Genomic Deoxyribonucleic Acid Research Articles

Correlations of analgesic dosage of morphine with SLC6A4 gene polymorphisms in patients with lung cancer.

To investigate the correlations of analgesic dosage of morphine with solute carrier family 6 member 4 (SLC6A4) gene polymorphisms in patients with lung cancer. A total of 200 lung cancer patients without cancer pain were selected as painless group, and another 200 lung cancer patients with cancer pain as cancer pain group. Visual Analogue Scale (VAS) was applied to grade the pain, the patients in cancer pain group were treated with morphine, and the dosage of morphine within 24 h was recorded. Then, the genomic deoxyribonucleic acid (DNA) was extracted from the peripheral blood of the research subjects, and the polymorphisms rs1042173 and rs7224199 of SLC6A4 gene were detected. There was a difference in the genotype distribution of SLC6A4 gene rs7224199 between painless group and cancer pain group (p=0.004), and the frequency of GG genotype was remarkably higher in cancer pain group [75 (0.375)]. The frequency of heterozygous model AC of rs1042173 and recessive model GT + TT of rs7224199 in cancer pain group was evidently lower than that in painless group (p=0.048, p=0.043). Besides, the lung cancer patients in cancer pain group had markedly lower frequency of AG haplotype (p=0.000), but notably higher frequency of AT (p=0.000) and CG (p=0.000) haplotypes of SLC6A4 gene rs1042173 and rs7224199 than those in painless group. No significant differences in genotypes of SLC6A4 gene rs1042173 (p=0.241) and rs7224199 (p=0.316) were detected among the degrees of cancer pain in cancer pain group. The analgesic dosage of morphine for the lung cancer patients was prominently correlated with the genotypes of SLC6A4 gene rs1042173 in cancer pain group. Moreover, in cancer pain group, there were significant differences in the dosage within 24 h (p=0.025), at 24 h after weight correction (p=0.001) and at 24 h after correction of weight and body surface area (p=0.000) among the genotypes, and the morphine dosage for the patients with CC genotype was significantly lower. Furthermore, the morphine dosage within 24 h (p=0.047), at 24 h after weight correction (p=0.042) and at 24 h after correction of weight and body surface area (p=0.031) were distinctly associated with the haplotypes of SLC6A4 gene in cancer pain group, of which the patients with CT haplotype were administered with a remarkably lower morphine dosage. The morphine dosage for analgesia has significant correlations with SLC6A4 gene polymorphisms in patients with lung cancer.

Association between C-myc and K-ras gene polymorphisms and non-Hodgkin lymphoma.

To explore the association between c-myc and K-ras gene polymorphisms and non-Hodgkin lymphoma (NHL). A total of 200 NHL patients in our hospital in the past 3 years were collected as disease group, while 200 healthy people were taken as control group. The genomic deoxyribonucleic acid (DNA) in the peripheral blood was extracted in both groups, amplified via Polymerase Chain Reaction (PCR) and sent to the company for the detection of c-myc and K-ras gene polymorphisms. The expressions of c-myc and K-ras were detected via Reverse Transcription-quantitative PCR (RT-qPCR), and the levels of clinical indexes hemoglobin (Hb), platelet (PLT) and lactate dehydrogenase (LDH) were determined in the Laboratory Department. The allele distribution at c-myc gene locus rs121918684 was different between control group and disease group (p=0.000), and the G allele frequency was 202 (0.505) in the control group and 263 (0.657) in the disease group. In the disease group, the GG genotype frequency at c-myc gene locus rs121918684 [97 (0.485)], the CC genotype frequency at rs775522201 [98 (0.490)], and the GA genotype frequency at K-ras gene locus rs1137188 [127 (0.635)] were all significantly higher than those in the control group (p=0.000, p=0.002, p=0.011). In the disease group, the frequency of recessive model GC+CC (p=0.003), heterozygous model GC (p=0.035), and homozygous model CC (p=0.037) at c-myc gene locus rs121918684 was significantly lower than that in the control group, and the frequency of recessive model CT+TT (p=0.046) at c-myc gene locus rs775522201 was also markedly lower than that in the control group. The haplotype frequency of c-myc CC (p=0.000), GC (p=0.000), and GT (p=0.018) in the disease group was different from that in the control group. Moreover, the CT genotype at c-myc gene locus rs775522201 was remarkably correlated with the c-myc gene expression, and the gene expression was markedly increased in the disease group. The TT genotype at K-ras gene locus rs12245 was correlated with the K-ras gene expression, and the gene expression was notably increased in the disease group. There was an association between GG genotype at c-myc gene locus rs121918684 and LDH level (p=0.000), between CT genotype at c-myc gene locus rs775522201 and PLT level (p=0.002), and between AA genotype at K-ras gene locus rs1137188 and Hb level (p=0.003). The c-myc and K-ras gene polymorphisms are associated with susceptibility to NHL, gene expression and levels of Hb, PLT, and LDH.

