Homozygous Wild Type Research Articles

SUN-117 Growth Hormone-Releasing Hormone (GHRH) Deficiency Promotes Inflammation Associated Carcinogenesis

The somatotropic axis, in addition to its well-known metabolic and endocrine effects, plays a pivotal role in modulation of inflammation. Additionally, GH-releasing hormone (GHRH) has been involved in the regulation of the growth of various human tumors. In this work, we aimed to investigate the consequences of GHRH and GH deficiency on the development of inflammation-associated colon carcinogenesis in a mouse model of isolated GH deficiency due to generalized ablation of the GHRH gene [GHRH knock out (GHRHKO)]. Homozygous GHRHKO (-/-) male mice and wild type (C57/BL6, +/+) male mice as control group were used.After azoxymetane (AOM)/dextran sodium sulfate (DSS) treatment GHRHKO-/- mice displayed higher Disease Activity Index (DAI) score, and more marked weight loss compared to +/+ animals. Additionally, -/- mice showed a significant increase in total colonic tumors, in particular of large size and predominantly localized in distal colon. In colonic tissue of AOM/DSS-treated -/- mice we found the presence of invasive adenocarcinomas, dysplasia and colitis with mucosal ulceration. Conversely, AOM/DSS-treated +/+ mice showed only presence of adenomas, without invasion of sub-mucosa. Treatment with AOM/DSS significantly increased prostaglandin (PG)E2 and 8-iso-PGF2α levels along with cyclooxygenase-2 (COX-2), tumor necrosis factor (TNF)-α, nuclear factor kappa B (NFkB) and inducible nitric oxide synthase (iNOS) gene expression, in colon specimens of both groups. However, the degree of increase of all these parameters was more marked in -/- than +/+ mice.In conclusion, generalized GHRH ablation increases inflammatory response and colon carcinogenesis in male mice. Whether this results from lack of GH or GHRH remains to be established.

Evaluation of two functional CD24 polymorphisms in primary sclerosing cholangitis

Background: Primary sclerosing cholangitis (PSC) is a progressive liver disease and characterized by chronic inflammation, sclerosis and strictures of bile ducts. Several genetic risk factors might contribute to pathogenesis. Functional single nucleotide polymorphisms (SNPs) in the CD24 gene have been associated with the development of autoimmune and autoinflammatory diseases and might contribute to the susceptibility for inflammatory bowel disease (IBD).Aim: This retrospective study aimed to evaluate the impact of two functional CD24 SNPs on clinical features and disease progression in patients with PSC.Methods: A C to T coding polymorphism (rs8734) and a TG deletion in the 3´- untranslated region (rs3838646) were genotyped. The study cohort comprises of 359 PSC patients for rs3838646 genotype and 335 PSC patients for rs8734 genotype. Clinical and laboratory parameters were collected by chart review.Results: For the rs8734 genotype, 175 patients (52.2%) were found to be homozygous wildtype (‘Ala/Ala’), 127 (37.9%) patients were heterozygous (‘Ala/Val’) and 33 patients (9.9%) were homozygous mutant (‘Val/Val’). The rs8734genotype was associated with a decreased risk for dominant strictures at first diagnosis of PSC (p = .04). For the rs3838646 genotype, 322 patients (89.7%) were found to be homozygous wildtype (‘TG/TG’); 37 showed the ‘TG/del’ genotype (10.3%). The ‘TG/del’genotype was associated with alower risk of IBD (p = .01).There was no influence of both CD24 SNPs with clinical end points or transplantation-free survival in our PSC cohort.Conclusion: Our results suggest a mild association of the rs8734 CD24 genotype with dominant strictures at first diagnosis of PSC. The rs3838646 CD24 genotype is associated with a lower rate of IBD. Both SNPs seem to modulate the clinical phenotype without major pathogenetic importance for disease progression in PSC.

Distribution of Anopheles mosquito species, their vectorial role and profiling of knock-down resistance mutations in Botswana.

Knowledge of vector species composition and monitoring their change over time is critical to evaluate malaria transmission and assess control interventions. This is especially important in countries such as Botswana, where malaria transmission is subjected to fluctuations due to climate variability. Another important aspect that impacts vector populations is the insecticide resistance. In order to assess species composition and the presence of mutations associated with insecticide resistance, Anopheles specimens from larval samplings and indoor pyrethrum spray sheet collections were analysed. A total of 349 Anopheles were screened by morphology and PCR as belonging to the An. gambiae complex and An. funestus group. Specimens were subsequently analysed for human blood meal and Plasmodium index. Finally, knock-down resistance polymorphisms were assessed. Anopheles arabiensis accounted for the majority of specimens collected through larval (96.7%) and pyrethrum spray sheet collection (87.4%) across all sampling sites, and this species was the only one found positive for human blood and for P. falciparum. Other Anopheles species were collected in small numbers by pyrethrum spray catches, namely An. quadriannulatus, An. longipalpis type C and An. parensis. The authors speculate on changing climate patterns and their possible impact on species composition. The kdr assay revealed that Anopheles mosquitoes were homozygous wild type for both L1014F (kdr-w) and L1014S (kdr-e) mutations. These results highlight the unique vectorial role of An. arabiensis in Botswana and indicated that even with prolonged use of pyrethroids and DDT, the mosquito population has not developed kdr mutations, despite some in vivo evidence of insecticide resistance.

