Hormone Gene Research Articles

Reclassifying tumour cell cycle activity in terms of its tissue of origin

Genomic alterations resulting in loss of control over the cell cycle is a fundamental hallmark of human malignancies. Whilst pan-cancer studies have broadly assessed tumour genomics and their impact on oncogenic pathways, analyses taking the baseline signalling levels in normal tissue into account are lacking. To this end, we aimed to reclassify the cell cycle activity of tumours in terms of their tissue of origin and determine if any common DNA mutations, chromosome arm-level changes or signalling pathways contribute to an increase in baseline corrected cell cycle activity. Combining normal tissue and pan-cancer data from over 13,000 samples we demonstrate that tumours of gynaecological origin show the highest levels of corrected cell cycle activity, partially owing to hormonal signalling and gene expression changes. We also show that normal and tumour tissues can be separated into groups (quadrants) of low/high cell cycle activity and propose the hypothesis of an upper limit on these activity levels in tumours.

Mapping Growth Hormone Gene of Body Weight Krui Cattle in Pesisir Barat Regency Lampung, Indonesia.

<b>Background and Objective:</b> The growth hormone (GH) gene plays a role in meat growth and has been shown to increase the growth rate and carcass composition after being given GH. For this function, this gene is used as a strong candidate for genetic markers for meat growth traits. The research objective was to map the growth hormone (GH) gene of the bodyweight of Krui cattle in the Pesisir Barat Regency. <b>Materials and Methods:</b> This research used 30 blood samples of 30 Krui cattle. The method used was by taking quantitative data and blood samples from adult Krui cattle in Pesisir Barat Regency and then the blood samples were analyzed by DNA isolation method. PCR amplification used was a pair of GH-Forward primers: 5 'ATC CAC ACC CCC TCC ACA CAGT 3' and GH- reverse: 5 'CAT TTT CCA CCC TCC CCT ACA G 3', as well as digestion using the RFLP method at the Laboratory of Animal Breeding and Genetics of Universitas Gadjah Mada, Yogyakarta. Association between genotype and body weight was analyzed descriptively. <b>Results:</b> The results showed that Krui cattle had polymorphic genes with three genotypes found, namely: CC, CT and TT. Cattle with CT genotype had the largest average body weight or meat production compared to those with other genotypes. <b>Conclusion:</b> These results indicated that the GH gene identifier has strong evidence that it can be used as a selection tool with the help of genotypes of body weight traits of Krui meat production in the Pesisir Barat Regency. Krui cattle with CT genotype can be developed further because it has high economic value with high average body weight and meat production.

An Insertion Variant in CRH Confers an Increased Risk of Central Serous Chorioretinopathy.

PurposeTo identify a novel corticotropin-releasing hormone (CRH) gene variant relevant in patients with central serous chorioretinopathy (CSC).MethodsWe performed a genetic study of CSC in families and sporadic cases with controls. Using whole-exome sequencing and linkage analysis, we identified a heterozygous insertion variant, Gln52insPro, in the CRH gene that cosegregated in two Chinese families with CSC. This variant was evaluated among an additional 1307 patients with CSC and 1438 ethnicity-matched control individuals from three independent Chinese cohorts.ResultsThe CRH variant was strongly associated with CSC in these cohorts of Chinese patients (Pmeta = 1.24 × 10−11; odds ratio, 3.01; 95% confidence interval, 2.15–4.21). The risk variant Gln52insPro decreased CRH gene expression.ConclusionsOur results implicate the hypothalamic–pituitary–adrenal stress response system in the pathogenesis of CSC and provide a novel rationale for therapeutic intervention.

Role of the silymarin on the prolactin gene expression in the pituitary, estrogen and progesterone hormones of western rats during lactation

Objective: To see how dose of Silymarin aqueous extracts affected prolactin hormone gene expression in pituitary glands and Androgen hormone in serum samples of female experimental rats throughout nursing. Methods: 24 adult healthy, Two sets of pregnant rats were created (12 each group) and treated as follows for ten days postnatal. G 1 were intubated orally distal water serving as control while, G 2 were intubated orally Silymarin (200 mg\kg\day). Results: The findings show that Silymarin had a strong influence on mRNA–PRL expression in pituitary glands, with a rise in Estrogen and a decline in Progesterone levels. Conclusion: The focus of this study mention a new evidence of the role of Silymarin on the prolactin gene expression for increasing the milk production and level of estrogen & progesterone hormones.