Molecular Identification and Genotyping of Atopobium Vaginae, 16S rRNA Gene from Bacterial Vaginosis Miscarriage Women in Al-Hillah City

This study was aimed to determine the Atopobium vaginae associated bacterial vaginosis (BV) in vaginosis women and women with miscarriage. Also other aim, the deoxyribonucleic acid (DNA) sequencing was performed for phylogenetic tree analysis of 16S rRNA gene in local A. vaginae isolates in comparison with NCBI-Genbank global A. vaginae isolates and finally submission of the present isolates in NCBI-Genbank database. One hundred fifty high vaginal swabs were collected from women with vaginosis(75 samples were taken from married vaginosis women without miscarriage and 75 samples from vaginosis women with miscarriage) from Babylon city hospital and private clinics. The age of patient 15 to 45 years. The sample was collected by disposable swabs, genomic DNA was extracted from these swabs. 16S rRNA gene detection by polymerase chain reaction technique . A. vaginae was isolated on Columbia blood agar supplemented with antibiotic for the first time in Iraq, the study confirmed that 9 (12.00%) and 5 (6.66%) of A. vaginae out of 150 swabs isolated from miscarriage and non-miscarriage vaginosis women respectively. According to the detection of the 16S rRNA gene, the study revealed that 69 (92.00%) and 47(62.66%) of A. vaginae out of 150 swabs obtained from miscarriage and non-miscarriage vaginosis women respectively. Basic Local Alignment Search Tool (BLAST) analysis showed that the 16S rRNA gene shared more than 98–99% sequence compatibility with the sequences of A. vaginae. Furthermore, the phylogenetic tree analysis of the 16S rRNA gene indicated that local A. vaginae (NO.1 and NO. 2 ) isolates shared higher homology with other A. vaginae isolates available in the GenBank. The homology of the nucleotides was between (99.17 and 98.75%) respectively.

Pathogenic mutation in a patient with Oguchi disease

Objective To identify the pathogenic mutation in a patient with Oguchi disease. Methods A Japanese patient with Oguchi disease was enrolled in this study, and underwent a comprehensive medical history assessment and multiple ophthalmic examinations, including BCVA, OCT, color fundus photography and full field electroretinogram. Genomic deoxyribonucleic acid (DNA) was extracted from peripheral blood samples for whole exome sequencing. The gene mutation was detected, and the analysis software was used to determine the conservation of the mutation and the possible structural changes. Results The patient, 71 years old, with consanguineous parents, complained of night blindness since early childhood. BCVA in both eyes was 0.7 and the golden-yellowish reflex appeared in the grey retina. The scotopic 0.01 ERGs showed a extinguished reaction in both eyes. The scotopic 3.0 ERGs showed a negative configuration with a significantly reduced a wave and a nearly absent b wave. A homozygous deletion mutation in the SAG gene (c.924delA, p.N309Tfs*12) in this patient was founded by DNA sequencing, which was predicted to generate prematurely truncated SAG protein and result in severe structural change. Homology analysis of the protein sequence indicated that the mutation resulted in an altered amino acid which was evolutionarily highly conserved among different species, strongly suggesting the potential pathogenicity of this homozygous mutation. Conclusion The mutation c.924delA(309Tfs*12) in SAG cause Oguchi disease in this patient. Key words: Retinal diseases/genetics; Frameshift mutation; Exome; Oguchi disease; SAG gene

A dysfunctional family environment and a high body fat percentage negatively affect telomere length in Mexican boys aged 8-10years.

