Protein Polymorphisms Research Articles

Influence of β-casein genotype on Cheddar cheese making and ripening

Milk characteristics and suitability for production of dairy products like cheese may be influenced by protein polymorphism; the possible influence of β-casein (β-CN) polymorphism on cheese manufacturing and cheese characteristics was examined using milk with three β-CN genotypes, A1A1, A1A2 and A2A2. The composition and rennet coagulation properties of raw and standardised cheese milk, cheese yield and their composition were examined, cheese meltability, texture and proteolysis were analysed at different ripening times. Milk with β-CN variant A2A2 was found to have poorer rennet coagulation properties following standardisation and pasteurisation. Cheese made from A1A1 milk had lower protein content, while that from A2A2 milk had lower fat content compared with the other two genotypes. There was no significant impact of genotype on proteolysis during cheese ripening. However, β-CN genotype was found to impact cheese texture; after 180-d ripening, the A1A1 cheese was the softest, and A2A2 cheese was the most fracturable.

A structural foundation for studying chlamydial polymorphic membrane proteins.

Polymorphic membrane proteins (Pmps) are chlamydial cell surface proteins with signature repeating FxxN and GGA (I, V, L) tetrapeptide motifs. These autotransporter proteins are key immune targets and mediators of host-microbe interactions but by unknown mechanisms. AlphaFold predictions reveal that Pmp passenger domains have two distinct β-helical parts: a C-terminal globular "Pmp middle region" that forms six β-helical rungs, and an N-terminal highly-regular "Pmp repeat region" that is from 9 to 25 rungs long. In the repeat region, paired FxxN and GGA (I, V, L) motifs - or close variants - are fully buried and packed together to form novel β-helical rungs from which variable-length loops extend out from one face. Similar motifs in proteins from diverse organisms are predicted to form analogous structures. In Pmps, the structures imply that the tetrapeptide motifs are not directly involved in host-cell adhesion, but position protein loops on a common face where they can interact with ligands important for infection. These structural descriptions provide a guide for mutagenesis studies to identify those parts of Pmps involved in infection.IMPORTANCEInfections by bacteria in the genus Chlamydia cause a range of widespread and potentially debilitating conditions in humans and other animals. We analyzed predicted structures of a family of proteins that are potential vaccine targets found in all Chlamydia spp. Our findings deepen the understanding of protein structure, provide a descriptive framework for discussion of the protein structure, and outline regions of the proteins that may be key targets in host-microbe interactions and anti-chlamydial immunity.

The role of HIV-1 polymorphism in the pathogenesis of the disease

High genetic variability is one of the defining HIV-1 properties. It contributes to the appearance of drug resistance and to the formation of various HIV variants. In the world, the different variants of HIV-1 are distributed unevenly. In Russia there are circulating unique virus variants which are characteristic for our country and some countries of the former USSR and which are difference from HIV-1 variants circulating in other countries of the world. For many years, the studies aimed on the analysis of the influence of various HIV-1 variants on pathogenesis have been conducted. Within the framework of these studies, the polymorphism mutations in different subtypes and recombinant forms have been detected. Currently the main objects of antiretroviral therapy are HIV-1 structural proteins, mainly enzyme proteins, thus polymorphic mutations in which could affect the degree of virus sensitivity to antiretroviral therapy. Non-structural HIV-1 proteins are involved in virus replication and virus protection from the host immune system, enter the bloodstream and tissues, causing the development of inflammation. The polymorphic mutations in non-structural proteins could affect the degree of HIV infection progression and the development of concomitant somatic diseases. Today, nonstructural proteins are considered as objects for the creation of therapeutic agents, thus the identification of polymorphic mutations in nonstructural proteins in different HIV-1 variants is a basis for such developments. Thus, the study of polymorphism of both structural and non-structural HIV-1 proteins is a promising area of research in the future.

Matrix Gla protein polymorphism rs1800802 is associated with atheroma plaque progression and with cardiovascular events in a chronic kidney disease cohort.

