NGF Genes Research Articles

The genetic characterization of germplasm and identification of the litter size trait associated candidate genes in Dexin mutton and fine-wool sheep.

Xinjiang is a major province of sheep breeding in China, which plays an important role in meeting people's needs for meat products, increasing farmers' income and sustainable development of animal husbandry. However, the genetic differentiation relationship between breeds was not clear, and most sheep had low fecundity, which seriously restricted the efficient development of sheep industry. Therefore, this study used the whole genome resequencing to detect the genetic variation of Dexin mutton and fine-wool sheep, explored the selected regions and important genes of the litter size traits, analyzed the genetic mechanism of reproductive traits, and provided new insights for the high fecundity breeding of sheep. A total of 5,236.338 G genome data and 35,884,037 SNPs were obtained. Furthermore, we identified 39 selection signals spanning candidate genes, 99 genes were significantly associated related to growth, reproduction and immunity, among which, BRIP1, BMPR1B, BMP4, NGF, etc. genes, and MAKP signaling pathway, Fanconi anemia pathway and Thyroid hormone signaling pathway and other signaling pathways were significantly correlated with litter size trait. Among them, we identified NGF, TrKA and BRIP1 genes was the important genes for sheep litter size traits and the mutation frequencies of 9 SNPs in BRIP1 gene were significantly different in domestic sheep in the world. The research provided new insights for the breeding of self-cultivated meat fine-wool sheep.

Effects of valproic acid on transcript levels in neurotrophin signaling pathway in mice hippocampus according to the implementation period

Abstract Background/Aim Valproic acid (VPA), is an antiepileptic drug and it has been known for a long time that exposure to VPA in the fetal period causes many behavioral, cognitive, and structural disorders. In this study, we aimed to evaluate the effects of the implementation period of VPA during pregnancy on both behavioral tests and the expression of 45 key genes belonging to the neurotrophin signaling pathway. Methods The study was designed to form four different groups of five female mice in each group. The experimental and control groups were divided into mature (n = 5) and immature (n = 5) subgroups, respectively. VPA (500 mg/kg dose) was subcutaneously administered to the experimental groups. All mice were first tested for behavioral changes, and then the hippo-campal tissues were used for RNA isolation. The expression levels of 45 transcripts associated with the neurotrophin signaling pathway were determined by real-time RT-PCR in the hippocampus. Results No differences were found between the groups in any of the behavioral tests. When gene expression was examined, there was a difference between the groups in the transcript levels of Ngf(neurotrophic growth factor), Ntrk2 (neurotrophic receptor tyrosine kinase 2), Shc1 (SHC adaptor protein 1), Chka (choline kinase alpha), and Mapk1 (mitogen-activated protein kinase 1) genes. Conclusions This study demonstrated the effects of VPA on transcript levels at different maturation stages. We believe that it is important to explain the structural and cognitive teratogenic side effects caused by VPA in human fetuses using genes whose expression changes.

Tempol maintained the cellular integrity of the cerebellar cortex by preserving neuron survival, autophagy, glial cells, and synapses after cisplatin exposure

Backgroundtempol is a free radical scavenger that passes through the blood-brain barrier. This study aimed to assess its ameliorative role in the cerebellar cortex after cisplatin injection. Methodsthe examined forty-eight adult male wistar albino rats were divided into four groups. The saline group received 0.5 ml of normal saline, the tempol group was injected with 0.5 ml of tempol (100 mg/kg/day), the cisplatin group was injected with cisplatin single dose (7 mg/kg) on the 8th day, and cisplatin + tempol group received combined treatment. All groups were dosed through intraperitoneal injections. Resultshistological examination revealed meningeal congestion and thickening with excess collagen, discrete vacuolated cortical cells, with pyknotic nuclei, and tissue loss after cisplatin. Immunohistochemical expression of NF-kβ and GFAP were enhanced, while LC3-II and synaptophysin expression decreased with cisplatin. Cisplatin treatment reduced mRNA expression of NGF, GLP-1, BDNF, PGC1-α, and PPAR-α while it boosted Caspase-3 expression. Moreover, it tripled the level of MDA and lowered levels of SOD, CAT, and GSH in cerebellar tissue homogenate. Tempol supplementation restored the meningeal and the normal histological structure of the cerebellar cortex. The immunohistochemical as well as mRNA expressions of different genes were highly normalized but still showed variable significant differences when compared to the control except for NGF, PGC1-α, and Caspase-3 genes. Great restoration of biochemical markers was evident with tempol, especially for CAT that showed no significant difference in comparison with the saline group. Conclusiontempol cisplatin combination can enhance neuronal survival, promote autophagy, decrease astrogliosis, and preserve synapses.