The aim of this study was to determine whether a direct relationship existed between absolute telomere length (aTL), obesity and familial functionality in a group of Mexican children. We recruited 134 children (52% boys) aged 8-10years during regular primary care check-ups in 2016 and evaluated physical activity (PA), feeding practices, anthropometrics, body fat percentage (BF%) and family dysfunction. Optimised quantitative PCR determined aTL from genomic deoxyribonucleic acid isolated from saliva samples. Boys with a healthy BF% showed a higher aTL than their high BF% counterparts (P<.01). aTL was higher in children who performed PA than their sedentary counterparts (P<.05). Alarmingly, 90% of the children belonged to dysfunctional families and a dysfunctional family was correlated with a higher BF% (r=-.57). Negative correlations between the BF% and aTL (r=-.1765) and the BF% and time dedicated to PA (r=-.031) were observed in boys. On the contrary, we found a positive correlation between the aTL and weekly PA (r=.1938). These correlations were not observed in girls. Telomere shortening was associated with a high BF% in boys, but not girls. Dysfunctional families were also a key factor. School PA programmes should be mandatory.

Correlation analysis between ADAMTS-13 gene polymorphism and hypertension-induced atrial fibrillation.

The aim of this study was to explore the relationships between ADAMTS-13 gene polymorphisms and hypertension-induced atrial fibrillation (AF). A total of 200 hypertensive patients without AF (hypertension group) and 200 hypertensive patients with AF (AF group) treated in our hospital were enrolled. Then, peripheral blood was drawn from these subjects enrolled, and the genomic deoxyribonucleic acids (DNAs) were extracted for analysis of ADAMTS-13 gene polymorphism. Next, Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) was employed to determine the expression of ADAMTS-13 gene, and the correlations of ADAMTS-13 gene polymorphism with ADAMTS-13 gene expression and clinical indicators were analyzed. Results revealed that there was a difference in the distribution of alleles of ADSMTS-13 rs3094374 (p=0.046) and rs34054981 (p=0.039) between AF group and hypertension group. The frequency of T allele of the locus rs3094374 and that of the locus rs34054981 in ADSMTS-13 gene was higher in AF group than that in hypertension group. The distribution of genotypes of ADSMTS-13 rs28503257 (p=0.047) and rs34054981 (p=0.013) in AF group were different from those in hypertension group, and AF group had lower frequency of GA genotype of ADAMTS-13 rs28503257 and higher frequency of CT genotype of ADAMTS-13 rs34054981 than hypertension group. Besides, a difference was found in the distribution of ADSMTS-13 rs3094374 between AF group and hypertension group in recessive model (p=0.043), and the frequency of TC + CC was higher in the recessive model. Moreover, the distribution of the haplotypes CAT (p=0.012) and CGT (p=0.031) in ADAMTS-13 gene showed a difference between AF group and hypertension group. The linkage disequilibrium of the loci rs3094374 and rs28503257 in ADAMTS-13 gene was relatively great (D'=0.293). In addition, the polymorphism of the locus rs34054981 in ADAMTS-13 gene had an association with ADAMTS-13 gene expression (p<0.05). The expression of ADAMTS-13 gene was lower in patients carrying genotype TT in AF group. Furthermore, the ADAMTS-13 rs3094374 polymorphism was related to international normalized ratio (INR) (p=0.034), and the ADAMTS-13 rs28503257 polymorphism was correlated with the levels of brain natriuretic peptide (BNP) (p=0.047) and D-dimer (p=0.033). ADAMTS-13 gene polymorphism is correlated with the susceptibility and procession of hypertension-induced AF.

