared to the higher doses of test compounds which was low. The results of different other studies also demonstrated that the decline of serum IgM levels was associated with ageing. The relationship between alcohol consumption and the concentration of serum Igs was also described in the report of different studies. These studies have shown that there was lower level of IgG in the blood serum of alcohol consumers compared to non-consumers. The study has also demonstrated a lower level of serum IgM with higher alcohol consumption and higher serum concentration with moderate beer consumption. Conclusion The trajectory of Igs’ immune response against different amounts of xenobiotics was highly associated with the trajectory of biological changes during ageing. These research findings might be the possible evidence to conclude that ageing is caused by the foodstuffs and non-foodstuffs we usually consume, the lifestyles we usually experience and the way of life we usually live in the environment which gradually defiling the natural processes of the body.

Histo-Morphological Effect of The Small, Large Intestines and Stomach of Animal Models Treated With Aqueous Extract of Abelmoschus Esculentus

This study investigates the effect of Aqueous extract of abelmoschus esculentus on the microanatomy of the small, large intestine and stomach and the body weight of Wister rats. Twenty-one adults male wistar rats weighing between 100-120 grams were assigned into three groups consisting of seven rats each; Group A (control), Group B (low dose), and Group C (high dose). The rats in the control group were fed with fed with feed and water only while the rats in groups B and C were treated with 0.1mg/kg body weight and 3.0mg/kg body weight of abelmoschus esculentus respectively for 14 days. At the end of administration, the final weights of all rats were recorded before sacrifice using cervical dislocation and the small, large intestine and the stomach were harvested, processed and stained using H&E stain. The results were revealed as significant (p<0.05) increased in the mean body weight compared with the weight in the control groups and experimental groups. The treated animal groups revealed increased cellularity, focal metaplasia of the mucosal cells with villous disruption in the small intestine and dysplasia of the mucosal with loss of epithelial shape in large intestine. The stomach histology showed gastric pits with goblet cells smooth muscles layer and surface epithelium in the control group. Sections from the low dose treated group showed deep epithelical gastric pit areas with marked depletion of pits and goblet cells while the high dose treated group revealed dysplasia of gastric pits, goblet cells and smooth muscles appear mildly eroded.

Biological Effects of High Radiofrequency Radiation on Wistar Rats: A Literature Review

Aim and Objective Despite the growing concerns about the relationship between exposure to radiofrequency radiation (RFR) and detrimental health effects due to the changes in biological processes of experimental animals, there is still ongoing debate on the significance of these findings in causing significant public health problems with the growing advancement in internet technology. The aim of this study is to review existing literature on the effects of high RFR on wistar rats. Method A search was conducted on Google scholar and PubMed to identify relevant peer-reviewed articles to be included into the review. Studies eligible for inclusion included free full text articles on wistar rats exposed to ≥ 2.45GHz RFR conducted in the past 5 years. Studies included in this review were written or transcribed in English language. From 286 titles, 36 eligible studies were included in the review and assessed for quality using the Strengthening the Report of Observational Studies in Epidemiology – Veterinary Extension (STROBE-Vet) quality assessment tool. Results Studies included in this review generally had good quality (>60%) based on the STROBE-Vet assessment. This review identified numerous biological changes in wistar rats exposed to high RFR including variations in biochemical, cholinergic, genetic, histopathologic, psychological, optical, and dermatological parameters. In this review, studies identified variations in protein and liver enzymes while high RFR was found to induce oxidative stress and cellular damage of exposed wistar rats compared to the unexposed groups. This was seen in the changes in protein, lipids, enzymatic and non-enzymatic antioxidants. Studies also identified changes in expression of genes and neurotransmitters with imbalance in hormones. In addition, this review identified structural changes of cells, tissues and organs indicative of apoptosis, damage and death. Exposed rats were identified to have behavioral changes indicative of anxiety and memory decline while studies identified optical and dermatologic changes in exposed rats compared to the unexposed. Conclusion With numerous biological changes identified in wistar rats exposed to high RFR, there is an increasing risk of detrimental health events giving the advancement in internet technology and limited regulations to control exposures to RFR. Therefore, studies should be conducted to identify strategies to mitigate human exposure to RFR while policies are developed and enforced to protect human health.

Computational Systemic Biology for Toxicity Studies: A Mini Review of Previously Published Articles

The strategy for safe drug discovery and development has limited clinical success as compared to wasted time and resources annually. This is due to the fact that the results of multiphase preclinical trials are less likely to make an accurate early prediction on the safety of test compounds to progress into the clinic as a valuable therapeutic agent. A lot of time and resources has been wasted in the multistage processes of drug discovery and development that does not work at the end of the procedure every year. During pre-marketing stage, for instance, the number of unsuccessful clinical trials are greater than the successful one because of safety issues. A toxicity study at different stages of preclinical and clinical trials is a routine procedure to investigate the undesirable side effects of test compounds being manifested on the natural processes of living things. It deals with the effect and mechanism of toxicity of test compounds that triggers different biological responses on different organ systems. The biological responses that would be manifested as a result of interaction between the receptors and active molecules of a test compound could be desirable pharmacological effect or undesirable side effect or both responses are manifested simultaneously depending on the selectivity or specificity of the molecule of a test compound for its receptor subtype which makes safe drug discovery and development very challenging. The response efficiency of the body (the net outcome of the body’s biological reaction against the side effect) would determine the potency of a test compound to manifest undesirable pharmacologic effect. In other words, the amount of a drug required to cause a biological harm or injury depends on the magnitude of the body’s biological reaction in which the immune response plays a great pharmacological role by neutralizing and harmonizing xenobiotics with the biological molecules. The dose of a test compound at 100 mg/kg body weight, for instance, could be lethal to some of the study animals while it is still non-lethal to some other study animals depending on the response efficiency of the body. The immune system is well connected to each and every biological systems of the body which allows it to detect undesirable side effects being manifested through immunoglobulins signalling and activation mechanisms. This complex communication network helps to localize the diverse side effects of a test compound being manifested on different organ systems into the immune system which makes a toxicity study relatively simple to monitor. The cellular immune system becomes active following the molecule-receptor interaction and start producing antibodies which is also known as immunoglobulins to protect bodily harm and destruction. Under normal biological circumstances, the amount of immunoglobulins produced by the cellular immune system following exposure to a test compound is proportional to the number of harmful molecules interacted with its receptor subtype. Thus, with the reference to the changes in the immune response against the administered dose, it would be able to deal with the diverse undesirable side effects of a test compound being manifested on treated study animals using computational systemic biology.

A Cancer Theory: The Central Nervous System’s Adaptive Changes Make Chronic Diseases Incurable

We examined special roles of the Central Nervous System (CNS) in an attempt to resolve the puzzle that chronic diseases cannot be cured in medicine. By exploring a skill-learning model, we found that the CNS is able to remember certain information reflecting biochemical and cellular (B&C) processes in the body. From the skill-using ability, we found that the CNS is able to control basic B&C processes that drive and power the skill. From the ability to adjust forces and moving direction of body parts, we infer that the CNS is able to adjust B&C processes that control physical acts. From this controlling capability, we inferred that the CNS must also store certain information on the baseline B&C processes, is able to up-regulate or down-regulate the B&C processes, and make comparisons in performing its regulatory functions. We found that chronic diseases are the results of deviated baseline B&C processes, the CNS plays a role in maintaining deviated baseline B&C processes, and protects the body state of a fully developed disease. The three CNS roles can explain that cancer progresses with increasing malignancy, cancer quickly returns after a surgery, cancer cells repopulate after chemotherapy and radiotherapy, cancer patients develop drug resistance inevitably, immune cells rebound after suppression, etc. We further showed that long-term exercises generally can correct part of the departures in B&C processes and thus help to reverse chronic diseases. Finally, we propose strategies for resetting the CNS’ state memory as an essential condition for curing chronic diseases and cancer.

Association of Serum Calcium Level with other Risk Factors of Ischaemic Stroke

Introduction Calcium (Ca2+) plays an important role in the pathogenesis of ischemic cell damage. Intracellular Ca2+ accumulation leads to neuronal damage by triggering the cycle of cytotoxic events, however the relationship of serum Ca levels and the pathways involved in ischemic injury is unclear. Aim of Study To investigate the relationship of serum Ca2+ levels with severity of acute ischaemic stroke, serum calcium (Ca2+) levels were measured within the first 48 hours and were compared with the clinical severity of acute ischaemic stroke. Material and Methods A hospital based cross sectional study was performed among 100 patients of acute ischaemic stroke who fulfilled the inclusion criteria. The Study was done from July 2020 to August 2021 in SPRC & Neurology Hospital Dhaka, Bangladesh And BSMMU Hospital Dhaka, Bangladesh. After hospitalization presenting complaints, physical findings of the patients were recorded. Severity of stroke was measured by NIHSS scale. Serum calcium level of every patient was measured. Calcium level was divided into 3 groups by weighted average. Statistical analysis was carried out by a non-parametric Ruska Wallis test. Results Among the 100 patients 59% were male. Among all patients 57% of patients were found to be smokers (98% male, 2% female). Among all patients 63% patients were found hypertensive and 21% of all patients (24% male, 17% female) were diabetic. Mean cholesterol level was 257.98mg/dl with standard deviation 55.49 which is above the reference range suggesting hypercholesterolemia, Triglyceride was borderline and LDL cholesterol was slightly higher and HDL cholesterol was slightly lower. Calcium level was divided into 3 groups and NIHSS score was calculated for every patient in each group. The median NIHSS score for group1 (calcium level ≤8.8 mg/dl) was 9(2-20), for group 2 (calcium level 8.9-9.6 mg/dl) was 6 (1-17) and for group 3 (calcium level ≥9.7mg/dl) was 4 (1-16). Conclusion Commonest risk factor of ischaemic stroke is hypertension. Other risk factors are smoking, diabetes mellitus and hyperlipidemia, cardiac disease. Higher serum calcium level is associated with less severity of ischaemic stroke.

Evolution of the Solid Human Tumor Cells Properties in Various Experimental Systems in Vitro

Background Human malignant cell models which reflect the structural and physiological complexity of tumor tissue are of great importance for preclinical research in oncology. Spheroids/tumoroids derived from solid tumors are of great interest as cellular models mimicking the first vascular-free growth phase of a tumor node. The fact of the identity between artificially created tumor multicellular aggregates and the real tumor tissue, however, needs to be specified, described and validated in order to see how closely the spheroids are biologically similar to the malignized tissues in vivo compared to the monolayer cell cultures traditionally used. We present here a comparison study of the characteristics of solid tumor cells of different histogenesis (melanomas, soft tissue sarcomas and bone sarcomas, epithelial tumors) cultured in two dimensions (monolayer culture) and three dimensional space (spheroid), namely: spatial organization, multiplication, metabolic activity. Patients and Methods For the creation of 2 D and 3D cell models the cells isolated from the patient's solid tumor fragments obtained intraoperatively were used. 15 samples of skin melanoma, 20 samples of soft tissue and osteogenic sarcomas (STBS), and 9 samples of epithelial tumors (ET). The tumor cells were all cultivated for at least 10 passages. We used phase contrast, confocal microscopy, and immunohistochemistry to investigate spheroids and monolayer cultures. The supernatants of tumor cells grown in 2D and 3D cultures were studied using ELISA and multiplex analysis for the production of a spectrum of chemokines and cytokines supporting the immunosuppression, invasion and metastasis processes. Results Tumor specimens received were predominantly of metastatic origin (75%). In 100% of cases 2D cultures were received, in 88.6% of cases (39 out of 44) we succeeded in obtaining spheroids. There was no direct correlation between the efficiency of tumoroid formation and the tumor's histogenetic origin and the stage of the cancer process (primary tumor, recurrence, metastasis). The median size of spheroids by 4-5 days of cultivation with a starting concentration of 10000 cells per well was 657.14 μm for melanoma (min 400 - max 1000 μm), 571.42 μm (min 400 - max 700 μm), 507.14 μm (min 300 - max 600 μm) for soft tissue sarcomas, 650.0 μm (min 400 - max 900 μm) for osteogenic sarcomas. Immunochemical analysis of Ki-67, GLUT1, and Ecadherin markers was carried out for tumor tissue samples, single-layer tumor cultures, and tumoroids of every patient. The distribution of the stained groups in the spheroids was distinct from the monolayer cultures and more in accordance with the distribution of such in the tissue tumor, the number of Ki-67+ cells was increasing in the spheroids. We detected no dependence of Ki-67+ and GLUT1+ cell localization grade on spheroid size. We identified E-cadherin in tumor tissue and tumoroids of breast carcinoma and one melanoma culture. Monolayer cultures did not express it. The increase in secretory cell activity of the solid tumor cells from 2D to 3D system was observed when CCL2, CCL3, CXCL1, CXCL16, MIF, IL10, MICA (p<0.01) were investigated. Conclusion The presence of patient-specific cells of solid tumors in a 3D environment causes activation of the proliferative and metabolic processes as compared to monolayer cultures, which makes these models approximate the real world clinical picture. The production of chemokines that can attract to the tumor various types of immune system cells, to include their immature versions, as well as production of cytokines and Immunosuppression factors that, when present in the tumor microenvironment in the high concentrations, contribute to the formation of immune cells having suppressive capacities occurs in the 3D cell system. Three-dimensional model of the initial tumor nodule formation stage thus demonstrates the forming process of tumor cells favorable for them microenvironment. Construction of three-dimensional models - spheroids of tumor cells of differing histogenesis demands individual approach and more thorough investigation.