Effect of thermal conditioning on serum electrolytes, metabolites, corticosterone and expression of CRH gene in selected chicken strains.

Early age thermal conditioning has been found to improve thermotolerance in birds. This study assessed the effect of perinatal thermal conditioning on serum parameters, corticosterone, free fatty acid, globulin and expression of corticotropin-releasing hormone (CRH) gene in five chicken strains; using fifty chicks per strain of Cobb 500 (C500), Ross 308 (R308), Shika Brown (SB), Normal Feathered Nigeria Indigenous (NF) and FUNAAB Alpha (FA). Twenty-five chicks per strain were conditioned at 40 ± 1°C for 3h on day 6. On day 10, both conditioned and unconditioned chicks were challenged acutely at 40 ± 1°C for 15min, without feed and water. Body weight and feed intake data were collected before and after the heat exposures. Blood samples were collected to determine serum electrolytes, metabolites and corticosterone levels. Brain tissue samples were collected from the 10-day-old conditioned and unconditioned chicks, from which RNA were extracted, synthesized into cDNA and subjected to qPCR. Serum parameters were significantly affected (p < 0.05) by strain, thermal conditioning and their interactions. Calcium and glucose concentrations were highest in NF while FA had highest in sodium. Calcium, glucose and phosphorus were higher in conditioned birds. NF had the highest free fatty acid while FA had the lowest. C500 had the highest globulin levels. Thermal conditioning significantly lowered corticosterone levels in conditioned birds. CRH was shown to be overexpressed in C500. From this research, it can be concluded that early age thermal conditioning affects body temperature regulation in chickens and enhances thermotolerance.

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Editorial introductions

Expression of the Growth Hormone Gene during Early Development of Tiger Shovelnose Catfish (Pseudoplatystoma Fasciatum Linnaeus, 1766)

The early development stage of fish is crucial in fish farming, especially catfish species with cannibal characteristics, such as the Tiger Shovelnose Catfish (Pseudoplatystoma fasciatum Linnaeus 1766). Growth plays an essential role in fish’s life, which is influenced by the mechanism of growth hormone (GH). This study analyzed the level of growth hormone (GH) gene expression at several early developmental stages of Tiger Shovelnose Catfish. Samples were taken from an embryo, three days post hatched (dph) larvae to 60 dph juvenile. Isolation of mRNA was carried from the pituitary gland located at the head of the fish. Gene expression analysis was performed using RT-PCR method that compared GH/b-actin genes from band thickness. The results of the analysis showed the highest expression at 60 dph juvenile. The most significant increase in GH gene expression occurred from the egg to three dph larvae (P &lt; 0.05), then decreased to 20 dph juvenile. The increased phenotypic growth was seen from the increased length and weight at 45 dph juveniles. This indicates the amount of growth hormone released at the juvenile stage of 20 dph, thus triggering rapid growth in the next stage at 45 dph juvenile. Growth hormones can act as growth biomarkers in Tiger Shovelnose Catfish. The novelty of this study was to determine the level of GH gene expression in Pseudoplatystoma fasciatum ornamental fish as the object of research. Suggestions for further research to explore genes related to growth mechanisms through transcriptomic and proteomic technologies.

Leptin Promotes Primordial Follicle Activation by Regulating Ovarian Insulin-like Growth Factor System in Chicken.

