Pit-1 Gene Research Articles

Polymorphism of allelic variants of genes in dairy cow’s breed in the conditions of mountain and plain dagestan

In the Republic of Dagestan, where livestock farming is a key area, dairy cattle breeds such as Red Steppe and Caucasian Brown have not been studied in terms of genetic identification. Variations in the PIT1, PRL and GH genes associated with milk productivity can provide important insights into the adaptation of these breeds to various environmental conditions of Dagestan - from plains to mountains and foothills. This can also reveal the unique allelic characteristics of these genes, which is the central focus of our study. The DNA used for the study was extracted from the blood of Caucasian Brown cows bred in mountain (n=74) and foothill (n=52) vertical zonality, as well as the Red Steppe breed bred in the conditions of lowland Dagestan (n=52). The studied cattle populations were genotyped by the polymerase chain reaction method – restriction fragment length polymorphism (PCR-RFLP) on a programmable fourchannel thermal cycler "Tercik" of the company "DNA-technology" (Russia). The polymerase chain reaction (PCR) was carried out using synthesized specific primers. The study of the Caucasian brown cattle breed showed differences in the genetic profiles of individuals living in different ecosystems. In mountainous areas, the allele of the pituitary transcription factor PIT-1A occurs with a frequency of 0.10, while the allele PIT-1B - with a frequency of 0.90. On the contrary, in lowland areas, the proportion of the allele PIT-1A increases to 0.18, and PIT-1B decreases to 0.82. In the mountain zone, the genotypes PIT-1AA, PIT-1BB and PIT-1AB are 2.0%, 82.0% and 16.0%, respectively. In the plain zone, the genotype PIT-1AA is not observed. Cattle raised in certain natural conditions demonstrate unique allelic combinations that are the result of adaptation to the local environment. The data obtained allow us to take these features into account and use them in further breeding work.

PIT1/HINFI POLYMORPHISM IN HOLSTEIN CATTLE IN AFYONKARAHISAR PROVINCE

Pit1 gene has been shown to be located on the centromeric region of chromosome 1 in cattle. This gene has responsible for development of pituitary gland and hormone expression in mammals. Pit1 deficiency causes a dwarfism phenotype in mice, dogs and fish. Previous studies has demonstrated to associated with polymorphisms of the Pit1 gene and growth and reproductive performance and milk, meat yield traits. In the cattle, a HinfI polymorphism on Pit1 gene (c.1178G>A ) have been reported that there is an association between milk and reproduction yield parameters. In this study, we aimed existence and distribution of Pit1/HinfI polymorphism in 81 head of Holstein cattle in Afyonkarahisar province. In our study, we found that there were 5 AA, 30 AG and 46 GG genotypes in our survey population. We also detected to frequency of A allele as 0.25 and the G allele as 0.75. We calculated to genetic index values such as PIC (0.3027418) and Heterozygosity (0.3742044) using by RStudio package. Chi-square value was found 0.049 and was exhibited to survey population in Hardy-Weinberg equilibrium. Considering of these findings, we denoted proper of Pit1/HinfI polymorphism using in marker assisted selection studies both of native and culture cattle breeds.