Cytokine Expression in Peripheral Blood Mononuclear Cell Cultures Obtained from Cattle with Different Stages of Natural Mycobacterium bovis Infection

In bovine tuberculosis (bTB), cellular, humoral, or both types of immune responses have been observed. The purpose of this study was to examine the immune status of tuberculous cows based on the differential cytokine gene expression associated with Th1 (IFN-γ, IL-2), or Th2 (IL-4, IL-10) responses. Twenty-three (23) cows belonging to a dairy herd located in a rural region of the State of Hidalgo, México, were selected for the study. Single Intradermal Comparative Cervical Tuberculin (SICCT) Test, Interferon-Gamma (IFN-γ) Release Assay (BOVIGAM), and Enzyme-Linked Immunosorbent Assay (ELISA) were used for detection of cattle infected by M. bovis. Thirteen cows were positive to all the tests (Group 1); ten cows were positive only to ELISA (Group 2), and the remaining Group (Group 3, control) included cows negative to all the tests. Peripheral blood mononuclear cells (PBMC) from animals were in vitro stimulated by bovin purified protein derivative (PPD), avian PPD, and Concanavalin A (Con A) mitogen for 72h. Changes in the levels of expression of mRNA of the respective cytokines was measured by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) using β-actin gene as internal control. In group 1, PPD bovis and Con A-stimulated cells exhibited high production of IFN-γ, IL-2 and IL-4, but not IL-10. In contrast, PPD avium-stimulated cells displayed a low production of cytokine transcripts. In group 2, cells showed a significant production of IL-10 in response to bovine PPD (P< 0.001). In the control group, a high production of IFN-γ and IL-2 was observed only in Con A-stimulated cells. Post-mortem examinations in animals of group 1 showed slight and medium lesions in lymph nodes, whereas in group 2, the lesions were more extensive. Results indicate differences on gene expression levels of cytokines considered to determine balance in Th1/Th2 response among the evaluated groups. In addition, high levels of antibodies against M. bovis and high IL-10 expression in PBMC together are indicators of progressive bTB when both tuberculin test and IFN-γ assay are negative in tuberculous anergic cattle. Inclusion of serology and IL-10 cytokine expression in in the diagnosis checklist improves detection of infected cattle to help control bovine tuberculosis.

Six Fractal Codes of Biological Life Unifying ATOMS, WAVES and INFORMATION: Perspectives in Exobiology, Cancers Basic Research and Artificial Intelligence Biomimetism Decisions Making

In this theoretical discovery of a law of Life, there is MATHEMATICS (Geometry, Bits and Numbers) that UNIFY 3 universes as complementary as ATOMIC MASS, WAVES, and INFORMATION (DNA, RNA and Amino Acids). The discovery of a simple numerical formula for the projection of all the atomic mass of life-sustaining CONHSP bioatoms leads to the emergence of a set of Nested CODES unifying all the biological, genetic and genomic components by unifying them from bioatoms up to 'to whole genomes. In particular, we demonstrate the existence of a digital meta-code common to the three languages ​​of biology that are RNA, DNA and amino acid sequences. Through this meta-code, genomic and proteomic images appear almost analogous and correlated. The analysis of the textures of these images then reveals a binary code as well as an undulatory code whose analysis on the human genome makes it possible to predict the alternating bands constituting the cariotypes of the chromosomes. The application of these codes to perspectives in astrobiology, cancer, and specifically in INFORMATION THEORY with the emergence of binary codes and regions of local stability (voting process), whose fractal nature we demonstrate, is illustrated. PREFACE by Professor Luc Montagnier Addendum by Robert Friedman M.D After the discovery of the DNA double helix structure allowing both the stable storage of genetic information and its transfer through messenger RNA to protein synthesis organelles themselves structured by RNA most abundant in cells, the ribosomal. This wonder of nature exists in ALL living beings from the virus to humans and is based on two codes, the linear sequence of nucleotides and that derived from codons where three nucleotides allow with a certain flexibility - synonymous codons - the choice in the twenty amino acids. But we are missing a third CODE the one governing at multicellular beings from the rotifer to human, the stabilized modulation of gene expression in a nutshell the differentiation of cells from the single cell of the fertilized egg. It is logical to think that this program which begins as soon as fertilization is written in the DNA. We are also prone to associate it with non-coding DNA sequences although they control gene expression. I introduce here the notion developed by Jean-Claude Pérez of mathematical harmony, a higher order present in all living beings and whose existence it finds in genomes, including those of viruses. Thus the natural evolution of variants of the genome of coronavirus Covid 19 tends towards increasingly long Fibonacci series. It remains to determine the Who, the How and the Why of such developments. I will bet with my mathematician colleague that waves and fractals play a role. Luc Montagnier ADDENDUM Jean-claude has given scientists a strong new direction for research. He has identified a unified field of science guided by the Golden Ratio and Fibonacci Sequence. By identifying an overall guiding principle that makes possible fractal-like nesting at all levels of biological manifestation, future researchers can begin with the "whole" instead of the "parts". If we know that complex systems are organized at varying levels by the Golden Ratio and Fibonacci Sequence, we can look for those universal patterns first and then fill in the gaps with small details to complete the picture. It's like having an overall view of a crossword puzzle before beginning to assemble the individual pieces. Without an overarching vision and guiding principle, completing the puzzle is infinitely more difficult. Once scientists and researchers realize and begin using this "SECRET IN HIDDEN IN PLAIN SIGHT," their discoveries will be orders of magnitude more fruitful.  Robert Friedman M.D

Attenuation of Microcystins Using Electron Beams and Gamma Radiation: A Study with Environment-Bound Conditions

Microcystins (MCs) are toxins profusely synthesized by cyanobacteria, causing livestock poisonings and endangering human health. We design and execute an experiment to investigate the attenuation (degradation) of microcystins by exposing them to gamma radiation and electron beams at doses of 0 (control), 3, 5, 10 and 15 kGy. The experimental conditions simulate microcystin contamination of aquatic environments; we thus consider (1) microcystins inside whole cells and extracellular dissolved in water, simulated by non-sonicated and sonicated cells, respectively, and (2) two acute microcystin concentrations within water. Toxicity tests of Microcystis aeruginosa detected immobilization (i.e., paralysis) of Ceriodaphniasilvestrii exposed to aqueous crude extracts of irradiated and non-irradiated M. aeruginosa (NPLJ-4 strain) at concentrations of 45 and 90 mg.L-1 (mg dry weight of freeze-dried material), and the results were analyzed using the Trimmed Spearman-Karber statistical program to obtain 48-h EC50, the average effective concentration causing immobility in 50% of organisms after 48 hours. We conclude that electron beams are effective physical agents for toxin attenuation (degradation) and reach 100% effectiveness at 5 kGy and above; their efficiency is two orders of magnitude greater than that of gamma radiation. This new body of information contributes to (1) remediating environmental water sources; (2) designing water/wastewater treatment facilities; (3) combatting chronic microcystin environmental contamination; and (4) inspiring further studies to promote the use of biomonitors (e.g., Cladocerans) to detect and evaluate microalgae contamination.

Liver Disease: A Retrospective Hospital Based Study in Addis Ababa-Ethiopia

Background Liver disease has caused significant morbidity and mortality worldwide. Its epidemiologic and clinical pattern, however, is not well characterized in sub-Saharan countries. Objective This study aimed to describe demographic, clinical characteristics, and patterns of liver disease in a community hospital in Addis Ababa, Ethiopia. Method A retrospective hospital-based study was conducted on patients with liver disease admitted at Ras Desta Damtew memorial hospital, in Addis Ababa-Ethiopia, from February 2015 to April 2020. Result Of the total 212 patients majority, 78.8% were male, 49.1% of patients were in the age range of 31-50 with a median age of 42. The most common initial clinical presentation was ascites (87.7 %), and more than half of patients (56.6%) had a history of alcohol misuse documented on their medical charts. Chronic liver disease (cirrhosis) was found in 177 (83.5%), and Hepatocellular Cancer accounted for 7.5% of the patients. Alcohol misuse caused 45% of chronic Liver Disease, followed by Hepatitis B virus infection. Conclusion Chronic liver disease is the most common form of liver disease, and the most affected were middle-aged men. The common cause of chronic liver disease was alcohol followed by hepatitis B virus infection.

Photobiomodulation, Depression, Anxiety, and Cognition

Background Anxiety and depression are key barriers to healthy aging and greatly heighten the risk for many negative health issues that seriously impact life quality. Aim This mini review examines the potential of low level laser treatments or photobiomodulation therapy for ameliorating severe anxiety and depression in older adults. Methods and Procedures Articles that adressed the current topic of interest extracted from PUBMED and Google Scholar were carefully and presented in narrative form. Results Photobiomodulation therapy appears to be a safe efficacious modality for ameliorating various degrees of anxiety and depression and for improving cognition, and is supported by several well established mechanisms of action at the molecular, cellular, and tissue levels. Conclusion More research to examine who might benefit most from this form of therapy, and in what respect in this area of growing global concern and few intervention options is strongly warranted.

Elasticity Profile of Skin, Neuronal, Cardiac, and Skeletal Muscle Cells after Treatment with the Biofield Energy Healing-Based Proprietary Test Formulation

The present study aimed to evaluate the effect of the Trivedi Effect®- Biofield Energy Treated/Blessed Test formulation/item (TI) composed of minerals (magnesium, zinc, copper, calcium, selenium, and iron), vitamins (ascorbic acid, pyridoxine HCl, alpha tocopherol, cyanocobalamin, and cholecalciferol), Panax ginseng extract, CBD isolates, and β-carotene on elasticity of skin, heart, muscle, and neuronal cells in the H9C2 (rat cardiomyocytes), C2C12 (mouse myoblast cells), HaCaT (human keratinocytes), and SH-SY5Y (human neuroblastoma cells) cell line in DMEM medium. The test formulation constituents were divided into two parts; one section was defined as untreated test formulation (UT), while the other portion of test formulation received Biofield Energy Healing/Blessing Treatment (BT) by a renowned Biofield Energy Healer, Mr. Mahendra Kumar Trivedi. The test items were treated with Biofield Energy Healing/Blessing Treatment and divided as Biofield Energy Treated/Blessed (BT) and untreated (UT) test items. MTT data showed that the test formulation in various concentrations was found as safe and nontoxic in the tested concentrations with viability range from 73% to 307%. Young’s modulus (YM) is a measure of cell stiffness, a decrease in YM value indicates increase elasticity of the cells and vice-versa. YM in H9C2 cells were decreased by 9.6% and 66.1% in the BT-DMEM + UT-TI group at 0.1 and 1 µg/mL respectively, as compared with untreated test group. However, C2C21 cells showed increased YM by 443.9% at 1 µg/mL in the UT-DMEM + BT-TI group, while 869.6% increased YM in the BT-DMEM + UT-TI group at 1 µg/mL as compared with untreated test group. However, 314% increased YM was reported in the BT-DMEM + BT-TI group at 1 µg/mL as compared with the untreated test group. However, the value of YM was significantly decreased in the HaCaT cell line by 247.7% (at 1 µg/mL), 225.8% (at 0.1 µg/mL), and 97.9% (at 1 µg/mL) in the UT-DMEM + BT-TI, BT-DMEM + UT-TI, and BT-DMEM + BT-TI group respectively, as compared with the untreated group. In addition, YM was significantly decreased in the SH-SY5Y cell line by 92.6%, 18.1%, and 26.6% at 1 µg/mL in the UT-DMEM + BT-TI, BT-DMEM + UT-TI, and BT-DMEM + BT-TI group respectively, as compared with the untreated group. Overall, the results showed the significant decreased YM among the SH-SY5Y, HaCaT, and H9C2 cells, while it was increased in the C2C21 cell line. Thus, the mechanical properties of cells such as cellular function, including shape, motility, differentiation, division, and adhesion to its surrounding extracellular matrix were improved. Overall, it can be useful in many disease progressions with improved cellular elasticity and its associated complications/symptoms.  