Leptin and insulin-like growth factor 1 (IGF-1) regulate follicle development and reproduction in vertebrates. This study investigated the role played by leptin and IGF-1 in primordial follicle activation in the ovary of 7-day-old chicks. Different doses of leptin were intraperitoneally administrated to female layer chicks, and further analyses were performed. While leptin administration did not affect hepatic leptin receptor (LEPR), growth hormone receptor (GHR), or IGF-1, the lower dose of leptin significantly increased the messenger RNA (mRNA) expression of IGF-1, IGF-1 receptor, and IGF-binding protein (IGFBP)-2 and attenuated anti-Müllerian hormone (AMH) gene expression in the ovary. Furthermore, the ovaries of the same age chicks were challenged with leptin and/or IGF-1 in vitro. Leptin at a lower dose increased the mRNA expression of IGF-1, LEPR, and leptin; 100 ng/mL leptin and 10 ng/mL IGF-1 alone or combined with leptin reduced IGFBP-2 mRNA expression. AMH gene expression was also reduced by all doses except 10 ng/mL leptin. Histological studies showed that a lower dose of leptin injection induced the primordial follicle growth in the ovary in vivo, and the number of primordial follicles was higher in all leptin treatments over control in vitro. Moreover, the luciferase assay revealed that leptin enhanced IGF-1 promoter activity in LEPR-expressing CHO-K1 cells. Collectively, these results indicate that leptin directly affects the IGF-1/IGFBP system and promotes primordial follicular growth in the ovary of early posthatch chicks. In addition, the follicular development by leptin-induced IGF-1 is, at least in part, caused by the suppression of AMH in the ovary.

Association of GH gene polymorphism with meat quality traits in meat and wool sheep

Increasing the productivity and effi ciency of lamb production is a key factor in the competitiveness of the meat sheep industry. Therefore, in order to conduct sheep breeding more effectively it is necessary to apply new approaches to increase and improve the quality of products.Considerable interest are modern technologies involving the use of DNA markers, which are widely used in national breeding programs of a number of countries with developed animal husbandry and have a signifi cant impact on improving productivity indicators. A significant number of genes associated with meat productivity and meat quality have been identifi ed. However, in sheep breeding, information about the main genes or loci that affect the growth characteristics and productive traits of sheep is relatively limited. Due to the growing interest in the production of young mutton and lamb, priority is given to the study of genes that control meat productivity. The greatest interests are studies on the evaluation of the polymorphism of the growth hormone (GH) gene, which controls growth and development, as well as infl uencing the meat productivity of sheep. In this regard, the purpose of this work was research aimed at fi nding associations between polymorphic variants of the growth hormone gene and the qualitative characteristics of meat in meat and wool sheep of the genotype ½ Poll Dorset × ½ North Caucasian Meat and Wool breed bred in the Stavropol Territory. The data obtained when studying the histostructure of the m. Longissimus dorsi, depending on the genotypes of GH gene, indicate that the muscle tissue in individuals with AB and BB genotypes was characterized by a large number of muscle fi bers by 5,7 and 6,4 %, their smaller diameter by 7,6 and 9,2 %, and a lower content of connective tissue compared to animals with the genotype AA of GH gene.

Age-associated changes in gene expression in the anterior pituitary glands of female Japanese black cattle.

Proper functioning of the anterior pituitary (AP) gland is imperative, however, is suppressed by aging via unclear mechanisms. Therefore, we identified differentially expressed genes (DEGs) in the AP glands of Japanese Black young heifers (approximately 22months old) compared to old cows (approximately 120months old) via deep sequencing of the transcriptome (RNA-seq) to characterize potentially important pathways. The young and old AP glands expressed 20,171 annotated genes. Of the total transcripts per million, approximately 41.6% and 35.5% were the sum of seven AP hormone genes in young and old AP glands, respectively, with difference observed in the sum between the young and old AP glands (P &lt; 0.05). Moreover, we identified 48 downregulated genes and 218 upregulated genes in old compared to young AP glands (P &lt; 0.01, fold change &gt; 120%). The DEGs included 1 cytokine (AIMP1), 3 growth factors (NRG2, PTN, and TGFB1), 1 receptor-associated protein gene (AGTRAP), and 10 receptor genes, including PRLHR and two orphan G-protein-coupled receptors (GPR156 and GPR176). Metascape analysis of the DEGs revealed "Peptide metabolic process," "Regulation of hormone levels," and "Peptide hormone processing" as enriched pathways. Furthermore, Ingenuity Pathway analysis of the DEGs revealed (1) a network of 24 genes (including GPR156 and PRLHR) named "Neurological disease, organismal injury and abnormalities, and psychological disorders", and (2) two canonical pathways (P &lt; 0.01), namely "Huntington's disease signaling", and "AMPK signaling". Thus, the findings of the current study revealed relevant DEGs, while identifying important pathways that occur during aging in AP glands of female cattle.