Chronic kidney disease (CKD) is associated with increased atherosclerotic burden and higher risk for cardiovascular events (CVE). Atherosclerosis has a significant genetic component and, in CKD, it is influenced by mineral metabolism alterations. Therefore, genetic modifications of mineral metabolism-related proteins could affect atherosclerosis in CKD patients. In the present study we investigated the role of single nucleotide polymorphisms (SNPs) of the matrix gamma-carboxy glutamic acid protein (MGP) on atherosclerosis progression and CVE in a CKD cohort. A total of 2187 CKD patients from the Observatorio Nacional de Aterosclerosis en Nefrologia (NEFRONA) study were genotyped for SNPs present in the matrix gamma-carboxy glutamic acid (Gla) protein (MGP) gene. Atheromatosis was detected by vascular ultrasound. Progression of atheromatosis, defined as an increase in territories with plaque, was assessed after 24 months. Patients were followed for 48 months for CVE. Association of SNPs with plaque progression was assessed by logistic regression and their capacity to predict CVE by Cox regression. Three SNPs of the MGP gene were analyzed. No association of the rs4236 or the rs1800801 SNPs was detected with any of the outcomes. However, patients homozygotes for the minor allele of the rs1800802 SNP showed higher adjusted risk for plaque progression [odds ratio 2.3 (95% confidence interval 1.06-4.9)] and higher risk of suffering a CVE [hazard ratio 2.16 (95% confidence interval 1.13-4.12)] compared with the rest of genotypes. No association of the SNP with total or dp-ucMGP levels was found in a subsample. The rs1800802 polymorphism of MGP is associated with plaque progression and CVE in CKD patients.

Recognition of polymorphic Csd proteins determines sex in the honeybee.

Sex in honeybees, Apis mellifera, is genetically determined by heterozygous versus homo/hemizygous genotypes involving numerous alleles at the single complementary sex determination locus. The molecular mechanism of sex determination is however unknown because there are more than 4950 known possible allele combinations, but only two sexes in the species. We show how protein variants expressed from complementary sex determiner (csd) gene determine sex. In females, the amino acid differences between Csd variants at the potential-specifying domain (PSD) direct the selection of a conserved coiled-coil domain for binding and protein complexation. This recognition mechanism activates Csd proteins and, thus, the female pathway. In males, the absence of polymorphisms establishes other binding elements at PSD for binding and complexation of identical Csd proteins. This second recognition mechanism inactivates Csd proteins and commits male development via default pathway. Our results demonstrate that the recognition of different versus identical variants of a single protein is a mechanism to determine sex.

Breast cancer resistance protein polymorphism ABCG2 c.421C>A (rs2231142) moderates the effect of valproate on lamotrigine trough concentrations in adults with epilepsy.

Valproate inhibits clearance of lamotrigine and greatly increases its concentrations. We assessed whether this effect was moderated by a polymorphism (ABCG2 c.421C>A) of the breast cancer resistance protein. In two consecutive independent studies in adults with epilepsy on lamotrigine monotherapy or cotreated with valproate: (i) Exposure to valproate was considered treatment, (ii) dose-adjusted lamotrigine troughs at steady state were the outcome, and (iii) ABCG2 c.421C>A genotype (wild-type [wt] homozygosity or variant carriage) was the tested moderator. We used entropy balancing (primary analysis) and exact/optimal full matching (secondary analysis) to control for confounding, including polymorphisms (and linked polymorphisms) suggested to affect exposure to lamotrigine (UGT1A4*3 c.142T>G, rs2011425; UGT2B7-161C>T, rs7668258; ABCB1 1236C>T, rs1128503) to generate frequentist and Bayesian estimates of valproate effects (geometric means ratios [GMR]). The two studies yielded consistent results (replicated); hence, we analyzed combined data (total N= 471, 140 treated, 331 controls, 378 ABCG2 c.421C>A wt subjects, 93 variant carriers). Primary analysis: in variant carriers, valproate effect (GMR) on lamotrigine (treated, n= 21 vs. controls, n= 72) was around 60% higher than in wt subjects (treated, n= 119 vs. controls, n= 259)-ratio of GMRs 1.61 (95%CI 1.23-2.11) (frequentist) and 1.63 (95%CrI 1.26-2.10) (Bayes). Similar differences in valproate effects between variant carriers and wt subjects were found in the secondary analysis (valproate troughs up to 364μmol/L vs. no valproate; or valproate ≥364μmol/L vs. no valproate). Susceptibility of the estimates to unmeasured confounding was low. Data suggest that polymorphism rs2231142 moderates the effect of valproate on exposure to lamotrigine.