9378 Knockdown Of The Cdh2 Gene In Zebrafish Determines Its Role In Terminal Hormone Differentiation

Abstract Disclosure: B.V. Fernandes: None. C. MacRae: None. L.R. Carvalho: None. M. Junqueira: None. Introduction: Congenital hypopituitarism (CH) is characterized by the deficiency of one or more pituitary hormones. Whole exome sequencing (WES) has allowed the identification of novel/rare variants in genetic diseases. Among CH patients, 85% lack a molecular diagnosis. WES approach in a patients born to consanguineous parents identified the homozygous variant (c.865G>A, p. Val289Ile) in the CDH2 gene that produce N-Cadherin protein responsible for cell-cell adhesion. This variant was predicted as likely pathogenic allelic variant by ACMG criteria was found in a patient with GH, TSH, ACTH, LF/FSH deficiencies associated with an ectopic neurohypophysis and pituitary hypoplasia. CDH2 analysis in a cohort of 219 patients with CH, followed in a single Brazilian center, identified 3 unrelated families with heterozygous variants and incomplete penetrance where 2 were novel, in addition to the p. Val289Ile. The deleterious effect of the homozygous variant on cell adhesion properties was confirmed in permanent transfection assays in L1 fibroblast cell lines. To determine the role of the cdh2 gene in pituitary development and hormone production and secretion, we selected zebrafish as an animal model due to its 75% genetic homology with humans. Knockout of cdh2 using Crispr/cas9 genomic edition resulted in 85% lethality of animals at 48 hours post-fertilization (hpf), precluding analysis of its role during pituitary development. Objective: To produce a cdh2 knockdown in zebrafish for phenotypic and molecular analysis. Materials and Methods: The pET28B-RfxCas13d-His plasmid was transformed, linearized, and followed by in vitro transcription to produce Cas13 mRNA. Three guide RNAs targeting the cdh2 gene were designed to guide Cas13 to the target. The Cas13/RNA guide complex was injected into single-cell embryos, and phenotypic analyses were performed under a stereomicroscope until 96 hpf. To evaluate cdh2, prop1, pit1 and gh gene expression by RT-PCR, embryos were collected at 24, 48, 72, and 96 hpf. Results: Knockdown animals exhibited microcephaly, cardiac edema, and microphthalmia during embryonic development. CDH2 expression was decreased by 75% in the first 24 hours of life and began to recover by 48 hours, gradually increasing until 96 hpf when it equaled to WT. gh expression was statistically decreased at 48h compared to WT, with an increased gh expression similar to cdh2 increasing reaching similar WT levels by 96 hpf. There was no statistically significant difference in prop1 and pit1 expression compared to WT in the analyzed periods. Conclusion: The cdh2 knockdown in zebrafish using Cas13 enzyme was effective with cdh2 decreased expression by 75% presenting a phenotype similar to the knockout model. Decreased gh expression at 48h suggests an important role of cdh2 in hormonal terminal differentiation, which should be corroborated by analysis of other hormone expressions. Presentation: 6/2/2024

Effects of polymorphisms in PIT1, H-FABP, PIK3C3 and CAST genes on chemical composition, amino acid content in meat of Vietnamese indigenous fatty pig breed “I”

This study aimed to evaluate the effects of polymorphisms of PIT1/RasI, H-FABP/HinfI, PIK3C3/Hpy8I, CAST/HinfI genes and gender on chemical composition, amino acid content in meat of Vietnamese indigenous fatty pig breed “I” raised under industrial condition at Dabaco Breeding Pig Company, Phu Tho Province, Vietnam. Samples of longissimus dorsi muscle from 23 “I” pigs (12 gilts and 11 barrows) at 8 months of age were used to measure chemical composition (dry matter, protein, lipids and ash) and content of 17 amino acids (aspartic, glutamic, serine, histidine, glycine, threonine, alanine, arginine, tyrosine, valine, methionine, phenylalanine, isoleucine, leucine, lysine, proline, cystine); while ear tissues were collected to identify polymorphisms of these genes. Chemical compositions were analyzed using ISO methods (1973, 1978, 1997, and 1998). The content of amino acids was quantified using high-performance liquid chromatography (HPLC) methods. The polymorphisms of four genes were identified by the PCR-RFLP technique. The results indicated that PIT1, H-FABP, PIK3C3, and CAST had no effects on chemical compositions and amino acid contents (P>0.05). Inversely, the contents of amino acids (aspartic, serine, threonine, arginine, phenylalanine, leucine, and lysine) were significantly different between gilts and barrows (P<0.05).

Associations Between BLG, CSN3, DGAT1, GH, PIT1, and PRL Gene Polymorphisms and Milk Production Traits in Holstein Dairy Cows: A Meta-Analysis.

