Normal Genotype Research Articles

Comparison of the effects of BDNF/TRKB signalling on metabolic biomarkers in the liver of sedentary and trained rats with normal and knockout BDNF genotypes.

Introduction: The effect of brain-derived neurotrophic factor (BDNF) on the modulation of metabolic processes in the liver is poorly understood. Therefore, the aim of this study was to investigate whether hepatic concentrations or activities of metabolic biomarkers depend on altered BDNF/TrkB content in the liver, resulting from different BDNF genotypes of rats. In addition, it was assessed whether 5-week moderate endurance training modifies the levels of BDNF/Trk-B signaling and studied hepatic markers. Methods: Experiments were performed on wild-type and heterozygous BDNF knockout (HET, SD-Bdnf) rats, which were divided into four groups: control with normal genotype (Bdnf+/+), control with BDNF knockout genotype (Bdnf+/-), trained with normal genotype (Bdnf+/+T) and trained with BDNF knockout genotype (Bdnf +/-T). BDNF/TrkB concentrations as well as selected metabolic biomarkers including lipids-total cholesterol (CHOL), low-density lipoprotein (LDL), triglycerides (TG); enzymes-alanine aminotransferase (ALAT), aspartate aminotransferase (ASAT), gamma-glutamyl transferase (GGT), lactate dehydrogenase (LDH), alkaline phosphatase (ALP); hormones-insulin (INS) and leptin (LEPT) as well as interleukin-6 (IL-6) as regeneration indicator were measured directly in liver homogenates. Results and Discussion: The study showed that Bdnf+/- rats exhibited reduced BDNF/TrkB signaling (BDNF, p < 0.0001; Trk-B, p = 0.0005), altered lipid levels (CHOL, p < 0.0001; LDL, p < 0.0001; TG, p = 0.0006) and reduced hepatic ALAT (p = 0.0004) and GGT (p < 0.0001) activity, which may contribute to hepatic steatosis and obesity, as well as indicate impairment of specific metabolic pathways in the liver. Interestingly, endurance training did not alter hepatic BDNF and TrkB content, but improved ALAT (p = 0.0366) and ASAT (p = 0.0191) activities and increased hepatic IL-6 (p = 0.0422) levels in Bdnf +/- rats, suggesting enhanced liver regeneration in animals with BDNF allele loss.

Molecular and immunohistochemical study of APC exon 16 and its possible role in colorectal carcinoma development

BackgroundColorectal cancer ranks second as a cause of cancer deaths. Mutations in the adenomatous polyposis coli (APC) gene, especially in exon 16, could contribute to colorectal carcinoma development. This study explored the correlations between APC gene exon 16 variations/expression and colorectal carcinoma progression. MethodsIn a case-control study, blood samples from 150 colorectal carcinoma patients and 50 healthy volunteers were analyzed by PCR and sequencing for APC exon 16 variations. The APC protein expression on tissue samples was evaluated by immunohistochemistry and statistical analyses were used to examine clinicopathological correlations. ResultsThe sequencing analysis revealed a mutation in exon 16 of the APC gene (rs459552) in 36 % of colorectal cancer cases while absent in all non-cancer controls. Subgroup analysis by tumor grade showed higher prevalence of mutant allele in Grade II and Grade III cases, with frequencies reaching 60.0 % and 69.2 %, respectively, compared to a substantially lower prevalence of 29.4 % in Grade I patients. Immunohistochemistry showed no significant correlation between this mutation and APC expression. APC positivity proportions were 25.5 % in Grade I tumors (n = 26/102) versus 17.1 % in Grade II (n = 6/35) and 46.2 % in Grade III (n = 6/13), showing a non-significant trend of reduced positivity in higher grade tumors (p>0.05). ConclusionsThe frequency of APC exon 16 mutation (rs459552) rose significantly with increasing tumor grade. Similarly, although not statistically significant, the percentage of APC positive staining increased with poorer tumor differentiation, rather than declining. Therefore, the APC exon 16 mutation and expression analysis provides insights into colorectal cancer progression, with the rs459552 mutation correlating with grade and may promoting aggression.

Pharmacogenomic testing for CYP2C19 and CYP2D6 in cyclic vomiting syndrome.

