Polymorphism Screening Research Articles

Genetic polymorphisms in the angiotensin converting enzyme, actinin 3 and paraoxonase 1 genes in women with diabetes and hypertension.

To study associations between polymorphisms in the angiotensin converting enzyme (ACE I/D), actinin 3 (ACTN3 R577X) and paraoxonase 1 (PON1 T(-107)C) genes and chronic diseases (diabetes and hypertension) in women. Genomic DNA was extracted from saliva samples of 78 women between 18 and 59 years old used for genetic polymorphism screening. Biochemical data were collected from the medical records in Basic Health Units from Southern Brazil. Questionnaires about food consumption, physical activity level and socioeconomic status were applied. The XX genotype of ACTN3 was associated with low HDL levels and high triglycerides, total cholesterol and glucose levels. Additionally, high triglycerides and LDL levels were observed in carriers of the TT genotype of PON1, and lower total cholesterol levels were associated to the CC genotype. As expected, women with diabetes/hypertense had increased body weight, BMI (p = 0.02), waist circumference (p = 0.01), body fat percentage, blood pressure (p = 0.02), cholesterol, triglycerides (p = 0.02), and blood glucose (p = 0.01), when compared to the control group. Both ACTN3 R577X and PON1 T(-107)C polymorphisms are associated with nutritional status and blood glucose and lipid levels in women with diabetes/hypertense. These results contribute to genetic knowledge about predisposition to obesity-related diseases.

Bioaccumulation of Chloropyrifos Organo-pesticide and Its Toxicogenic Association with Antioxidant GSTP1 in Pakistani Pest Control Workers.

Synthetic pesticides are employed to enhance agricultural production. Chronic exposure to organophosphate (OP) pesticides may be a source of health problems. The present study was designed to examine an association of GSTP1 (rs1695) polymorphism with OP pesticide chronic exposure. A case-control study was recruited with 250 subjects comprising exposed (n = 100) and controls (n = 150). A survey was conducted to determine the pesticide type to which workers had exposed. According to recorded survey assessment, two compounds of OP pesticides chloropyrifos and malathion were investigated in the blood samples of exposed study subjects using high-performance liquid chromatography (HPLC). For screening of genetic polymorphism in GSTP1 (rs1695) polymerase chain reaction, restriction length polymorphism (PCR-RFLP) and agarose gel electrophoresis were performed. Statistically, data were analyzed using SPSS v. 20.0 and MedCal© software. Total chrom© navigator programmer was used for detection of OP residues in serum and local pesticide solution. chloropyrifos-OP pesticide residues were detected in serum of estimated chronically exposed subjects at 206nm HPLC optimal conditions. The pattern of GSTP1 (rs1695) genotypic frequencies depicted that heterozygous genotype was higher in Chloropyrifos exposed subjects (0.56) when compared with controls (0.44). Statistical outcomes showed an insignificant association with GSTP1 (rs1695) polymorphism and chloropyrifos-OP pesticide toxicity (Fisher's exact test 1.0, p = 0.25). An insignificant allelic investigation reflected a protective effect of mutant allele G against chloropyrifos-OP pesticide toxicity in exposed subjects. Findings may be helpful in identifying bioaccumulated pesticide residues, but in studied Pakistani exposed workers, no significant association of GSTP1 (rs1695) variant with chloropyrifos-OPs was demonstrated.

Encapsulated nanodroplet crystallisation: expanding solution-phase crystallisation methodologies for statistically directed polymorph screening

Encapsulated nanodroplet crystallisation: expanding solution-phase crystallisation methodologies for statistically directed polymorph screening

Emerging microfluidic platforms for crystallization process development

The microfluidic platform offers an exclusive environment compatible with different analytical tools to investigate crystallization processes. Moreover, microfluidic devices allow precise and convenient control of crystallization conditions at nano or meso scales, making them suitable for pharmaceutical and energy applications. These miniaturized screening devices can be effective during drug discovery stages for screening active ingredients available on micro/milligram scales. In this review, we discuss the advancements and applications of microfluidics platforms in the development of crystallization processes, including measurement of crystal growth and nucleation rates, screening of co-crystals, polymorphs and morphology, synthesis of nanocrystals, and determination of metastable zones and solubility. In addition, the latest microfluidics setups for high-throughput screening and lab-on-chip enhancements with sensors and automation are presented in this review. Lastly, an outlook on how microfluidic platforms can be adapted to improve workflow in crystallization process development is presented.