Evaluation of Immunomodulatory Effect of a Novel Test Formulation in D-Galactose-Induced Aging Dysfunction in Sprague Dawley Rats

The aim of the study was to evaluate the immunomodulatory activity of the Biofield Treated/Blessed proprietary test formulation consisting of essential ingredients viz. minerals (zinc, magnesium, iron, and copper) and vitamins (B6, B12, and D3) in male Sprague Dawley rats. Each ingredient of the test formulation was divided into two parts. One part was denoted as the control without any Biofield Energy Healing Treatment/Blessing, while the other part was defined as the Biofield Energy Treated/Blessed sample, which received the Biofield Energy Healing Treatment/Blessing by a renowned Biofield Energy Healer, Mr. Mahendra Kumar Trivedi remotely. Additionally, three group of animals were also received Biofield Energy Treatment per se (at day -15) under similar conditions. The parameters were assessed such as immune biomarkers (IgM, IgG, IgA, IgE, CD4+, CD8+, and CD28+), biochemistry and hematology and histopathology. The experimental results showed IgG level was significantly increased by 10.70% and 8.03% in the G6 (Biofield Energy Treatment per se at day -15) and G8 (Biofield Treatment per se to animals plus Biofield Treated test formulation from day -15) groups, respectively as compared with untreated test formulation (G4). Additionally, CD8+ count was significantly increased by 20.67% in the G8 group, while CD28+ count was significantly increased by 11.70%, 8.32%, and 9.82% in the G7 (Biofield Energy Treated test formulation at day -15), G8, and G9 (Biofield Treatment per se (day -15) to animals plus untreated test formulation) groups, respectively after Biofield Energy Treatment to the animals as compared with the untreated test formulation. In hematological analysis, platelet count was increased in the G5, G6, G7, G8, and G9 groups by 40.69%, 27.95%, 26.67%, 38.58%, and 28.28%, respectively compared with the disease control (G2) group. Biochemical parameters showed significant decrease in the level of creatinine by 32.14% in the G9 group as compared with the G2 group. Further, animal body weight, feed intake, relative organ weight, and histopathological findings of all the tested groups did not show any abnormal findings with respect to the safe and non-toxic treatment strategies. Overall, the experimental data concluded that the Biofield Energy Treated/Blessed test formulation showed considerable improved cellular and humoral immune response as compared with the untreated test formulation. Thus, the Trivedi Effect®-Biofield Energy Healing Treatment per se and the test formulation has the significant capacity for immunomodulatory effect, stress management and anti-aging by improving overall health.  

SARS-CoV-2 affected cells Pathogeny and Therapy

There were compared mechanisms infecting a human organism by different viruses in relation to interaction between human diploid cellular cycle mechanisms and coronaviruses haploid genomic mechanism. Besides there were described mechamism forming combined haploid-diploid cellular cycle of viral affected cells due to interactions between human cellular cycle mechanisms and coronaviruses genomic mechanism. Further there were considered infected way of SARS-CoV-2 from mechanism maintenance stability Internal Energy of an organism’s able-bodies cells and transmutation them into viral affected cells leading to death of affected cells of high respiratory level in nose-trachea-bronchi with transiting coronaviruses through dead cells‘ wall and infecting lungs‘ cells. Taking into account great searches of methods treatments Coronaviruses infected disease, we offered to approved through detail clinical Trial of new efficient method of treatment ill patients with SARS-CoV-2 disease which can rescue of still alive lungs‘ cells. Moreover there was reviewed offered therapy of SARS-CoV-2 induced disease.

NNC55-0396 Reduces SH-SY5Y Cell Damage Induced by Bupivacaine Hydrochloride

Background Local anesthetic neurotoxicity is a common complication in clinical anesthesia, which can cause permanent nerve damage in severe cases. The T-type calcium channel is an important channel for regulating the excitability of neurons. Normally, extracellular calcium ions enter the cell through the T-type calcium channel to change the excitability of neurons. When the intracellular calcium is overloaded, it can cause cell damage. Aims To investigated the roles of T-type calcium channel in the SH-SY5Y cells injury induced by the bupivacaine. Methods The SH-SY5Y cell culture model was used to observe the effect of T-type calcium channel blocker NNC55-0396 on the neurotoxicity of bupivacaine hydrochloride by MTT methold,flow cytometry, Western blotting and other methods. Results The results show that NNC55-0396 can block the T-type calcium channel of SH-SY5Y cells, improve the decrease of cell viability caused by bupivacaine hydrochloride, reduce the level of intracellular calcium ion, reduce the expression of Cleavedcaspase-3, and reduce cell apoptosis. Conclusion The above results indicate that the T-type calcium channel is involved in the SH-SY5Y cell damage caused by bupivacaine hydrochloride, and blocking the T-type calcium channel can reduce the neurotoxicity of bupivacaine hydrochloride.

Cloning, Expression and Characterization of the α-glucuronidase from the Hyperthermophile DictyoglomusturgidumDSM 6724Ô

Conversion of biomass into fermentable sugars is a major requirement for successful and cost-effective biofuels production. The conversion of xylan to sugars requires multiple enzymes including α-glucuronidase. Here we report the cloning, expression, purification and characterization of the α-glucuronidase from Dictyoglomusturgidum(DtuAgu). DtuAgu is an intracellular protein of 685 amino acids and a predicted molecular weight of 79.4 kD. Enzymatic activity was optimum between pH 7.0 and 8.0 and at 85°C. The specific activity of the enzyme was 10 u/mg when measured using mixed aldouronic acids. The specific activity on isolated glucuronoxylan was approximately 20% of the value obtained with xylooligosaccharides. DtuAgu significantly improved xylan conversion to xylose when evaluated using two mixtures of thermostable bacterial enzymes and two sources of xylan. DtuAgu has the potential to be a key player in thermostable enzyme cocktails for the conversion to biomass to biofuels.α

Investigation of Neoplastic Cells in the Bone Marrow of Female Dogs with Mammary Gland Tumors

Background The mammary glands are the second most common tumor development site in female dogs. One of the ways of staging such tumors is to evaluate the presence or absence of distant metastasis, including in bone marrow. Such findings in human medicine are associated with poor survival of women with breast tumors. However, in veterinary medicine, this clinical staging is used more for patients with lymphomas and mastocytomas. Studies using bone marrow biopsies as a staging method for mammary tumors are scarce. Objectives The present study was to evaluate mammary lesions and bone marrow in 23 female dogs, searching for disseminated tumor cells or metastatic foci. Results: Grade I carcinoma in mixed tumors was the type most observed (22.4%), and there was no statistical difference in relation to tumor size or presence of metastasis in lymph nodes. In the bone marrow of one female dog with carcinosarcoma (4.35%), there was cytoplasmic marking of a probable disseminated tumor cell of epithelial origin, and immunohistochemical evaluation showed presence of cytokeratin-19 antibodies. None of the female dogs presenting reduced cellularity or medullary fibrosis, confirmed through Masson’s trichrome technique, had cell marking in immunohistochemical analyses. Conclusions Bone marrow evaluation can be used as a staging method for mammary gland tumors in female dogs, since disseminated tumor cells present the potential to become secondary lesions and to disseminate to distant foci, thereby causing tertiary metastases over an indeterminate period of time.

Effects of Aluminium Sulphates on Weight and Structure of Adrenal Gland of Adult Wistar Rats

This study evaluated the effects of aluminum sulphate exposure on the histology of adrenal gland of Wistar rats. Thirty adult Wistar rats were used for this study. The Wister rats were divided into three groups; group A was the control animals and tagged C, group B animals received 10g of alum dissolved in 1000cm3 of distilled water and were tagged T1, group C animals received 50g of alum dissolved in 1000cm3of distilled water and were tagged T2, via drinking water for duration of four weeks. Twenty-four hours after the last administration, the rats were sacrificed by cervical dislocation. The adrenal gland was excised and preserved in 10 % formosaline after which it was routinely processed for hematoxylin and eosin staining (H&E). Histological observations showed normal cell distribution in the control group but treated group revealed evidences of cellular obliteration& hemorrhagic necrosis. The results obtained from this study suggest that aluminum sulphate has a damaging effect on the structure of the adrenal gland.

Use of Immune Modulator Interferon-Gamma to Support Combating COVID-19 Pandemic

While the COVID-19 pandemic has raised concerns about the future of people worldwide, it has made it necessary to take measures with high economic costs, including quarantine. We consider it is more logical for some scientists to investigate time-saving treatment options until vaccination studies, which are started to be studied rapidly, are accomplished or specific antiviral agents are found. In this context, treatment combinations of one or more of the immune modulators known as cytokines, which can stimulate or accelerate the immune system, should be tried. In our opinion, although such options are not as effective as specific treatments such as vaccines, such options will offer highly effective alternatives in times of emergency. For this reason, we found it appropriate to make a reminder by preparing a broad review about interferon gamma, which is an antivirus and is an immunomodulator and which plays a critical role in humoral and cellular immunity. 

Resveratrol Prevents Cataract Formation by Inhibiting Pro-inflammatory Mediator-induced Dysregulation of Lens Calcium

The effect of resveratrol, a free radical scavenger, during cataract development was evaluated in the Wistar rat pup model. This study investigated the possible free radical scavenging potential of resveratrol at 40 mg/ kg body wt dose in selenite-induced cataract in rat pups. Intraperitoneal injection of sodium selenite (15 µm mol/ kg body wt) in 8 to 10 day old rat pups lead to severe oxidative stress in the tissues evidenced by decreased antioxidants and increased lipid peroxidase, nitric oxide, superoxide anion, hydroxyl radical generation, inducible nitric oxide synthase (iNOS) as well as nuclear factor kappa B (NF-kB) expression levels that probably led to cataract formation. Selenite exposure also caused an increase in total calcium in the eye lens and significantly inhibited the activity of Ca2+ ATPase but not Na+/ K+ ATPase or Mg2+ ATPase. However, both pre- and co-treatments with resveratrol, but not post-treatment, led to an increase in antioxidant levels with a concomitant reduction in oxidative stress and also rescued the selenite-mediated increase in lens Ca2+ and inhibition of Ca2+ ATPase activity in the eye lens. The results of this study demonstrate antioxidants decrease and increase in free radical generation triggered by selenite causes the inactivation of lens Ca2+ ATPase leading to a rise in intracellular Ca2+ level. Resveratol treatment was able to prevent selenite-induced oxidative stress and in turn the inhibition of lens opacification. Thus, resveratrol has the potential to function as an anti-cataractogenic agent, possibly by preventing free radical-mediated accumulation of Ca2+ in the eye lens.

The Mechanism of Decline of Senescent Skeletal Muscle Satellite Cell Self-Renewal and Regenerative Proliferation: The Role of Heparan Sulfate-FGF-2--FGFR1-p38αMAPK Axis, Sprouty1, miR-1, miR-133 and miR-29a

Aging mammalian skeletal muscle satellite cells (MuSCs) undergo a decline of stem cell/progenitor cell proliferative and regenerative capacity, and the development of a physiological milieu characteristic of a state of chronic sterile inflammation. p38αMAPK and ERK1/2 are two major signaling pathways that regulate the age-associated decline of MuSC proliferative capacity. In this review we propose the following mechanism that links the p38αMAPK pathway to the decline of self-renewal and regenerative capacity of aged MuSCs: a) the HS-FGF-2-FGFR1-p38αMAPK-Axis, a tightly linked homeostatic signaling complex, is in synchrony with the autoinhibition of FGFR1; b) autoinhibition contributes to the Axis’ regulation of the homeostasis of P-p38αMAPK activity in juvenile MuSC; c) this combination of protein-protein interactions is characteristic of a juvenile cytoplasmic milieu of beneficial P-p38αMAPK activity and d) includes Sprouty1 inhibition that supports the stimulation of FGF-2 --> miR-29a; e) the miR29a dismantles the basement membrane in preparation for the initiation of replication; f) an age-associated impaired, dysregulated, over-sulfated heparan sulfate ligand (HS)-FGF-2 fails to activate FGFR1 in aged MuSCs; g) this uncouples its regulation of p38αMAPK and ERK1/2 pathways and results in desensitization of FGFR1; h) desensitization of FGFR1 and Sprouty1 interaction in aged MuSC uncouples their regulation of P-p38αMAPK in the aged MuSCs; i) this enables a state of chronic sterile inflammation to promote and sustain an increased level of P-p38αMAPK activity; and, j) the increased activity of P-p38αMAPK in aged MuSC stimulates the production of cell cycle inhibitors, miR-1 and miR-133, thereby attenuating the expression of the cell cycle regulators, SP1 and cyclin D1, resulting in a G1/S arrest; j) the increased level of p38αMAPK activity promotes the apoptosis of the aged activated MuSCs. This mechanism involves the synergistic interactions of HS-FGF2-FGFR-1, Sprouty (spry1), miR-1, miR-133 and miR-29a that unify the extracellular niche and intracellular milieu for the juvenile vs age-associated regulation of proliferative capacity of the MuSC. Our hypothesis unifies these interactions with the role of the extracellular niche and intracellular milieu in the stimulation of juvenile proliferation vs age-associated decline of skeletal muscle satellite cell self-renewal and regenerative proliferation. Word Count = 344

Chondroplasty Efficacy of Bone Matrix

Aim To study the chondroplasty efficacy of the bone matrix obtained using an original technology in restoring cartilage defect of the knee joint. Material and Methods Marginal defects were modeled on the surface of the distal end of the femur in 40 adult male Wistar rats. The bone matrix obtained using an original technology was implanted in the damaged area in animals of the experimental group. Material was investigated by means of light microscopy, transmission and scanning electron microscopy, and electron probe X-ray microanalysis. Results It was found that the bone matrix implanted did not cause an immune rejection reaction, activated reparative chondrogenesis for a prolonged period. In the area of articular cartilage lesion, the regenerate acquiring cellular and histochemical characteristics of the hyaline cartilage tissue was formed. The chondroinductive properties for the bone matrix were ensured by localized growth factors and morphogenetic proteins released during osteoclastic resorption. Conclusion The application of the bone matrix as a stimulator of chondrogenesis is theoretically reasonable and has a good perspective in treatment of damages and diseases of the articular cartilage.