The Effects of Regular Sport and Nanofibrous Conduit on Peripheral Nerve Repair: An In Vitro and In Vivo Study

In the current research, the effects of a regular sport program was investigated on peripheral nerve regeneration in a rat model. The critical sized peripheral nerve gap was created in a rat model and the nerve injury gap was bridged by an electrospun polycaprolacton/collagen-based neural conduit. After bridging the nerve gap, the animals received a regular exercise three times a week until week 8. Study showed that the regular exercise program significantly augmented the level of sciatic nerve repair as evidenced by histopathological examinations and functional recovery experiments. Gene expression studies showed that rats treated with regular exercise had higher expression levels of BDNF, NGF, and b-FGF genes.

The Effects of Temperature and Humidity Index on Growth Performance, Colon Microbiota, and Serum Metabolome of Ira Rabbits

This study investigates the effects of different THI values on growth performance, intestinal microbes, and serum metabolism in meat rabbits. The results showed that there were significant differences in THI in different location regions of the rabbit house. The high-THI group (HG) could significantly reduce average daily gain and average daily feed intake in Ira rabbits (p < 0.05). The low-THI group (LG) significantly increased the relative abundance of Blautia (p < 0.05). The HG significantly increased the relative abundance of Lachnospiraceae NK4A136 group and reduced bacterial community interaction (p < 0.05). The cytokine–cytokine receptor interactions, nuclear factor kappa B signaling pathway, and toll-like receptor signaling pathway in each rabbit’s gut were activated when the THI was 26.14 (p < 0.05). Metabolic pathways such as the phenylalanine, tyrosine, and tryptophan biosynthesis and phenylalanine metabolisms were activated when the THI was 27.25 (p < 0.05). Meanwhile, the TRPV3 and NGF genes that were associated with heat sensitivity were significantly upregulated (p < 0.05). In addition, five metabolites were found to be able to predict THI levels in the environment with an accuracy of 91.7%. In summary, a THI of 26.14 is more suitable for the growth of meat rabbits than a THI of 27.25, providing a reference for the efficient feeding of meat rabbits.

Identification of founder and novel mutations that cause congenital insensitivity to pain (CIP) in palestinian patients

BackgroundCongenital insensitivity to pain (CIP) is a rare autosomal recessive disorder characterized primarily by an inability to perceive physical pain from birth, resulting in the accumulation of bruising, inflammation, and fractures that affect patient’s life expectancy. CIP has different forms including CIP and CIPA. CIP with Anhidrosis (CIPA) is the most common type of CIP, which is caused mainly by mutations in NTRK1 and NGF genes, and is characterized by mental retardation and the inability to sweat (Anhidrosis). Because of high consanguinity rates in Palestine, this rare disease appears to have a higher frequency than in other communities. However, there were no systematic studies to address the genetic factors that cause CIP in the Palestinian community.MethodsIn our study, we used Sanger and Whole exome sequencing to genotype members of five CIP-affected Palestinian families.ResultsOur results confirm the presence of the founder c.1860-1861insT mutation in the NTRK1 gene of Palestinian Bedouin CIPA patients. Furthermore, one CIPA family carried a missense c.2170 G > A (G724 S) mutation in exon 16 of the NTRK1 gene. Finally, a novel nonsense c.901 A > T mutation (K301*) was detected in exon 7 of the SCN9A gene in CIP without anhidrosis family.ConclusionsOur study revealed three mutations that cause CIP and CIPA in the Palestinian community, which can help in improving the process of diagnosis and genetic counseling and establishing protocols for the diagnosis and follow-up for the affected individuals. This is especially important given that early diagnosis and medical care interference can prevent unpleasant CIP and CIPA complications.