Organic Crystal Packing Is Key to Determining the Photomechanical Response.

Organic photomechanical crystals have great promise as molecular machines, but their development has been hindered by a lack of clear theoretical design principles. While much research has focused on the choice of the molecular photochrome, density functional theory calculations here demonstrate that crystal packing has a major impact on the work densities that can be produced by a photochrome. Examination of two diarylethene molecules reveals that the predicted work densities can vary by an order of magnitude across different experimentally known crystal structures of the same species. The highest work densities occur when molecules are aligned in parallel, thereby producing a highly anisotropic photomechanical response. These results suggest that a greater emphasis on polymorph screening and/or crystal engineering could improve the work densities achieved by photomechanical engines. Finally, an inherent thermodynamic asymmetry is identified that biases photomechanical engines to exhibit higher work densities in the forward stroke direction.

Single Nucleotide Polymorphism of LDLR Gene as Atherogenesis Markers on Macaca fascicularis and Macaca nemestrina

Long-tailed macaques (Macaca fascicularis) and pig-tailed macaques (M. nemestrina) are non-human primate species that most commonly used as animal model in the study of atherosclerosis. Genetic variation in the low density lipoprotein receptor gene (LDLR) has been associated with normal variations in plasma lipid profile and the risk of coronary heart disease (CAD) in humans. In this study, the screening of nucleotide polymorphisms on LDLR genes as molecular markers of atherogenesis in M. fascicularis and M. nemestrina was performed. The LDLR gene of exon region 6 is amplified with a specific primers. The nucleotide sequences of the amplicons is determined by the sequencing technique and the results were bioinformatic analyzed. Analysis of exon 6 region LDLR gene in M. fascicularis and M. nemestrina revealed no SNP in this exon. Based on the alignment results, the entire sample has a type of haplotype I. The type of haplotype owned by the six animals is related to the hyper-repons. Both of this species are a potential species for animals model in the study of atherosclerotic disease. The use of exon 6 as a genetic marker of M. fascicularis and M. nemestrina on a fat cholesterol diet makes the animal selection of hypo- from hyper-responder are more efficient.

Statistically guided high-throughput polymorph screening of ROY

Statistically guided high-throughput polymorph screening of ROY

Screening of interleukin 17F gene polymorphisms and eight subgingival pathogens in chronic periodontitis in Libyan patients

ABSTRACT Background: Chronic periodontitis (CP) is triggered by periodontal pathogens influenced by genetic and environmental factors. Recent studies have suggested that anti-inflammatory cytokines such as interleukin 17 (IL−17) play a prominent role in the pathogenesis of CP. Aim:This study aimed to investigate the association between eight sub-gingival pathogens and interleukin 17F (IL−17F) gene single nucleotide polymorphisms with CP among Libyans. Materials and Methods: A case–control study was conducted on 100 individuals between the ages of 25–65 years. Species-specific 16S rRNA primers for each pathogen were used in a multiplex PCR reaction to detect sub-gingival pathogens from a paper point sample. DNA was also extracted from buccal swab samples and IL−17F polymorphisms were detected by Sanger sequencing. Results: A highly significant association between the seven sub-gingival pathogens and CP, (p-value 0.0001) and a high prevalence of P. intermedia (100%), T. forsythia (96%), T. denticola and E. corrodens (92%), P. gingivalis (82%), C. rectus (74%), P. nigrescens (72%), A. actinomvcetcmcomitans (40%) were found in the case group compared with control group. A novel variant in the c. *34 G>A in IL−17F gene caused a change in glutamic amino acid to lysine amino acid, position on chromosome number (6) in the third exon, mRNA/genomic position 597, found in 14.6% of CP patients (p-value = 0.010) while the IL−17F (rs763780) SNP showed no association with CP (p-value = 0.334). Conclusion: P. intermedia appear as keystone pathogen for CP in the Libyan population. A novel variant in the IL−7F gene may be related to the severity of CP.