Metabolome and transcriptome profiling reveal regulatory network and mechanism of flavonoid biosynthesis during color formation of Dioscorea cirrhosa L.

Dioscorea cirrhosa is a plant that is used as a dye as well as in medicine. Many metabolites with pharmacological activity exist in the tubers of D. cirrhosa. However, little is known about the mechanism regulating biosynthesis in these metabolites. In this study, transcriptome and metabolome profiling were performed in four color tubers. A total of 531 metabolites, including 62 flavonoids, were identified. Epicatechin and proanthocyanin B2 were the key metabolites that exhibited high content levels in the four tubers. These metabolites were divided into nine classes with distinct change patterns. A total of 22,865 differentially expressed genes (DEGs) were identified by transcriptome analysis. Among these DEGs, we identified 67 candidate genes related to the flavonoid biosynthesis pathway and three genes that played pivotal roles in proanthocyanin (PA) synthesis. A weighted gene co-expression network analysis (WGCNA) revealed that the two modules, “MEblue” and “MEblack,” were two key gene sets strongly associated with phenylpropanoid and flavonoid biosynthesis. We also found that the plant hormone signal transduction biological process exhibited activity in the late stage of tuber color formation. Additionally, we identified 37 hub transcript factors related to flavonoid biosynthesis, of which 24 were found to be highly associated with flavonoid pathway genes. In addition to the MYB-bHLH-WD40 (MBW) genes, we found that the plant hormone gene families exhibited high expression levels. This study provides a reference for understanding the synthesis of D. cirrhosa tuber metabolites at the molecular level and provides a foundation for the further development of D. cirrhosa related plant pigments as well as its further use in the pharmaceutical industry.

P-778 High altitude significantly affects pregnancy, implantation, live birth, and miscarriage rates in patients subjected to in vitro fertilization procedures with donated oocytes

Abstract Study question Is high altitude an important factor that affects the clinical outcomes in patients subjected toin vitrofertilization with donated oocytes? Summary answer Pregnancy, implantation, and live birth rates are significantly impacted by paternal chronic hypoxia in IVF procedures involving egg donors. What is known already High altitude is characterized by low temperatures, high UV radiation, and a hypoxic environment that significantly affect human physiology, including endocrine, cardiovascular, metabolic, and reproductive functions. Hypoxia can affect male reproduction differently, such as at the hormonal level, during spermatogenesis and gene expression. More importantly, hypoxia can affect key events during early embryo development, with time of exposure being the determining factor in the stage of fertilization and embryonic development in mammals. Study design, size, duration The following work is a retrospective nonrandomized study based on data from 85 IVF donor egg cycles performed at high altitude (Huancayo-Peru, 3,300 meter above sea level;n = 42) and at sea level (Guayaquil-Ecuador;n = 43) between July 2019 and December 2020. The same protocols of culture and embryo classification were used in both assisted reproduction laboratories. Participants/materials, setting, methods Anonymous oocytes donors (18-29 years old) underwent physical, gynecological and psychological examinations, with no family histories of hereditary or chromosomal diseases reported. According to seminal samples, IVF or ICSI was elected as the insemination procedure. Extended embryo culture until blastocyst stage was done in all procedures. Pregnancy, implantation, live birth, and miscarriage rates were compared between groups. Main results and the role of chance Out of a total of 983 included oocytes, 529 corresponded to high level patients; and 454, to sea level patients. Fertilization rates (95.1% vs 92.1%) and good-quality embryos at day 3 (79.7% vs 83.8%) were similar between high altitude and sea level groups (P &amp;gt; 0.05). The number of transferred embryos in recipients at high altitude was significantly higher compared to those transferred in recipients at sea level (1.90±0.29 vs 1.62±0.49) (P &amp;lt; 0.05). Blastocyst formation rates (39.3% vs 44.3%), pregnancy rates (61.9% vs 71.4%), implantation rates (48.8% vs 64.7%) and live birth rates per embryo transfer (46.3% vs 65.7%) were significantly lower in the patients at high altitude compared to those from sea level (P &amp;lt; 0.05). Significantly more miscarriages were observed at high altitude (23.1% vs 6.5%) (P &amp;lt; 0.05). Seminal parameters were similar between both evaluated groups. Limitations, reasons for caution The retrospective nature of this study and the small sample size may be reason for caution. Wider implications of the findings The negative effect of high altitude on reproduction has been shown previously in men, hypoxia could cause a degeneration in the germinal epithelium that affects spermatogenesis. We show that paternal chronic hypoxia affects preimplantation embryonic development, pregnancy, implantation, live birth, and miscarriage rates in IVF procedures using donated oocytes. Trial registration number not applicable