The current meta-analysis was employed to combine the results of multiple studies into a single estimate related to B-LG, CSN3, DGAT1, PRL, GH, and PIT1 gene polymorphisms and their effects on milk production traits. The purpose of this meta-analysis was to investigateassociations between B-LG, CSN3, DGAT1, GH, PIT1, and PRLgene polymorphisms with milk production traits in Holstein dairy cows. An extensive search was done from 2002 to 2022year. Statistical analyses were performed by using Stata 11.2 software. Genetic models viz codominant (AA vs. AB, AA vs. BB, and AB vs. BB), dominant (AA + AB vs. BB), recessive (AA vs. AB + BB), and completely over-dominant (AA + BB vs. AB) were applied. The results of meta-analysis of association between B-LG genotypes with milk yield where found a significant (P < 0.05) and with fat and protein contents (P < 0.01). In CSN3 polymorphisms of A/A and A/B genotypes had a significant effect on fat yield (P < 0.05) and protein content (P < 0.01). DGAT1 polymorphisms had a significant effect on milk yield, fat yield, protein yield (P < 0.05), with fat and protein contents showed high effect (P < 0.01). No significant association was detected between GH and milk traits (P > 0.05). PIT1 genotype polymorphisms had a significant effect on milk yield (P < 0.05) and protein content (P < 0.01). PRL genotype polymorphisms were significantly associated with milk yield (P < 0.05), fat content and protein yield (P < 0.01). The B-LG, DGAT1,CSN3 and PRL gene polymorphisms could be utilized as good markers to improve milk production traits in the Holstein cattle breed.

Inhibiting Lysyl Oxidases prevents pathologic cartilage calcification

Lysyl oxidases (LOX(L)) are enzymes that catalyze the formation of cross-links in collagen and elastin fibers during physiologic calcification of bone. However, it remains unknown whether they may promote pathologic calcification of articular cartilage, an important hallmark of debilitating arthropathies. Here, we have studied the possible roles of LOX(L) in cartilage calcification, related and not related to their cross-linking activity. We first demonstrated that inhibition of LOX(L) by β-aminoproprionitrile (BAPN) significantly reduced calcification in murine and human chondrocytes, and in joint of meniscectomized mice. These BAPN’s effects on calcification were accounted for by different LOX(L) roles. Firstly, reduced LOX(L)-mediated extracellular matrix cross-links downregulated Anx5, Pit1 and Pit2 calcification genes. Secondly, BAPN reduced collagen fibrotic markers Col1 and Col3. Additionally, LOX(L) inhibition blocked chondrocytes hypertrophic differentiation (Runx2 and COL10), pro-inflammatory IL-6 release and reactive oxygen species (ROS) production, all triggers of chondrocyte calcification. Through unbiased transcriptomic analysis we confirmed a positive correlation between LOX(L) genes and genes for calcification, hypertrophy and extracellular matrix catabolism. This association was conserved throughout species (mouse, human) and tissues that can undergo pathologic calcification (kidney, arteries, skin). Overall, LOX(L) play a critical role in the process of chondrocyte calcification and may be therapeutic targets to treat cartilage calcification in arthropathies.

ПОРОДНІ ОСОБЛИВОСТІ АЛЕЛЬНОГО ПРОФІЛЮ ГЕНІВ PIT1 ТА GH УКРАЇНСЬКОЇ АБОРИГЕННОЇ ЛЕБЕДИНСЬКОЇ ПОРОДИ КОРІВ

The preservation of genetic diversity is a paramount concern in addressing global challenges today. The foundation of biological diversity hinges on its genetic components, and the loss of species and genetic variability poses a genuine threat to the biosphere. This is because the sustainability of natural ecosystems and agroecosystems depends directly on their genetic adaptability to changing environmental conditions.&#x0D; In cattle, genes associated with milk yield play a crucial role, including genes related to milk proteins and hormones. Among these, the pituitary-specific transcription factor (PIT1) and growth hormone (GH) genes are of particular significance. In this study, we investigated the polymorphism of these genes in the Lebedyn breed of cattle.&#x0D; A total of 2 gene polymorphisms, PIT1 and GH, were examined through molecular and genetic research conducted at the genetics laboratory of the Institute of Animal Breeding and Genetics named after M. V. Zubets, part of the National Academy of Agrarian Science of Ukraine. The study utilized 32 DNA samples extracted from the venous blood of Lebedyn breed cows using the 'DNA Sorb-B' kit from AmpliSens. Genotyping was carried out through polymerase chain reaction (PCR-RFLP) analysis of restriction fragment length polymorphism, employing the “Tertsyk” amplifier for PCR. The resulting DNA restriction analysis products were assessed via electrophoresis in a 2–3% agarose gel containing ethidium bromide, with DNA fragment lengths determined using the GeneRuler TM 50 bp DNA Ladder and Thermo Scientific™ GeneRuler 1 kb Plus DNA Ladder markers.&#x0D; For the PIT1 gene analysis, a 451 bp amplified fragment was treated with the enzyme HinfI, yielding the following allele frequencies: A (0.38) and B (0.62). The heterozygous AB genotype was the most prevalent (0.45), while homozygotes for the A allele constituted 0.15 of the population. In the case of the GH gene fragment (223 bp), hydrolysis was performed using the restriction enzyme AluI. Analysis of allele frequency distribution indicated a high frequency of the L allele (0.85), with no animals exhibiting the homozygous GHVV genotype, and the GHLL genotype was present in 0.7 of the population.&#x0D; These findings reveal unique features of the allelic spectrum of the PIT1 (V-0.62) and GH (L-0.85) genes specific to the Ukrainian indigenous Lebedyn breed of cows. Selectionally significant genotypes include PIT1AA (15%) and GHLL (70%), while heterozygote carriers, which contribute to genetic diversity within the population, account for 45% and 30%, respectively.&#x0D; This study’s results hold particular value due to the alarming decline in the population of local indigenous breeds and the imminent threat to the preservation of agricultural genetic resources.