Type-2 Diabetes and Gene Therapy: The Promise of CRISPR Gene Therapy in type-2 Diabetes Mellitus

Gene therapy has entered a new era with the dawn of CRISPR/Cas9 technology which though were always available in nature but rediscovered to tame into a real-tlife genome editing tool. With the modernization upsurge and changes in ways the “homo sapiens” survived on this planet from hunger to current era of exuberance has led to multiple metabolic issues like type-2 diabetes. Notwithstanding the rapid emergence of medication to suppress the hyperglycemia and insulin resistance associated with this menace, need has definitely emerge to find more personalized and curative dimensions to therapeutics of type-2 diabetes mellitus. Gene therapy is one more addition to Type-2 Diabetes Mellitus (T2DM) therapy, where multiple options have emerged in the shape of microRNA, direct knocking out of cellular structures like proteins and enzymes and very recently the precision nucleases associated with CRISPR technologies. This mini-review attempt to summarize some of the recent examples of gene therapy with major focus on CRISPR/Cas technologies.

Long Non-Coding RNAs Emerging as Potential Epigenetic Biomarkers for Tobacco and/or Alcohol-Induced Head and Neck Cancer

Head and Neck cancer (HNC) is one of the most prevalent and lethal cancer globally. The incidence of tobacco-induced HNC is gradually increasing in low and middle income countries. Among the various causative factors associated with HNCs, tobacco and alcohol play synergistic effect and are frequently associated with the risk of HNC. Tobacco-induced HNCs show distinct genetic and epigenetic alterations leading to different clinical outcomes in comparison to HPV-infected HNCs. Tobacco-induced HNCs are often associated with tumor aggressiveness, poor prognosis and low or nil prevalence of HPV infection. Apart from carcinogenic effects of these causative factors (use of tobacco products, alcohol intake and HPV or EBV infections), recent studies show that exposure to these factors alter/disrupt the regulation of non-coding RNAs including the long non-coding RNAs (lncRNAs). Altered lncRNA regulation is brought about by signalling networks that regulate cellular differentiation, apoptosis, angiogenesis and inflammatory pathways which play key functions in the genesis of different cancers including HNCs. There are numbers of studies supporting the emerging role of lncRNAs in development of HNC; however, reports connecting lncRNAs expression and addiction habits in HNC are still preliminary and sparse. Therefore, identification and characterization of lncRNAs that are differentially expressed upon exposure to risk-factors can serve as unique therapeutic targets and potential biomarker(s) for effective treatment of HNC subtypes. In this short review, we briefly reviewed the emerging role of lncRNAs in tobacco and alcohol induced HNCs.

In Vitro Cell-Based Biomarkers Study of Vital Organs: Impact of the Biofield Energy Based Test Formulation

The present study was undertaken to evaluate the impact of Biofield Energy Treated test formulation using multiple cell-lines. The test formulation and cell media (Med) was divided into two parts; one part was untreated (UT) and other part received Biofield Energy Treatment remotely by a renowned Biofield Energy Healer, Krista Joanne Callas, USA and labeled as Biofield Energy Treated (BT) test item (TI)/Med. Based on cell viability, test formulation was found safe. Cytoprotective action of test formulation showed significant restoration of cell viability by 89.9% and 106.4% in human cardiac fibroblasts cells (HCF) cells, while improved restoration of cell viability by 77.3% and 69% in HepG2 cells compared to untreated. Cellular restoration in A549 cells was also improved by 141.2% and 157.1% compared to untreated. ALP activity was significantly increased by 118.7% and 140.7% in UT-Med + BT-TI and BT-Med + UT-TI, respectively at 0.1 µg/mL than untreated. Percent cellular protection of HCF (heart) cells (decreased of LDH activity) was significantly increased by 89.9% and 106.4% in UT-Med + BT-TI and BT-Med + BT-TI, respectively than untreated. HepG2 cells protection (decreased ALT activity) was increased by 59.8% in BT-Med + BT-TI than untreated. Superoxide dismutase (SOD) level was increased by 22.8% in BT-Med + BT-TI than untreated. Serotonin level was significantly increased by 361.7% and 197.6% in BT-Med + UT-TI and BT-Med + BT-TI, respectively than untreated in human neuroblastoma cells (SH-SY5Y). However, relative quantification (RQ) of vitamin D receptor (VDR) was significantly increased by 116.5%, 214.7%, and 241.5% in UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI, respectively than untreated in MG-63 cells. Overall, data showed a significant improvement of organ-specific functional enzyme biomarkers. Thus, Biofield Energy Treated Test formulation (the Trivedi Effect®) would be useful for multiple organs health that can be beneficial against coronary artery disease, arrhythmias, congenital heart disease, cardiomyopathy, cirrhosis, liver cancer, hemochromatosis, asthma, chronic bronchitis, cystic fibrosis, osteoporosis, etc.

Impact of Biofield Energy Treatment Based Test Formulation on Vital Organ Health Specific Biomarkers Using Cell Line Study

Multiple organ dysfunction syndrome or failure is one of the major concerns against healthcare services in order to maintain the normal function. The present study aimed to explore the impact of the Biofield Energy Treated test formulation on the function of vital organs such as bones, heart, liver, lungs, and brain using standard activity parameters in specific cell-based assays. The test formulation and cells medium was divided into two parts, one untreated (UT) and other part received the Biofield Energy Treatment remotely by a renowned Biofield Energy Healer, Ariadne Esmene Afaganis, Canada and was labeled as the Biofield Treated (BT) test formulation/media. The test formulation was tested for cell viability, and the data suggested that the test formulation was found safe and non-toxic against all the cell lines. Cytoprotective activity among the experimental groups showed a significant improved activity by 94.4% at 1 µg/mL in untreated medium (UT-Med) + Biofield Treated Test Item (BT-TI) group in human cardiac fibroblasts cells (HCF) cells, while 84.4% at 10 µg/mL in BT-Med + BT-TI groups in human hepatoma cells (HepG2), and 124% increased cytoprotective action at 1 µg/mL in UT-Med + BT-TI group in adenocarcinomic human alveolar basal epithelial cells (A549) cells as compared with the untreated test group. ALP activity in MG-63 cells was significantly increased by 85.9% at 10 µg/mL in the UT-Med + BT-TI group, while in Ishikawa cells showed maximum increased ALP activity by 59.2% at 0.1 µg/mL in BT-Med + BT-TI groups as compared to the untreated group. The percent protection of HCF (heart) cells (decreased of LDH activity) was significantly increased by 53% and 40.5% at 1 and 10 µg/mL concentrations respectively, in UT-Med + BT-TI group, while BT-Med + UT-TI group showed increased protection by 68.5%, 70.7%, and 16.8% at 0.1, 1, and 10 µg/mL respectively, and 86.5%, 62.5%, and 34.2% improved cellular protection at 0.1, 1, and 10 µg/mL respectively, in BT-Med + BT-TI group as compared to the untreated test group. The percent protection of HepG2 (liver) cells (decreased of ALT activity) was reported by 33.5%, 63.2%, and 99.2% at 10 µg/mL in the UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI groups, respectively compared to the untreated group. Cellular protection of A549 (lungs) cells (increased of SOD activity) in terms of percentage was increased by increased by 39.8% (at 10 µg/mL), 44% (at 25.5 µg/mL), and 59.7% (at 25.5 µg/mL) in the UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI groups, respectively compared to untreated group. Serotonin level was significantly increased by 59.2% (at 0.1 µg/mL), 190.3% (at 0.1 µg/mL), and 201% (at 1 µg/mL) in the UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI groups, respectively compared to untreated in human neuroblastoma cells (SH-SY5Y). However, the relative quantification (RQ) of vitamin D receptor (VDR) was significantly increased by 159.1% (at 50 µg/mL), 212.7% (at 1 µg/mL), and 278.3% (at 10 µg/mL) in the UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI groups, respectively as compared to the untreated in MG-63 cells. Thus, the present data concluded that the overall multiple organ health using various standard biomarkers in specific cell lines were significantly improved with respect to health of bones, heart, liver, lungs, and brain after treatment with the Biofield Energy treated test formulation (The Trivedi Effect®). Thus, it can be used as a complementary and alternative therapy approach against many multiple organ disorders such as coronary artery disease, arrhythmias, congenital heart disease, cardiomyopathy, cirrhosis, liver cancer, hemochromatosis, asthma, chronic bronchitis, cystic fibrosis, osteoporosis, etc.

A Case Series of Resolving Vitreous Haemorrhage in the Setting of Diabetic Retinopathy

This case series presents 2 patients (67 and 58 year old females) with resolving vitreous hemorrhage in the setting of diabetic retinopathy. Both patients had around 50% of the retina still visible and the hemorrhage was confined at the posterior vitreous cavity near the retina, hence, a partial pan retinal photocoagulation (PRP) was feasibly applied. On B-scan, both cases showed a liquified vitreous with posterior vitreous detachment (PVD). For both patients, vitreous hemorrhage resolution occurred at less than 4 months from onset and consequently PRP was completed. Vitreous hemorrhage differs with bleeding in other tissues due to the presence of vitreous collagen matrix which promotes rapid clotting and hinders resolution of hemorrhage by preventing passive diffusion and delay in inflammatory cellular response. The prognosis for clearing hemorrhage was better when the fundus reflex was brighter and the retina adjacent to the ora serrata was visible. Clearing occurred sooner when hemorrhage was retrohyaloid rather than in the vitreous gel. In diabetic eye, the vitreous undergoes metabolic derangements resulting in premature liquefaction and abnormal vitreoretinal adhesion leading to traction and membrane formation. Partial PVD has also been associated with higher rate of diabetic retinopathy progression since it serves as scaffold for growth of neovascular tufts.