Self-cleaning photoactive cotton fabric modified with nanocrystalline TiO2 for efficient degradation of volatile organic compounds and DNA contaminants

In this paper, we describe the chemical engineering aspects of the preparation of highly active and stable photocatalytic textiles and show several examples of the application of these materials for the efficient degradation of harmful volatile organic compounds (VOCs) and adverse macromolecules, namely, deoxyribonucleic acid (DNA) contaminants. The photoactive textiles were obtained by the impregnation method. Titanium(IV) oxysulfate and titanium(IV) isopropoxide were used as the precursors of TiO2 and were hydrolyzed under different conditions. The effect of the preparation conditions on the photocatalytic activity of the materials was investigated. The addition of nanocrystalline TiO2 to the impregnating compound during the synthesis substantially increased the photocatalytic activity of the materials. As a result, we proposed a technique for the modification of cotton fabric using titanium(IV) isopropoxide and nanocrystalline TiO2, which allowed the production of textiles with high stability toward washing and with photocatalytic activity similar to the activity of powdered TiO2 photocatalysts. The complete oxidation of VOCs with the formation of carbon oxides and water as the final oxidation products was detected on the surface of synthesized textiles under UV light. In the case of an S-containing pollutant, deactivation of the material was observed during long-term oxidation, but as an advantage over powdered photocatalysts, the textile material could be easily reactivated by simple washing. Additionally, in this paper, we show using the polymerase chain reaction (PCR) technique the ability of photoactive textiles to decompose human genomic DNA contaminants under UV light.

Identification and Quantification of Nonviable Lactobacillus pentosus Cells in a Health Food Product.

The lack of formal protocol to verify the nonviable cell probiotic product authenticity blocks its registration and supervision process. To develop a protocol for identification, enumeration, and purity determination of a Lactobacillus pentosus nonviable cell product. The 16S ribosomal RNA (rRNA) sequencing, 16S rRNA metagenomic analysis, whole-genome sequencing, matrix-assisted laser desorption ionization-time of flight MS (MALDI-TOF-MS), and FACSMicroCount™ system were applied to establish a protocol of identification, enumeration and purity determination of a Lactobacillus pentosus nonviable cell product. The 1530 bp of 16S rRNA sequence could only identify the bacteria at genus level, but the MALDI-TOF-MS could identify both the nonviable cell and fresh culture to species level with high confidence. Metagenomic analysis of the 16S rRNA amplicon could recognize Lactobacillus as the dominant genus in the nonviable cell product. The total number of matching k-mers between the nonviable cell product and the L. pentosus BGM48 in the GenBank was the highest. The 95% confidence interval of the nonviable cell concentration in the product was determined as 4.31-4.68 × 1010 cells/g through the BD FACSMicroCount system. This validation protocol offers an executable approach that can verify microbial contents in nonviable cell products and ensure the compliance with label claims. The established validation protocol could determine the nonviable cell species through MALDI-TOF-MS, the concentration through FACSMicroCount system, and the purity and strain level identification through metagenomic analysis of 16S rRNA and the genomic deoxyribonucleic acid.

Effect of Vitamin K Epoxide Reductase Complex 1 Polymorphism on Warfarin Dose Requirement among Patients in Tertiary Care Hospital.