Meta-analysis of the association between RNF213 polymorphisms and clinical features of moyamoya disease in Asian population

BackgroundWe performed this study to explore the relationship between ring finger protein 213 (RNF213) gene polymorphisms and clinical features in moyamoya disease (MMD). MethodsElectronic databases (PubMed, Google Scholar, Embase, Scopus, Cochrane Library) were conducted from inception to May 15th, 2022. Odds ratios (ORs) with 95 % confidence intervals (CIs) were generated as effect size for binary variants. Subgroup analyses were performed by the RNF213 polymorphisms. Sensitivity was used to examine the robustness of associations. ResultsA total of 16 articles and 3061 MMD patients were included and the association of five RNF213 polymorphisms on 9 clinical features of MMD were identified. Patients under 18 years of age at onset, familial MMD, cerebral ischemic stroke and posterior cerebral artery involvement (PCi) were significantly more common in mutant type compared with wild type of RNF213. Compared with each wild type, subgroup analysis showed that rs11273543 and rs9916351 remarkably increased risk of MMD on early onset, but rs371441113 evidently delayed the onset of MMD. Rs112735431 in mutant type was significantly higher than wild type in patients with PCi. Subgroup analysis in mutant type showed that rs112735431 conspicuously decreased intracerebral/ intraventricular hemorrhage (ICH/IVH) risk and yet rs148731719 obviously increased the risk in ICH/IVH. ConclusionMore attention should be paid to patients on whom the ischemic MMD occurs younger than 18 years old. RNF213 polymorphism screening and cerebrovascular imaging examination should be performed to evaluate intracranial vascular involvement, to achieve early detection and early treatment and avoid more serious cerebrovascular events.

A study on the immunological vitality of an inflammatory biomarker explored with rs5743708 polymorphism in TLR2 gene among Saudi women confirmed with polycystic ovarian syndrome

IntroductionPolycystic ovary syndrome (PCOS) is an ovarian health condition as well as a long-term endocrine dysfunction that affects reproductive-aged women. Toll-like receptor 2 (TLR2) gene was linked to PCOS and chronic inflammation, and the prevalence of obesity was rising in Saudi women. Previous studies on rs5743708 polymorphism were documented in the obesity as well as in PCOS women. AimIn this study, we investigated the molecular role of rs5743708 polymorphism in TLR2 gene among Saudi women diagnosed with PCOS using the Rotterdam criteria. MethodsBlood samples were collected from 220 Saudi women in this hospital-based case-control study; 110 were PCOS women and remaining 110 were non-PCOS (control women). Biochemical analysis was performed on serum samples, and molecular analysis was performed on EDTA blood. Genotyping for rs5743708 polymorphism was performed with polymerase chain reaction-restriction fragment length polymorphism analysis. ResultsIn both groups, clinical data was calculated using t-test, which revealed both positive (p < 0.05) and negative (p > 0.05) associations. HWE analysis supported the rs5743708 polymorphism (p < 0.05). In the rs5743708 polymorphism, none of the genotypes, genetic models, or allele frequencies were found to be associated with PCOS and non-PCOS women. However, both ANOVA and regression analyses revealed a positive relationship in PCOS with weight and BMI (p < 0.0001). ConclusionThe rs5743708 polymorphism was not associated to PCOS in Saudi women. One of the predictions could be that 42.7% of PCOS and 73.6% of non-PCOS women were obese, and the rs5743708 polymorphism has been linked to both obesity and PCOS in the previous studies. This study suggests screening for additional polymorphisms with a large sample size.