Catha Edulis Extract Loaded Nanofibrous Neural Channels Augment Sciatic Nerve Injury Repair Trough Upregulation of BDNF and NGF Genes

The study investigated the impact of regular treadmill exercise and catha edulis extract-loaded nanofibrous scaffolds on rats with sciatic nerve injury. The findings indicated that PC-12 cells survived on the membranes and exhibited greater metabolic activity on the catha edulis extract-loaded nanofibrous membranes. Furthermore, the combined approach improved sciatic nerve healing, as confirmed by histopathological examinations and functional recovery in vivo. Additionally, gene expression studies revealed an increase in BDNF and NGF genes due to this approach.

The Sirtuin 2 Inhibitor AK-7 Leads to an Antidepressant-Like Effect in Mice via Upregulation of CREB1, BDNF, and NTRK2 Pathways.

Depression is one of the most important and serious health problems in developing countries which affects millions of people. It is associated with the decrease of the quality of life as well as suicides and mortality. The disease may show recurrent episodes in some patients. Obviously, not all the patients with depression could be treated properly, because some individuals are drug-resistant and the options for the therapy are limited. Therefore, it is crucial to investigate new molecules and pathways that may have possible antidepressant activity. Sirtuin (SIRT), known as a class III histone deacetylase, which is regulated by nicotinamide adenine dinucleotide (NAD +), is one of these molecules. In the current study, we investigated the possible antidepressant-like effect of SIRT2 inhibitor AK-7. For this purpose, behavioral tests were performed in chronic AK-7-treated mice, and the expression levels of BDNF, NGF, NTF3, CREB, NTRK2, ERK1, ERK2, and GAP43 genes were evaluated by qRT-PCR analysis in brain tissues. Protein levels for BDNF, CREB1, and NTRK2 were determined by western blot. Our data showed that AK-7 significantly decreased immobility time and showed antidepressant-like effect. In addition, AK-7 treatment significantly increased mRNA levels of CREB and NTRK2 and protein levels of CREB1, BDNF, and NTRK2. Finally, our results suggest that SIRT2 and AK-7 may have a potential role in the cellular mechanisms of depression.

Effect of Azithromycin on Sciatic Nerve Injury in the Wistar Rats.

After a severe peripheral nerve injury, complete functional recovery is rare. Modulating the inflammatory response could be an effective way to enhance peripheral nerve regeneration. The present study aimed to determine the effect of azithromycin on functional recovery following sciatic nerve crush in Wistar rats. 40 male Wistar rats were used in four groups, including: the negative control, sham, and two groups of azithromycin (15 and 150mg/kg/day) (n = 10).The rats' right sciatic nerve was crushed using a non-serrated clamp. In experimental groups, animals were treated with azithromycin (15 and 150mg/kg/day) for 7 days. Then, sensory-motor functions were evaluated over eight weeks. Real-time PCR was used to measure the expression of NGF and BDNF genes. At the end of the 4th week, the sensory recovery accelerated in the azithromycin-treated rats so that the reaction times in the groups treated with 15mg/kg and 150mg/kg doses of azithromycin reached 5.14s and 6.61s, respectively, which were significantly lower than the 12s in the negative control group (P < 0.05).Eventually, the mean SFI values in the negative control and both azithromycin-treated groups recovered to preoperative levels in the 8th week, with no significant difference between the sciatic lesion groups. Findings showed a seven-day course of azithromycin administered immediately after a sciatic nerve crush could accelerate regeneration and improve motor and sensory function recovery compared to negative controls. These significant effects were observed in both the azithromycin 15mg/kg and the azithromycin 150mg/kg treatment groups. Azithromycin treatment upregulated the expression of NGF and BDNF genes in crushed sciatic nerve. Our findings suggest that a seven-day treatment of azithromycin after a sciatic nerve injury could accelerate the regeneration process and improve functional recovery.

Network pharmacology identify intersection genes of quercetin and Alzheimer's disease as potential therapeutic targets.