Cell-Based Vital Organs Specific Biomarkers Assessment using Biofield Energy Based Novel Test Formulation

The aim of the present study was to determine the impact of Biofield Energy Treated test formulation using six differentcell-lines. The test formulation/item (TI) and cell media (Med) was divided into two parts; one part was untreated (UT) and other part received Biofield Energy Treatment remotely by a renowned Biofield Energy Healer, Janice Patricia Kinney, USA and labeled as Biofield Energy Treated (BT) test item (TI)/media. Based on cell viability assay, test formulation was found as safe at tested concentrations. Cytoprotective activity of test formulation showed a significant restoration of cell viability by 60.6% (10 µg/mL), 67.5% (63.75 µg/mL), and 117.5% (63.75 µg/mL) in UT-Med + BT-TI, BT-Med + UT-TI, BT-Med + BT-TI, respectively compared to untreated in human cardiac fibroblasts cells (HCF) cells. Moreover, restoration of cell viability was improved by 64% and 127.3% in UT-Med + BT-TI and BT-Med + UT-TI, respectively at 1 µg/mL compared to untreated in human liver cancer (HepG2) cells. Cellular restoration in A549 cells was improved by 314% and 112.3% at 1 µg/mL in BT-Med + UT-TI and BT-Med + BT-TI, respectively than untreated. ALP activity in Ishikawa cells was significantly increased by 175.5%, 547.2%, and 220.8% in UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI, respectively at 0.1 µg/mL as compared to untreated. Additionally, in MG-63 cells showed increased ALP activity by 76.9%, 78.4%, and 79% in UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI, respectively at 50 µg/mL compared to untreated. The percent cellular protection of HCF (heart) cells (decreased of LDH activity) was significantly increased by 60.6% (10 µg/mL), 67.5% (63.75 µg/mL), and 117.5% (63.75 µg/mL) in UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI, respectively as compared to untreated. An improved HepG2 cells protection (represents decreased ALT activity) by 115.1% (1 µg/mL), 42.5% (25.5 µg/mL), and 60.8% (10 µg/mL) in UT-Med + BT-TI, BT-Med + UT-TI, BT-Med + BT-TI, respectively as compared to untreated. Percentage cellular protection of A549 (lungs) cells (represents increased of SOD activity) was significantly increased by 191.1% and 81.4% at 0.1 µg/mL in UT-Med + BT-TI and BT-Med + BT-TI, respectively as compared to untreated. Serotonin level was significantly increased by 31.8% (10 µg/mL) and 56.9% (25.5 µg/mL) in UT-Med + BT-TI and BT-Med + BT-TI, respectively compared to untreated in human neuroblastoma cells (SH-SY5Y). Relative quantification (RQ) of vitamin D receptor (VDR) was significantly increased by 304.3% (0.01 µg/mL), 128.4% (0.1 µg/mL), and 240% (0.1 µg/mL) in UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI, respectively compared to untreated in MG-63 cells. Thus, Biofield Energy Treated test formulation (The Trivedi Effect®) significantly improved organ specific functional biomarkers and would be useful for multiple organs health related to coronary artery disease, arrhythmias, congenital heart disease, cardiomyopathy, cirrhosis, liver cancer, hemochromatosis, asthma, chronic bronchitis, cystic fibrosis, osteoporosis, etc.

On the Role of Cholecystokinin (CCK) in Fear and Anxiety: A Review and Research Proposal

Cholecystokinin (CCK) is found in high concentrations in cortical and limbic structures including the amygdala of rodents, and evidence has been gathered supporting a role for CCK in the neurobiology of anxiety. A variety of animal models have been used to study a central state of fear or anxiety, state that appears to produce a complex pattern of behaviors highly correlated with each other. It is now well established that the amygdala in particular is a critical link in the pathway through which sensory stimuli come to acquire fear evoking properties. The purpose of the proposed experiments is to study the role of the putative neurotransmitter CCK in fear and anxiety in vivo by means of a methodology coupling electrochemical and electrophysiological measurements in various brain areas. Indeed, the association of in vivo differential pulse voltammetry (DPV) with in vivo extracellular single unit recording could be able to provide concomitant physiological and neurochemical indications and to relate them to behavioral events. To further study and support the initial observations pharmacological experiments will also be performed by means of CCK receptor agonists and antagonists. This may eventually lead to development of more effective pharmacological strategies for treating clinical anxiety disorders.

Yeast Species Mediated Bioprocesses and Bio-Products for Biotechnological Application

Yeast as unicellular organism, has shown multiple application due to exhibition of noble ability in its cells. And engineered yeast has found more suitability in bioprocesses application as well as adverse conditions adaptation. Different types of yeast strains showed their best capability to adapt the salt and sugar rich environment with their optimal growth capability. These strains, used as suitable and novel cell factories for production of value added bio-products (via utilization of fermentation processes) and also for different types of bioprocesses. Application of yeast species in biotechnology field, enhanced in current periods, due to conversion of its wild to engineer strain, suitable for bioprocesses utilization and also for different types of biochemical synthesis. Different yeast species identified due to known their genetic, regulatory mechanism and also competitive metabolic pathways. In this regards, different type of engineering approaches (for genetic or pathways modification), applied to construct the optimal and suitable cell factories for different types of bioprocesses as utilized in different sectors (foods with mineral or protein rich, bread, brewing, cosmetics, chemical, agriculture, pharmacy and distillation industries) via improving the quality of bio-products. Further, in silico designed based metabolic engineering technique showed the improvement in performance of yeast strain. System and synthetic biology with engineering approaches applied to further improve the yeast mediated bioprocesses as well as biochemical products formation for industrial or biotechnological application. Some bio-products such as functional bio-molecule, different types of alcoholic biofuels, organic acids and enzymes etc are good examples of yeast mediated biochemicals products, utilized more frequently in our life. Author will focus recent research and development on bio-product formation or bioprocesses with their regulatory control mechanism in different yeast strains.

The use of Dermacell® in Fingertip Injury

Matrices or tissue scaffolds provide a collagen structure for tissue remodelling while the removal of viable cells aims to minimize or prevent inflammatory or immunogenic response. Allograft collagen scaffold can support the patient’s own cellular ingrowth, ingeneered to minimize an immune response and to yeld a bio-compatible matrix and support incoming cellular growth. The decellyularized dermis retains its growth factors, native collagen scaffold, and elastin, thanks to a LifeNet Health proprietaryprocessin technology.

The Current Immunoassays and Emerging Immunogenomic Approaches for Immunomonitoring Cancer and Infectious Diseases

As remarkable advances have been made in immunotherapies, the overall goal of immunotherapy has become the selection of patients and evaluating the benefits of treatment. One of the major obstacles to develop immunotherapies is the lack of effective immune monitoring. Monitoring of key changes in the immune system during immunotherapy (immunomonitoring) provides important insights into efficacy as well as the immune mechanisms of response at the molecular and cellular levels. Immunomonitoring techniques include traditional immunoassays that use specific antibodies to recognize the analytes of interest, new high-throughput immunoassays that target immune cells and nucleic acids, and less classical immunogenomic approaches that rely on genome-wide profiling and computational analysis on various types of clinical samples. Substantial progress has been made in the application of immunomonitoring strategies to pre-clinical and clinical studies, especially for patients with cancer and infectious diseases. Current and emerging immunoassays performed in clinical practice will be examined herein, and immunogenomic approaches that complement these techniques will be highlighted and compared with traditional methods. Finally, we will discuss several new computational methods for analyzing gene signatures for immunomonitoring, including gene expression data profiling by microarray, the nCounter technique, regular RNA-seq, and single-cell RNA-seq. Novel immunomonitoring techniques, especially immunogenomic approaches, will continue to be developed to facilitate assessment of immunotherapeutic response and predict patient outcomes in cancer and infectious disease.  

How Knowledge on Microbiota may be Helpful to Establish an Optimal Diet for Health Maintenance

In the last few years, gut microbiota has been identified to be an essential mediator in health and disease. In fact, it interacts with various organs and systems in the body, including brain, lung, liver, bone, cardiovascular system, and others. Microbiota-derived metabolites such as the short chain fatty acid (SCFA) butyrate are primary signals, which link the gut microbiota and physiology. Then, the findings on the roles of microbiota profoundly change not only the key concepts of biology and medicine, but also of nutrition. In fact, it is currently evident how the main task of nutrition is not to nourish us, but to maintain a comfortable environment for the intestinal microbiota. In this way, it works in symbiosis with us, correctly controlling the functioning of the organs, the physiological parameters and the cellular regenerative processes. It is also evident that the strength of reparative processes correlates with the ability of digestive system to process complex foods, which increases during weaning, a period of time in which the diversity of bacterial strains increases. Therefore, a task of food is to keep trained the digestive system, to which it corresponds an high microbiota diversity. Elderly leads to reduced microbiota diversity to which corresponds an intestinal frailty, responsible for the frailty of the elderly. In conclusion, a correct diet may not only keep us in good health but may also guarantee us longer longevity.

Action Mechanisms and Therapeutic Targets of Renal Fibrosis

Renal fibrosis was a chronic and progressive process affecting kidneys in chronic kidney disease (CKD), regardless of cause. Although no effective targeted therapy yet existed to retard renal fibrosis, a number of important recent advances have highlighted the cellular and molecular mechanisms underlying the renal fibrosis. The advances including TGF-β/Smad pathway, oxidative stress and inflammation, hypoxia and gut microbiota-derived from uremic solutes were highlighted that could provide therapeutic targets. New therapeutic targets and strategies that are particularly promising for development of new treatments for patients with CKD were also highlighted.

Biological Networks: An Introductory Review

All aspects of life activities in living cells are mediated/executed and regulated by a vast number of networks, comprising a wide spectrum of components, starting with simple biomolecules and ending with the whole organism, and functioning within a precisely organized tight framework. Proper mediation of cellular activities necessitates their inclusion within the context of structured and organized network systems capable of regulating/coordinating and synchronizing the countless numbers of biological processes occurring within living cells. The number of biological networks and pathways within the living cell is considerably huge, being dependent on the structural complexity and functional capabilities of the cell. Pathogenesis and progression of human diseases result from functional disturbances of biological networks within the cell as disturbed network function leads to deleterious effects on physiological processes dependent on, and mediated by, affected network(s). Ensuing pathological processes, defined by the nature of disturbed networks and the specific organs or tissues affected, pave the way for the development of pathognomonic and characteristic disease entities. As most network functions are dependent on relatively small number of key regulatory biomolecules, i.e. enzymes/proteins and signal transducing factors, it follows that functional disturbances of biological networks and pathogenesis of disease states can be attributed, in most instances, to quantitative and/or qualitative abnormalities of these key regulatory molecules. Study and analysis of the structural designs and the functional mechanisms of biological networks would have crucial and important impacts on many theoretical and applied aspects of biology, in general, and of medical sciences in particular. Meticulous study of biological networks represents an important and integral aspect in study of biology. Interpretation and analysis of key information deduced from observing and analyzing structural designs and functional characteristics and dynamics of biological networks discloses and defines the basic framework within which life activities in living cells are initiated, adapted to physiological requirements, maintained, and terminated upon completion of their aims. More important, however, is the contribution of this information to proper understanding of the different mechanisms responsible for regulating and synchronizing the functions and performances of the vast spectrum of different network categories within the cell. In addition to its vital scientific significance, discovering and defining the key pivotal structural and regulatory molecules within life-mediating networks, and along different pathways responsible for controlling functional dynamics of the network, represent an indispensable diagnostic approach insistent for designing proper therapeutic approaches to diseases caused by network defects.

Changes in Rats’ Liver Structure Induced by Zinc Oxide Nanoparticles and the Possible Protective Role of Vitamin E

Background: Oral ingestion of zinc oxide nanoparticles (ZnONPs) may lead to serious liver injury. Vitamin E (VE) is an important antioxidant factor that can reduce such damage. Aim: This study aimed to evaluate the possible changes that could take place in the liver of adult male albino rats after oral ingestion of ZnONPs and elucidate the potential protective role of VE against such damage. Material and Methods: Forty eight male albino rats were divided into four groups of 12 animals each. Group (1) served as control group and received normal saline. Group (2) “VE-treated” received 100 mg/kg/day of VE dissolved in normal saline by oral gavage for 21 days. Group (3) “ZnONPs-treated” received a daily dose of ZnONPs dispersed in the fresh sterilized physiological saline solution 1mg/kg for 5 constitutive days. Group (4) “concomitant ZnONPs and VE-treated” was pretreated with VE 100 mg/kg/day for 14 days followed by the same dose of ZnONPs as in group (3) for 5 days. The extent of hepatic damage was evaluated by histological and immunohistochemical examination of liver samples and serological analysis of liver enzymes. Results: Body weights and liver weights showed very highly significant decrease (P <0.001) in the ZnONPs-treated group. The histological results in ZnONPs-treated group revealed congested dilated central veins and blood sinusoids, loss of normal arrangement of hepatocytes and most of hepatocytes showed marked vacuolated cytoplasm with darkly stained nuclei. Portal area affection was in the form of congested dilated portal veins with bile duct hyperplasia and cellular infiltration. There was an increase in the mount of blue stained collagen fibers around central veins together with strong positive reaction for Caspase 3 in ZnONPs-treated group. Similarly biochemical analysis indicated that the levels of serum aminotransferase (AST &ALT) significantly increased in ZnONPs-treated group when compared with other groups. Rats pretreated with VE showed improvement of the histological findings and biochemical parameters. Conclusion: Ingestion of ZnONPs could be associated with serious liver affection and pretreatment with VE is suggested to induce some improvement of such deleterious changes.