Background:Warfarin, anticoagulant is used for thromboembolic disorders. Inter-individual variation in clinical response to warfarin is due to various factors, including polymorphism of Vitamin K epoxide reductase complex 1 (VKORC1)-1639G>A. The aim of our study was to evaluate the effect of VKORC1 polymorphism on the maintenance dose of warfarin.Materials and Methods:Cross-sectional study conducted by the departments of Pharmacology, Cell Biology and Molecular Genetics on patients attending cardiology clinic, receiving warfarin for at least 2 months. Genomic deoxyribonucleic acid was extracted and genotyping was done by Polymerase Chain Reaction - Restriction Fragment Length Polymorphism. The correlation between VKORC1 gene polymorphism and warfarin maintenance dose was analyzed.Results:A total of 102 patients with a mean age of 47.72 ± 10.31 years, of which 58 (56.86%) were male. The frequency of VKORC1 G>A for GG, GA, and AA genotypes was 74.51%, 19.61%, and 5.88%, respectively. Variant allele AA was less frequent than the wild type. Mean weekly warfarin dose was 23.12 ± 8.08, 22.93 ± 8.21, and 15.6 ± 5.35 mg in patients with GG, GA, and AA genotypes, respectively. Patients with GG genotype required therapeutic dose compared to variant type (P = 0.001). Multiple stepwise regression model showed 26.3% variability in warfarin dose was due to VKORC1 genotype (R = 0.513, R2 = 0.263, adjusted R2 = 0.256, P = 0.0001).Conclusion:VKORC1 polymorphism alone influence 26.3% variability in warfarin dose and AA genotype patients required lower dose.

Isolation of genomic DNA from groundnut plant by modified, rapid and efficient protocol

DNA isolation is prerequisite to study the crop at molecular level. However, DNA (Deoxyribonucleic Acid) isolation is difficult in groundnut due to presence of large amount of polyphenols and polysaccharides. The age and growth stage of the plant also affect the DNA purity during isolation. In this study, newly expanded leaves and stems of five days old seedlings and matured leaves of 20 days old seedlings were used for optimization of extraction protocol. High quality DNA was obtained from all plant materials by this modified CTAB (Cetyltrimethylammonium bromide) protocol without separate purification. Optimized protocol was used to isolate DNA from newly expanded leaves of 5 days old seedlings of 100 groundnut genotypes. DNA obtained ranged from 6.3-22.1 µg/ g of sample when quantified by Nano Drop spectrophotometer and was able to amplify with Simple sequence repeats (SSR) primers during Polymerase Chain Reaction (PCR).

Human Epidermal Growth Factor Receptor-2 gene polymorphism and breast cancer risk in women from the Northeastern region of Brazil

OBJECTIVES:In the Human Epidermal Growth Factor Receptor-2 (HER2) rs1136201 variant, the presence of the G allele may promote cellular alterations and increase breast cancer risk, in addition to enhanced cellular proliferation, tumor aggressiveness, and metastases. The aim of this study was to investigate the presence of the single-nucleotide polymorphism (SNP) variant, rs1136201, within the HER2 gene in women from the Northeastern region of Brazil and breast cancer risk.METHODS:The study included 140 women who were divided into two groups, case (breast cancer) and control (without breast cancer), with 70 women in each group. Peripheral blood of each woman was drawn for the study of genomic Deoxyribonucleic acid (DNA) extracted from leukocytes using the genotyping technique by real-time polymerase chain reaction.RESULTS:The GG genotype occurred in 1 woman in both groups (1.4%) (p=0.32), while the AG genotype occurred in 19 (27.2%) and 13 (18.6%) women in the case and control (p=1.00) groups, respectively. No statistically significant difference in GG and AG genotypes was observed between the case and control groups in premenopausal women (p=1.00). Furthermore, no significant difference in genotypes was observed between the groups, among postmenopausal women (p=0.14).CONCLUSION:In this study, the HER2 rs1136201 polymorphism did not show any statistically significant association with breast cancer, both in premenopausal and postmenopausal women. Nevertheless, further studies with a larger sample size should be performed to assess the association of HER2 polymorphism with breast cancer risk in women from the Northeastern region of Brazil.