Polymorphic variants of the PIT-1 gene in simmental and red steppe cattle of the Republic of Kazakhstan

The search and identification of genes responsible for economically useful traits of cattle is relevant. Dairy productivity as a complex quantitative trait, is controlled by a large number of genes and their transcription factors taking into account the physiology of the animal. The dynamics of dairy productivity depends on a number of factors, since the secretory activity of the mammary gland is controlled by a complex of hormones, genes, transcription factors and enzymes. The studied gene Pit-1 (POU1F1) is an informative marker in determining milk and meat productivity. The genotype of an individual determines its productivity potential and the norm of reaction to environmental conditions. Genotypes of the studied gene are identified by polymerase chain reaction and restriction fragment length polymorphism (PCR-PDRP) analysis. When analyzing the data, the chi-square test is used to determine the genotype, allele frequencies and Hardy-Weinberg equilibrium. The object of research – DNA of blood of red steppe, simmental breeds of cattle of Pavlodar region of Kazakhstan. According to the results of amplification of Pit-1 gene fragment the PCR product with the size of 451 bp was obtained. According to the results of amplicon splitting the genotypes AA (451 bp), AB (451, 207, 244 bp) and BB (207, 244 bp) were obtained. In the considered farms of the region genotypes BB, AB prevail. In the studied groups of animals of the region high frequency of allele B is observed; in red steppe – 0.689, simmentals – 0.549; frequency of allele A – 0.312; 0.451 respectively. Genetic equilibrium is not disturbed; the criteria of reliability of the empirical distribution to the theoretical one in red steppe breed – 0.04, in simmentals – 0.16. The revealed associations of Pit-1 gene polymorphism can be used for selection and selecting parental pairs at directed breeding.

THU049 Growth Hormone Treatment Reverses Deficits In POU1F1 Gene Mutation

Abstract Disclosure: S. Al-Samerria: None. J. Phelan: None. H. Xu: None. A. Negron: None. F. Wondisford: None. S. Radovick: None. Mutations in the Pit 1 gene, also known as POU1F1, have been linked to the deficient development of somatotrophs and the synthesis of growth hormone (GH) in the anterior pituitary gland. In a previous study, we reported a point mutation in the PIT1 gene caused by a substitution of a lysine (K) with a glutamic acid (E) at position 216 (K216E) on one allele in a patient presenting with combined pituitary hormone deficiency (CPHD). This patient had undetectable levels of GH, TSH, very low PRL, and severe mental retardation associated with short stature (SS). To study the effects of GH treatment on growth and metabolism in CPHD, we created a unique mouse model with the K216E mutation in the PIT1 gene using the CRISPR/Cas9 genome editing system. In this study, 4-week-old male and female homozygous mice were randomly divided into two groups. The treatment group received subcutaneous injections of GH (5 mg/g/day) for 5 consecutive days per week for 4 weeks, while the control group received saline injections on the same schedule. GH treatment significantly improved the growth and body composition of male and female mice bearing the K216E. GH treatment also increased metabolism as measured by oxygen consumption, carbon dioxide production, and energy expenditure. These effects were observed both during treatment and 4 weeks post-treatment, although some reversal of the changes in body composition was observed 4 weeks post-treatment. Our findings demonstrate the utility of this mouse model in studying the effects of GH deficiency and treatment on growth and metabolism. This model holds great promise in advancing our understanding of the underlying mechanisms of hypopituitarism and CPHD, potentially serving as a valuable tool in identifying metabolic biomarkers for these conditions. Presentation: Thursday, June 15, 2023