Novel Prion Protein Gene Polymorphisms in Awassi Sheep in Three Regions of the Fertile Crescent.

Scrapie is a fatal, neurodegenerative disease that affects sheep and goats, and genetic susceptibility to scrapie in sheep is associated with polymorphisms in the prion protein (PRNP) gene. The aim of this study is to identify PRNP polymorphism in Awassi sheep from Türkiye, the Palestinian Authority, and Saudi Arabia. A total of 150 healthy sheep were genotyped for PRNP, using Sanger sequencing. There were seven alleles and eleven genotypes observed based on codons 136, 154, and 171 of PRNP. The ARQ allele was predominant in all populations. The most resistant allele to scrapie, ARR, was present in all three regions. The VRQ allele, associated with the highest susceptibility to scrapie, was detected only in Türkiye at a low frequency. In this study, twenty-seven amino acid substitutions were found. Eight of them (R40Q, G65E, H88L, S98T, A118P, S138T, V192F and L250I) have not been previously reported. These data indicate that sheep breeds close to the sheep domestication center have maintained high genetic diversity in the PRNP region. Our findings on PRNP will provide valuable insights for sheep breeding programs, aiding in the selection of genotypes resistant to scrapie in Türkiye, the Palestinian Authority, and Saudi Arabia.

First report of a novel polymorphism and genetic characteristics of the leporine prion protein (PRNP) gene.

Transmissible spongiform encephalopathies (TSEs) have been reported in a broad spectrum of hosts. The genetic polymorphisms and characteristics of the prion protein (PRNP) gene have a vital impact on the development of TSEs. Notably, natural TSE infection cases have never been reported in rabbits, and genetic variations of the leporine PRNP gene have not been investigated to date. To identify leporine PRNP gene polymorphism, we performed amplicon sequencing in 203 rabbits. We report a novel single nucleotide polymorphism on the leporine PRNP gene. In addition, we performed a comparative analysis of amino acid sequences of prion protein (PrP) across several hosts using ClustalW2. Furthermore, we evaluated the effect of changes of unique leporine PrP amino acids with those conserved among various species using Swiss-Pdb Viewer. Interestingly, we found seven unique leporine amino acids, and the change of unique leporine amino acids with those conserved among other species, including S175N, Q221K, Q221R, A226Y, A230G, and A230S, was predicted to reduce hydrogen bonds in leporine PrP.

Evaluation of freshwater heavy metals accumulation effect on oxidative stress, Metallothionein biosynthesis and histopathology of Procambarus clarkii (Girard,1985) collected from three locations in the Delta region, Egypt