Space of Cross-Disciplinary Researches of Life, Nature and Society

Journal of Biosemiotic Research is a new periodical devoted to a young, actively developing science. A review of recent scientific publications shows that in the broad scientific space of biosemiotics contemporary questions and "eternal themes" interact, not finding an answer in the private sciences - anthropology, semiotics of culture and philosophy1,2. To solve them, the fundamental foundations of science and new achievements, the opportunities of the latest technologies and scientific communications are attracted. Like all young sciences, biosemiotics has many definitions. We give here the most famous ones. "Biosemiotics: (bios, life + semion = sign) is an interdisciplinary field of theoretical and empirical research, analyzing communication and signification in living systems. Signed processes, ranging from molecular to ecological and evolutionary, have been studied throughout the history of biology; however, very often descriptions of information and communication aspects of living systems were considered only metaphorical, believing that the essence of them can be understood with the help of physical and chemical descriptions. In biosemiotics, on the other hand, information sign processes are considered as the primordial, basic system of phenomena of life, requiring a new understanding..."3. "Biosemiotics explores sign systems of various levels: molecular biological (genetic code), intracellular (signal peptides), intercellular (mediators, immune interactions), intraorganism (hormones, conditioned reflex reactions) and interorganism (telergons, pheromones, attractants) ... In addition, biosemiotics covers all the problems associated with the problem of the existence of language and thinking in animals." However, today we can go further and add to the analysis the next stage of evolution, standing between animals and modern human (Homo sapiens sapiens).

Breast Cancer, Chemokines, And Metastasis: A Search for Decoy Ligands of the CXCR4 Receptor

Breast cancer (BC) is the leading cause of cancer-related deaths in young to middle-aged women worldwide. Moreover, the survival rate in BC-patients is only 20% when associated with metastatic disease. The high mortality rate observed in BC women with metastatic disease has precipitated a major challenge revealing an unmet need to develop new therapeutic strategies in treating metastatic cancer. One such approach has involved utilization of chemokines and their receptors as therapeutic targets for cancer metastasis. It has been established that a definitive correlation exists between overexpressed CXCR4 malignant cell receptors and cancer cell growth, invasion, and migration. It is also widely accepted that the CXCR4 receptor, complexed to its CXCL12 ligand, plays a major role in establishing migratory pathway gradients for cancer cells migrating to distant tissues/organ sites. It would follow that chemokine decoy ligands, such as peptide antagonists and inhibitors, could serve to induce receptor blockade and impede subsequent intracellular signaling. Such ligands, synthetic and natural, reportedly contribute to reducing cancer cell growth, invasion, adherence, and migration. The present commentary describes several existing synthetic CXCR4 receptor-ligand peptide antagonists and presents a strategy to develop naturally-occurring human protein-derived peptide candidates.

Molecular and Cell Biological Considerations in the Initiation and Development of Sporadic Non-Hereditary Solid Cancers  

This paper reviews the state of cancer research in the post-mutation era. It presents cancer as a highly complex disease viewed differently by scientists from various research fields. Histopathologists considered cancer as a disease of cell differentiation, cancer cell biologists overestimated the causal role of accumulated DNA mutations. More recently molecular biologists have focused on driver genes and driver mutations, regulatory gene networks and deregulation of the genomic balance between unicellular and multicellular gene sets (UG/MG balance). From a developmental biological standpoint, there is a clear analogy between the reproductive life cycles of cancer and protists. The key player of both analogous life cycles is the polyploid cyst, the atavistic cyst-like structure aCLS (PGCC). In the analogy to protists, we assume that the first aCLS initiating cancer originates from a mitoticly blocked cell (cell of origin of cancer, protoprecursor) that escapes death entering an atavistic reproductive process of polyploidisation and depolyploidisation; it forms the atavistic cyst-like structure aCLS and numerous daughter cells (microcells). The microcell progeny develops a multi-lined cell lineage containing stem cells as well as somatic and reproductive cells and clones. Subsequent aCLSs are formed sequentially by committed daughter cells or occasionally by stressed somatic cells. Accordingly, cancer initiation occurs by genomic changes leading to the amitotic cell state and reactivation of an atavistic life cycle. In humans, atavistic life cycles and hyperpolyploidisation (n >16) are mostly repressed by stable gene regulatory networks – but not in cancer. The permanent UG/MG gene conflict and robust ancient surveillance mechanisms trigger a cascade of molecular lesions leading to genomic heterogeneity and aberrant cancer cell states.

Statins and Lung Cancer: A Review of Current Literature

Cardiovascular disease and lung cancer are two of the most common causes of death in the United States. The cardioprotective benefits of statin class drugs is predominantly mediated through the inhibition of 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase, decreasing available mevalonate, and thus limiting in vivo cholesterol biosynthesis. Mevalonate and its metabolites have significant roles in cellular membrane synthesis, which is dysregulated during tumorigenesis, and is therefore a potential source for anti-tumor effects of statins. Similarly, dysregulation of cellular signaling is a hallmark of tumorigenesis. In vitro studies of EGFR, RAS, and AKT signaling pathways in cancer cells can all be reformed back to states more indicative of normally functioning cells when treated with statins. Statins have also been shown to exert beneficial properties in the presence of chemotherapeutic medications and radiation therapies by modulating the deleterious effects of reactive oxygen species, decreasing tumor cell resistance, and minimizing damage to surrounding native tissues. There is abundant of in vitro evidence to support the beneficial effects of statins on lung cancer patients. Prospective studies to determine the value of statin therapy on lung cancer prevention could lead to a significant change in lung cancer treatment.

Towards Precision Rheumatology?

The possibility of tailoring treatment on specific characteristics of patients – i.e. personalized medicine – has received attention in the field of rheumatic diseases since biological DMARDs targeting a unique pathway have become available. However the idea of personalized rheumatology has advanced slowly, at different paces in different disease groups, and it is only now surfacing in the recommendations for assessment and treatment of rheumatoid arthritis (RA). Many of the difficulties encountered stem from the recognition that many rheumatic diseases are not a single entity but encompass different subsets identified on the basis of genetic traits, cellular and molecular characterization both in blood and in tissues, laboratory markers and clinical manifestations (most notably in SLE). These differences suggest a multiplicity of pathogenetic triggers, whose various combination results in slightly or very diverse presentations. Developments in companion diagnostics and the identification of distinct subsets within complex syndromes are going to allow the definition of predictive biomarkers able to reduce poor treatment outcome, thus ensuring that we are treating “the right patient with the right drug”.

Substrate Stiffness Influences the Time Dependence of CTGF Protein Expression in Müller Cells

Following ocular trauma and retinal detachment, gliotic changes in the retina may develop over the subsequent month, a process known as PVR (proliferative vitreoretinopathy). There have been no successful therapeutic interventions to inhibit PVR. The protein CTGF (Connective Tissue Growth Factor) has been associated with retinal PVR and other fibrotic diseases of the retina in clinical studies but the mechanistic link between different pathologies and retinal gliosis has not been determined. In addition, CTGF has been previously noted to be associated, in some cases, with YAP/TAZ (Yes-associated protein and Tafazzin protein complex), transcriptional regulatory proteins that change subcellular localization in response to mechanical cues, such as the stiffness of the underlying material. We have previously shown that the mRNA for CTGF is markedly (100-fold) upregulated in retinal Müller cells grown on soft substrates. In order to evaluate if the mechanism by which mechanotransduction modulating CTGF production in retinal Müller cells involves the YAP/TAZ complex, this study tests the influence of substrate stiffness on the time dependence of CTGF protein expression, as well as subcellular localization of YAP/TAZ using a conditionally-immortalized mouse retinal Müller cell line plated on laminin-coated, polyacrylamide substrates of varying elastic modulus. Changes were assayed using immunohistochemistry and ELISA (Enzyme-Linked ImmunoSorbent Assay). In retinal Müller cells, the relationship between elastic modulus and the pattern of CTGF protein expression was bimodal, with CTGF levels rising more rapidly for cells on hard substrates and more slowly for cells grown on soft substrates. In addition, nuclear localization of YAP/TAZ corresponded directly to the maximum CTGF expression.

Acute and Chronic Wound Fluid Inversely Influence Wound Healing in an in-Vitro 3D Wound Model

If a wound progressively heals or the healing process is impaired is basically influenced by the surrounding milieu. This is reflected by the wound fluid. Its specific composition triggers the migration, proliferation and differentiation of dermal and epidermal cells which so far was not sufficiently examined in 2D cell culture models. The influence of the different wound entities was analyzed on a newly implemented three dimensional in-vitro model, which improved the transferability to the in-vivo situation. The influence of pooled wound fluids from patients suffering from acute or chronic wounds were investigated within a time period of 10 days after wound application. Histological and immunohistochemical analyses were performed addressing the impact of AWF and CWF on regeneration, such as cell proliferation, fibroblast activity and cell migration. AWF slightly stimulated fibroblast migration while CWF inhibited their activation and migration. The CXCR4- immunopositive population was continuously decreased compared to the control and AWF treatment. The expression of FAP was enhanced under AWF and medium. In keratinocytes CWF massively stimulated cell proliferation initiating on day six after injury. The presence of 10% CWF inhibited fibroblast activation and migration and induced the degradation of the collagen matrix. Keratinocytes were stimulated to proliferate, resulting in healing inhibiting hyperplasia. Transferred to human wounds, no effective wound closure would be achieved because of the de-regulation of pro-proliferative and migration-stimulating factors and a degraded extracellular matrix. This newly implemented 3D study model represents a novel appropriate in-vitro system for studying healing mechanisms and potential therapeutic applications.

Signal Transduction of hCG Induces Decidualization and Uterine Receptivity

All independent experimental data on epithelial and glandular cells lines of human endometrium support the evidence for a rapid production of eicosanoids from the LH/hCG receptors when exposed to the hCG hormone. Prostaglandins rapidly act on the surrounding endometrial stromal cells throughout the adenylyl cyclase enzyme leading to very large amounts of cAMP and angiogenic factors (VEGF) production. The cAMP is the most important intracellular second messenger and along with progesterone accomplishes the full process of decidualization and acquisition of receptivity after estrogenic priming of the endometrium. The status of uterine receptivity lasts few days only and timing for successful embryo-signal transduction system activation by the endometrium is probably short. In absence of in vivo embryonic signals it is impossible to predict, on individual bases, how the intensity of all the complex interlinked molecular changes of decidualization might ever be in case of exposure to native hCG. In other terms, amount of prostaglandins and cAMP produced in response to variably glycosylated hCG are all, in vivo, not measurable variables and should be viewed as a “wave” of biochemical chain reactions. Embryonic hCG is secreted in form of multiple isomers having an unpredictable variable level of glycosylation and control of this variable remains elusive. During cycles of ovarian stimulation many drugs (FSH, LH, HCG) interact with different G-protein coupled receptors (GPCRs) making it possible to alter the prostaglandins-mediated decidualization process ready to be elicited only by hCG of pregnancy. Since the molecules (cAMP and progesterone) controlling endometrial stromal cells differentiation into decidual cells are critical for successful implantation and placenta formation, the evidence of fast eicosanoids production associated with endometrial LH/hCG receptors exposure to hCG and the potential by human endometrium to produce, in response, very large amounts of cAMP has biological and clinical relevance.

Regulation of Expression of Reactive Oxygen Intermediates During Plasmodium Infection to Reduce Immunopathology Provides a Possible Antioxidant Adjuvant to Enhance Anti-Malarial Drug Therapy

Malaria is a mosquito-transmitted infectious disease caused by intracellular protozoan parasites of the genus Plasmodium. In the absence of prompt and appropriate treatment contraction of primary infection by a human being often represents a medical emergency since it may progress rapidly to life-threatening complications. Exposure to parasites activates the immune system resulting in, among other effects, the release of reactive oxygen intermediates (ROI). This has the potential to induce oxidative damage, thereby causing cellular destruction, and hence to have a severe effect on vital organs of the body. Overexpression of ROI leads to immunosuppression and is a causal factor in the development of malaria-related disease symptoms. However, the body possesses various defence mechanisms, notably including the production of antioxidants, which are capable of reducing the cellular effects of ROI. Antioxidants are either sourced exogenously from the diet or synthesized through different intracellular mechanisms. Antioxidants that include glutathione peroxidase, catalase, EDTA and vitamin C suppress the initial production of ROI. Others such as uric acid, superoxide dismutase and vitamin E may also inhibit potentially damaging products of ROI metabolism. Current anti-malarial drugs often have damaging side-effects, as exemplified by memory impairment following treatment for cerebral malaria. Recent studies have explored the potential use of antioxidants alone or in combination with anti-malarials as a therapeutic means to negate Plasmodium-induced oxidative stress and its associated metabolic complications. It is indicated that when utilized in an adjuvant capacity antioxidants of natural and synthetic origin may improve anti-malarial therapy by causing less damage to the host during malaria infection.