Amitriptyline (AT) is recommended as first-line prophylactic therapy in patients with cyclic vomiting syndrome (CVS). However, significant side effects limit its use and dosing is based on trial and error. Though the Clinical Pharmacogenetic Implementation Consortium (CPIC) Guidelines recommend dosing for AT based on CY2D6 and CYP2C19 genotype profile, this is not followed in clinical practice. This pilot study determined CYP2C19 and CYP2D6 genotypes and ascertained its association with adverse drug reactions (ADRs), clinical response, and serum concentration of AT and nortriptyline in a well-characterized cohort of adults with CVS. Of 46 subjects with CVS, age 33 ± 12 years, 61% female, 85% Caucasian, a third (33%) had normal CYP2C19 metabolizer status, while 4% were poor, and 43% were ultrarapid metabolizers. Most (61%) had normal CYP2D6 genotype while 9% were poor and 2% were ultra-rapid metabolizers. There was no statistically significantassociation between genotype and ADRs, clinical response or serum drug concentration. There was a trend towards significance between genotype and clinical response, with 64% of responders having normal CYP2D6 metabolism versus 36% of nonresponders (p = 0.06). ADRs were encountered in 46% of patients with 28% discontinuing the medication as a result. A subset of patients with CVS have dysfunctional alleles of CYP2C19 and CYP2D6. Larger prospective studies to evaluate the clinical impact of pharmacogenomic testing in CVS are needed. This has the potential to optimize clinical management, predict ADRs and allow for personalized therapy.

Serum matrix metalloproteinase-9 (MMP9) and amyloid-beta protein precursor (APP) as potential biomarkers in children with Fragile-X syndrome: A cross sectional study

IntroductionFragile-X syndrome(FXS) is a neurological disease caused by abnormal repeats in the 5′untranslated region of the FMR1 gene leading to a defective fragile-X-messenger-ribonucleoprotein-1 (FMRP). Although relatively common in children, it is usually under-diagnosed especially in developing countries where genetic screening is not routinely practiced. So far, FXS lacks a laboratory biomarker that can be used for screening, severity scoring or therapeutic monitoring of potential new treatments. Methods110 subjects were recruited; 80 male children with suspected FXS and 30 matched healthy children. We evaluated the clinical utility of serum matrix metalloproteinase-9(MMP9) and amyloid-beta protein precursor(APP) as potential biomarkers for FXS. ResultsOut of 80 suspected children, 14 had full mutation, 8 had the premutation and 58 children had normal genotypes. No statistically-significant difference was detected between children with different genotypes concerning age of onset(P = 0.658), main clinical presentation(P = 0.388), clinical severity-score(P = 0.799), patient’s disease-course(P = 0.719) and intellectual disability(P = 0.351). Both MMP9 and APP showed a statistically significant difference when comparing different genotype subgroups(P = 0.019 and < 0.001, respectively). Clinically, MMP9 levels were highest in children presenting with language defects, while APP was highest in children with neurodevelopmental delay. In receiver operating curve analysis, comparing full and premutation with the normal genotype group, MMP9 has an area-under-the-curve of 0.701(95 % CI 0.557–0.845), while APP was marginally better at 0.763(95 % CI 0.620–0.906). When combined together, elevated MMP9 or APP had excellent sensitivity > 95 % for picking-up FXS cases in the clinical setting. ConclusionsScreening for circulating biomarkers in the absence of FXS genetic diagnosis is justified. Our study is the first to evaluate both MMP9 and APP in FXS suspected children in a clinical setting and to assess their correlation with disease presentation and severity.

Study of glutathione-S-transferase (GST) T1 and M1 genes in aluminum industry workers with comorbid cardiovascular pathology