BackgroundCurrently, there are no efficient therapies for Alzheimer’s disease (AD) among the elderly, although it is the most common etiology of dementia among the elderly. Quercetin, which has a variety of therapeutic properties, may pave the way for novel approaches to AD treatment. In the AD patients’ frontal cortex, current study aims to identify the potential mechanisms of quercetin’s pharmacological targets.Materials and methodsThe pharmacological targets of quercetin have been studied from DrugBank and SwissTarget. In order to distinguish AD-associated genes targeted by quercetin (Q-ADGs), we utilized an integrated intersection of gene expressions of the frontal cortex in combination with transcriptome analysis. To detect cortex-related Q-ADGs and immune-related Q-ADGs, a drug screening database and the immune infiltration analysis was utilized. The Q-ADGs were then linked with the AD severity scores (MMSE scores) to find severity-associated Q-ADGs. In addition, the miRNA-seq datasets were examined to identify severity-associated Q-ADG-miRNAs. Twelve genes, more frequently related to AD by previous studies among all the genes identified in the present study, were subjected to the verification of qRT-PCR in AD cell model.ResultsIn the frontal lobe of AD, 207 Q-ADGs were discovered and found that axonogenesis, glial differentiation, and other biological processes had been enriched. There were 155 immune-related Q-ADGs (e.g., COX2, NOS2, HMGB1) and 65 cortex-related Q-ADGs (e.g., FOXO1, CXCL16, NOTCH3). Sixteen Q-ADGs (e.g., STAT3, RORA, BCL6) and 28 miRNAs (e.g., miR-142-5p, miR-17-5p) were found to be related to MMSE scores. In the qRT-PCR results, six out of twelve genes were significantly regulated by quercetin. DYRK1A, FOXO1, NOS2, NGF, NQO1, and RORA genes were novel target of quercetin in AD. DYRK1A, NOS2, and NQO1 genes targeted by quercetin have benefits in the treatment of AD. However, FOXO1, NGF, and RORA genes targeted by quercetin might have a negative impact on AD.ConclusionThe role of quercetin in AD appears to be multifaceted, and it can affect patients’ frontal cortex in a variety of pathways, such as axonogenesis, immune infiltration, and glial cell differentiation. DYRK1A, NOS2, and NQO1 might be potential novel effective drug targets for quercetin in AD.

A Prognostic Model for Predicting Tumor Mutation Burden and Tumor-Infiltrating Immune Cells in Bladder Urothelial Carcinoma.

Tremendous progress has been made in development of immunotherapeutic approaches for treatment of bladder urothelial carcinoma (BLCA). However, efficacy and safety of these approaches remain unsatisfactory, necessitating further investigations for identification of indicators for predicting prognosis and efficacy. In this study, we downloaded transcriptomic and clinical data of BLCA patients from The Cancer Genome Atlas (TCGA) database, and identified differentially expressed genes (DEGs) between tumor and normal tissues. We incorporated these DEGs in an intersection analysis with immune-related genes (IRGs) obtained from the Immunology Database and Analysis Portal (ImmPort) database, and identified immune-related DEGs. These genes were subjected to Cox and least absolute shrinkage and selection operator (LASSO) regression analyses, then a prognostic model containing AHNAK, OAS1, NGF, PPY and SCG2 genes was constructed, for prediction of prognosis of BLCA and efficacy of immunotherapy. Finally, we explored the relationship between the prognostic model and tumor mutational burden (TMB), abundance of tumor-infiltrating immune cells (TICs) and immunotherapeutic targets, and found that patients with higher risk score (RS) had poorer prognosis and significantly lower levels of TMB. Patients in the low-RS group exhibited higher numbers of lymphoid cells, whereas those in the high-RS group exhibited higher proportions of myeloid cells. However, patients with high-RS tended to respond better to immunotherapy relative to those in the low-RS group. The constructed prognostic model provides a new tool for predicting prognosis of BLCA patients and efficacy of immunotherapy, offering a feasible option for management of the disease.

Detection of Species DNA in Chicken Meatball Products Using NGF Genes as Molecular Markers

Species DNA detection in chicken meatball products using the NGF gene as a molecular marker was carried out to see if the NGF gene could be used to test species DNA detection in chicken meatball processed food products. The test method used in this study is the SYBR green test method which is read using real-time PCR and the extraction technique used is the centrifuge column technique. The test results in this technique can be in the form of Ct (Cycling) and Tm (Melting Temperature) values. The results showed that the DNA isolation carried out showed that the concentration and purity of the isolated DNA were in the range of 51,100 ng/µL – 52,300ng/µL with an average of 51,883 ng/µL. As for the value of purity is the absorption value measured at wavelength A260/A280, the results are obtained in the range between 1,850 – 1,920 with an average of 1,880. The results of the real-time PCR analysis obtained the Ct value of the chest sample at 21.10, the Ct LOD value at 25.20 and the positive control Ct value at 20.30. For the Tm value of the busty sample at 78.20, the Tm LOD value at 78.80 and the positive control Tm value at 79.10. Based on the results of this study, it can be concluded that the genetic markers of the NGF gene can be used to test specific DNA detection of chicken species in processed chicken meatball products so that this test method can be used to detect species DNA.