Detection of Growth Hormone (GH) Gene Polymorphism in Norduz Sheep

Background: The Norduz sheep is a local breed of sheep bred for its meat, milk, progeny and fleece and is one of the genetic resources in the rural areas of Norduz, a district of the Turkish province of Van. The PCR-based method involves amplification of a conserved DNA region and digestion of the PCR products with DNA restriction endonucleases throughout the genome. Based on the resulting polymorphisms, statistical methods can be used to identify the desired genotypes among the alleles found. The objective of this study was to identify the polymorphisms of the growth hormone gene and determine their association with six growth traits in Norduz sheep. Methods: Out of 50 Norduz sheep, three different loci, GHY1, GHY2 and GHY3 (with the length of 599, 690 and 679 bp respectively) of GH gene on chromosome 11 of sheep genome were selected to identify polymorphisms among individuals using the PCR-RFLP technique (GH/MspI). Result: In this research study, no polymorphisms were detected after digesting three loci of the GH gene with the enzyme MspI. The 50 individuals of Norduz sheep had only AB genotypes for all three loci, indicating that no polymorphisms were present in the regions of the GH genes studied.

Can dietary phytogenic mixture improve performance for growth, digestive enzyme activity, blood parameters, and antioxidant and related gene expressions of Nile tilapia, Oreochromis niloticus?

The form of dietary phytogenic inclusion and its physiological causal mechanisms for growth promotion and immune stimulation in fish remain unknown. The study examined the effects of dietary phytogenic mixture extracted from lemon (Citrus limon), onion (Allium cepa), and garlic (Allium sativum) (LOG) on Nile tilapia performance, digestive enzymes, haemato-biochemical indices, oxidative stress, and associated gene expression for 70 days. In this experiment, diets were supplemented with 0, 10, 20, and 30 ml LOG kg−1 in isonitrogenous and isoenergetic diets and fed to tilapia with an average initial body weight (4.23 ± 0.09 g). Compared to the control diet, the dietary LOG at 20 ml kg−1 elicited the highest final body weight (FBW, 35.50 g fish−1), weight gain (WG, 31.2 g fish−1), specific growth rate (SGR, 3.02%, day fish−1), and survival rate (99.33%). Significant quadratic differences in chymotrypsin, trypsin, amylase, and lipase were shown with increasing LOG supplementation. There was a quadratic response in hematology parameters of fish with increasing LOG levels. Significant linear decreases in ALT, AST, cholesterol, and triglyceride were shown with the increased LOG inclusion in the diets. A polynomial correlation in total protein, albumin, and globulin was found under different inclusion levels of LOG while significant quadratic increases in SOD, CAT, and Gpx and significant quadratic decrease in MDA was found with increasing LOG supplementation. The IGM-2, SOD, and CAT gene expressions were quadratically improved; the highest relative expression was obtained by fish received 20 ml LOG kg−1 diet. Growth hormone gene expression was quadratically modulated in the liver and pituitary of fish fed diverse doses of dietary LOG compared with the control. The phytogenic of LOG at 20 ml kg−1 elicited the best tilapia performance and hematological indices, increased antioxidant and digestive enzyme activities, and gene expressions of growth, immunoglobulin and superoxide dismutase.