Modular microfluidic platform for solubility measurement, nucleation statistics and polymorph screening of active pharmaceutical ingredients: Irbesartan, Rimonabant, Aripiprazole and Sulfathiazole

Drug efficacy strongly relies on the solid state of the active pharmaceutical ingredient. Classical solid-state screening methods involve different solvent compositions and supersaturations. Moreover, the many repeated experiments needed to address the stochasticity of nucleation make this approach costly. This paper presents a newly developed modular microfluidic platform that provides a universal and flexible plug-and-play tool for crystallisation studies without use of surfactants. By dissolving a powder, our set-up generates saturated solutions that can be used for solubility measurements or distributed in microdroplets. Here, we describe solubility measurements performed on different forms, stable and metastable, of pharmaceutical molecules (Irbesartan, Rimonabant and Aripiprazole) in organic and aqueous solvents. In addition, we provide nucleation statistics obtained for Sulfathiazole in water and in acetonitrile. Reporting polymorph screening on Sulfathiazole and statistics for nucleated forms, we find that the cooling rate influences both nucleation and polymorphism results, reflecting the competition between thermodynamics and kinetics. Three unknown forms were discovered, with XRD patterns and Raman spectra that do not match any referenced form. We also demonstrate the limitations of microfluidics for crystallisation by cooling: reducing the crystalliser volume considerably increases nucleation induction time.

Formation of Hemihydrate Crystal form Overcomes Milling Issue Induced by Exposed Functional Groups on Cleavage Plane for a Y5 Receptor Antagonist of Neuropeptide Y

This study aimed to investigate the crystal forms of an originally designed Y5 receptor antagonist of neuropeptide Y. Polymorphic screening was performed via solvent evaporation and slurry conversion using various solvents. The obtained crystal forms α, β, and γ were characterized by X-ray powder diffraction analysis. Thermal analysis determined that forms α, β, and γ were hemihydrate, metastable and stable forms, respectively; the hemihydrate and the stable forms were candidates. To arrange the particle size, forms α and γ were subjected to jet milling. However, form γ could not be milled because of powder stiction to the apparatus, whereas form α could be. To investigate this mechanism, single-crystal X-ray diffraction analysis was performed. The crystal structure of form γ was characterized by two-dimensional hydrogen bonding between neighboring molecules. This revealed that the functional groups forming hydrogen bonds were exposed on the cleavage plane of form γ. The three-dimensional hydrogen-bonding network with water stabilized the hemihydrate form, α. These results indicate that the hydrogen bondable groups exposed on the cleavage plane of form γ should result in stiction of the powder and adherence to the apparatus. It was concluded that crystal conversion is a method to overcome the milling issue.

Screening of single nucleotide polymorphisms within HLA region related to hematopoietic stem cell transplantation using MassARRAY technology

A growing number of studies showed that single nucleotide polymorphisms (SNPs) in the human leukocyte antigen (HLA)-related genes were associated with the outcome of hematopoietic stem cell transplantation (HSCT). Thus, other SNPs located nearby the classical HLA genes must be considered in HSCT. We evaluated the clinical feasibility of MassARRAY by comparing to Sanger sequencing. The PCR amplicons with each one of the 17 loci that were related to the outcomes of HSCT published by our previous study were transferred onto a SpectroCHIP Array for genotyping by mass spectrometry. The sensitivity of MassARRAY was 97.9% (614/627) and the specificity was 100% (1281/1281), where the positive predictive value (PPV) was 100% (614/614) and the negative predictive value (NPV) was 99.0% (1281/1294). MassARRAY is high-throughput, which can accurately analyze multiple SNPs at the same time. Based on these properties, we proposed that it could be an efficient method to match the genotype between the graft and the recipient before transplantation.

Alignment of human aquaporin 4 and ß-amyloid proteins may indicate involvement of ß-amyloid in brain water homeostasis and prevention of brain edema.

Alignment of human aquaporin 4 and ß-amyloid proteins may indicate involvement of ß-amyloid in brain water homeostasis and prevention of brain edema.