EFFECT OF G.13700A&gt;G AND G.25783C&gt;T IN PIT-1 GENE ON GROWTH TRAITS OF DUROC, LANDRACE, YORKSHIRE PIG

Pituitary specific transcription factor-1 (Pit-1) gene is responsible for pituitary development and growth hormone expression, which is considered a pivotal candidate gene for growth in pig. This study aims to detect novel single nucleotide polymorphisms (SNPs) in porcine Pit-1 gene, investigate its effect on growth traits in the Duroc, Landrace and Yorkshire pigs. Two novel polymorphisms were detected, one located in the intron 2 (g. 13700 A&gt;G) and another located in exon 4 (g.25783 C&gt;T) which caused an amino acid change from threonine to methionine (Thr36Met). At g.13700 A&gt;G site, the frequency of A allele was higher than G allele in Duroc, Landrace and Yorkshire. At the g.25783 C&gt;T site, it was remarkable differences in allelic frequencies and genotypic frequencies among the three pig breeds. This study demonstrated the significant genotype effect of Pit-1 on body weights at 70 d, age when reached 30kg, 50kg, 100kg and average daily gain from 30kg to 100kg. According to our results, the novel Pit-1 polymorphisms may be useful in pig selection as molecular markers and required in future studies. Keywords: Pit-1, pig, growth traits, polymorphisms.

Unveiling the Etiopathogenic Spectrum of Hypophysitis: A Narrative Review.

Hypophysitis, a rare inflammatory disorder of the pituitary gland, has seen an uptick in reported cases in recent years. Our objective is to summarize the most recent research on the etiopathogenesis, molecular mechanisms, and genetics of both primary and secondary hypophysitis. Primary autoimmune hypophysitis (PAH): During the acute phase of the disease, the pituitary gland in enlarged due to the infiltration of T and B lymphocytes. The chronic phase is characterized by progressive and irreversible pituitary atrophy. APA may play a role in the management, diagnosis, and prognosis of PAH. Specific autoantibodies such as anti-GH, anti-PIT-1, and anti-T-PIT have been found in patients with hypophysitis and hypopituitarism. A recent study suggested that a mechanism of escaping clonal deletion and mounting an immune response against self antigens can explain the unusual nature of the immune response observed in PAH patients. A cytokine array shows the presence of gamma-interferon and interleukin-17. Patients carrying mutations in the PIT1 or PROP1 genes may present PAH. Individuals carrying the HLA DQ8 haplotype are four times more likely to develop PAH. Immune checkpoint inhibitors induce hypophysitis (IIHs): IIHs is an increasingly frequent toxicity of in patients on treatment with inhibitors targeting cytotoxic T-lymphocyte antigen 4 (CTLA-4) and programmed cell death-1 (PD-1). ICIs inhibit the CTLA-4 pathway, leading to overactivation of T lymphocytes. The binding of PD-1/PD-L1 suppresses the activity of T cells, promotes the conversion of T-helpers into T-regulatory cells, and activates pro-survival signaling pathways in cancer cells. Cytokines play a crucial role in IIHs. B-cell infiltration has been observed in IIHs, suggesting that antibody-mediated pituitary injury may contribute. Genetic polymorphisms of CTLA-4 and PD-1 genes can increase the risk of IIHs. HLA alleles may also be involved in the onset of IIHs; this HLA association presents a possible alternative mechanistic hypothesis. IIHs may also be linked to a paraneoplastic syndrome triggered by ectopic expression of pituitary specific antigens. SARS-CoV-2-related hypophysitis: Recently, the literature has reported occurrences of hypophysitis associated with the SARS-CoV-2 virus; long COVID-19 may also present as infundibulo-neuro-hypophysitis. The virus enters the central nervous system because of its distinct interaction with angiotensin-converting enzyme receptors via spike proteins binding the capillary endothelium, and it directly damages the pituitary cells. The effect of SARS-CoV-2 can occur indirectly through inflammation and the release of cytokines. The exact mechanism remains ambiguous. The available data on endocrine complications associated with the SARS-CoV-2 vaccine are scant. Nonetheless, isolated cases of hypophysitis have been documented. Treatment of hypophysitis: Glucocorticoids are the cornerstone in managing primary hypophysitis, given their targeted action on inflammation. A better understanding of the etiopathogenesis and molecular mechanism of hypophysitis can lead to more effective and personalized treatment strategies.