Association of VDR (rs2228570, rs731236, rs7975232, rs1544410) and DBP (rs7041) genes polymorphisms with chronicity of hepatitis B in Iranian patients

BackgroundHost genetic factors and single nucleotide polymorphism (SNP) in host genes play important roles in the progression of disease stages and response to treatment of viral infections, including hepatitis B infection. Vitamin D plays a key role in a wide range of biological activities such as the immune response, proliferation and cellular differentiation. Since the biological effects of this vitamin are affected by the Vitamin D receptor (VDR) and the vitamin D-Binding Protein (DBP), in this study we investigated the association between VDR and DBP SNPs with the risk of chronic hepatitis B (CHB) infection in Iranian patients. MethodsOur case-control study consisted of two groups including 100 patients with CHB and 100 healthy controls. SNP of ApaI (rs7975232), TaqI (rs731236), BsmI (rs1544410) and FokI (rs2228570) VDR-related and DBP (rs7041) gene loci were determined by PCR-RFLP method. Serum ALT and AST levels and HBV virus load were determined and its association with SNP was investigated. Statistical analysis was performed using SPSS software (V22.0). ResultsThe results of this study showed that the TT (Homozygous Mutant) genotype of FokI locus (C>T; rs2228570) compared with CC (Homozygous Wild Type) and CT (Heterozygous) genotypes may be associated with an increased risk of CHB infection (P; 0.012, OR 2.674 (1.233–5.800), 95% CI). The T allele was also significantly associated with CHB infection (P; 0.009, OR 1.687 (1.136–2.506), 95% CI) compared to the C allele FokI. In this study, no significant relationship was found between CHB infection and rs7975232, rs731236, rs1544410 and rs7041 polymorphisms (P > 0.05). Also, there was no significant relationship between serum ALT and AST levels and HBV Viral load in patients with rs2228570 polymorphism (P > 0.05). DiscussionAccording to the results, VDR FokI (rs2228570) polymorphism has an impact on the chance of chronicity of hepatitis B in Iranian patients.

Role of Toll Like Receptor 2 and 4 (TLR2 and TLR4) in Susceptibility and Resistance to Mycobacterium Tuberculosis Infection among Sudanese Patients

Background: Tuberculosis (TB), an infectious disease caused by Mycobacterium Tuberculosis (MTB). Toll-like receptors (TLRs) are the best-studied class of PRRs, and they recognize specific pathogen-associated molecular patterns (PAMPs) from various microorganisms. Objectives: This study aimed to determine and measure cellular TLR2 and TLR4 in active TB, latent TB and healthy individuals. Also aimed to define correlation between gene expression and present of TLR4 and TLR2 in study volunteers. Materials & Methods: This Multi-site, prospective unmatched age and sex case-control, hospital and community-based study. 244 Consented volunteersor patients diagnosed as active tuberculosis, diagnosed latent TBI and a community controls volunteers with negative both TST and INF γ release assay were enrolled. Three-color Flow cytometry using Heparinized blood to study surface expression of TLR4 (CD284) and TLR2 (CD282), by Mean channel fluorescence intensity (MFI). EDTA blood was used for mRNA expression of both receptors using Quantitative RT-PCR. Results: TLR2, TLR4 cell surface expression by Flow cytometry was decreased in all study volunteers with no significance differences (p.value 0.24).mRNA expression of TLR2 were increased in active TB compared to Latent TBI and healthy individuals,while mRNA TLR4 decreased in active TB and showed increased pattern in latent and low level of expression in healthy volunteers, associated with no significant difference in the expression of TLR-2 mRNA between patients and close contacts with LTBI (p. value 0.045), compared with significance correlation between cellular TLR2 and expression and BCG status (p. value 0.00) respectively. There was highly significance correlation between TST indurations and level of both TLR2 and TLR4 (p. value 0.00). Also there was significant correlation between mRNA expression of TLR2 and disease types (p value 0.01). Single Nucleotide Polymorphisms (SNPs) in TLR2and TLR4,in active Tb Patients Showed three alleles,group one were heterozygous allele, homozygous wild type in group two, while homozygous mutant present in group three with statistical significant with disease stages, (P.value 0.02), while LTBI volunteers had only homozygous mutant present and group two had undetermined alleles, and four of them develop active TB infection, while healthy controls had similar alleles. Conclusion: Up-regulation of TLR-2 and TLR-4 due to SNPs may be involved in the process of tuberculosis infection in Sudanese individuals.

The correlation between total immunoglobulin-E levels and interleukin-13 gene polymorphism in asthmatic children treated with inhaled corticosteroids or montelukast

Background: The aim of this research was to study the possible correlation between single-nucleotide polymorphism of interleukin (IL)-13-1112 C/T with specific parameters in asthmatic children (total serum immunoglobulin-E [TSIgE] levels and asthma severity) treated with montelukast or inhaled corticosteroid. Methods: The study included one hundred asthmatic patients attending Karbala Teaching Hospital for children and similar numbers of healthy unrelated age-matched controls from the same locality of Iraq. TSIgE levels were determined by ELISA technique. DNA was extracted and processed by the allele-specific polymerase chain reaction technique for characterization of genetic variants of IL-13-1112 C/T gene polymorphisms. Results: Iraqi children with asthma showed a lower frequency of the IL-13-1112 CC genotype (13% vs. 16%) for control cases with a higher frequency of the heterozygous IL-13-1112 CT genotype (78% vs. 75%). IL-13-1112 showed significantly negative association with asthma in the dominant, codominant, and overdominant models of inheritance. On the other hand, comparing genotypes of subgroups related to gender, asthma severity, and asthma control showed a nonsignificant difference (P > 0.05). Homozygous wild types of IL-13-1112 TT might be considered protective to Iraqi asthma children. Conclusion: The IL-13 gene may be associated with the level of control in treated asthmatic patients. There is no correlation between TSIgE and the SNP.