Molecular Cloning, Screening of Single Nucleotide Polymorphisms, and Analysis of Growth-Associated Traits of igf2 in Spotted Sea Bass (Lateolabrax maculatus)

The insulin-like growth factor 2 gene (igf2) is thought to be a key factor that could regulate animal growth. In fish, few researchers have reported on the single nucleotide polymorphisms (SNPs) located in igf2 and their association with growth traits. We screened the SNPs of igf2 from the spotted sea bass (Lateolabrax maculatus) by Sanger sequencing and made an association between these SNPs with growth traits. The full-length complementary (c) DNA of igf2 was 1045 bp, including an open reading frame of 648 bp. The amino acid sequence of Igf2 contained a signal peptide, an IGF domain, and an IGF2_C domain. Multiple sequence alignment showed that the IGF domain and IGF2_C domain were conserved in vertebrates. The genome sequence of igf2 had a length of 6227 bp. Fourteen SNPs (13 in the introns and one in one of the exons) were found in the genome sequence of igf2. Four SNPs located in the intron were significantly associated with growth traits (p < 0.05). These results demonstrated that these SNPs could be candidate molecular markers for breeding programs in L. maculatus.

Rs205764 and rs547311 in linc00513 may influence treatment responses in multiple sclerosis patients: A pharmacogenomics Egyptian study.

Multiple sclerosis (MS) is characterized by a complex etiology that is reflected in the lack of consistently predictable treatment responses across patients of seemingly similar characteristics. Approaches to demystify the underlying predictors of aberrant treatment responses have made use of genome-wide association studies (GWAS), with imminent progress made in identifying single nucleotide polymorphisms (SNPs) associated with MS risk, disease progression, and treatment response. Ultimately, such pharmacogenomic studies aim to utilize the approach of personalized medicine to maximize patient benefit and minimize rate of disease progression. Very limited research is available around the long intergenic non-coding RNA (linc)00513, recently being reported as a novel positive regulator of the type-1 interferon (IFN) pathway, following its overexpression in the presence of two polymorphisms: rs205764 and rs547311 in the promoter region of this gene. We attempt to provide data on the prevalence of genetic variations at rs205764 and rs547311 in Egyptian MS patients, and correlate these polymorphisms with the patients' responses to disease-modifying treatments. Genomic DNA from 144 RRMS patients was isolated and analyzed for genotypes at the positions of interest on linc00513 using RT-qPCR. Genotype groups were compared with regards to their response to treatment; additional secondary clinical parameters including the estimated disability status score (EDSS), and onset of the disease were examined in relation to these polymorphisms. Polymorphisms at rs205764 were associated with a significantly higher response to fingolimod and a significantly lower response to dimethylfumarate. Moreover, the average EDSS of patients carrying polymorphisms at rs547311 was significantly higher, whereas no correlation appeared to exist with the onset of MS. Understanding the complex interplay of factors influencing treatment response is pivotal in MS. One of the factors contributing to a patient's response to treatment, as well as disease disability, may be polymorphisms on non-coding genetic material, such as rs205764 and rs547311 on linc00513. Through this work, we propose that genetic polymorphisms may partially drive disease disability and inconsistent responses to treatment in MS; we also aim to draw attention towards genetic approaches, such as screening for specific polymorphisms, to possibly direct treatment choices in such a complex disease.

ENOS Glu298Asp (rs1799983) and AGT Met235Thr (rs699) polymorphisms in adverse cardiometabolic phenotypes: A study among Bhil tribal population of India