Direct Solvent-Free Modification of the Inner Wall of the Microchip for Rapid DNA Extraction with Enhanced Capturing Efficiency

Nucleic acid (NA) extraction and purification are one of the crucial steps for NA-based molecular diagnosis. However, the currently developed methods are still suffering from many issues including long process time, complicated steps, requirement of trained personnel and potential inhibition caused by chaotropic agents and/ or residual reagents. Herein, a surface-modified NA extraction microchip (SNC) is newly fabricated by introducing poly(2-dimethylaminomethyl styrene) (pDMAMS) film engaged directly on the microchip surface via initiated chemical vapor deposition (iCVD) process. The positively charged SNC inner surface could directly capture the negatively charged NA efficiently and its performance is confirmed by fluorescence microscopy and X-ray photoelectron spectroscopy. The developed SNC exhibits the deoxyribonucleic acid (DNA) capture efficiency higher than 92% regardless of initial DNA concentration in range of 20 ng/µL to 0.01 ng/µL. With this versatile DNA-capturing surface, the genomic DNAs of Escherichia Coli O157:H7 (E. coli O157:H7) is successfully extracted directly from cell lysate in the SNC with higher than 90% of efficiency within 30 min. The extraction time can be reduced to at least of 10 min maintaining extraction efficiency higher than 50%. Furthermore, the genomic DNAs are directly extracted using the SNC without loss from various real samples including juice, milk and blood serum. The proposed SNC enables us to perform an one-step NA extraction for molecular diagnosis and has the potential to be integrated into a micro-total analysis in the fields of point-of-care diagnosis.

Distinguishing genomic DNA of Brassica juncea and Arabidopsis thaliana using surface‐enhanced Raman scattering

AbstractThe emergence of the surface‐enhanced Raman scattering (SERS) technique as a prospective approach for the label‐free detection and fingerprinting of deoxyribonucleic acid (DNA) of microorganisms motivated us to detect traces of genomic DNA (gDNA) of Brassicaceae plant species, namely, Brassica juncea and Arabidopsis thaliana. DNA from the young leaf tissue of germinated B. juncea was extracted by modified cetyl trimethyl ammonium bromide extraction method and added to silver (Ag) and gold (Au) nanoparticles (NPs). The reduction in the absorbance of the surface plasmon resonance band of the metal NPs along with the appearance of a red‐shifted band indicated that DNA gets adsorbed on the metal NPs. The change in helicity of gDNA upon interaction with metal NPs was confirmed from circular dichroism studies. Dynamic light scattering measurements indicated that extent of aggregation is more in gDNA functionalized Au NPs than in Ag NPs. The transmission electron microscopy results corroborated the dynamic light scattering data. The marginal change in zeta potential value of Ag and Au NPs after binding to gDNA indicated good stability of the functionalized metal NPs. The SERS spectra revealed that adenine and deoxyribose were the active sites of gDNA that interacted with the metal NPs. From SERS, the limit of detection of gDNA was found to be ~10 and 1 ng μl−1 with Ag and Au NPs. The wavelength‐dependent Raman study showed that the charge transfer effect contributed to the SERS enhancement in addition to the electromagnetic effect. The SERS spectra of gDNA of A. thaliana functionalized Ag NPs were found to be different as compared with the gDNA of B. juncea probably due to the difference in their genome length and biochemical variability. Moreover, the Raman spectral signatures might differ due to changes in the adsorption pattern of the nucleotide bases on metal NPs surfaces.

Comparison of Methods for Genomic Deoxyribonucleic Acid (DNA) Extraction Suitable for Whole-Genome Genotyping in Traditional Varieties of Rice

The utilization and conservation traditional rice genotypes have attracted global attention. Optimization of DNA isolation protocol for genetic characterization of plants is a necessary and primary step. Over the recent years, next generation sequencing and microarray technologies have revolutionized scientific research, with their applications to high-throughput analysis of biological systems. Isolation of high quantities of pure, intact, double stranded, highly concentrated DNA is prerequisite for successful and reliable large-scale genotyping analysis. Therefore, standardization of DNA isolation is a basic requirement. Here we employed three methods of DNA isolation namely, Dellaporta, Hi-purA and modified CTAB techniques for isolation of genomic DNA from 25 indigenous rice genotypes. From the results, it was found that genomic DNA isolated by modified CTAB method to be the most appropriate for extracting high quality and maximum quantity DNA suitable for genotyping. The quality and quantity of the differentially extracted DNA was subsequently assessed by spectrophotometric measurements and gel electrophoresis.