The Effect of Valproic Acid on the Transcriptional Activity of Ngf and Bdnf Genes of in Vitro Cultured Neurons Under Oxidative Stress Conditions

Brain-derived neurotrophic factor (BDNF) is a secretory molecule that promotes peripheral neurons synaptic transmission and plasticity by TrkB receptor activation. This is shown in cultured central nervous system (CNS) neurons, including hippocampal and cortical cholinergic, dopaminergic and serotonergic neurons. Hypotheses suggesting that BDNF may play a potential role in the pathophysiology of schizophrenia are based on the key role of BDNF in the synaptic plasticity and, consequently, regulation of cognitive functions. In the schizophrenia treatment valproic acid is used in complex combined therapy regimens. Treatment of schizophrenia patients with valproate increases the BDNF level. Since it is not yet clear whether the BDNF protein levels measured in serum samples and in the brain correlate, we investigated valproate effects on the cultured neurons Bdnf transcription level. The primary neuron-glia culture was obtained from the cerebellum of 8-9-day-old Wistar rats. Valproic acid was added to the neurons (at a concentration of 50 µg/ml), oxidative stress was stimulated by 40 µMof H2O2, and injury was caused by mechanical damage to the neuron culture. It was shown that valproic acid in 3-24 hours increases the transcriptional activity of the Bdnf and Ngf (nerve growth factor) genes 2–2.5-fold (p&lt;0.01) and approximately 1.5-fold (p&lt;0.01), respectively. Mechanical trauma, unlike oxidative stress, activates the transcriptional activity of the Ngf and Bdnf genes (p&lt;0.01). However, under oxidative stress and mechanical damage to neurons, the effect of valproic acid on the Ngf and Bdnf genes expression was insignificant. Fluorescence microscopy analysis using specific antibodies to neurons (anti-Map-2) showed that in the presence of valproic acid, the number of neuronal processes and contacts between them significantly increased. Evidently, valproate addition to antipsychotics can be effective for the overall clinical response. Relatively little research has been done on the signaling pathways in neurons that are activated by the valproic acid. However, we have obtained evidence of activation of the Ngf and Bdnf genes transcription in cultured neurons in vitro. We also found that in the presence of valproic acid, the number of neuronal processes and contacts between them significantly increased. However, we have also found that the oxidative stress accompanying the schizophrenia can significantly reduce the valproic acid effect on the Ngf and Bdnf genes expression. The results of the study may be potentially useful for new schizophrenia therapy strategies development.

Ovarian Expression Profile of KISS1 and NGF Genes in Indian Goat

Background: Litter size is one of the most important economic traits that translates into higher returns for farmers. KISS 1 and NGF genes plays significant role in female reproduction. The purpose of the study was to evaluate the potential variation in the of expression levels of KISS1 and NGF genes in ovarian tissue of two different goat breeds with diverse prolificacy. Methods: Phenotypic records pertaining to litter size of Barbari and Jamunapari goats were collected. Total RNA was extracted from both breeds' ovarian tissues for real-time PCR quantification of the KISS1 and NGF genes, using GAPDH and β-actin as endogenous controls. The target gene’s expression levels were measured and the fold-change was determined. Result: The phenotypic recording revealed that Barbari goats are more prolific in nature than Jamunapari goats. The expression levels of KISS1 and NGF genes were higher in Barbari goat and lower in Jamunapari goat (P less than 0.05). The results of this study can be used to further discover the crucial role of KISS1 and NGF genes in reproduction for improving the prolificacy in goats.

Expression of neurotrophic factor genes by human adipose stem cells post-induction by deprenyl.