BackgroundCardiometabolic adversities constitute an enormous public health challenge. Along with lifestyle/socio-cultural determinants, disease outcome is modulated by genetic variants as well. Genetic polymorphisms like eNOS Glu298Asp (rs1799983) and AGT Met235Thr (rs699) have been identified as significant contributors to cardiometabolic phenotypes (obesity, hypertension and dyslipidemia) due to their role in vascular homeostasis, endothelial function, cardiac performance, etc. However, the association of these two genetic polymorphisms with cardiometabolic phenotypes among Indian Bhil tribal population have not yet been determined. MethodsThis was a cross-sectional study conducted among Bhil tribal population from India. The study recruited 297 participants aged 25–75 years through door-to-door household survey. Data on demographic and lifestyle variables were collected using a pre-tested modified interview schedule. Anthropometric (height, weight, waist circumference and hip circumference) and physiological (systolic and diastolic blood pressure) parameters were measured to define obesity and hypertension. Fasting blood samples were collected and serum was separated for lipid profiling (TC, TG, HDL-C, LDL-C and VLDL-C) and DNA was extracted from whole blood for genetic analysis (PCR-RFLP). Multivariable logistic regression was employed to identify association of genetic polymorphisms with cardiometabolic phenotypes. ResultsThe frequency of GG, GT and TT genotypes of eNOS rs1799983 were 52.86%, 45.45% and 1.68%, respectively and 0.24 minor allele frequency (MAF). The frequency of TT, TC and CC genotypes of AGT rs699 were 37.71%, 41.41% and 20.88%, respectively and 0.42 MAF. GT genotype and T allele (GT + TT genotypes) of rs1799983 were inversely associated with high WC, prehypertension, low HDL-C, and high LDL-C (all adjusted OR values <1 and p < 0.05). Conversely, TC and C allele (TC + CC genotypes) of rs699 were significantly associated with dyslipidemia, specifically high TC, TG and VLDL-C (all adjusted OR values >1.6 and p < 0.05). Also, individuals with the combination of high TC and TC genotype or C allele of rs699 appeared to have survival disadvantage at older age. ConclusionWith high frequency of AGT rs699 mutant C allele and significant association with dyslipidemia, our study suggests screening of candidate gene polymorphisms for cardiometabolic diseases among tribal populations to identify risk genotypes or alleles, which may pave way for future pharmacogenetic therapy and precision medicine.

Polymorph screening at surfaces of a benzothienobenzothiophene derivative: discovering new solvate forms.

The discovery of new polymorphs opens up unique applications for molecular materials since their physical properties are predominantly influenced by the crystal structure type. The deposition of molecules at surfaces offers great potential in the variation of the crystallization conditions, thereby allowing access to unknown polymorphs. With our surface crystallization approach, four new phases are found for an oligoethylene glycol-benzothienobenzothiophene molecule, and none of these phases could be identified via classical polymorph screening. The corresponding crystal lattices of three of the new phases were obtained via X-ray diffraction (XRD). Based on the volumetric considerations together with X-ray fluorescence and Raman spectroscopy data, the phases are identified as solvates containing one, two or three solvent molecules per molecule. The strong interaction of dichloromethane with the oligoethylene glycol side chains of the molecules may be responsible for the formation of the solvates. Temperature-dependent XRD reveals the low thermal stability of the new phases, contrary to the thermodynamically stable bulk form. Nevertheless, the four solvates are stable under ambient conditions for at least two years. This work illustrates that defined crystallization at surfaces enables access to multiple solvates of a given material through precise and controlled variations in the crystallization kinetics.

Liquid–liquid phase separation and crystallization of aspirin polymorphs from a water–acetonitrile mixed solvent medium through a swift cooling process

Screening of form-I and form-II polymorphs of the pharmaceutical solid aspirin in the vicinity of liquid–liquid phase separation in water–acetonitrile solvent mixtures through a novel swift cooling crystallization process is successfully attained.

Solvatomorphism and first-time observation of acid-acid catemer in 4-phenylamino-benzoic acids.

To investigate the polymorphism in 4-phenylamino-benzoic acids (4-PABAs) in general, and the effect on the polymorphism of these compounds exerted by substitution in particular, a series of 4-PABAs (1-8) varying in the substitution position and pattern were synthesized, and their polymorphic behavior was investigated for the first time. A relatively comprehensive polymorph screening led to the discovery of two forms, one solvent-free and the other solvate, for compounds 1, 3 and 8, and one form for the other compounds. The crystal structures were determined by single-crystal XRD. All the 4-PABAs in the crystal structures are highly twisted, and all the solvent-free crystals are based on the conventional acid-acid dimer motif, except for 2, which has a rarely observed acid-acid catemer motif. Two of the solvates (1-S and 8-S) have pyridine in the lattice while the other (3-S) has dichloromethane. The observation indicates that neither conformational flexibility or substitution alone nor the combination of both leads to polymorphism in these compounds, which is in dramatic contrast to the polymorphism of fenamic acids. The thermal properties of each system were investigated by differential scanning calorimetry and desolvation of the solvates was studied by thermogravimetric analysis. Hirshfeld surface analysis and molecular dynamics simulation were performed to study the mechanism of polymorphism and the intermolecular interactions contributing to the formation and stability of each crystal form.