BackgroundIn this study, the effect of heavy metals accumulation influence was evaluated on adult crayfish Procambarus clarkii (Decapoda, Astacidea) collected from three different Governmental locations (Kafr El-Shaikh, El-Menofya, and El-Gharbiya) of the Egyptian Delta. The activity of super oxidase dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) of gills, hepatopancreas, and muscle tissue were measured. SDS Polyacrylamide gel electrophoresis (SDS-PAGE) and West blotting technique were performed to detect MT Protein expression.ResultsThe results revealed that Kafr El-Shaikh reflected the highest Superoxide dismutase (SOD), Catalase, and Glutathione S-transferase (GST) activity levels (97.2 u/100 mg, 28.5 u/100 mg, and 8.3 nmol mg (-1) protein min (-1) respectively. Superior protein polymorphism % (30%) remarked collected Freshwater crayfish P. clarkii from Kafr El-Shaikh location. Varied protein polymorphism % was shown between collected crayfish from El-Menofya, and El-Gharbiya locations (5.5 and 6.2 respectively) Increasing Metallothioneins intensity (15.4%) for collected Freshwater crayfish Procambarus clarkii from Kafr El-Shaikh Location.ConclusionHeavy metal stress influences antioxidant status and also induces increasing Metallothioneins intensity, especially samples that were collected from the Kafr El-Shaikh area.

ANALYSIS OF THE INTERCONNECTION OF STALLION SEMEN INDICATORS WITH GENETIC MARKERS OF PROTEINS

The purpose of the research was to study the associations of genetic polymorphism of proteins and enzymes of blood serum (albumin, transferrin, and esterase) on the quality of freshly obtained and cryopreserved semen of breeding stallions, taking into account their breed characteristics. The authors divided horses into four breed groups, taking into account their characteristics – riding horses, Orlov trotters, prize-winning trotters, and draft horses. A comparative assessment of 154 stallions revealed no significant differences in the quality of semen from different breed groups. For the first time, the influence coefficient (Fisher coefficient) of different genetic variants of proteins and enzymes on the semen parameters of stallions was analyzed and calculated. At the same time, the authors revealed the multidirectional effect of the same alleles of albumin, transferrin, and carboxylesterase on semen activity in different breeds. Thus, the presence of the albumin allele B (ALBB) had a positive effect on the mobility of fresh semen of stallions of riding breeds but a negative effect on stallions of Orlov trotters and draft horses. The heterozygous state of alleles revealed a positive effect on semen parameters. The analysis of variance of all examined stallions showed the highest level of the esterase locus (ESI) on the mobility of fresh semen and the albumin locus (ALBA) – on the mobility of frozen-thawed semen. The reliability of it is close to a statistically significant value. The results obtained suggest that specific gene complexes and associations with productive traits are formed in breeds under the influence of breeding. It must be taken into account as additional criteria for controlling the breeding process.

Characterization of prolamin storage protein profiles in Tunisian pearl millet landraces toward better utilization and improvement

AbstractBackground and ObjectivesThis study reports for the first time the variation of prolamin profiles among 51 pearl millet landraces representing seven populations prospected from contrasted bioclimatic areas of Tunisia.FindingsA total of 27 prolamins bands were detected, ranging from 11 to 245 kDa. Among them, 14 were polymorphic, resulting in a polymorphism rate of 51.85%. The bands with lower molecular weights are the most frequent. The biochemical marker M25.4 showed the highest value of Nei genetic diversity (h = 0.500), Shannon information index (I = 0.693), and effective number of alleles per locus Ne (2). At the population level, the polymorphism rates ranged between 50% (Mahdia) and 92.86% (Hammem Jebli, Kairouan, Medenine). The population of Kairouan showed the highest genetic diversity indices (I = 0.511 ± 0.054, h = 0.345 ± 0.041), while the lowest levels were recorded for the population of Mahdia (I = 0.268 ± 0.076, h = 0.177 ± 0.052). Analysis of molecular variance analysis indicated that the highest amount of genetic diversity was encompassed within the studied populations (89%). This was supported by low genetic differentiation (PhiPT = 0.110, p = .029) and Nei genetic distance values. The classification pattern using ward dendrogram and factorial analyses revealed the distinction of two clusters with a low genetic structure based on the geographic origins of landraces.Significance and NoveltyThe detected biochemical protein polymorphism could be a highly effective tool for the genetic resources characterization of pearl millet landraces, which will be further utilized in breeding investigations and taxonomic surveys of this cereal crop.