Human adipose stem cells (hASCs) were introduced as appropriate candidate due to advantages like ease of isolation, in vitro expansion and lack of immune response. Deprenyl (Dep) was used to induce bone marrow stem cells into neuron-like cells. We investigated the Dep effect on neurotrophin genes expression in hASCs and their differentiation into neuron-like cells. The cells were isolated from small pieces of abdominal adipose tissue and subjected to flow cytometry to confirm purification. The osteogenic and adipogenic differentiation were identified. The proliferation rate and neurotrophin genes expression of treated cells were evaluated by MTT, TH immunostaining and RT-PCR. hASCs had positive response to CD44, CD73, CD90, CD105 markers and negative response to CD34 and CD45 markers and differentiated into adipocytes and osteocytes. Exposure to 10–7 M of Dep for 24 hours caused a significant increase of viable cells and BDNF, NTF-3 genes expression as compared to cultured cells in serum free medium and had no effect on the expression of NGF and GDNF genes. Based on our results, Dep is able to induce BDNF, NTF-3 and NTF-4 genes expression and neroun-like morphology in hASCs.

Effects of Eight Weeks of Resistance Exercises on Neurotrophins and Trk Receptors in Alzheimer Model Male Wistar Rats.

Introduction:This study evaluates the effects of 8 weeks of resistance exercises on the expression of neurotrophins and Trk receptors in Alzheimer model male Wistar rats.Methods:For this purpose, 32 mature male Wistar rats with a mean weight of 230–280 g were chosen and divided into Alzheimer and Sham groups. The rats in the sham group received normal saline, while the ones in the Alzheimer group received streptomycin via intraventricular injection. These rats were then divided into the following four subgroups: 1) resting sham, 2) exercising sham, 3) resting Alzheimer, and 4) exercising Alzheimer. The two exercising rat subgroups exercised three times a week for 8 weeks. A weight was attached to their tails, and they had to carry it on a 26-step ladder in each cycle. Resting groups were handled every day to minimize the effects of stress. At the end of the eighth week and 24 hours after the last exercise session (to avoid the effects of the last exercise session), the rats were put under deep anesthesia and beheaded. Hippocampus tissues were precisely extracted, and samples were sent to the laboratory for molecular and cellular tests. To investigate gene expression, quantitative RTPCR was used.Results:The tests for comparing the mean values of BDNF, NT3, NGF, TrkA, and TrkB in two rat groups showed that with error levels of less than 5%, there is a significant difference in the amounts of BDNF, NT3, NGF, TrkA, and TrkB between exercising rats and resting ones. These amounts were much higher in the exercising Alzheimer rats group.Conclusion:Eight weeks of resistance exercises increased the expression of BDNF, NT3, and NGF genes and TrkA and TrkB receptors in Alzheimer model Wistar rats.

Fingolimod increases oligodendrocytes markers expression in epidermal neural crest stem cells

Epidermal neural crest stem cells (EPI-NCSCs) are propitious candidates for cell replacement therapy and supplying neurotrophic factors in the neurological disorders. Considering the potential remyelinating and regenerative effects of fingolimod, in this study, we evaluated its effects on EPI-NCSCs viability and the expression of neurotrophic and oligodendrocyte differentiation factors. EPI-NCSCs, extracted from the bulge of rat hair follicles, were characterized and treated with fingolimod (0, 50, 100, 200, 400, 600, 1000, and 5000 nM). The cell viability was evaluated by MTT assay at 6, 24 and 72 h. The expression of neurotrophic and differentiation factors in the cells treated with 100 and 400 nM fingolimod were measured at 24 and 120 h. Fingolimod at 50–600 nM increased the cells viability after 6 h, with no change at the higher concentrations. The highest concentration (5000nM) induced toxicity at 24 and 72 h. NGF and GDNF genes expression were decreased at 120 h, but on the contrary, brain derived neurotrophic factor (BDNF) and neurotrophin 3 (NT3) were increased by both concentrations at both time points. Oligodendrocyte markers including platelet-derived growth factor receptor A (PDGFRα), neuron-glial antigen 2 (NG2) and growth associated protein 43 (GAP43) were elevated at 120 h, which was accompanied with reduce in stemness markers (Nestin and early growth response 1 (EGR1)). Fingolimod increased the expression of neurotrophic factors in EPI-NCSCs, and guided them to oligodendrocyte fate. Therefore, fingolimod in combination with EPI-NCSCs, can be considered as a promising approach for demyelinating neurological disorders.