The study of the strength and significance of four biological parameters on the body weight of goose.

Alternative products such as those from high-value protien animals have increased the demand for the production of high-quality chicken meat in past few years. This study examines the impact of two distinct feeding types on goose body-weight, as well as the genetic variation of growth hormone (GH) and pituitary-specific transcription factor (Pit-1) genes in ten goose populations using single nucleotide polymorphism (SNP) markers and PCR-RFLP analysis. Both genes were seen as very important for productivity, especially in light of the COVID-19 and its effect on poultry industry at the time. The findings suggest that employing genetic indicators in these two genes in conjunction with a high-fat diet may be a feasible strategy for goose selection programme aiming to increase marketing body weight, as the high-fat diet outperformed the balanced diet. The study investigates the effect of gender, 2 types of diets, breeds and the genetic variation of the two genes, four SNPs were reported to be found: two at the GH gene exons C123T and C158T, and two at the Pit-1 gene exons G161A and T282G. Certain genotypes were found to have a substantial effect on the marketing body-weight of goose, which varied depending on the tested breeds. However, in terms of gender, males report higher and better performance levels than females. Diet, breeds and genotype interaction, and breeds, gender and genotype interaction were found to have a minor effect on goose body weight. However, diet, breeds, gender, SNP locus, diet and breeds interaction, and breeds and gender interaction were found to have a significant effect on goose body weight, as indicated by the effect size results.

The relationships between Pit-1 gene polymorphism and some performance traits in simmental and brown swiss breeds

It was aimed to investigate the genotypic structures of the Pit-1 gene from genomic DNA samples taken from Simmental and Brown Swiss breeds and determine the distribution of genotype and allele frequencies of bovine breeds and to determine the relationships between the detected genotypes and some performance traits. Pit-1/HinfI gene polymorphisms were determined using the PCR-RFLP method from genomic DNA obtained from Brown Swiss and Simmental breed cattle used in the study. According to the Hardy-Weinberg genetic test, the distribution of genotype frequencies of the studied population was found to be in equilibrium (p˃0.05) in both breeds. Genotype frequencies of AA, AB, and BB alleles of the Pit-1 gene were determined as 5.73%, 36.42%, and 57.85%, respectively, for Simmental breed, while the genotype frequencies for Brown Swiss breed were determined as 9.43%, 42.86%, and 48.01%, respectively. The actual milk yield for Simmental breed was determined as 5953 ± 364.2, 5212 ± 993.4, 5507 ± 889.5 kg, the 305-day milk yield as 5953 ± 276.8, 5642 ± 782.7, 5427 ± 246.7 kg, the lactation period as 307 ± 47, 293 ± 29, 303 ± 22 days, the daily milk yield as 19.53 ± 6, 18.49 ± 8, 17.81 ± 2 kg for AA, AB, and BB genotypes, respectively. The actual milk yield for the Brown Swiss breed was determined as 3606 ± 253, 3558 ± 530, 3999 ± 099 kg, the 305-day milk yield as 4920 ± 217, 4462 ± 900, 4635 ± 870 kg, the lactation period as 222 ± 36, 240 ± 04, 258 ± 37 days, the daily milk yield as 17.62 ± 2, 15.49 ± 2, 16.04 ± 3 kg for AA, AB, and BB genotypes, respectively. In the statistical analyses, the relationships between the performance traits studied and Pit-1 genotypes in Simmental and Brown Swiss cattle were not prove to be significant statistically.