Analysis of null deletion polymorphism of glutathione S-transferase theta (GSTT-1), associated with anti-GSTT-1 antibodies development in transplantation.

Glutathione S-transferase theta 1 (GSTT1) is an enzyme involved in phase II biotransformation processes and a member of a multigene family of detoxifying and clearing reactive oxygen species. GSTT1 is polymorphic like other biotransforming enzymes, allowing variability in hepatic conjugation processes. Immunological recognition of the GSTT1 alloantigen, as evidenced by donor-specific antibodies formation, has previously been observed in recipients lacking GSTT1 protein (called GSTT1-, GSTT*0, null phenotype or homozygous for the GSTT1 deletion) who receive liver or kidney transplants from GSTT1+ donors and is a risk factor for the development of de novo hepatitis following liver transplants from a GSTT1 expressing donor. Antibodies against GSTT1 are demonstrated in patients who are GSTT1 null and received a transplant from a GSTT1+ donor. Understanding the local population frequency of the GSTT1 deletion is of value in understanding the potential clinical risk of developing post-transplant complications, which can be attributed to the nonexpression of GSTT1. A population of 173 healthy donors of the Murcia Region in Southeast Spain was evaluated for a null allele of GSTT1 (n=173). DNA was extracted, and GSTT-1 null allele detection was performed by real-time polymerase chain reaction. The frequency of the null GSTT1 genotype (nonexpression or deletion of the homozygous polymorphism of the GSTT1 protein) was 17.9% (n=31 null allele GSTT1/173 total individuals). Our data suggest that the frequency of null GSTT1 mutations in our population in Southeast Spain is 17.9%, lower than in other Caucasoid populations. This would convert our recipient population into more susceptible to nonlocal potential organ donors and less susceptible to local donors. All recipients bearing this GSTT1 deletion homozygous would be without the protein and triggering an alloantigen in the case of transplantation with a donor without deletion.

Evaluation of the influence of polymorphisms of the transporter genes (<i>RFC1</i>, <i>MDR1</i>) and <i>GGH</i> on the efficacy of methotrexate in rheumatoid arthritis

The efficacy of methotrexate (MT) in patients with rheumatoid arthritis (RA) may be determined by genetic factors.Objective: to evaluate the isolated and combined effects of single nucleotide polymorphisms (SNPs) of membrane transporter proteins (RFC1 80G>A and MDR1 3435C>T) and the GGH -401C>T gamma-glutamyl hydrolase enzyme genes on the efficacy of MT in patients with RA.Material and methods. The study group consisted of 85 patients with a confirmed diagnosis of RA, who received therapy with MT starting at 10 mg/week and increasing in dose to a maximum of 25 mg/week. Efficacy was assessed after six months of treatment using the dynamics of the DAS28 index, identifying patients who responded and those who did not respond to MT therapy.Genotyping of RFC1, MDR1 and GGH gene polymorphisms was performed by real-time polymerase chain reaction. Three different approaches were used to analyze the results: 1) analysis for each of the genes; 2) logistic regression; and 3) multifactor dimensionality reduction (MDR).Results and discussion. Single gene analysis was used to determine the most likely predictors of non-response to therapy: 1) for GGH-401C>T, TT genotype (odds ratio, OR 5.09; 95% confidence interval, C11.11—23.3); 2) forMDR13435C>T, the TT genotype (OR 2.38; 95% CI0.89-6.37); 3) for RFC180G>A, not - AA genotype (OR 1.87; 95% CI 0.93-3.76).The logistic regression model showed a significant effect of homozygous genotype GGH -401TT on the efficacy of MT with low sensitivity of the method. The multifactorial dimensionality reduction results show a significant synergistic effect of the MT transport genes (MDR1, RFC1) and the GGH enzyme encoding the conversion of MT to the elimination form.Conclusion. Using various statistical methods, the following results were obtained: Single gene analysis revealed the most likely predictors of nonresponse to MT therapy: GGH -401C>T - TT genotype, MDR1 3435C>T - TT genotype, RFC1 80G>A - not-AA genotype; the method of multiple logistic regression allowed to determine the significant effect of GGH -401ТТ genotype on the effect of the drug with a low sensitivity of the method; the isolated effect of polymorphisms is probably less pronounced than their combined effect on the effectiveness of MT. SNP synergism is a major contributor to the development of treatment resistance. MDR is a promising method that can be used in the future to assess the impact of SNPs.