549-P: Transplantation of Human Dental Pulp Stem Cells Ameliorates Diabetic Polyneuropathy via Increasing Angiogenic and Neurotrophic Gene Expression in the Transplanted Muscles

Transplantation of cryopreserved DPSCs ameliorated diabetic neuropathy equal to freshly isolated DPSCs. Human DPSCs (hDPSCs) can be isolated from teeth extracted at a young age and can be cryopreserved until use. The aim of this study is to clarify the therapeutic mechanism of hDPSCs transplantation on diabetic polyneuropathy. We collected human impacted third molars from adult at Aichi-Gakuin University hospital. Written informed consent was obtained from each donor. Identification of hDPSCs was analyzed by FACS and differentiation capabilities. Diabetes was induced by injection of STZ in 6 week-old BALB/cAJcl-nu/nu mice. Eight weeks after STZ injection, hDPSCs were transplanted into unilateral hindlimb skeletal muscles of normal and diabetic mice. Four weeks after transplantation, sciatic blood flow (SNBF), sciatic motor/sensory nerve conduction velocity (MNCV/SNCV) and current perception threshold (CPT) were assessed. To elucidate the therapeutic effects of hDPSCs, diabetic mice were treated with neutralizing antibody soon after transplantation. Immunohistological and gene expression analysis of hindlimb skeletal muscles were also performed at the end of the experiments. Diabetic mice showed significant reductions in SNBF, MNCV and SNCV and increase in CPTs in the control side compared with normal mice. Transplantation of hDPSCs significantly ameliorated the impaired SNBF, MNCV, SNCV and CPTs in the hDPSCs-injected side. The transplanted hDPSCs were located around the muscle bundles and expressed the human VEGF and NGF genes. Capillary/muscle bundle ratio was also increased in the hDPSCs-injected side. Furthremore, the effects of hDPSCs transplantation were cancelled by neutralizing antibodies of VEGF and NGF. These results suggest that cell therapy using hDPSCs may be useful for treatment of diabetic neuropathy via the angiogenic and neurotrophic factor by hDPSCs secretion. Disclosure M. Hata: None. M. Omi: None. N. Nakamura: None. M. Miyabe: None. M. Ito: None. E. Makino: None. S. Kanada: None. T. Ono: None. Y. Imanishi: None. T. Himeno: None. J. Nakamura: Research Support; Self; Astellas Pharma Inc., Boehlinger Ingelheim Japan Co., Ltd., Daiichi Sankyo, Eli Lilly Japan K.K., Japan Tobacco Inc., Kaken Pharmaceutical Co., Ltd., Kowa Company, Ltd., Kyowa Hakko Kirin Co., Ltd., Mitsubishi Tanabe Pharma Corporation, MSD K.K., Novartis Pharma K.K., Novo Nordisk Pharma Ltd., Ono Pharmaceutical Co., Ltd., Otsuka Pharmaceutical Co., Ltd., Pfizer Japan Inc., Sanofi K.K., Sanwa Kagaku Kenkyusho, Shionogi &amp; Co., Ltd., Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited. Speaker’s Bureau; Self; Abbott Japan Co., Ltd., ARKRAY, Astellas Pharma Inc., AstraZeneca K.K., Boehlinger Ingelheim Japan Co., Ltd.,, Daiichi Sankyo, Eli Lilly Japan K.K., Fukuda Denshi, Kissei Pharmaceutical Co., Ltd., Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation, MSD K.K., Mylan, Novartis Pharma K.K., Novo Nordisk Pharma Ltd, Ono Pharmaceutical Co., Ltd., Sanofi, Sanwa Kagaku Kenkyusho, Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited, Terumo Medical Corporation. H. Kamiya: Speaker’s Bureau; Self; Astellas Pharma Inc., AstraZeneca K.K., Boehringer Ingelheim K.K., Daiichi Sankyo, Eli Lilly Japan K.K., Fukuda Denshi, Kissei Pharmaceutical Co., Ltd., Kowa Company, Ltd., Kyowa Hakko Kirin Co., Ltd., Mitsubishi Tanabe Pharma Corporation, MSD K.K., Novartis Pharma K.K., Novo Nordisk Pharma Ltd., Ono Pharmaceutical Co., Ltd., Sanofi K.K., Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited. S. Ozawa: None. J. Takebe: None. T. Matsubara: None. K. Naruse: None.