Introduction. Employees of the main professions of aluminum production have a high level of professional and production-related pathology, primarily diseases of the circulatory system associated with the long-term influence of vasotoxic factors on the body. The pathogenetic course of angiopathies does not proceed in isolation, it is predetermined by chronic fluoride intoxication and hereditary component. The establishment of associations determined on the basis of studying the polymorphism of glutathione-S-transferase genes with the risk of developing comorbid cardiovascular pathology makes it possible to use genetic analysis in its early diagnosis.&#x0D; The study aims to explore the polymorphism of GST genes in aluminum industry workers with comorbid cardiovascular pathology.&#x0D; Materials and methods. The authors have carried out the study using clinical and molecular genetic diagnostic methods in workers of the main professions engaged in the production of aluminum, with chronic occupational intoxication with fluorine compounds and persons with individual signs of fluoride exposure to the skeleton (comparison group). By detecting extended deletions in the loci of the GSTT1 and GSTM1 genes, the researchers have assessed the association of the studied genotypes with the risk of chronic intoxication and atherosclerosis.&#x0D; Results. In the cohort of patients (36.58%) and in the comparison group (16.98%), the authors have revealed a combined form of cardiomyopathy complicated by coronary atherosclerosis, coronary heart disease. Also, scientists have established significant differences in the frequency distribution of genotypes of GST genes. Homozygous polymorphism GSTT1 0/0 is a risk factor for the development of occupational pathology against the background of the predominance of the normal genotype GSTM1+. Predictors (GSTT1 0/0, GSTM1 0/0) of the risk of atherosclerosis associated with hypertension in persons with chronic fluoride intoxication were determined.&#x0D; Conclusion. The association of GST genes with the development of comorbid cardiovascular pathology was determined in the structure of professional and production-related pathology in aluminum industry workers.&#x0D; Ethics. We have conducted the study of employees at the aluminum plant on a voluntary basis with the written consent of the examined persons on the basis of a specialized clinic of the Research Institute of Complex Problems of Hygiene and Occupational Diseases in accordance with the Ethical principles of conducting scientific and medical research with human participation ("Rules of Clinical Practice in the Russian Federation", Order of the Ministry of Health of the Russian Federation No. 266, 2003; Helsinki Declaration of the World Medical Association "Ethical principles of conducting scientific medical research with human participation", 2013).

Y-27632 acts beyond ROCK inhibition to maintain epidermal stem-like cells in culture.

Conditional reprogramming is a cell culture technique that effectively immortalizes epithelial cells with normal genotypes by renewing epidermal stem cells. Y-27632, a compound that promotes conditional reprogramming through an unknown mechanism, was developed to inhibit the two Rho-associated kinase (ROCK) isoforms. We used human foreskin keratinocytes (HFKs) to study the role of Y-27632 in conditional reprogramming and learn how ROCKs control epidermal stem cell renewal. In conditional reprogramming, Y-27632 increased HFK adherence to culture dishes, progression through S, G2 and M phases of the cell cycle, and epidermal stem cell marker levels. Although this correlated with ROCK inhibition by Y-27632, we generated CRISPR-Cas9-mediated HFK ROCK knockouts to test the direct role of ROCK inhibition. Knockout of single ROCK isoforms was insufficient to disrupt ROCK activity or promote HFK propagation without Y-27632. Although ROCK activity was reduced, HFKs with double knockout of ROCK1 and ROCK2 still required Y-27632 to propagate. Y-27632 was the most effective among the ROCK inhibitors we tested at promoting HFK proliferation and epidermal stem cell marker expression. Thus, the ability of Y-27632 to promote an epidermal stem cell state in conditional reprogramming not only depends upon ROCK inhibition but also acts via as-yet-unidentified mechanisms. Epidermal stem cell renewal might in part be regulated by ROCKs, but also involves additional pathways.

Novel insights into genetic susceptibility for colorectal cancer from transcriptome-wide association and functional investigation.

Transcriptome-wide association studies have been successful in identifying candidate susceptibility genes for colorectal cancer (CRC). To strengthen susceptibility gene discovery, we conducted a large transcriptome-wide association study and an alternative splicing transcriptome-wide association study in CRC using improved genetic prediction models and performed in-depth functional investigations. We analyzed RNA-sequencing data from normal colon tissues and genotype data from 423 European descendants to build genetic prediction models of gene expression and alternative splicing and evaluated model performance using independent RNA-sequencing data from normal colon tissues of the Genotype-Tissue Expression Project. We applied the verified models to genome-wide association studies (GWAS) summary statistics among 58 131 CRC cases and 67 347 controls of European ancestry to evaluate associations of genetically predicted gene expression and alternative splicing with CRC risk. We performed in vitro functional assays for 3 selected genes in multiple CRC cell lines. We identified 57 putative CRC susceptibility genes, which included the 48 genes from transcriptome-wide association studies and 15 genes from splicing transcriptome-wide association studies, at a Bonferroni-corrected P value less than .05. Of these, 16 genes were not previously implicated in CRC susceptibility, including a gene PDE7B (6q23.3) at locus previously not reported by CRC GWAS. Gene knockdown experiments confirmed the oncogenic roles for 2 unreported genes, TRPS1 and METRNL, and a recently reported gene, C14orf166. This study discovered new putative susceptibility genes of CRC and provided novel insights into the biological mechanisms underlying CRC development.