LA PREDICCIÓN IN SILICO DETERMINA LA PRESENCIA DE EPÍTOPOS INMUNOGÉNICOS ALTAMENTE ESPECÍFICOS EN FRAGMENTOS DE LA PROTEÍNA DE LA MEMBRANA POLIMÓRFICA (PMP17G) DE Chlamydia abortus

<p><strong>Background.</strong> Ovine Enzootic Abortion is a contagious infectious disease caused by a Gram negative and obligate intracellular bacterium, <em>Chlamydia abortus</em>. For field diagnosis, commercial serological tests are used; however, some of these tests show low sensitivity and specificity rates, due to the cross-reactions that the antigens used have against other pathogens. For the most accurate diagnosis, it is necessary to develop tests with more specific antigens such as polymorphic membrane proteins (Pmp's), that allow to determine the presence of specific epitopes using new technologies. <strong>Objective.</strong> To determine <em>in silico</em> the presence of epitopes with specific immunogenic potential against <em>Chlamydia abortus</em> of two fragments of the PMP17G protein. <strong>Methodology.</strong> The cloning and sequencing of the fragments was carried out from a field isolate of <em>Chlamydia abortus</em>, and from the analysis of these sequences, with the help of two bioinformatics software’s. <strong>Results.</strong> Several epitopes from <em>Chlamydia abortus</em> were found, rPOMP90-3 (eight epitopes) and rPOMP90-4 (one epitope). <strong>Implications.</strong> Bioinformatics analysis indicated that both fragments of the protein have the capacity to activate the immune system, which would be useful for the development of diagnostic kits and immunogens. <strong>Conclusions.</strong> The <em>in silico</em> analysis allowed to efficiently predict and identify specific epitopes against <em>Chlamydia abortus</em> in both fragments of the protein. </p>

Tandem mass spectrometry tag proteomic analysis of milk and colostrum of Guanzhong dairy goat

Goat milk is an indispensable part of China's dairy industry. Milk in different lactation periods has different nutritional values. To explore the protein differences of Guanzhong dairy goat milk in different lactation periods, this experiment used Tandem Mass Tags technology to analyse the differences between colostrum group and mature milk group of Guanzhong dairy goats. The results showed that 241 differential proteins were screened in colostrum and mature milk, of which 80 proteins were significantly up-regulated, 161 proteins were significantly down-regulated, and goat colostrum exhibited more metabolic pathways related to the immune system, while mature milk exhibited more pathways related to angiogenesis and bone development. This research result promoted the study of protein polymorphism in goat milk and provided a theoretical basis for further revealing the nutritional value of goat milk at different lactation stages.

Marcadores genéticos y séricos relacionados con una rápida progresión clínica de la arteriosclerosis coronaria

Introduction and objectivesPatients with clinically evident coronary artery disease differ in their rate of progression, which impacts prognosis. We aimed to characterize serum and genetic markers in patients with rapid clinical progression (RCP) of coronary artery disease vs those with long standing stable (LSS) disease. MethodsRetrospective study of cases (RCP) and controls (LSS) (1:2). Patients requiring ≥ 2 revascularizations due to atherosclerotic progression in the 10 years after a first angioplasty were considered to be RCP and those without events during the same period after the first angioplasty were considered to have LSS disease. After patient selection, we analyzed serum values, mRNA expression and genetic polymorphisms of inflammatory markers, including interleukin-6, C-reactive protein, and tumor necrosis factor (TNF)-a, and atherogenic markers consisted of proprotein convertase subtilisin/kexin type 9 (PCSK9), low-density lipoprotein receptor, sterol regulatory element binding transcription factor 2, and apolipoprotein-B. ResultsThe study included 180 patients (58 RCP and 122 LSS). Demographic characteristics, classic risk factors and the extent of coronary disease were similar in the 2 groups. Patients with RCP showed higher serum levels of interleukin-6 and PCSK9 and higher TNF mRNA expression. Interleukin-6 rs180075C, TNF rs3093664 non-G and PCSK9 rs2483205 T alleles conferred a risk of RCP (P<.05 in all cases). Among patients with RCP, 51.7% had all 3 risk alleles vs 18% of those with LSS (P<.001). ConclusionsWe suggest the existence of specific phenotypic and genotypic markers associated with RCP of coronary artery disease that could help to individualize the type and intensity of treatment.