Thermal Conditioning Can Improve Thermoregulation of Young Chicks During Exposure to Low Temperatures

The risk of climate change is increasing year by year and changing environmental temperatures will increasingly have effects on productivity in the poultry industry. Thermal conditioning is a method of improving thermotolerance and productivity in chickens (Gallus gallus domesticus) that experience high ambient temperatures. Thermal conditioning involves exposure of chickens to high temperatures at an early age. This conditioning treatment can affect tolerance to other type of stress. However, the effect of thermal conditioning on tolerance of low temperatures has not been investigated. Therefore, in this study we investigated the effect of thermal conditioning in chickens on thermoregulation during exposure to low temperatures. Three day-old female broiler chicks were exposed to high ambient temperatures (40°C for 12 h) as a thermal conditioning treatment. A control group of chicks was kept at 30°C. At 7 days-old, both groups of chicks were exposed to low temperatures (16 ± 0.5°C) for 3 h. Thermal conditioning treatment reduced the decrease in rectal temperature during cold exposure that occurred in control chicks. In addition, hypothalamic mRNA expression of brain derived neurotrophic factor, thyrotropin-releasing hormone and arginine vasotocin genes was higher in thermal conditioning treated chicks than control chicks. The mRNA expression of avian uncoupling protein in the liver was also higher in thermal conditioning chicks. These results suggest that thermal conditioning treatment can improve thermoregulatory mechanisms of chicks under low temperature environments.

Ovarian expression of follicle stimulating hormone and activin receptors genes in a prenatally-androgenized rat model of polycystic ovary syndrome in adulthood.

The expression of genes involved in basic pathways, such as folliculogenesis and steroidogenesis may be affected following prenatal androgen exposure. Besides, exposure to androgens during prenatal life plays a central role in developing polycystic ovary syndrome (PCOS) in females in later life. In the present study, we aimed to examine the expression of the follicle stimulating hormone receptor (FSHR) and activin receptor (actR) genes in ovarian granulosa cells (GCs) of a prenatally-androgenized rat model of PCOS in adulthood. In the adult rat model of PCOS and their controls (n = 8 in each group), different phases of the estrous cycle were determined by vaginal smear. Total RNA was extracted from the ovarian GCs using the TRIzol protocol, a reverse transcription kit was used for complementary DNA (cDNA) synthesis, and the expression of FSHR and actR genes was measured by SYBR-Green Real-Time PCR. GraphPad Prism was used for statistical analysis of data, and the t-Student'stestwasused to compare the results between the two groups. PCOS rats had longer and irregular estrous cycles compared to controls. The expression of FSHR and actR genes were significantly decreased in the rat model of PCOS compared to control rats. In PCOS rats, genes expression ratios for FSHR and actR were 0.91 ± 0.11 times (P = 0.008) and 0.42 ± 0.13 times (P = 0.048) less than controls, respectively. Reduced expression of the FSHR and actR genes in ovarian GCs may be one of the mechanisms mediating PCOS-related disorders, especially abnormal ovarian folliculogenesis and ovulation dysfunction, following exposure to androgens during fetal life.

Bioactive, full-length parathyroid hormone delivered using an adeno-associated viral vector.

Delivering the parathyroid hormone (PTH) gene has been attempted preclinically in a handful of studies, but delivering full-length PTH (1-84) using adeno-associated viral (AAV) vectors has not. Given the difficulty in achieving therapeutic levels of secreted proteins using gene therapy, this study seeks to determine the feasibility of doing so with PTH. An AAV vector was used to deliver human PTH driven by a strong promoter. We demonstrate the ability to secrete full-length PTH from various cell types in vitro. PTH secretion from hepatocytes was measured over time and a fluorescent marker was used to compare the secretion rate of PTH in various cell types. Potency was measured by the ability of PTH to act on the PTH receptors of osteosarcoma cells and induced proliferation. PTH showed potency in vitro by inducing proliferation in two osteosarcoma cell lines. In vivo, AAV was administered systemically in immunocompromised mice which received xenografts of osteosarcoma cells. Animals that received the highest dose of AAV-PTH had higher liver and plasma concentrations of PTH. All dosing groups achieved measurable plasma concentrations of human PTH that were above the normal range. The high-dose group also had significantly larger tumors compared to control groups on the final day of the study. The tumors also showed dose-dependent differences in morphology. When looking at endocrine signaling and endogenous bone turnover, we observed a significant difference in tibial growth plate width in animals that received the high-dose AAV as well as dose-dependent changes in blood biomarkers related to PTH. This proof-of-concept study shows promise for further exploration of an AAV gene therapy to deliver full-length PTH for hypoparathyroidism. Additional investigation will determine efficacy in a disease model, but data shown establish bioactivity in well-established models of osteosarcoma.