RS17817449 FTO gene variation associated with familial disease burden rather than individual risk for breast cancer

Introduction: Breast cancer (BC) is the most common malignancy in the female population globally. Obesity is associated with an increased risk of postmenopausal BC, BC recurrence, and mortality. Fat mass and obesity-associated (FTO) gene polymorphisms have attracted the most attention due to several single-nucleotide polymorphisms (SNPs) that may have an impact on obesity and different types of cancer. The primary goal of our work was to assess the association of the SNP rs17817449 FTO, physical status/metabolic changes, and dietary habits with the occurrence of BC. Methods: We conducted research as a population-genetic study including 93 women with a diagnosis of BC during their lifetime. Genomic DNA was extracted from the swabs of the buccal mucosa. Genotyping was achieved by polymerase chain reaction-restriction fragment length polymorphism. The IBM SPSS Statistics program v. 23.0 was used for statistical analysis. All values of p &lt; 0.05 were considered statistically significant. Results: The risk genotype of the FTO gene (rs17817449) GG was detected in 16 subjects (17.2%), the heterozygous TG in 46 subjects (49.5%), while the normal genotype TT was recorded in 29 subjects (31.2%). We found no statistically significant difference in the body mass index values of the three genotype groups, p = 0.72, χ2 = 2.1 and no significant relationship between the allelic or genotypic frequencies of the rs17817449 FTO gene polymorphism and other variables examined in our study. Analysis of the distribution of hereditary diseases in the family according to the molecular subtype of BC showed statistically significant p-values, p = 0.02. Conclusion: While previous research has suggested a potential link between FTO gene polymorphism, obesity, and BC, our study did not find a statistically significant association between the aforementioned variables. Future studies with a larger number of subjects in different populations should confirm the role of the FTO genotype in the risk of BC.

Laboratory methods for prenatal diagnosis

This article is aimed at discussing prenatal diagnosis of fetal genetic abnormalities as an important step in the detection and prevention of birth defects and genetic syndromes. The authors show the multi-vector nature of this problem, which requires an integrated approach and the participation of a multidisciplinary team of specialists, such as gynecologists, radiologists, neonatologists, clinical geneticists, and pediatricians. Among the many methods that allow diagnosing congenital genetic pathology, this publication highlights fluorescent hybridization in situ. In comparison with other immunogenetic methods, this technique allows assessing the genetic status of an individual cell and detecting several etiopathogenetically significant abnormal cells among thousands of others with a normal genotype. This is its advantage over PCR, in which the DNA of all cells is mixed and the result is averaged. The article provides indicators for the selection of patients for screening for fetal pathology. Prenatal screening pathways, as nowadays in most countries consist of similar tests. This article is meant to be an introduction into more detailed ethical discussions about prenatal screening. A new approach of prenatal testing (PNT) will be useful given the currently available diagnostic tests. Genetic tests and general trend of individualization in healthcare policies are directions for establishing prenatal diagnosis with consideration of ethical policies.

Cord Blood Hematological Parameters of Fetuses Detected Different Thalassemia Genotypes in the Second Trimester of Pregnancy

Hemoglobinopathies are the most common inherited diseases in humans resulting from impaired globin chain synthesis of hemoglobin. The progression of thalassemia rates is prevented with prenatal screening methods. To evaluate the hematological parameters of α- and β-thalassemia and normal fetuses aged 17-25 weeks of gestation. A cross-sectional study. Pregnant women who underwent cordocentesis in the second trimester because of the risk of having a baby with thalassemia were included in the study. Hematological indices and molecular DNA methods were analyzed from the cord blood samples of 129 women who were 17-25 weeks into pregnancy. The HPLC method was used for Hb fraction analysis. Amplification refractory mutation system, restriction enzyme analysis, multiplex polymerase chain reaction, and sequencing methods were used for the molecular analysis. Maternal contamination was eliminated by the short tandem repeat method. In total, 112 of the fetuses carry α- and β-thalassemia heterozygous or homozygous (α: 37, β: 58, mixed: 17) and 17 fetuses had a normal genotype for thalassemia. Significant differences in adult hemoglobin (HbA), fetal hemoglobin (HbF), Hb Barts, MCV, MCH, and RDW were detected in three groups compared with the normal group (p < 0.001, except for RBC, Hb, HCT, and MCHC). Differences in HbF, Hb Barts, MCV, MCH, and RDW were observed in the α-thalassemia groups compared with the normal group (p < 0.001). Among the five β-thalassemia subgroups, only HbA and RDW were different from the normal group (p < 0.001). This study could be a good reference for future studies and prenatal diagnostic applications in emphasizing the importance of changes in the blood parameters of fetuses before molecular genotyping. These hematological data give valuable information to clinicians about the fetus to enlighten families in making appropriate decisions during prenatal diagnosis.