Angiographic dynamics of coronary flow state after percutaneous coronary intervention in carriers of polymorphic RS1800470 variants of the TGF-β1 gene

Aim. To study the relationship between the angiographic dynamics of the state of the coronary bed in patients undergoing percutaneous coronary intervention (PCI) due to myocardial infarction (MI) and the carriage of polymorphic rs1800470 variants of the transforming growth factor beta-1 gene (TGF-β1).Material and methods. The study included 89 men with MI of the Caucasian race aged 32 to 65 years (52 [45,0-58,0]). Genomic deoxyribonucleic acid (DNA) was extracted from venous blood by phenol-chloroform technique. The rs1800470 polymorphism of the TGF-β1 gene was tested using real-time polymerase chain reaction (PCR) (TaqMan probes, AB 7900HT). Assessment of the severity of coronary lesions was carried out initially according to the standard coronary angiography (CAG) protocol with the calculation of the Gensini score. CAG was also conducted in the dynamics after 5-103 months (42,3±29,5 months) of the study beginning.Results. In male carriers of the rs1800470 A allele of the TGF-β1 gene, the mean values of the Gensini score statistically significantly increased (47,5±34,1 (CAG-1) and 64,5±35,5 (CAG-2), p &lt;0,001) in comparison with carriers of the homozygous GG rs1800470 variant of the TGF-β1 gene (43,5±21,1 (CAG-1) and 46,2±23,2 (CAG2), p=0,066). In patients who had rs1800470 A allele of the TGF-β1 gene, a 20%decrease in the Gensini score was observed after 7 months (p=0,013), and 30% — after 5 months (p=0,003) of the initial CAG. The development of late stent restenoses in carriers of the rs1800470 A allele and the homozygous risk genotype AA of the TGF-β1 gene was noted at an earlier date — 8 (p=0,047) and 12 months (p=0,002), respectively.Conclusion. Currently, the conduct of CAG in dynamics in patients undergoing PCI is recommended as clinically indicated. The exception is the group of patients who underwent endoprosthesis replacement of the left coronary artery or have uncorrected stenosis of a different location from 3 to 12 months after PCI. The presented data show a possible predictor role of rG1800470 polymorphism of the TGF-β1 gene in relation to the progression of coronary atherosclerosis and the development of late stent restenoses.

Acute human bocavirus 1 infection in child with life-threatening bilateral bronchiolitis and right-sided pneumonia: a case report

BackgroundHuman bocavirus 1 is a commonly detected human parvovirus. Many studies have shown human bocavirus 1 as a pathogen in association with acute respiratory tract infections in children. However, because human bocavirus 1 persists in the upper airways for extensive time periods after acute infection, the definition and diagnostics of acute human bocavirus 1 infection is challenging. Until now, detection of human bocavirus 1 exclusively, high viral load in respiratory samples, and viremia have been associated with a clinical picture of acute respiratory illness. There are no studies showing detection of human bocavirus 1 messenger ribonucleic acid in the peripheral blood mononuclear cells as a diagnostic marker for acute lower respiratory tract infection.Case presentationWe report the case of a 17-month-old Latvian boy who presented in intensive care unit with acute bilateral bronchiolitis, with a history of rhinorrhea and cough for 6 days and fever for the last 2 days prior to admission, followed by severe respiratory distress and tracheal intubation. Human bocavirus 1 was the only respiratory virus detected by a qualitative multiplex polymerase chain reaction panel. For the diagnosis of acute human bocavirus 1 infection, both molecular and serological approaches were used. Human bocavirus 1 deoxyribonucleic acid (DNA) was detected simultaneously in nasopharyngeal aspirate, stool, and blood, as well as in the corresponding cell-free blood plasma by qualitative and quantitative polymerase chain reaction, revealing high DNA-copy numbers in nasopharyngeal aspirate and stool. Despite a low-load viremia, human bocavirus 1 messenger ribonucleic acid was found in the peripheral blood mononuclear cells. For detection of human bocavirus 1-specific antibodies, non-competitive immunoglobulin M and competitive immunoglobulin G enzyme immunoassays were used. The plasma was positive for both human bocavirus 1-specific immunoglobulin M and immunoglobulin G antibodies.ConclusionsThe presence of human bocavirus 1 genomic DNA in blood plasma and human bocavirus 1 messenger ribonucleic acid in peripheral blood mononuclear cells together with human bocavirus 1-specific immunoglobulin M are markers of acute human bocavirus 1 infection that may cause life-threatening acute bronchiolitis.