Shotgun Label-Free Proteomic Analyses of the Oyster Parasite Perkinsus Marinus

Perkinsus marinus is an intracellular parasitic protozoan that is responsible for serious disease epizootics in marine bivalve mollusks worldwide. Despite all available information on P. marinus genomics, more baseline data is required at the proteomic level. Our aim was to study the proteome profile of in vitro cultured P. marinus isolated from oysters Crassostrea spp. using a label-free shotgun UDMSE approach. A total of 4073 non-redundant proteins were identified across three biological replicates with stringent identification. Proteins specifically related to adaptive survival, cell recognition, antioxidants, regulation of apoptosis and others were detected. Important virulence factors of P. marinus were identified including serine protease and iron-dependent superoxide dismutase. Other proteins with involvement in several pathogens invasion strategies were rhoptries, serine-threonine kinases, and protein phosphatases. Interestingly, peptides corresponding to retroviruses polyproteins were identified in all replicates. The interactomic analysis of P. marinus proteins demonstrated extensive clusters network related to biological processes. In conclusion, we provide the first comprehensive proteomic profile of P. marinus that can be useful for further investigations on Perkinsus biology and virulence mechanisms.

The Role of Heparin in Lung Cancer

Non-small cell lung cancer is a major health problem worldwide. Surgery is still the mainstay of treatment especially in early stages of the disease. Despite the fact that surgery is the potentially curative treatment, the recurrence and mortality rates are still high specifically with more advanced stages of cancer. Heparin has been suggested to have a positive impact on the outcome of various cancers through its anticoagulants properties and; in some instances; due to their antitumor activity. Recently, the molecular mechanisms of tumor cell spreading have been recognised. Metastasis is a complex process that could be therapeutically affected wherever certain extra-cellular matrix proteins could play an important role in prevention of tumor cell migration and invasion. Experimental studies have shown decreased metastases development after heparin use in rat models. We have reviewed the literature to study the role of anticoagulants in cancer patients in general and in patients with Non Small Cell Lung Cancer (NSCLC) specifically.

The Assessment of Alchornea Cordifolia Activity in Aflatoxicosis Reduction in Rat.

Introduction: Aflatoxins are cytotoxic andserve as one of the key risk factors of hepatocellular carcinoma. Currently, plants and extract are widely used as potential scavenging substances for the detoxification of mycotoxins. Thus, this study aims to investigate the activity of the crude ethanolic leaves extract from Alchorneacordifolia in aflatoxicosis prevention. Material and Methods: The phytochemical screening was performed through qualitative analysis based on coloring and/or precipitation reactions. Groups of rats were treated daily with a mixture dose of aflatoxin B1 (AFB1) at 150 µg/kg and the crude extract of Alchorneacordifolia at doses of 50, 100, and 300 mg/kg for 21 days. The body weight, biochemical, and histological assessments were determined. Results: The phytochemical screening revealed the presence of polyphenols, flavonoids, sterols and terpenoids, quinoid compounds, tannins catechic and alkaloids. AFB1 treatmentcaused a significant increase of transaminases, urea, and creatinine abundances but reduced the rates of albumin and total proteins. Alchorneacordifolia administration alleviated biochemical parameters and body weight gain compared with the AFB1 group (p<0.05). The histological lesions of organs (liver and kidney) caused by AFB1 were significantly improved after administration of the extract at a dose of 300 mg/kg. Conclusion: This plant plays a beneficial role in AFB1-induced injury and may be used in the treatment of aflatoxicosis.

Rescuing Canavan Disease by Redirecting Metabolic Processing: Support for the Astrocyte Hypothesis of Canavan Disease Generation and A Possible Human Cure

Canavan disease (CD) is a globally occurring but rare human spongiform leukodystrophy that is associated with inborn errors affecting the activity of aspartoacylase (ASPA), an enzyme highly expressed in oligodendrocytes that hydrolyzes N-acetylaspartate (NAA). Lack of ASPA activity is associated with the inability of oligodendrocytes to build or maintain axon-enveloping myelin sheaths. The primary source of NAA in brain is neurons, cells that synthesize but cannot catabolize it. Neurons also synthesize N-acetylaspartylglutamate (NAAG) from NAA and glutamate but cannot catabolize this substance as well. For their metabolism, these substances are released to extracellular fluid and are metabolized by oligodendrocyte ASPA and astrocyte NAAG peptidase respectively. A hypothesis developed suggested that the cause of the leukodystrophy component in CD was due to release of NAAG by neurons at white matter nodes of Ranvier, its catabolism by astrocytes forming NAA and increased osmotic-hydrostatic pressure as a result of its buildup at these nodes due to the lack of ASPA activity. In this communication, we provide evidence supporting this hypothesis and comment on the cause and possible cure for human CD.

Dual Effects of Neuroprotection and Neurotoxicity by General Anesthetics on Neural Stem Cells: Role of Autophagy

General anesthetics (GAs) are widely used for various essential surgical or medical procedures. Recent studies implicate the GAs has dual effects of neuroprotection and neurotoxicity on neurogenesis with unclear mechanisms. This minireview summarizes recent studies on GAs mediated effects on neurogenesis and proposed mechanisms, with focus on autophagy regulation and intracellular calcium homeostasis.

Methionine and Antioxidant Potential

Methionine (Met) is a nutritionally essential amino acid and has been widely demonstrated to improve cellular oxidative balance and mediate oxidative stress. Met targets reactive oxygen species (ROS) directly by being oxidized to Met sulfoxide (MetO) 1. Met can be metabolized to cysteine (Cys) through transsulfuration pathway, which is further metabolized to glutathione (GSH), taurine, and hydrogen sulfide (H2S). All these metabolites exhibit antioxidant functions in various models (reviewed at 2). More recently, Met also has been demonstrated to enhance cellular oxidative tolerance via pentose phosphate pathway (PPP) 3, which contributes to the balance of cellular reducing power and accelerates the reduction reaction of MetO and GSH oxidative product GSSH back to Met and GSH.

Family with Sequence Similarity 213, Member A is Associated with the Fertility Selection Index in Holsteins

Female fertility is an economically important trait in the dairy industry, and the fertility selection index has been developed as a method of including female fertility in the breeding goals of this industry. This index considers a combination of factors, including days open, number of inseminations per lactation, success after first insemination, and pregnancy within 70 d, 90 d, and 110 d after parity. Based on a genome-wide association study of the fertility selection index using 442 Holsteins, we found that the index is influenced by a variation in the thioredoxin fold region of the family with the sequence similarity 213, member A (FAM213A) protein. FAM213A is a CXXC motif-containing peroxiredoxin 2-like protein that regulates cellular redox status. A replacement of isoleucine with valine in FAM213A was associated with poor fertility in cows. The overexpression of FAM213AVal in bovine endometrial epithelial cells reduced reactive oxygen species to a lesser extent relative to the overexpression of FAM213AIle and caused a decrease in cyclooxygenase-2 expression. Downregulation of cyclooxygenase-2 led to a decline in prostaglandin E2, which is critical for implantation because it protects the conceptus from the maternal immune system. Cows with FAM213AVal showed lower levels of prostaglandin E2 than did cows with FAM213AIle, suggesting that cows with FAM213AVal are less fertile than cows with FAM213AIle because of their reduced uterine environment. Thus, the present study found that FAM213A unexpectedly modulates female fertility in cattle.

Evidence that N-acetylaspartylglutamate is the Astrocyte-Targeted Neurovascular Coupling Agent that Regulates Slow Tonic Control of Brain Blood Flow

N-acetylaspartylglutamate (NAAG) is the highest concentration dipeptide present in brain. It is found primarily in neurons but its function is unclear. NAAG is synthesized by neurons from N-acetylaspartate and glutamate (Glu), maintained at mM concentrations and is released non-synaptically to extracellular fluid (ECF). NAAG is a non-excitatory form of Glu, and is targeted to the metabotropic group II Glu receptor 3 (mGluR3) on the surface of astrocytes. After docking with the receptor, Glu is released by the action of NAAG peptidase. Previously, it was shown for the first time that an NAAG-peptidase inhibitor reduced global cerebral blood flow (CBF) in mouse brain but did not affect their physical performance. Recently, it has been demonstrated that there are two separate systems involved in neurovascular coupling by astrocytes, one is a rapid focal phasic response providing energy for stimulation-induced neuronal activity, and the other a slower global tonic response providing energy for ongoing metabolic activities. Many neurovascular coupling mechanisms are known that regulate phasic changes in CBF, but how the brain accomplishes tonic control is unknown. In this paper we bring together two separate lines of inquiry, the decades’ long search for the function of NAAG, and the more recent search for the mechanism of tonic neurovascular control. Herein, we present evidence that NAAG is the neurovascular coupling agent that regulates tonic changes in CBF via the astrocyte mGluR3-NAAG peptidase connection.

An Update on Hemocytes in Biomphalaria Snails

The hemocyte is a major immunological cell of molluscs. Much of the immunological phenomena associated with molluscan immunology can be attributed to cellular immunity associated with these cells suspended in the hemolymph. These cells are often referred to as amoebocytes or hemocytes. Such cells are of great importance to immune mechanisms associated withBiomphalaria snails. The Biomphalaria snail is the main vector of the important trematode parasite Schistosoma mansoni. This is a waterborne parasite that affects about 200 million people globally and puts countless other millions at risk of infection. Larval stages of the parasite are released from the snail in tainted waters and the larval cercarial stage actively penetrates the skin of humans and other vertebrates. Larvae migrate via the venous system to vital organs associated with the heaptic portal and mesenteric blood vessels. Larvae develop into sexually mature male and female adult worms that live in major venous blood vessels. The worms mate and produce eggs that lodge in major organs such as the spleen, liver, and intestines. Eggs produce extensive granulomas that cause cirrhosis and other pathological conditions in the affected organs.

Rapidly Progressing Visual Loss Associated with Optic Nerve Head Drusen: Is there A Role For Lowering Intraocular Pressure?

Background: Optic nerve head drusen are acellular hyaline deposits located anterior to the lamina cribrosa, frequently associated with visual field defects. Sometimes rapid worsening of vision may occur due to complications such as acute vascular events, choroidal neovascularization, or serous maculopathy. Case Presentation: Although there are no proven treatments for Optic nerve head drusen associated field loss, we present the case of a patient with Optic nerve head drusen and bilateral rapid progression of visual field loss that has stabilized on intraocular pressure lowering medication. This suggests a role for IOP-mediated retinal ganglion cell loss in this individual. The mechanism of progressive Optic nerve head drusen associated field loss is poorly understood, however experimental glaucoma models and human in vivo imaging studies have shown that structural differences within the optic nerve head are likely to contribute to individual susceptibility to IOP-mediated damage. Conclusion: We propose that eyes with Optic nerve head drusen may be less able to dampen IOP mediated stress, contributing to loss of retinal ganglion cells in some patients.

The Chromosomal and Functional Clustering of Markedly Divergent Human-Mouse Orthologs Run Parallel to their Compositional Features

It was, previously, reported that the specific pattern of the compositional features of particular human-mouse orthologs defining in human two clusters, named C2 and C5, are present in different clusters in mouse. Since, thus, these orthologs can harbor a significant number of nucleotide differences a large sample of human-mouse orthologs having in human the C2 and C5 compositional features were collected in order to identify the orthologs that have been conserved or diverged during speciation. From the collection, 945 and 1051 orthologs had in human the C2 and C5 profile, respectively, while in mouse only 77 and 125, respectively, had these profiles. We further analyzed whether or not the frequency-usage of trinucleotides having the same gross composition computed from the reading of all nearest-neighbors of the DNA sequence might convey a layer of biological information in terms of chromosomal topology and function. In human, more than 50% of the C2 and C5 genes were found distributed in six chromosomes and preferentially located in GC-rich bands of chromosomes 11, 16 and 19. It was, also, found that 80% of the entire set of genes of band 19p13.3 had the C2 and C5 profile. The data shown also indicate that the proteins codified by the C5 genes have a bias towards nucleus and cytoplasm and specific post-translational modifications while the proteins codified by the C2 genes are mainly located in the cellular membrane or secreted to the external cellular milieu and particular post-transcriptional modifications

Ciliary Neurotrophic Factor Activated Signaling Pathways in Retinal Müller Cells

Ciliary neurotrophic factor (CNTF) is a well-tested, neuroprotective agent that has been shown to retard photoreceptor degeneration in several animal models of retinitis pigmentosa. The molecular mechanisms underlying CNTF-mediated neuroprotection are currently not understood. CNTF could act directly on photoreceptors or it could act indirectly by stimulating Müller glial cells to produce photoreceptor neuroprotective agents. To better characterize CNTF action on Müller cells, we have studied signaling pathways activated by CNTF using an established retinal Müller cell line, rMC-1. RNA was isolated from CNTF-treated cultures, and suppressor of signal transducer and activator of transcription (SOCS3) and Glial fibrillary acidic protein (GFAP) transcript levels were assessed by quantitative real-time PCR. Immunoblotting was used to examine activation ofmitogen activated protein kinase (ERK1/2/MAPK) and phosphoinositide 3-kinase (PI3-K)/Aktpathways in response to CNTF. Additionally, the level of5' AMP-activated protein kinase (AMPK), an enzyme that plays a key role in cellular energy homeostasis levels, was determined by immunoblotting. CNTF treatment resulted strong upregulation of SOCS3 and GFAP transcripts that were blocked by expression of a dominant-negative STAT3 mutant. CNTF treatment also resulted in transient activation of ERK1/2/MAPK but not PI3K/Akt signaling pathway. There was no change in activation of AMPK. We conclude that CNTF treatment leads to stimulation of JAK-STAT and MAPK signaling pathways but not the PI3K/AKT pathway, associated with cell death, in Müller cells.