Inheritance Pattern of Huntington’s Disease, a Multiplayer Game

Evolutionary game theory (EGT) is one of several major developments of game theory. EGTcovers ecology and population genetics, among other fields in biology. Most studies in EGTwere on a two-player game but non-linearities in biology often occur that need to be considered.Huntington's disease (HD), named after the person who wrote the first detailed description ofthe disease in 1872, is a neurodegenerative disease that is inherited. This is a case in populationgenetics, which follows the inheritance pattern called the dominant lethal. In this study, wepresented this disease as a multiplayer game among the alleles of the HD gene. We utilizedGokhale and Traulsen’s model, wherein a payoff matrix for a four-player game was reducedinto a payoff matrix for a two-player game. Depending on the fitness values of each genotype,we have determined that populations consisting of both Huntington and normal alleles mayconverge to either a purely Huntington, a purely normal, or a mixed composition where bothtypes of genes coexist. If the normal genotype produces more surviving offspring than the othergenotypes, then even if a small frequency of normal alleles is injected into a purely Huntingtonpopulation, the population will be replaced by the normal genotype over time. Such a result wasobtained using replicator dynamics and analysis of the stability of equilibrium points. Similaranalyses on other genotypes were provided in relation to the inheritance pattern of HD.

Fruit color and firmness in tomato hybrids as a function of the alleles rin, nor, norA, ogc and hp

The objective of this work was to quantify the effects of tomato ripening mutant alleles rin, nor and norA in fruit shelf life and in the internal and external fruit color, and the possible effects of their joint deployment with alleles that enhance fruit color ogc and hp. A randomized complete block design trial with three replications was used to test twelve tomato genotypes (all in genotypic background Floradade), which comprised four inbred lines (Floradade-normal genotype, TOM-559-norA/norA, TOM-613-nor/nor and TOM-614-rin/rin) and eight hybrids [F1(TOM-559 x TOM-613)-nor/norA, F1(FloraDade x TOM-559)-nor+/norA, F1(FloraDade x TOM-613)-nor+/nor, F1(FloraDade x TOM-614)-rin+/rin, F1(TOM-559 x TOM-614)-rin+/rin nor+/norA, F1(TOM-613 x TOM-614)-rin+/rin nor+/nor, F1(TOM-591 x TOM-614)-rin+/rin nor+/norA ogc+/ogc and F1(TOM-589 x TOM-614)-rin+/rin nor+/norA ogc+/ogc hp+/hp]. Our results indicate that deployment of the double heterozygous combination rin+/rin nor+/norA is effective towards improving fruit shelf life, not only with relation to the normal genotype, but also relatively to the single heterozygous genotypes rin+/rin or nor+/norA. Possible deleterious effects of the double combination rin+/rin nor+/norA on external and on internal fruit color are similar to those presented by the single heterozygous genotype rin+/rin (widely deployed in commercial hybrids), and may be attenuated by simultaneous deployment of alelles ogc and hp in the combination rin+/rin nor+/norA ogc+/ogc hp+/hp.

A Pilot Study on the Prevalence of Transcription Factor 7-Like 2 Gene (TCFL2), Rs290487 in Ethnic Groups with Type 2 Diabetes Mellitus in Bayelsa State of Nigeria

The negative impact of type 2 diabetes mellitus (T2DM) on individuals, families, health system and the economic development as a whole, is an important justification for research in this field. The present study is a hospital-based case-control type to ascertain the genetic susceptibility of T2DM among a sample population of various ethnic groups resident in Bayelsa State, Nigeria. Also evaluated was the relationship between transcription factor 7-like 2 (TCFL2) rs12255372 and rs290487 genetic polymorphisms with development of T2DM. Genotyping of TCFL2 rs12255372 and rs290487 were carried out using the polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method. Among the indigenous groups, the allelic frequencies determined for the abnormal (CC) and the normal (TT) genotypes were 94.4 and 5.6 (%), respectively; whereas, the values 91.7 and 8.3 (%), in that other, were determined for the non-indigenous groups. A convincing significant difference was found between subjects with T2DMne and their counterpart controls within indigenous people of Bayelsa State with respect to confounders, including age (t=8.046, p&lt;0.00001), BMI (t =2.628, p&lt;0.0190), waist circumference (t=2.800, p=0.0091) and fasting blood sugar (t=3.212, p&lt; 0.0006). This study verified the association of TCFL2 rs290487 with the development of T2DM in indigenous and non-indigenous people in Bayelsa State. The risk conferred by the homozygous CC genotype was higher than that of the heterozygous TC state – a fact indicative of an additive model of inheritance.