Identification of the coagulation properties of Chinese Holstein bovine milk: Effects of milk compositions, milk protein polymorphism, and phosphorylation levels on milk coagulation ability

Milk coagulation ability plays critical roles in cheese production. To assess the effects of milk composition, protein polymorphisms, and phosphorylation levels on milk coagulation properties, milk samples were collected from 3625 Chinese Holstein cows and milk was pooled based on their coagulation properties. The results showed that good coagulating milk had shorter rennet coagulation time and higher gel firmness with tighter connections within the formed gel. Multiple factors were observed to differ among milk with different coagulation ability. For example, lower milk pH, higher amounts of proteins and casein, were related to good coagulation properties. The salt distributions also influenced milk coagulation ability. Especially the calcium content has a large influence on milk coagulation. This may be due to its influence on the aggregation of paracasein micelles in the secondary phase of rennet coagulation. Protein polymorphisms and phosphorylation degrees were found being higher in non-coagulating milk. The proteomics results showed that lower amounts of αS1-casein may result in non-coagulation. Differences of milk fat globule membrane proteins were also observed among milk samples with different coagulation ability, possibly being related to milk protein and fat synthesis, in turn influencing the coagulation process. In addition, Ser76 of αS2-CN was found being highly phosphorylated in non-coagulating milk which may be related to its binding with other caseins during the second phase of rennet coagulation. Of these factors, salt distribution, mainly calcium and magnesium, may be the most important factor affecting casein structures and thereby milk coagulation. Importantly, these factors can affect each other while forming the cheese matrix, resulting in a complex relation with milk coagulation.

Expression comparisons of defense-related genes in resistant and susceptible chickpea cultivars in response to Ascochyta rabiei

Ascochyta blight caused by Ascochyta rabiei is a destructive disease affecting chickpea crops worldwide. Numerous studies have sought to identify resistance genes involved in the defense response against Ascochyta blight, but the function of potential resistant genes across different chickpea genetic backgrounds remains poorly understood. This study evaluated the potential role of 16 genes involved in defense responses in partially resistant (ILC482) and susceptible (Sarı98) chickpea cultivars. The leaf samples were collected from 2-week-old seedlings at 6, 12, 24, 48, and 72 h post-inoculation (hpi) with A. rabiei and analyzed for differential expression of defense genes using qRT-PCR. Twelve out of 16 genes were differentially regulated between chickpea cultivars. The qRT-PCR analysis indicated that the expression of defense genes was significantly higher in the partially resistant cultivar than that in the susceptible cultivar. The time point of the highest upregulation ratio for defense genes was variable between the cultivars. The expression of glutathione S-transferase, serine/threonine-protein kinase, WRKY gene (CaWRKY16), and wall associated-receptor kinase genes reached maximum levels in the resistant cultivar at 6 hpi, while antimicrobial peptide precursor (SNAKIN2), polymorphic antigen membrane protein, ethylene receptor-like sequences (CaETR1), and two nucleotide-binding site-leucine-rich repeat genes showed the highest activity at 24 hpi. In particular, CaWRKY16, CaWRKY50, glutathione S-transferase, CaETR1, and wall-associated-receptor kinase genes provided valuable information for assessing chickpea cultivars with different resistance levels. The results indicated that the expression changes of these genes involved in defense pathways play a crucial role in enhanced resistance of chickpea plants against Ascochyta blight and, therefore could be potential candidate genes for future breeding studies.