Identification of Pituitary-Specific Transcription Factor-1 (PIT-1) Genotype in Bali Cattle

This study aimed to identify the genotype of the gene Pituitary-specific transcription factor 1 (Pit-1) in Bali cattle (Bos sondaicus). A total of 20 cows Bali (13 males and 7 females) were used in this study. DNA samples isolated from blood samples using phenol-chloroform extraction method (Sambrook et al., 1989). This study using PCR-RFLP method. Amplification of specific DNA fragments (size 451 bp, intron regions stretching from 4 to exon 6) of the Pit-1 gene Bali cattle samples was performed using a pair of specific primers (forward: 5'-AAA-CCA-TCA-TCT-CCC-TTC-TT -3 ', and reverse: 5'-AAT-GTA-CAA-TGT-GCC-TTC-TGA-G-3'). Based on the results of RFLP analysis/HinfI was found that all of Bali cattle research resulted banding pattern uniform (monomorphic), the band size 244 bp and 207 bp, and the band patterns thus identified as genotype BB, so it is known that the frequency of A and B alleles at 0 and 1. The frequency of A allele was not found in Bali cattle (Bos sondaicus) showed a significant difference with other cattle breeds (Bos taurus and Bos indicus).

Analysis of ProP1 Gene in a Cohort of Tunisian Patients with Congenital Combined Pituitary Hormone Deficiency.

Background: Non-syndromic combined pituitary hormone deficiency (CPHD) occurs due to defects in transcription factors that govern early pituitary development and the specification of hormone-producing cells. The most common mutations are in the Prophet of Pit-1 (ProP1) gene. This work aims to (1) report findings of genetic analyses of Tunisian patients with non-syndromic CPHD and (2) describe their phenotype patterns and their evolution through life. Methods: Fifteen patients from twelve unrelated families with variable clinical phenotypes were included after excluding autoimmune and acquired forms of non-syndromic CPHD. Detailed pedigree charts and auxological, hormonal, radiological, and therapeutic details were recorded. Sanger sequencing was performed, and sequences were analyzed with a specific focus on coding and splice site regions of the ProP1 gene. Retained variants were classified using several in silico pathogenicity prediction tools and the VarSome platform. Results: We identified the common p.Arg73Cys mutation in seven patients from four unrelated pedigrees. We found a novel homozygous mutation (c.340C>T) in one sporadic case. This mutation generates a truncated ProP1 protein, predicted to be non-functional, lacking the last 112 codons (p.(Gln114Ter)). We confirmed by polymerase chain reaction (PCR) the absence of large exon deletions or insertions in the remaining sporadic patients (7/8). Conclusions: We report two mutations {one newly identified [p.(Gln114Ter)] and one previously reported (p.Arg73Cys)} in five unrelated Tunisian families with non-syndromic CPHD. This work is of clinical importance as it reports the high frequency of the p.Arg73Cys mutation in Tunisian CPHD families. Our study also illuminated the involvement of novel gene(s) in the emergence of non-syndromic CPHD.

Connection between gene markers with milk production traits of Ukrainian dairy cows

The purpose of this study was to obtain information on the genotyping of cows of Ukrainian dairy breeds for the presented loci capa-casein gene (CSN3), thyroglobulin (TG-5) gene, leptin (LEP), pituitary-specific transcription factor (Pit-1), and beta-lactoglobulin (BLG) by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and to test their association with milk production. The influence of the genotype on milk yield, fat and protein content in the milk of cows of Ukrainian selection has been established. The highest levels of milk productivity traits were expected from animals with the CT genotype for the LEP gene, AA for the CSN3 gene, CC for the TG5 gene, and AA and BB for the Pit-1 gene. But at the same time, milk obtained from cows with CSN3 BB genotype were characterized by the best cheese suitability. The effect of the homo- or heterozygous state of BLG on the signs of milk production was less noticeable, and the homozygous TT genotype for the TG5 gene was not found in any of the studied breeds. The results obtained can be used in the practical work of breeding farms along with traditional methods of selection, control and preservation of the genetic diversity of specific herds at an optimal level.