The XRCC1 and TP53 gene polymorphisms are associated with advanced-stage disease and early distant metastasis at diagnosis in non-small cell lung cancer.

Studies on single nucleotide polymorphisms (SNPs) in non-small cell lung cancer (NSCLC) suggest that DNA repair capacity may have prognostic implications for disease recurrence and survival. However, there is no study investigating the relationship between SNPs and the risk of metastasis at the time of initial diagnosis in patients with NSCLC. This study aimed to investigate the potential predictive value of SNPs in detecting the risk of metastasis at the time of initial diagnosis and poor prognosis in patients with NSCLC. In this prospective cohort study, we evaluated 275 patients with NSCLC. Analysis of SNPs from peripheral blood cells was performed by a polymerase chain reaction. Excision repair cross-complementing group 1 (ERCC1)- Asn118Asn, excision repair cross-complementing group 2 (ERCC2)-Lys751Gln, X-ray repair cross-complementing group 1 (XRCC1)-Arg399Gln, and tumor protein 53 (TP53)-Arg72Pro polymorphisms were evaluated in conjunction with the development of metastasis. The ERCC1 normal genotype, ERCC2 heterozygote genotype, XRCC1 normal genotype, and TP53 normal genotype were associated with a higher stage and more advanced-stage disease at the time of initial diagnosis (P = 0.027, 0.005, <0.001, and 0.006, respectively). Also, XRCC1 normal genotype and TP53 normal genotype were associated with the risk of metastasis at the time of initial diagnosis (P = <0.001 and 0.002, respectively). Moreover, the XRCC1 normal genotype was associated with the risk of brain metastasis at the time of initial diagnosis (P = 0.031). We showed that SNPs are related to a higher stage and more advanced-stage disease at the time of initial diagnosis in patients with NSCLC, and XRCC1 and TP53 gene polymorphisms are associated with the risk of metastasis. These results may contribute to the identification of high-risk groups and may help to earlier diagnosis and treatment in patients with NSCLC.

Fabry disease screening in young patients with acute ischemic stroke in Korea

Background: Fabry disease is an X-linked lysosomal storage disorder that results from a mutation in the α-galactosidase A (GLA) gene. It shows multiple organ involvement, including cerebrovascular disease. Since Fabry disease has a prevalence of approximately 4% in young patients with cryptogenic stroke, screening for this condition is recommended for young stroke patients. This study aimed to investigate the prevalence of Fabry disease in young acute ischemic stroke patients in Korea, the distribution of GLA gene mutations, and the subtypes of ischemic stroke. Methods: This study included 211 young patients with acute ischemic stroke or transient ischemic attack. To screen for Fabry disease, α-galactosidase A (α-Gal A) enzyme activity was measured and DNA sequencing analysis of the GLA gene was performed. Results: None of the patients exhibited low α-Gal A enzyme activity or had a pathogenic GLA mutation, but 18 nonpathogenic GLA gene variants were detected, including c.-10C&gt;T in 16 patients, c.-33C&gt;T in one patient, and c.196G&gt;C in one patient. The mean α-Gal A enzyme activity in 14 male patients with the c.-10C&gt;T variant was 5.17±1.19, which was significantly lower than that of male patients with the normal genotype (7.47±3.48, P&lt;0.05). The distribution of stroke subtypes in patients with GLA gene polymorphisms was not significantly different from that in patients with a normal genotype.Conclusions: This study demonstrates that Fabry disease is rare in young patients with ischemic stroke or transient ischemic attack in Korea, and we suggest that routine screening for Fabry disease may not be necessary for ischemic stroke patients.