NGF (\u2212198C\u2009>\u2009T, Ala35Val) and p75NTR (Ser205Leu) gene mutations are associated with liver function in different histopathological profiles of the patients with chronic viral hepatitis in the Brazilian Amazon

BackgroundsNeural growth factor (NGF) is a neurotrophin that can interact with the p75NTR receptor and initiate a cascade of reactions that determines cell survival or death, and both are associated with the physiology of liver tissue. Single nucleotide polymorphisms (SNPs) in the NGF and p75NTR genes have been investigated in different pathologies; however, there are no studies that have analyzed their biological roles in the hepatic microenvironment. In the present study, we evaluated the impact of SNPs in these genes on the maintenance of liver function at different stages of inflammation and fibrosis in patients with chronic viral liver disease in the Brazilian Amazon.MethodsThe SNPs -198C > T, Arg80Gln, Val72Met, Ala35Val, Ala18Ala and Ser205Leu were genotyped by real-time PCR in samples from patients with chronic viral hepatitis stratified by stage of inflammation and liver fibrosis. Histopathological, viral load (VL), liver enzyme and comorbidities data were obtained from updated medical records. Other aspects were highlighted by applied epidemiological questionnaires.ResultsThe -198C/T and Ala35Val polymorphisms in NGF were associated with changes in histopathological profiles, VL and liver enzymes. Ser205Leu polymorphism in p75NTR was associated only with changes in VL and liver enzymes. Polymorphic frequencies were variable among different ethnic populations, mainly for biologically relevant polymorphisms. A multifactorial network of interactions has been established based on genetic, virological, behavioral and biochemical aspects.ConclusionMutations in the NGF (−198C > T, Ala35Val) and p75NTR (Ser205Leu) genes, within the list of multifactorial aspects, are associated with liver function in different histopathological profiles of patients with chronic viral liver disease in the Brazilian Amazon.

Oxytocin/Osteocalcin/IL-6 and NGF/BDNF mRNA Levels in Response to Cold Stress Challenge in Mice: Possible Oxytonic Brain-Bone-Muscle-Interaction.

Oxytocin (Oxt), osteocalcin (Ost), and NGF/BDNF have a role in bone homeostasis, reproduction, and cognition. Oxt/Ost is required for muscle repair. We investigated gene response of muscle and the inter-organ communication following cold stress (CS). The mRNA quantity of Ngf, Ost, Oxt, Bdnf, p75ntr, Ntrk1, Gprc6a, Oxtr, Ntrk2, UCP1, and Il-6 genes in bone, brain, soleus (SOL), and tibialis anterior (TA) muscles from adult mice following CS were investigated. The myosin heavy-chain Mhc2b, Mhc1, Mhc2x, and Mhc2a gene expression were investigated. Mice were maintained at T = 23°C or 4°C for 6 h and 5-days (5d). CS mice did not show signs of muscle degeneration. An upregulation of Ucp1 and Ngf genes by 2 and 1.5 folds, respectively, in TA after 6 h CS and Ntrk1 by 4 and 22 folds in SOL muscle after 6 h and 5d CS, respectively, was observed; while after 6 h CS p75Ntr was downregulated in either muscle. Bdnf was unaffected, while after 5d CS Ntrk2 was upregulated in TA. Ost was downregulated in SOL by 0.9-folds at 5d. Following 5d CS, Oxtr and Il-6 genes were upregulated, respectively, by 1 and 1.5 folds in SOL. A downregulation of Mhc2b, respectively, by 0.96 and 0.88-folds after 6 h and 5d CS in SOL and Mhc2a was also downregulated by 0.88-fold after 5d CS in TA. Mhc1 and Mhc2x were not affected. Changes in the expression levels of genes in TA and SOL muscles, bone, and brain following CS were regulated by IL6 and Oxt. CS potentiates the slow-twitch phenotype of SOL which is in line with the metabolic need of this muscle, and the potentiation of the slow-twitch phenotype in TA. Oxt and IL6 coordinate a phenotype-dependent tonic effect of slow-twitch muscle and Oxt regulates the inter-organ interaction between brain and SOL muscle. Muscle tropism is maintained by NGF signaling following CS.