Association of HSP70 gene with milk yield and milk quality of Friesian Holstein in Indonesia

The aim of this study was to identify the polymorphism of the ScrFI locus of HSP70 and the association of genetic variant of HSP70 with milk yield and milk quality of Holstein-Friesian (HF) cows in Indonesia. A total of 115 heads of HF lactating cows which come from different locations of lowland, medium, and highland. Genotyping of HSP70 genes was performed by nested PCR-RFLP method. The variant of HSP70 was polymorphism: 47 and 49 bp for homozygous wild type (CC) genotype and 47, 49 and 96 for heterozygous cytosine deletion mutant (C-) genotype. Dairy cows with CC genotypes were higher 4%FCM, protein, fat, milk total solid and lactose (P <0.05) compare to C-genotype. It could be concluded that dairy cows with CC genotypes produce higher 4% FCM and better milk quality than dairy cows with C-.

Genotype of Potassium Inwardly Rectifying Channel, Subfamily J, Member 11 (KCNJ 11) Gene and Glycaemia Control in Diabetic Patients: A Narrative Review

There are several factors that may affect the body response to the medicine and the drug response. One of the factors is the genetic variations. Some genes are predicted to have significant roles in oral antidiabetic response. This review is intended to define the role of Potassium inwardly rectifying channel, subfamily J, member 11 (KCNJ11) genotypes to the glycemia control in Diabetes melitus type 2 (DMT2) patients. We conducted literature review using keywords “genotype description, kcnj11 gene, blood sugar level, diabetes melitus type 2” in Pubmed, Science Direct, BMC, PMC and Google Scholar database. Among the fourteen articles, we found that the most alleles studied was: rs5219 and rs5215. The homozygous wildtypes of rs5219 have no correlation with the risk of DMT2. However, the correlation between the allele of rs5219 and the medication response are still unclear. Furthermore, the correlation of alleles of rs5215 and the risk of DMT2 and medication response are still contradictive. The future studies are still needed to confirm the correlation between allele of rs5219, rs5215 and the risk of DMT2 and medication response. How to Cite this Article Pubmed Style Perwitasari DA, Faridah IN, Dania H, Irham LM, Salsabila FV, Maliza R. of Potassium Inwardly Rectifying Channel, Subfamily J, Member 11 (KCNJ 11) Gene and Glycaemia Control in Diabetic Patients: A Narrative Review. SRP. 2020; 11(9): 627-631. doi:10.31838/srp.2020.9.92 Web Style Perwitasari DA, Faridah IN, Dania H, Irham LM, Salsabila FV, Maliza R. of Potassium Inwardly Rectifying Channel, Subfamily J, Member 11 (KCNJ 11) Gene and Glycaemia Control in Diabetic Patients: A Narrative Review. http://www.sysrevpharm.org/?mno=15263 [Access: March 29, 2021]. doi:10.31838/srp.2020.9.92 AMA (American Medical Association) Style Perwitasari DA, Faridah IN, Dania H, Irham LM, Salsabila FV, Maliza R. of Potassium Inwardly Rectifying Channel, Subfamily J, Member 11 (KCNJ 11) Gene and Glycaemia Control in Diabetic Patients: A Narrative Review. SRP. 2020; 11(9): 627-631. doi:10.31838/srp.2020.9.92 Vancouver/ICMJE Style Perwitasari DA, Faridah IN, Dania H, Irham LM, Salsabila FV, Maliza R. of Potassium Inwardly Rectifying Channel, Subfamily J, Member 11 (KCNJ 11) Gene and Glycaemia Control in Diabetic Patients: A Narrative Review. SRP. (2020), [cited March 29, 2021]; 11(9): 627-631. doi:10.31838/srp.2020.9.92 Harvard Style Perwitasari, D. A., Faridah, . I. N., Dania, . H., Irham, . L. M., Salsabila, . F. V. & Maliza, . R. (2020) of Potassium Inwardly Rectifying Channel, Subfamily J, Member 11 (KCNJ 11) Gene and Glycaemia Control in Diabetic Patients: A Narrative Review. SRP, 11 (9), 627-631. doi:10.31838/srp.2020.9.92 Turabian Style Perwitasari, Dyah Aryani, Imaniar Noor Faridah, Haafizah Dania, Lalu Muhammad Irham, Fathia Vikri Salsabila, and Rita Maliza. 2020. of Potassium Inwardly Rectifying Channel, Subfamily J, Member 11 (KCNJ 11) Gene and Glycaemia Control in Diabetic Patients: A Narrative Reviews in Pharmacy, 11 (9), 627-631. doi:10.31838/srp.2020.9.92 Chicago Style Perwitasari, Dyah Aryani, Imaniar Noor Faridah, Haafizah Dania, Lalu Muhammad Irham, Fathia Vikri Salsabila, and Rita Maliza. Genotype of Potassium Inwardly Rectifying Channel, Subfamily J, Member 11 (KCNJ 11) Gene and Glycaemia Control in Diabetic Patients: A Narrative Review. Systematic Reviews in Pharmacy 11 (2020), 627-631. doi:10.31838/srp.2020.9.92 MLA (The Modern Language Association) Style Perwitasari, Dyah Aryani, Imaniar Noor Faridah, Haafizah Dania, Lalu Muhammad Irham, Fathia Vikri Salsabila, and Rita Maliza. Genotype of Potassium Inwardly Rectifying Channel, Subfamily J, Member 11 (KCNJ 11) Gene and Glycaemia Control in Diabetic Patients: A Narrative Review. Systematic Reviews in Pharmacy 11.9 (2020), 627-631. Print. doi:10.31838/srp.2020.9.92 APA (American Psychological Association) Style Perwitasari, D. A., Faridah, . I. N., Dania, . H., Irham, . L. M., Salsabila, . F. V. & Maliza, . R. (2020) of Potassium Inwardly Rectifying Channel, Subfamily J, Member 11 (KCNJ 11) Gene and Glycaemia Control in Diabetic Patients: A Narrative Reviews in Pharmacy, 11 (9), 627-631. doi:10.31838/srp.2020.9.92