Genetic analysis and reproductive intervention of 7 families with gonadal mosaicism for Duchenne muscular dystrophy

To explore the genetic basis for 7 families with gonadal mosaicism for Duchenne muscular dystrophy (DMD). For the 7 families presented at the CITIC Xiangya Reproductive and Genetic Hospital from September 2014 to March 2022, clinical data were collected. Preimplantation genetic testing for monogenic disorders (PGT-M) was carried out for the mother of the proband from family 6. Peripheral venous blood samples of the probands, their mothers and other patients from the families, amniotic fluid samples from families 1 ~ 4 and biopsied cells of embryos cultured in vitro from family 6 were collected for the extraction of genomic DNA. Multiplex ligation-dependent probe amplification (MLPA) was carried out for the DMD gene, and short tandem repeat (STR)/single nucleotide polymorphism (SNP)-based haplotypes were constructed for the probands, other patients, fetuses and embryos. The results of MLPA showed that the probands and the fetuses/probands' brothers in families 1 ~ 4, 5, 7 had carried the same DMD gene variants, whilst the probands' mothers were all normal. The proband in family 6 carried the same DMD gene variant with only 1 embryo (9 in total) cultured in vitro, and the DMD gene of the proband's mother and the fetus obtained through the PGT-M were normal. STR-based haplotype analysis showed that the probands and the fetuses/probands' brothers in families 1 ~ 3 and 5 have inherited the same maternal X chromosome. SNP-based haplotype analysis showed that the proband from family 6 has inherited the same maternal X chromosome with only 1 embryo (9 in total) cultured in vitro. The fetuses in families 1 and 6 (via PGT-M) were both confirmed to be healthy by follow up, whilst the mothers from families 2 and 3 had chosen induced labor. Haplotype analysis based on STR/SNP is an effective method for judging gonad mosaicism. Gonad mosaicisms should be suspected for women who have given births to children with DMD gene variants but with a normal peripheral blood genotype. Prenatal diagnosis and reproductive intervention may be adapted to reduce the births of further affected children in such families.

Composition, functionalities, and digestibility of proteins from high protein and normal pea (Pisum sativum) genotypes

AbstractAlthough a lot of research has been focused on the applications of pea protein, the effects of genotypes on protein applications have not been sufficiently investigated. Three high protein genotypes and four normal genotypes were included in this study. The results showed that the pea proteins from these seven genotypes differed widely in 11S/7S ratio. P1141 and Lacombe had the highest 11S/7S ratio while P1142, P0540, and Cooper had the lowest. Since the three high protein genotypes were selected from different parent lines, they had different 11S/7S ratios, which may partially explain their various functionalities. This demonstrated the potential of using breeding as a tool to manipulate the 11S/7S ratio of pea protein. The 11S/7S ratio may play a more important role in determining the protein functionalities than high/normal protein level in the seed. The solubilities of all seven samples were pH dependent. At pH 7, Lacombe and P1141 had the lowest solubility among all the tested samples, which may be a result of their high 11S/7S ratio. The proteins from all genotypes showed comparative water and oil holding capacities. P1141, Lacombe and Earlystar showed excellent emulsifying and foaming capacity at all tested pHs, which may be attributed to their relatively higher 11S/7S ratio. The in vitro digestibility varied among genotypes regardless of high protein or normal genotypes but all higher than 75%. The results of this study demonstrated that breeding could manipulate the pea protein content and composition, which further determined its functional properties and applications in food products.

Accelerating Cervical Cancer Screening With Human Papillomavirus Genotyping

Selective utilization of human papillomavirus (HPV) genotyping in cervical cancer screening can accelerate clinical management, leading to earlier identification and treatment of precancerous lesions and cancer. Specifically, immediate colposcopy (instead of 1-year return) is recommended in persons with normal cytology and HPV genotypes 16 and/or 18, and expedited treatment (instead of colposcopy) is recommended in persons with high-grade squamous intraepithelial lesion (HSIL) cytology and HPV genotype 16. The effects of implementing HPV testing and genotyping into a screening program are largely unknown. Average-risk persons aged 30-65 years screened for cervical cancer in the National Breast and Cervical Cancer Early Detection Program from 2019 to 2020 were included (N=104,991). Percentage HPV genotyping test positivity was estimated within cytology result categories. Analyses were performed in 2022. The most common abnormality was positive high-risk HPV testing with normal cytology, representing 40.1% (7,155/17,832) of all abnormal test result categories; HSIL cytology represented 3.0% (530/17,832) of all abnormal test result categories. In high-risk HPV‒positive persons with normal or high-grade cytology, HPV genotyping could accelerate management (immediate colposcopy and expedited treatment) in 5.4% of all persons with abnormal screening test results; if HPV genotyping had been performed in all high-risk HPV‒positive persons with normal or HSIL cytology, approximately 13.1% could have accelerated management. HPV genotyping in human papillomavirus‒positive persons with normal or HSIL cytology could accelerate management in a sizable percentage of persons with abnormal test results and may be particularly useful in populations with challenges adhering to longitudinal follow-up.