Influence of Garlic (Allium sativum) Clove-Based Selenium Nanoparticles on Status of Nutritional, Biochemical, Enzymological, and Gene Expressions in the Freshwater Prawn Macrobrachium rosenbergii (De Man, 1879).

Selenium (Se) is one of the essential micronutrients for performing vital body functions. This study aims at examining the influence of dietary supplementation of garlic clove-based green-synthesized selenium nanoparticles (GBGS-SeNPs, 48-87nm) on carcass minerals and trace elements, and growth, biochemical, enzymological, and gene expression analyses in the freshwater prawn, Macrobrachium rosenbergii post larvae (PL). The 96h LC50 of this GBGS-SeNPs to M. rosenbergii PL was 52.23mgL-1. Five different artificial diets without supplementation of GBGS-SeNPs (control, 0.0mgkg-1) and with supplementations of GBGS-SeNPs starting from 100 times lower than the LC50 value (0.5, 1.0, 1.5, and 2.0mgkg-1) were prepared and fed to M. rosenbergii PL for 90days. A dose-dependent accumulation of Se was observed in the carcass of experimental prawns. GBGS-SeNPs, up to 1.5mgkg-1 significantly influenced the absorption of other trace elements (Ca, Cu, and Fe) and mineral salts (K, Mg, Na, and Zn). GBGS-SeNPs-supplemented diets showed efficient food conversion ratio (FCR) of 1.32g against 2.71g, and therefore enhanced the survival rate (85.6% against 78.8% in control) and weight gain (WG) of 1.41g against 0.46g of control prawn. GBGS-SeNPs significantly elevated the activities of protease, amylase, and lipase, and the contents of total protein, essential amino acids (EAA), total carbohydrate, total lipid, monounsaturated fatty acids (MUFA), polyunsaturated fatty acids (PUFA), and ash. These indicate the growth promoting potential of GBGS-SeNPs in prawn. The insignificantly altered activities of glutamic oxaloacetate transaminase (GOT), glutamic pyruvate transaminase (GPT), superoxide dismutase (SOD), and catalase, and the content of malondialdehyde (MDA) up to 1.5mgkg-1 suggest its acceptability in prawn. Moreover, a respective down- and upregulated myostatin (MSTN) and crustacean hyperglycemic hormone (CHH) genes confirmed the influence of GBGS-SeNPs on the growth of prawn. In contrast, 2.0mgkg-1 GBGS-SeNPs supplementation starts to produce negative effects on prawn (FCR, 1.76g; survival rate, 82.2%; WG, 0.84g against respective values of 1.32g, 85.6%; and 1.41g observed in 1.5mgkg-1 of GBGS-SeNPs-supplemented diet fed prawn). This study recommends a maximum of 1.5mgkg-1 GBGS-SeNPs as dietary supplement to attain sustainable growth of M. rosenbergii. This was confirmed through polynomial and linear regression analyses.