Growth hormone-releasing hormone (GHRH) deficiency promotes inflammation-associated carcinogenesis

The somatotropic axis, in addition to its well-known metabolic and endocrine effects, plays a pivotal role in modulation of inflammation. Moreover, growth hormone (GH)-releasing hormone (GHRH) has been involved in the development of various human tumors. In this work we aimed to investigate the consequences of GHRH deficiency on the development of inflammation-associated colon carcinogenesis in a mouse model of isolated GH deficiency due to generalized ablation of the GHRH gene [GHRH knock out (GHRHKO)]. Homozygous GHRHKO (-/-) male mice and wild type (C57/BL6, +/+) male mice as control group, were used.After azoxymetane (AOM)/dextran sodium sulfate (DSS) treatment -/- mice displayed higher Disease Activity Index (DAI) score, and more marked weight loss compared to +/+ animals. Additionally, -/- mice showed a significant increase in total tumors, in particular of large size predominantly localized in distal colon. In colonic tissue of AOM/DSS-treated -/- mice we found the presence of invasive adenocarcinomas, dysplasia and colitis with mucosal ulceration. Conversely, AOM/DSS-treated +/+ mice showed only presence of adenomas, without invasion of sub-mucosa.Treatment with AOM/DSS significantly increased prostaglandin (PG)E2 and 8-iso-PGF2α levels along with cyclooxygenase-2 (COX-2), tumor necrosis factor (TNF)-α, nuclear factor kappa B (NF-kB) and inducible nitric oxide synthase (iNOS) gene expression, in colon specimens. The degree of increase of all these parameters was more markedly in -/- than +/+ mice.In conclusion, generalized GHRH ablation increases colon carcinogenesis responsiveness in male mice. Whether this results from lack of GH or GHRH remains to be established.

Trade-Off Between Triadimefon Sensitivity and Pathogenicity in a Selfed Sexual Population of Puccinia striiformis f. sp. Tritici.

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most destructive diseases of wheat and has been largely managed using demethylation inhibitor (DMI) fungicide triadimefon in China. To determine the sensitivity of Pst, a Chinese Pst isolate and its sexually produced progeny isolates were tested with triadimefon using the detached leaf method. The half maximal effective concentration (EC50) values varied greatly among the progeny isolates, ranging from 0.06 mg L–1 to 7.89 mg L–1. Twenty-six of the 56 tested progeny isolates were less sensitive to triadimefon than the parental isolate. A single-nucleotide mutation at the 401 position resulting in an amino acid change from tyrosine (Y) to phenylalanine (F) in the 134th codon (Y134F) of the cytochrome P450 sterol 14a-demethylase enzyme (CYP51), the target gene of DMI fungicide, was identified in the parental isolate. The 87 tested progeny isolates segregated into 19 homozygous wild type (AA), 40 heterozygous (AT), and 28 homozygous mutant (TT) genotypes, fitting a 1:2:1 ratio (χ2 = 2.43; P = 0.30). The mutant isolates had higher EC50 values than the wild type isolates. Significant differences in logEC50 were found between the mutant isolates and the wild type isolates (P = 2.2e–16). However, homozygous and heterozygous mutant isolates were not significantly different (P = 0.21), indicating dominant mutation. Twenty-two progeny isolates were used to inoculate a susceptible wheat variety, and latency period and lesion growth were recorded to compare wild type and mutant isolates for the pathogenicity fitness components. A moderate but significant negative correlation was detected between lesion growth and sensitivity to triadimefon (r = −0.53; P = 0.01). No significant variation in lesion growth was found between homozygous and heterozygous mutant isolates (P = 0.83). In the case of latency period and triadimefon sensitivity, no significant correlation was found (P = 0.17). These results are useful for understanding reduced sensitivity in the pathogen population and improving stripe rust management.