Mesothelioma-associated fibroblasts enhance proliferation and migration of pleural mesothelioma cells via c-Met/PI3K and WNT signaling but do not protect against cisplatin

BackgroundPleural mesothelioma (PM) is an aggressive malignancy with poor prognosis. Unlike many other cancers, PM is mostly characterized by inactivation of tumor suppressor genes. Its highly malignant nature in absence of tumor driving oncogene mutations indicates an extrinsic supply of stimulating signals by cells of the tumor microenvironment (TME). Cancer-associated fibroblasts (CAFs) are an abundant cell type of the TME and have been shown to drive the progression of several malignancies. The aim of the current study was to isolate and characterize patient-derived mesothelioma-associated fibroblasts (Meso-CAFs), and evaluate their impact on PM cells.MethodsMeso-CAFs were isolated from surgical specimens of PM patients and analyzed by array comparative genomic hybridization, next generation sequencing, transcriptomics and proteomics. Human PM cell lines were retrovirally transduced with GFP. The impact of Meso-CAFs on tumor cell growth, migration, as well as the response to small molecule inhibitors, cisplatin and pemetrexed treatment was investigated in 2D and 3D co-culture models by videomicroscopy and automated image analysis.ResultsMeso-CAFs show a normal diploid genotype without gene copy number aberrations typical for PM cells. They express CAF markers and lack PM marker expression. Their proteome and secretome profiles clearly differ from normal lung fibroblasts with particularly strong differences in actively secreted proteins. The presence of Meso-CAFs in co-culture resulted in significantly increased proliferation and migration of PM cells. A similar effect on PM cell growth and migration was induced by Meso-CAF-conditioned medium. Inhibition of c-Met with crizotinib, PI3K with LY-2940002 or WNT signaling with WNT-C59 significantly impaired the Meso-CAF-mediated growth stimulation of PM cells in co-culture at concentrations not affecting the PM cells alone. Meso-CAFs did not provide protection of PM cells against cisplatin but showed significant protection against the EGFR inhibitor erlotinib.ConclusionsOur study provides the first characterization of human patient-derived Meso-CAFs and demonstrates a strong impact of Meso-CAFs on PM cell growth and migration, two key characteristics of PM aggressiveness, indicating a major role of Meso-CAFs in driving PM progression. Moreover, we identify signaling pathways required for Meso-CAF-mediated growth stimulation. These data could be relevant for novel therapeutic strategies against PM.

Effect of α+ Thalassemia on the Severity of Plasmodium falciparum Malaria in Different Sickle Cell Genotypes in Indian Adults: A Hospital-Based Study

There is a paucity of literature on the association of α+-thalassemia, sickle-cell hemoglobin disorders, and malaria in India. This study aimed to understand the effect of α+-thalassemia on the severity of Plasmodium falciparum malaria in adults with respect to sickle-cell genotypes. The study subjects were categorized into ‘severe-malaria’ and ‘uncomplicated-malaria’ and age-gender matched ‘control’ groups. Sickle-cell and α+-thalassemia were investigated in all the recruited subjects. The effect of α+-thalassemia on the severity of malaria was analyzed in HbAA and sickle-cell genotypes (HbAS and HbSS) separately. The prevalence of α+-thalassemia in various groups ranged from 41.5% to 81.8%. The prevalence of α+-thalassemia was lower (OR = 1.64; p = 0.0013) in severe malaria (41.5%) as compared to healthy controls (53.8%) with HbAA genotype. In contrast, in HbAS genotype, the prevalence of α+-thalassemia was higher (OR = 4.11; p = 0.0002) in severe malaria (81.8%) compared to controls (52.2%). In severe malaria with HbAA genotype, there was a significantly higher hemoglobin level and low MCV and MCH level in patients with α+-thalassemia compared to the normal α-globin genotype. Further, the incidence of cerebral malaria, hepatopathy, and mortality was lower in patients (HbAA) with α+-thalassemia as compared to normal α-globin genotype (HbAA). In severe malaria with either HbAS or HbSS genotype, only a few parameters showed statistical differences with respect to α+-thalassemia. Low prevalence of α+-thalassemia in severe malaria with HbAA genotype compared to healthy controls with HbAA genotype indicates the protective effect of α+-thalassemia against severe malaria. However, the high prevalence of α+-thalassemia in patients with HbAS genotype depicts its interference in the protective effect of sickle-cell against severe malaria.