Genetic polymorphism of Pit-1 and CSN3 genes in Holstein calves and its associations with calf birth weight

The aim of this study was to examine the polymorphic structures of Pit-1 and CSN3 genes of Holstein calves bred in Gümüşhane province of Türkiye, to determine the distribution of genotype and allele gene frequencies, as well as examine the effects of determined polymorphisms on birth weight of calves. HinfI polymorphisms of Pit-1 and CSN3 genes were identified in DNA isolated from blood samples of 100 Holstein calves used in the study, using the PCR-RFLP method. According to the Hardy–Weinberg genetic equilibrium test, it was observed that the distribution of genotype frequencies of HinfI polymorphisms of Pit-1 genes in the studied population was in equilibrium, but not in equilibrium in terms of CSN3 gene location. The AA, AB, and BB genotype frequencies of the Pit-1 gene in the population were 13.4 %, 40.2 %, and 46.3 %, respectively; the frequency of the A allele was 0.34, while for B allele it was 0.66. The AA, AB, and BB genotype frequencies of the CSN3 gene were found to be 24.5 %, 36.7 %, and 38.8 %, respectively; the frequency of the A allele was 0.43 and the frequency of the B allele was 0.57. According to the Hardy–Weinberg genetic equilibrium test, the distribution of genotype frequencies was in equilibrium in the Pit-1/HinfI polymorphism, but not in the CSN3/HinfI polymorphism. A statistically significant relationship was not found between the genotypes of both polymorphic regions and calf birth weight.

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Editorial introductions

Relationship of RS645439735, RS66142748 and RS670830003 mutations in the PIT-1 gene to the productive and reproductive performance of local goat

The current study included 85 does with the same management and feeding conditions belonging to two breeds (Local and Shami goats), located in the west of Baghdad. 4–5 years old, multiparous, lactating and in their first to fourth lactation. PIT-1 gene polymorphism was indicated and results showed a significant effect (P≤0.05) PIT-1 gene polymorphism (rs645439735) on protein rate in Cyprus breed, the highest value was recorded in the individuals with hetero genotype (AC) namely 3.16 % compared with 2.92 and 2.80% in both wild and mutant genotypes respectively. Results showed a significant effect (P≤ 0.05) of rs661742748 SNP on some of milk traits in Cyprus breed, the milk peak was higher in hetero genotypes compared with the wild type namely, 64.0 and 34.46 day respectively. Fat rate was increased significantly in hetero type (5.6%) compared with wild type (3.11%). Results showed a significant effect of rs670830003 SNP on weaning weight and body Weight gain from birth to weaning in the local goat breed, the highest values were 16.83 and 14.04 Kg was recorded in individuals with hetero genotype (TG) while the lowest values were in individuals with the wild type namely, 13.81 and 11.18 Kg respectively.

Associations among the TREM-1 Pathway, Tau Hyperphosphorylation, Prolactin Expression, and Metformin in Diabetes Mice

Introduction: Diabetes mellitus (DM) is a risk factor for Alzheimer’s disease (AD). Increasing evidence indicates that the triggering receptor expressed on myeloid cells (TREM)-1 amplifies chronic inflammation, as well as the roles of prolactin (PRL) and metformin (MET) in tau hyperphosphorylation. However, the associations among TREM-1, tau hyperphosphorylation, PRL expression, and MET in DM remain unclear. Methods: Streptozotocin was used to induce experimental DM in C57BL/6N mice. MET was orally administered at a dose of 400 mg/kg body weight for 6 weeks prior to hippocampal collection in DM mice. Various parameters pertaining to the TREM-1 pathway, tau hyperphosphorylation, PRL, and related factors were analyzed. Results: Quantitative polymerase chain reaction and Western blot analysis demonstrated that the expression levels of TREM-1, DAP12, casp1, interleukin-1β, Cox2, inducible nitric oxide synthase, pituitary transcriptional factor-1 (Pit-1), and PRL were significantly increased in the hippocampus of DM mice; the expression levels of these pro-inflammatory mediators, PRL receptor (PRLR) short or long (PRLR-S and PRLR-L), and PRL regulatory element-binding (Preb) protein in DM mice treated with MET (DM + MET) were significantly decreased compared with those in control (CON) mice. The levels of p-Tau and glycogen synthase kinase-3 in the DM group were significantly higher than those in the CON group and significantly lower than those in the DM + MET group. Conclusion: We confirmed the therapeutic potential of MET for both DM and neurodegeneration. Our findings shed new light on the effects of DM on the pathophysiology of AD via the TREM-1 pathway and PRL expression. Thus, an improved understanding of the TREM-1 pathway in hyperglycemic conditions, as well as PRL, Preb, Pit-1, PRLR-L, and PRLR-S gene expression in the liver, brain, and other sites, may help unravel the pathogenesis of insulin resistance and neurodegeneration.