840-P: Proof of Concept of I.P. Administered Neo-Islet (NI) T1DM Biotherapy: Omentum Becomes the New Endocrine Pancreas by Spontaneous Redifferentiation of In Vitro Dedifferentiated Islet Cells

We published that treatment with allogeneic, species specific NIs, organoids of Mesenchymal Stromal Cells (MSCs) and culture expanded islet cells (ICs) functionally cured/improved the diabetic state in NOD mice, insulin dependent pet dogs, and NOD/SCID mice treated with dog or human NIs without requiring antirejection drugs or encapsulation devices (see also abstract this meeting) . Fig 1 summarizes the mode of action of NI therapy and experimental detail: Dedifferentiated ICs (∼ 6 PDLs) are incorporated into NIs and implanted into diabetic animals. They redifferentiate in vivo and normalize blood glucose levels as demonstrated by gene expression analysis on NIs retrieved from euglycemic diabetic mice at 21 weeks post administration. As seen in the right portion of the graph, gene expression levels of islet endocrine hormones in retrieved NIs are not significantly different from those of freshly isolated islets (log10 (RQ) &amp;lt; ± 2) . These data demonstrate that although culture expansion of islet cells decreases endocrine gene expression as a function of PDLs, incorporation of ICs into NIs and implantation into a diabetic subject results in redifferentiation of the ICs, reestablishment of euglycemia, and restoration of islet hormone gene expression. Disclosure A.Gooch: Board Member; SymbioCellTech, LLC., Employee; SymbioCellTech, LLC., Stock/Shareholder; SymbioCellTech, LLC. S.S.Chowdhury: Employee; SymbioCellTech, LLC. C.Westenfelder: None.

249-LB: PAX4 Loss of Function Alters Human Endocrine Cell Development and Influences Diabetes Risk

In an effort to identify disease-causing mechanisms, numerous genome-wide studies across different ancestries have discovered genetic variants influencing type 2 diabetes (T2D) risk. Amongst identified diabetes-associated loci, a PAX4 coding variant (p.Arg192His) is uniquely and reproducibly associated with altered T2D-risk in East Asian populations. PAX4 encodes a transcription factor, and in mice Pax4 is essential for beta cell development. Whether PAX4 plays a similar role in humans and how diabetes associated PAX4 variants impact beta cell development and/or function is unknown. We combined in vivo studies in human variant carriers with disease modelling in human stem cell models to address these questions. Carriers of the PAX4 p.Arg192His allele have reduced islet cell function compared to non-carriers (AIRg: p.Arg192 756.9 vs. p.His192 530.8 p&amp;lt;0.002) . Deletion of PAX4 by CRISPR-Cas9 genome-editing in isogenic human induced pluripotent stem cell (hiPSC) -derived beta-like cells caused de-repression of alpha cell gene expression, suggesting decreased function in carriers of PAX4 variants may be due to altered beta cell development. hiPSCs derived from carriers of either the PAX4 p.Arg192His or a novel p.Tyr186X allele identified in a proband with early onset diabetes, had increased polyhormonal endocrine cell formation and reduced insulin content following in vitro differentiation towards beta-like cells; correction of the diabetes-associated PAX4 alleles reversed these phenotypic changes. Our in vitro and in silico molecular studies of diabetes associated alleles support reduced PAX4 expression and/or function. PAX4 knockdown in the EndoC-βH1 human beta cell model altered hormone gene expression, reduced total insulin content, and impaired insulin secretion. In conclusion, we have demonstrated the importance of PAX4 for human endocrine cell development and provide a potential mechanism for diabetes-associated alleles through de-repression of alpha cell gene expression. Disclosure N. Krentz: None. S. Hoon: None. D. Gardner: None. S. Kao: None. E. Tai: Advisory Panel; Amgen Inc., Merck Sharp &amp; Dohme Corp., Novartis AG, Novo Nordisk, Stock/Shareholder; Abbott, AbbVie Inc. A. L. Gloyn: Other Relationship; Genentech, Inc., Roche Pharmaceuticals. A. Teo: Stock/Shareholder; BetaLife Pte Ltd. H. Lau: None. F. Abaitua: Employee; Vertex Pharmaceuticals Incorporated. M. Perez-alcantara: None. J. Chan: None. J. N. Ajeian: None. B. Champon: None. H. Sun: None. A. Jha: None. Funding Wellcome (200837) ; National Institutes of Health (U01-DK105535, U01-DK085545, UM1DK126185, U01DK123743, U24DK098085, P30DK116074) ; Lee Foundation Grant (SHTX/LFG/002/2018) Paris-NUS 2021-06-R/UP-NUS (ANR-18-IDEX-0001)