Polymorphic enzymes and proteins in forensic science

The population is evolving with time and with evolution there are certain genetic changes going on in humans, and this forms the foundation of polymorphism. In biological sciences the polymorphism implies that there are different allelic variants of one gene in different individuals. Therefore, the meaning of polymorphism directs us to link it to the forensic investigations. In this review we will start with the basics of polymorphism. The various research studies related to different polymorphic enzymes have been explored that have supported or would aid further in forensic investigation. Proteins are an essential part of the humans and other species; therefore, they also are found to have significance in forensic investigations. Examples of proteins are mentioned with suitable examples and their applications in various fields of forensic science. Forensic proteomics is a valuable tool in the field of forensic science, providing a wealth of information that can be used to identify suspects, link evidence to crime scenes, and determine the cause of death. Polymorphic enzymes and proteins have been extensively used in forensic science, as they can provide valuable information for individual identification and determination of biological relationships. The review explores how the polymorphic enzymes such as CYP family of enzymes, Red cell enzymes, phosphoglucomutase can be used to establish individuality and to determine biological stains' origin. Moreover, discussion about how polymorphisms in blood group systems, such as ABO are employed for the identification of individuals and the determination of biological relationships, including paternity testing. The use of these polymorphic markers in forensic science significantly improves the accuracy of individual identification and provide crucial evidence in many criminal investigations. Two case studies are discussed that give good example of how enzyme polymorphism had a pivotal role where even other biological and genetic methods could not help. However, it is observed that the interpretation of the results obtained from these markers must be done with care, considering factors such as population frequency and sample quality, to avoid incorrect conclusions.

MOLECULAR GENETIC MARKERS IN GOAT BREEDING

Introduction. The study of the genetic structure of the goat population by candidate genes associated with indicators of animal development and milk productivity is a promising direction, because the developing industry requires the introduction of advanced research methods. The use of methods of molecular genetic analysis can supplement breeding work to create optimally productive herds. In this connection, research is actively being conducted to study the influence of milk protein and hormone gene polymorphisms on milk productivity.&#x0D; Materials and methods of research. This work was carried out as a search for molecular genetic markers of productive traits in goat breeding, based on research in the world scientific literature on this topic.&#x0D; Research results. Goat breeding is an important branch of world animal husbandry. Goats are bred in all parts of the world, but the distribution of breeds in terms of productivity varies depending on the consumption tradition. For example, Europe is characterized by a predominance of dairy breeds, in Asia combined breeds, and in Africa meat breeds of goats are most often bred. The largest population of goats is kept in Asia and Africa. Significant producers of goat milk in the world are India, Bangladesh, Pakistan and Sudan. Global production of goat meat has increased by 41.66% over the past few years. Asia has the largest contribution to total meat production (70.7%). The leader in meat production is China, which produces 35.89% of goat meat from the entire world production. Goat breeding is widespread due to the ability of small cattle to easily adapt to different management systems and the ability to adapt to various climatic conditions and features of the terrain. A review of the world scientific literature confirms the fact that countries with significant demographic growth are most interested in selection work in goat breeding, taking into account genotyping by allelic variants of candidate genes for productive traits.&#x0D; Conclusions. As a result of the work, two proteins associated with quality indicators of milk were selected as candidate genes for productive traits: kappa-casein and beta-lactoglobulin, and two hormones that indirectly affect the growth and development of animals: leptin and somatotropin. The article briefly describes their functions in the body and the localization of the corresponding loci in the genome of animals. These markers are widely used for researching populations of cattle, goats and sheep in the world. This search for molecular genetic markers is aimed at carrying out similar studies in Ukraine to promote selection work in goat breeding.