Cholesterol-Conjugated siRNA Accumulates in the Different Hematopoietic and Lymphoid Cells.

Small interfering RNA (siRNA) based drugs for overcoming multiple drug resistance of hematological malignancies could solve the problem of poor response to the chemotherapy and hight relapse rate. The main factor that significantly limits biomedical application of siRNA is inefficient delivery to target cells and tissues. The attachment of siRNA to molecules, which enter into the cell by natural transport mechanisms, can improve cellular uptake of siRNA. In current study the carrier-free cellular uptake of siRNA containig cholesterol residues conjugated to the 5’-end of the sense strand via oligomethylene linker of various length (here and after Ch-siRNA) was explored. The data demonstrate that cholesterol residue increase the accumulation of siRNA in all tested cell lines and the primary cells. The efficiency of Ch-siRNA accumulation in K562 cells depends greatly on the leangth of the linker connecting cholesterol and siRNA: Ch-siRNAs with linker of 10 - 12 methylene units accumulate the most efficiently in this cells. It was found that Ch-siRNA effectively accumulates in MOLT-3 (acute lymphoblastic leukemia, ALL), HL-60 (acute myelogenous leukemia, AML), K562 (chronic myelogenous leukemia CML) and primary peripheral blood mononuclear cells (PBMC) from patient with non-Hodgkin lymphoma (NHL) or healthy donor resulting in near 100% of transfected cell when used at 1 mM concentration.

Oxidative Telomere Attrition, Nutritional Antioxidants and Biological Aging

Telomeres are strings of DNA that are not themselves genes but that extend every chromosome beyond its last gene. Terminal telomeres are sacrificed during every mitotic event in human cells (“telomere attrition”), preserving the functional genome despite the “end replication problem.” However, the “telomeric theory of biological aging” suggests that when an individual cell has reproduced itself a sufficient number of times (the “Hayflick limit”), some the its telomeres have become critically shortened (“telomeric crisis”) and cannot completely “cap off” a chromosome, and any further attempts to replicate such a chromosome would produce damaged DNA and a dysfunctional cell (“cellular aging”). As cells enter telomeric crisis, they usually initiate intracellular signaling cascades that arrest DNA replication and mitotic activity, converting biologically active cells into inactive cells (“cellular senescence”). The progressive accumulation of senescent cells impairs the healthy functioning of tissues and produces “biological aging.” Oxidative stress damages telomeres and accelerates telomere attrition and biological aging. Premature biological aging is associated with degenerative diseases and diminished quality of life. Reducing the level of systemic oxidative stress can ease the oxidative drive toward cellular senescence and premature biological aging. Increased intakes of antioxidant-rich foods and specific antioxidant nutrients (such as fruits and vegetables, α -lipoic acid, astaxanthin, eicosapentaenoic acid, docosahexaenoic acid, trans-resveratrol, N-acetylcysteine, methylsulfonylmethane, lutein, vitamin C, vitamin D, vitamin E, and γ-tocotrienol) may decrease cellular and systemic oxidative stress and decelerate biological aging.

Correlation of Oncotype DX Recurrence Score with the Expression of Insulin Receptor Substrate Proteins in Estrogen Receptor + Breast Cancer

Insulin receptor substrate (IRS) 1 and 2 are downstream signaling proteins that influence breast pathophysiology. IRS-1 promotes carcinoma cell proliferation; whereas IRS-2 regulates cell motility, invasion, and glycolysis. Our lab has shown that distinct cellular localization of IRS-2 also plays a role in carcinoma cell function. Oncotype DX (Genomic Health) (ODX) is a 21-gene expression profile used to classify carcinomas with low, intermediate, and high risk recurrence scores (RS). Our aim is to correlate expression and cellular localization of IRS proteins in breast carcinomas with their ODX RS. 97 breast carcinomas sent for ODX testing from 2006-2009 were collected and grouped according to their RS (low, intermediate or high). Immunohistochemistry for IRS-1/-2 was performed. Specific criteria were used to evaluate IRS staining patterns. Follow-up data, ranging from 3-6 years, was available. Statistical analysis was performed to correlate staining patterns of IRS-1/-2 with the three RS groups. IRS-1 staining, predominantly nuclear, did not significantly correlate with RS (P=.5645). IRS-2 expression patterns did show statistical significance amongst the three RS groups (P=.0371). Tumors with intermediate and low RS were more likely to exhibit punctate and diffuse cytoplasmic expression of IRS-2, and cell membrane expression was uncommon in this group. Expression and cellular localization of IRS proteins play an important role in breast cancer cell biology, and expression patterns for IRS-2 do demonstrate a significant correlation with ODX RS. Further studies are required to elucidate the significance of cellular localization of IRS-1/-2 proteins in breast carcinoma cells and their relationship to ODX scores.

Phytohemagglutinin and Light-Induced Charge Density Effects on Plasma Membrane of PlectonemaBoryanum

Effects of the plant lectin phytohemagglutinin M (PHA-M) and illumination on the electrophoretic mobility (EPM) and 90°-light scattering of suspension of spheroplasts obtained by lysis of the unicellular diazothrophic cyanobacteriumPlectonemaboryanumwere studied. Cells cultures were grown on media with different iron content: an iron-deficient (denominated as “Fe-starved” culture), an iron-sufficient (“Fe-sufficient”), and a twentyfold iron excess medium (“Fe-excess”). Lectin addition led to a decrease of the membrane charge density and aggregation of the vesicles. Our results indicate significant influence of the Fe2+ and illumination on the lectin-induced decreasing of EPM and light scattering intensity.

Carbofuran Abrogates the Membrane Ca++-Atpase Activity in Liver: The Role of Cholesterol

Carbofuran is a broad spectrum pesticide used in agricultural fields and domestic places throughout the world. It is one of the deadly toxic carbamate pesticide that kills the pest by inhibiting the crucial enzyme of nervous system known as acetyl cholinesterase. In the present study, we report how carbofuran increases the different spectrum of cholesterols, including free cholesterol and esterified cholesterol in the fish hepatocytes. It is observed that induced-cholesterol can inhibit the enzymatic activity such as Ca++-ATPase, which is a critical protein for maintaining the calcium homoeostasis in the cellular microenvironment. Carbofuran integrates into human body through foods and drinks. As trace of carbofuran is identified in our daily food and drinks, we examined the homology of Ca++-ATPase between the fish and human, so our data can illuminate the effects of carbofuran on this crucial enzyme. While studying the homology with the help of bioinformatics, we recognized that there is around 70% homology in the protein sequence of Ca++-ATPase between fish Heteropneustesfossilisand human (Homo sapiens), which appears as sufficient to simulate our fish-model data in human. This study demonstrates that carbofuran affects our day-to-day life by inhibiting Ca++-ATPase through modulation of lipid synthesis, a critical regulatory system that controls overall homeostasis in our body.

The Role of FIP-2 (Optineurin) in Regulation of the Chemokines and Kinases

FIP-2 is a multifunctional protein which is involved in various cellular processes. Using different approaches we investigated its regulatory activity. The microarray analysis has shown that FIP-2 substantially altered the expression of 75 genes (35+/40-) from different functional groups with maximal presentation in “Signal transduction” and “Transcription regulation”. Real time RT-PCR indicated significant elevation in the transcription of chemokines, particularly IL-8 (CXCL8). Production of IL-8 in HEK293 cells dramatically increased with FIP-2 overexpression. We also demonstrated that FIP-2 induced activation of IL-8 promoter activity through NF-kB binding site. Additionally, we showed that FIP-2 could interact with PAK3 and increase its kinase activity. Overall, we demonstrated the role of FIP-2 in the regulation of chemokines (IL-8, MPIF-1, MCP-1) and kinases (PAK3, ALK).

Evaluating the Efficacy of Gene Silencing in Dopaminergic Neuronal Cells In-Vitro using Gold Nanorods (GNR) with Different Surface Properties Complexed to DARPP-32 siRNA.

Gold nanorods (GNRs) are plasmonic nanostructures by virtue of their size-dependent optical properties, offer a bionanotechnology platform in areas of bioimaging, drug delivery etc for disease diagnosis, prognosis, and therapy. GNRs are more sensitive to changes in local environments, and offer strong scattering and absorption efficiencies thus providing opportunities to integrate multiple imaging modes and therapeutic strategies. The hydrodynamic size of these GNR under physiological condition is <100 nm, making them ideal as intracellular delivery agents. RNA interference using small inhibitory RNA (siRNA) has become a powerful tool to downregulate mRNA levels by cellular nucleases that become activated when a sequence homology between the siRNA and a respective mRNA molecule is detected. siRNA is used to silence genes involved in the pathogenesis of various diseases and holds a promising option for the development of novel therapeutic strategies in neurological dysregulation such as that observed in drug addiction. However, a major challenge in gene therapy continues to be effective delivery of siRNA and its sustained release at targeted sites. Previously, we have shown the GNR coated with poly (diallyldimethyl ammoniumchloride) (GNR-PDDAC) electrostatically complexed to the dopamine- and cAMP-regulated neuronal phosphoprotein (DARPP-32) siRNA forming a GNR-nanoplex that was able to effectively silence the DARPP-32 gene expression in dopaminergic neuronal (DAN) cell cultures in- vitro. The current report, explores if modification of the surface coating properties of the GNRs with different surface coatings namely, amino terminated polyethylene glycol (GNR-PEG), polyethyleneimine (GNR-PEI) and Chitosan (GNR-CIT) alters their stability, cytotoxicity and DARPP-32 gene silencing efficiency in-vitro dopaminergic neuronal (DAN) cell cultures with the goal of determining the most suitable surface coating for the GNR that would provide a GNR-nanoplex with the most stability, least cytotoxicity and most efficacious gene silencing.

Differences in the Alveolar Macrophage Proteome in Transgenic Mice Expressing Human SP-A1 and SP-A2

Surfactant protein A (SP-A) plays a number of roles in lung host defense and innate immunity. There are two human genes, SFTPA1 and SFTPA2, and evidence indicates that the function of SP-A1 and SP-A2 proteins differ in several respects. To investigate the impact of SP-A1 and SP-A2 on the alveolar macrophage (AM) phenotype, we generated humanized transgenic (hTG) mice on the SP-A knockout (KO) background, each expressing human SP-A1 or SP-A2. Using two-dimensional difference gel electrophoresis (2D-DIGE) we studied the AM cellular proteome. We compared mouse lines expressing high levels of SP-A1, high levels of SP-A2, low levels of SP-A1, and low levels of SP-A2, with wild type (WT) and SP-A KO mice. AM from mice expressing high levels of SP-A2 were the most similar to WT mice, particularly for proteins related to actin and the cytoskeleton, as well as proteins regulated by Nrf2. The expression patterns from mouse lines expressing higher levels of the transgenes were almost the inverse of one another – the most highly expressed proteins in SP-A2 exhibited the lowest levels in the SP-A1 mice and vice versa. The mouse lines where each expressed low levels of SP-A1 or SP-A2 transgene had very similar protein expression patterns suggesting that responses to low levels of SP-A are independent of SP-A genotype, whereas the responses to higher amounts of SP-A are genotype-dependent. Together these observations indicate that in vivo exposure to SP-A1 or SP-A2 differentially affects the proteomic expression of AMs, with SP-A2 being more similar to WT.

Optimization and ZSPORE Analysis of Affinity Purification Coupled with Tandem Mass Spectrometry in Mammalian Cells

Defining protein-protein interactions is essential for understanding the mechanisms by which cells regulate basic functions, such as metabolism, transcription, and signal transduction. Affinity purification followed by tandem mass spectrometry (AP-MS) has application for discovery of new interactors regulating various cellular processes. Here we optimize the purification method for AP-MS and develop a simplified unbiased analytical tool, Z-score plus prey occurrence and reproducibility (ZSPORE) for data analysis. Using this pipeline we achieve a higher efficiency of AP-MS and enhanced identification of high confidence interacting proteins (HCIP) in mammalian cells. When applied to analysis of the innate immune interactome, these methods enhanced HCIP identification. In addition, we investigated the GRB2 complex, which is associated with signal transduction and cell growth. Twenty-four known GRB2 interacting proteins were identified plus 26 new GRB2 binding partners. Thus, these straightforward methods recapitulate known protein interactions, discover novel complexes, and allow mapping of protein interaction networks.