Field Performance of High Oleic Soybeans with Mutant FAD2‐1A and FAD2‐1B Genes in Tennessee

AbstractSoybean [Glycine max (L.) Merr.] oil with high oleic acid (&gt;75%) has increased oxidative stability and health benefits that are valuable for food, fuel, and industrial products. It has been determined that two naturally occurring mutations in genes FAD2‐1A and FAD2‐1B can combine to produce high oleic soybeans. The objective of this study was to test the effect of these mutant alleles on seed yield and oil and protein concentration. Molecular markers assisted in the creation of a population of 48 BC3F2:4 lines (93.75% expected genome commonality). Each line was classified into one of four genotypic groups where both FAD2‐1A and FAD2‐1B genes were either homozygous wild type or mutant, respectively. Twelve lines for each genotypic group were evaluated in three replications at six locations across Tennessee. There was no seed yield difference between the high oleic genotypic group and the other groups (P &lt; 0.05). On the other hand, there were differences in fatty acid profiles and oil and protein concentrations. In combination, the mutant FAD2‐1A and FAD2‐1B alleles produced a mean of 803.1 g kg−1 oleic acid. This is, on average, approximately 500 g kg−1 more oleic acid compared to soybean lines with only one mutant FAD2‐1 allele. The high oleic double mutant group had more total oil (228.0 g kg−1) and protein (401.0 g kg−1) compared to all other genotypic groups (P &lt; 0.05). Overall, this specific combination of mutant FAD2‐1A and FAD2‐1B alleles appears to generate conventional high oleic soybeans without a yield drag.

Short communication: Lethal mutations in Vorderwald cattle through Montbéliarde incrossings

Vorderwald cattle are a dual-purpose cattle breed with high migrant contributions from Montbéliarde bulls in the recent past. Through the wide use of Montbéliarde bulls, undesirable alleles were also disseminated into the Vorderwald population. Haplotypes on bovine chromosome 19 (MH1) and 29 (MH2), supposed to harbor lethal mutations, were identified in Montbéliarde cattle. A study in French Montbéliarde cattle identified the PFAS:g.28511199C>T (rs455876205) variant as the most likely MH1 embryonic lethal mutation. The objective of the present study was to determine whether the PFAS:g.28511199C>T variant was introduced into Vorderwald cattle through Montbéliarde bulls and disseminated in this population. The present study expands on previous work on the deleterious SLC37A2 variant (ss2019324563) of the MH2 locus. Herein, we traced the ss2019324563 variant back to the Montbéliarde bull, which was the most likely source for this deleterious mutation in Vorderwald cattle. We genotyped 354 Vorderwald cattle for the PFAS variant, resulting in 41 heterozygous individuals and a T allele frequency of 0.058. An aborted fetus homozygous mutant for SLC37A2 from our previous study on the MH2 locus in Vorderwald cattle was wild type for the PFAS variant. Both lethal mutations were segregating independently of each other, and we found no indications of joint occurrence in a larger number of animals. Neither SLC37A2 nor PFAS double heterozygous mutants were lethal. The earliest animal with a heterozygous PFAS genotype was 1 of 5 migrant Montbéliarde bulls, and this bull was the most likely origin of the deleterious PFAS allele in Vorderwald cattle. All Vorderwald cattle under study born before introgression of this Montbéliarde bull were homozygous wild type. In addition, all 41 heterozygous Vorderwald cattle had genetic contributions from this Montbéliarde bull, whereas in 74 Vorderwald cattle without genes from Montbéliarde bulls, the PFAS T allele was not observed. In a sample of actual German Fleckvieh the PFAS T allele could be found at a very low frequency. Our study demonstrated the introgression of lethal variants through Montbéliarde bulls into a traditional cattle breed highly adapted to harsh local conditions. These findings underline the need to screen bulls for lethal mutations before their wide use in breeding, particularly in breeds with a focus on preservation of their genetic uniqueness.

Mutations in the Progesterone Receptor (PROGINS) May Reduce the Symptoms of Acute Hepatitis E and Protect Against Infection.

BackgroundMutations in the progesterone receptor (PR) gene, PROGINS, have been studied in relation to hepatitis E virus (HEV) infection. Patients with the PROGINS gene may develop a worse clinical course of hepatitis E. The aim of our study was to evaluate the influence of PROGINS on the susceptibility to and the clinical course of HEV infection in HIV patients.MethodsThis study included patients with HIV who were evaluated in previous prospective studies for the prevalence and incidence of HEV. The following three groups of patients were studied: (i) never infected, (ii) past infections, and (iii) recently infected. We determined the PR genotype to evaluate the proportion of patients who were homozygous for PROGINS according to HEV infection. We also compared the proportion of PROGINS carriers with a recent HEV infection according to their symptomatology.ResultsIn this study, 311 patients infected with HIV were included. Of those patients, 198 were homozygous wild type (63.7%), 91 were heterozygous (29.3%), and 22 were homozygous PROGINS (7.1%). We found that the homozygous PROGINS genotype in women was associated with a lower HEV seroprevalence. In addition, in patients with a recent HEV infection, none of those homozygous for PROGINS presented symptoms.ConclusionThe PROGINS mutation plays a protective role against HEV infection and is associated with subclinical infection in HIV-infected patients, particularly women.

The influence of Tumor Necrosis Factor-alpha gene polymorphism on oxidative stress in patients with oral precancerous lesions and oral cancer

ObjectiveThe aim of our study was to determine the association of Tumor Necrosis Factor alpha (TNF-α) single nucleotide polymorphism (SNP) characterized by the substitution of guanine (G) by adenine (A) at the position -238 (TNF-α-238G/A) and position -308 (TNF-α-308G/A) in the promoter region of TNFα with oral precancerous lesions and oral cancer and its influence on oxidative stress and antioxidant enzymes. DesignThe study group consisted of 125 subjects were included for the study among which 40 subjects each of oral cancer and oral precancerous lesions respectively and 45 were normal healthy controls. Genotyping was done by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) and antioxidant enzymes like catalase, superoxide dismutase (SOD), glutathione reductase (GR), and oxidative stress markers like malondialdehyde (MDA) were estimated in blood plasma. ResultsSignificant association of TNF-α-308G/A single nucleotide polymorphism with oral precancerous lesions and oral cancer. A significant difference in the catalase activity was observed in the TNFα-308GA genotype as compared to the GG genotype in oral cancer. Superoxide dismutase and glutathione reductase activity was reduced in TNFα−238AA subgroup of oral cancer patients and in TNFα-308 GA genotype in both the groups of patients. A significant increase in malondialdehyde level was observed in the TNF-α-238 rare genotype as compared to the homozygous wild type. ConclusionOxidative stress in patients with oral precancerous lesions and oral cancer correlates with the presence of TNFα−238AA and TNFα-308GA genotypes which could help in designing new therapeutic strategies for oral cancer treatment.

Relationship Between Single Nucleotide Polymorphisms and the Toxicy and Side Effects of Paclitaxel and Platinum-Based Chemotherapy in Patients with Malignant Tumors

Objective: Investigating the relationship between single nucleotide polymorphisms (SNPs) and the toxic and adverse effects of paclitaxel and platinum-based chemotherapy in patients with malignant tumors, to provide recommendations for individualized treatment. Methods: Determinate 17 patients with malignant tumor DNA site and analysis. Results: 1. All 17 selected specimens’ fluorouracil related genes 18DPYD*2A(476002G&gt;A)GG type、153DPYD*13(1679T&gt;G)TT type 154DPYD(2846A&gt;T)TT type, and the synthesis of DPYD enzyme activity. 21GSTP1(313A&gt;G) polymorphism site mutation rate was 25.0%(4 cases), 29XRCC1(16323944T&gt;C) polymorphism site mutation rate was 90.9%(10 cases), 62ABCB1(3435C&gt;T) polymorphism site mutation rate was 52.9%(9 cases), and 68MTHFR(677C&gt;T) polymorphism site mutation rate was 50.0%(8 cases). 2. Fluorouracil related genes 18DPYD*2A(476002G&gt;A)GG type, 153DPYD*13(1679T&gt;G)TT type, 154DPYD(2846A&gt;T)TT type ,and the synthesis DPYD enzyme activity is normal. 3. Paclitaxel related genes 62ABCB1(3435T&gt;C) CC type has a lower incidence of hematotoxicity and neurotoxicity, than CT type and TT type. 13ABCB1(2677T&gt;G)GG type has a higher rate of drug resistance than TT type. 14CYP1B1*3(C&gt;G)CC type has a higher progression-free survival. Platinum-related genes 21GSTP1(313A&gt;G)AA is homozygous wild type and has a higher incidence of hematotoxicity than GA type. 29XRCC1(1196T&gt;C)CC is homozygous mutant and has a higher risk of serious neutropenia than CT type. 62ABCB1 (3435T&gt;C)CC is homozygous mutant and has a higher risk of lymphatic metastasis than TC type and TT type. 68MTHFR(677C&gt;T)TT type is homozygous mutant and has a higher mucosal toxicity and toxic and side effects than CT type and CC type. Conclusion: Single nucleotide polymorphism is related to the toxic and side effects of chemotherapy，the detection of SNP to predict the toxicity risk of drug users can be an important reference index to guide clinical individualized treatment.

Vascular Endothelial Growth Factor Receptor Polymorphisms and im Treatment Response in Chronic Myeloid Leukemia Patients

Abstract Background Imatinib mesylate (IM) is the gold standard drug for chronic myeloid leukemia (CML) treatment. However, intrinsic and acquired resistance against IM has become a problem in CML treatment. Vascular endothelial growth factor receptor 2 (VEGFR2) plays a pivotal role in leukemia associated angiogenesis. Its expression is a good predictor of poor survival in CML. Genetic polymorphisms in VEGFR2 result in varied VEGFR2 expression and may lead to inter individual variation in disease progression and outcome to IM therapy. Aim of this study was to investigate the frequencies of VEGFR2 polymorphisms rs1531289 and rs2305948 in Malaysian CML patients and to determine the impact of these SNPs on IM treatment response in these CML patients. Methods DNA extracted from 230 CML patients (121 IM resistant and 109 IM good response) were genotyped using restriction fragment length polymorphism analysis and genotypes were categorized into homozygous wildtype, heterozygous and homozygous variants. The difference in genotype frequencies of the two SNPs, between the two groups of CML patients were compared using x2 test and the association between genotypes and treatment response was assessed using logistic regression analysis and deriving ORs with 95% CI. Results The SNP rs1531289 did not show any significant association with IM resistance. For rs2305948, the heterozygous variant genotype (CT) and variant allele (T) showed significant association with IM resistance (OR, 2.38; CI,1.33-4.25, p = 0.003 and OR,1.48; CI, 1.02-2.14,p=0.039 respectively). Haplotype analysis of the two SNPs did not show any association with treatment response. Conclusion VEGFR2 polymorphism rs2305948 which is associated with cytogenetic response and development of resistance to IM therapy might be a potential prognostic predictor of IM treatment response in CML patients. This study was financially supported by Research University Grant of Universiti Sains Malaysia, 2017.

Role of polymorphism (rs1024611) in monocyte chemoattractant protein-1 gene in diabetic foot ulcer of Iraqi patients

BackgroundDiabetic foot ulcer (DFU) is one of the frequent diabetic problems, and the main reason for hospitalization and lower extremity amputation, among patients with type 2 diabetes mellitus (T2DM). The factors correlated with vascular functions and angiogenesis may be involved in the development and prevalence of DFU. Monocyte chemoattractant protein-1 (MCP-1) is a pro-inflammatory cytokine, playing a principal role in the inflammatory process. The influence of MCP-1 –2518A/G polymorphism and its association with the MCP-1 expression level had hardly been reported among patients with DFU. ObjectiveOur objective was to study the risk of genetic variation of MCP-1 -2518A/G in DFU and its association with the MCP-1 expression level and some biomarkers in serum of DFU patients. Subjects and methodsA cross-sectional study was performed on 235 subjects, including 135 individuals with DFU and another 100 with T2DM but without foot ulcer. Phenotypic data, including serum MCP-1 level, lipid profile, blood glucose, and HOMA-IR was investigated. DNA was extracted from the whole blood and the genotyping of the rs1024611 was determined, using tetra amplification refractory mutational system-polymerase chain reaction (tetra ARMS-PCR), followed by electrophoresis on 1.5% agarose gel. ResultsThe amplification of the MCP-1 gene gives three genotypes as indicated by (255 bp 379 bp) bands for those with homozygous wild type (AA), homozygous mutant (GG) genotypes and heterozygous (255 bp, and 379 bp) respectively, in addition to control with 594 bp band. Genotype frequencies of the rs1024611 polymorphism were consistent with Hardy–Weinberg equilibrium (HWE). Allele frequencies were 17%, 60% and 22% for AA, AG and GG, respectively, in DFU group. While the frequencies in the non-DFU group were 36%, 52% and 12% for AA, AG and GG, respectively. The homozygous genotype (GG) significantly increased the risk of DFU with respect to the AA wild type (OR = 3.35, CI 95% = 1.3–8.62, P = 0.01). The AG genotype significantly raised the risk of DFU (OR = 2.8, CI 95% = 1.42–5.7, P = 0.002). Co-dominant genotypes of the rs1024611 polymorphism exhibited significant association with high serum MCP-1 level, insulin resistance and lipid profile, in DFU group. ConclusionAccordingly, we conclude that the rs1024611 polymorphism was significantly correlated with increased DFU susceptibility in the Iraqi Patients, and the frequency of AG and GG genotypes was higher in DFU, compared to T2DM without DFU.

ITPA, TPMT, and NUDT15 Genetic Polymorphisms Predict 6-Mercaptopurine Toxicity in Middle Eastern Children With Acute Lymphoblastic Leukemia.

Background: Acute lymphoblastic leukemia (ALL) is the most common cancer seen in children worldwide and in the Middle East. Although there have been major advances in treatment approaches for childhood ALL, serious toxicities do occur but with significant inter-individual variability. The aim of this study is to measure the frequency of polymorphisms in candidate genes involved in 6-Mercaptopurine (6-MP) disposition in a combined cohort of Middle Eastern Children with ALL, and evaluate whether these polymorphisms predict 6-MP intolerance and toxicity during ALL maintenance therapy. Methods: The study includes children treated for ALL on two treatment protocols from two cohorts; one from Lebanon (N = 136) and another from Kurdistan province of Iran (N = 74). Genotyping for the following six candidate genetic polymorphisms: ITPA 94C > A (rs1127354) and IVS2+21A > C (rs7270101), TPMT*2 238G > C (rs1800462), TPMT*3B 460G > A (rs1800460) and *3C 719A > G (rs1142345), and NUDT15 415C > T (rs116855232) was performed and analyzed in association with 6-MP dose intensity and toxicity. Results: As expected, TPMT and NUDT15 variants were uncommon. As for ITPA, both polymorphisms were more common in the Lebanese as compared to the Kurdish cohort with a minor allele frequency of 0.05 for 94C > A and 0.14 for IVS2+21A > C in the Lebanese only (N = 121), and of 0.01 for either ITPA polymorphism in Kurds. The most significant toxic effects were depicted with the NUDT15 polymorphism with a median 6-MP dose intensity of 33.33%, followed by 46.65% for TPMT*3A polymorphism, followed by 65.33% for two ITPA risk allele carriers and 74% for one ITPA risk allele carriers, in comparison to a median of 100% for the homozygous wild type in the combined cohort (P < 0.001). In addition, the onset of febrile neutropenia was significantly higher in variant allele carriers in the combined cohorts. Conclusions: These data confirm the predictive role of TPMT, NUDT15, and ITPA in 6-MP intolerance in Middle Eastern children with ALL. Given the relatively high frequency of ITPA variants in our study and their significant association with 6-MP dose intensity, we recommend that physicians consider genotyping for ITPA variants in conjunction with TPMT and NUDT15 prior to 6-MP therapy in these children.

AKNA Frameshift Variant in Three Dogs with Recurrent Inflammatory Pulmonary Disease.

We investigated three related Rough Collies with recurrent inflammatory pulmonary disease. The clinical symptoms were similar to primary ciliary dyskinesia (PCD). However, the affected dogs did not carry any known pathogenic PCD variants. Pedigree analysis suggested a recessive mode of inheritance. Combined linkage and homozygosity mapping in three cases and seven non-affected family members delineated 19 critical intervals on 10 chromosomes comprising a total of 99 Mb. The genome of one affected dog was sequenced and compared to 601 control genomes. We detected only a single private homozygous protein-changing variant in the critical intervals. The detected variant was a 4 bp deletion, c.2717_2720delACAG, in the AKNA gene encoding the AT-hook transcription factor. It causes a frame-shift introducing a premature stop codon and truncates 37% of the open reading frame, p.(Asp906Alafs*173). We genotyped 88 Rough Collies consisting of family members and unrelated individuals. All three available cases were homozygous for the mutant allele and all 85 non-affected dogs were either homozygous wildtype (n = 67) or heterozygous (n = 18). AKNA modulates inflammatory immune responses. Akna−/− knockout mice die shortly after birth due to systemic autoimmune inflammatory processes including lung inflammation that is accompanied by enhanced leukocyte infiltration and alveolar destruction. The perfect genotype-phenotype association and the comparative functional data strongly suggest that the detected AKNA:c.2717_2720delACAG variant caused the observed severe airway inflammation in the investigated dogs. Our findings enable genetic testing, which can be used to avoid the unintentional breeding of affected puppies.

Abstract 2940: 4-Hydroxytamoxifen enhances sensitivity of estrogen receptor α-positive breast cancer to docetaxel in an estrogen and ZNF423 SNP-dependent fashion

Abstract Background: In early stage, ERα-positive breast cancer, the concurrent use of endocrine therapy and chemotherapy has not been shown to be superior to sequential use. We hypothesized that genetic biomarkers can aid in selecting patients who would benefit from chemo-endocrine therapy. Our previous genome-wide association study (GWAS) and functional studies revealed that a single nucleotide polymorphism (SNP) in ZNF423, rs9940645, determines tamoxifen response. Specifically, 4-hydroxytamoxifen (4-OH-TAM) increases the gene expression of ZNF423, a transcription factor for BRCA1, in cells that are carry the rs9940645 variant, but not the wildtype, genotype. In this study, we set out to identify additional genes regulated by ZNF423, which, upon identifying a relationship between ZNF423 and cell cycle genes, led us to investigate taxane response in a SNP- and tamoxifen-dependent fashion. Method: Gene correlation analysis using The Cancer Genome Atlas (TCGA) breast cancer dataset was used to identify genes with expression correlated with that of ZNF423. Quantitative reverse transcription PCR, chromatin immunoprecipitation, and luciferase reporter assays were used to validate the gene associations. To test ZNF423 rs9940645-specific phenotypes, we used ZR75-1 cells, which are homozygous for the variant SNP, and ZR75-1 cells that were generated to be homozygous wild type (WT) using CRISPR-Cas9 technology, in addition to Hs578T breast cells with ERα overexpression (WT) and HCC1500 cells (variant). Cell cycle was assessed by propidium iodide staining. Cytotoxicity was assessed using a colorimetric assay. Results: Mitosis-related genes VRK1 and PBK, which encode histone H3 kinases, were experimentally validated to be regulated by ZNF423. Specifically, ZNF423 knockdown resulted in decreased VRK1 and PBK expression and activity. Additionally, ZNF423 knockdown resulted in enhanced docetaxel-induced G2/M arrest and cytotoxicity which could be rescued by VRK1 or PBK overexpression. Lastly, breast cancer cells carrying the rs9940645 variant SNP genotype had increased G2-M arrest and decreased cell viability when treated with docetaxel in combination with estradiol and 4-OH-TAM. Conclusion: We identified ZNF423regulated genes involved in the G2/M phase of the cell cycle. We found that 4-OH-TAM sensitized ERα-positive breast cancer cells to docetaxel treatment in a ZNF423 SNP-dependent manner. Our findings suggest that patients with rs9940645 variant genotype may benefit from concurrent tamoxifen endocrine therapy and docetaxel chemotherapy. This would impact a substantial proportion of patients given the minor allele frequency of 0.47 for this SNP. Citation Format: Gen Wang, Sisi QIN, Jacqueline Zayas, James N. Ingle, Mohan Liu, Richard Weinshilboum, Kunwei Shen, Liewei Wang. 4-Hydroxytamoxifen enhances sensitivity of estrogen receptor α-positive breast cancer to docetaxel in an estrogen and ZNF423 SNP-dependent fashion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2940.