DNA Fingerprinting of Resistant Mycobacterium tuberculosis Isolates in Iran by IS6110-RFLP Method

The objective of the research was to identify resistant Mycobacterium tuberculosis strains and recognize their molecular epidemiology and how they are disseminated in order to determine the cause and the effect of drug resistance and the process of its development.&#x0D; Materials and methods. Genomic deoxyribonucleic acid obtained from 37 drug-resistant Mycobacterium tuberculosis isolates were examined and fingerprinted by the IS6110- restriction fragment length polymorphism method. The data obtained were then analyzed using SPSS statistics software.&#x0D; Results. The mean patients’ age was 51 ± 15.5 years. There were 46% of male patients, 67.6% of the patients from urban areas, 86.5% of Iranians, 21.6% of relapsed cases, 8.1% of human immunodeficiency virus-positive patients, and 10.8% of the patients with a history of contact with tuberculosis patient. Based on IS6110 - restriction fragment length polymorphism, 30 different genetic types were observed which indicated a significant variation of this pathogen in Markazi province, Iran. The number of cluster genotypes was determined by 6 clusters; the number of unique types was 24. There were no relationships between age, gender, nationality, residence, close contact with tuberculosis patients, recurrence of tuberculosis, positive human immunodeficiency virus status and clustered or non-clustered strain genotype.&#x0D; Conclusions. Considering the high genetic diversity in Mycobacterium tuberculosis strains, it can be concluded that, based on IS6110 - restriction fragment length polymorphism, about 65% of cases occurred due to reactivation, and 35% of the cases were due to recent transmission. The information obtained through the molecular typing method can be very effective in future planning for tuberculosis control in Iran.

Long Noncoding RNAs of the Arterial Wall as Therapeutic Agents and Targets in Atherosclerosis.

Long noncoding ribonucleic acids (lncRNAs) have been defined as transcripts which are > 200 ribonucleotides in size and are not translated into protein. Recent work has shown that many lncRNAs do have specific molecular functions and biological effects, and are involved in a growing number of diseases, including atherosclerosis. As a consequence, lncRNAs are also becoming interesting targets for therapeutic intervention. Here, we focus on lncRNAs which are expressed in the arterial wall, and describe potential RNA therapeutic approaches of atherosclerosis by manipulating lncRNAs without affecting genome deoxyribonucleic acid content: Starting out with an overview of all lncRNAs that have so far been implicated in atherosclerosis by in vivo studies, we describe methodologies for their activation, inactivation, and RNA sequence manipulation. We continue by addressing how artificial (nonnative) therapeutic lncRNAs may be designed, and which molecular functions these designer lncRNAs may exploit. We conclude with an outlook on approaches for chemical lncRNA modification, RNA mass production, and site-specific therapeutic delivery.

Influence of Nucleic Acids on the Synthesis of Crystalline Ca(II), Ba(II), and Sr(II) Silica–Carbonate Biomorphs: Implications for the Chemical Origin of Life on Primitive Earth

The Precambrian era is associated with the origin of life on primitive Earth. It was, in fact, in that era that ribonucleic acid (RNA) was first acquired. From this evolved the genomic deoxyribonucleic acid (DNA) that later generated the biomolecules that formed the first cell. In this way, with the ongoing changes in environmental factors, the first cells evolved to give rise to higher multicellular organisms, i.e., plants and animals. Radiolarians and diatoms are organisms that have been conserved since the Precambrian era. However, although these organisms are alive (they can be considered as living fossils), they do not suffice to explain the origin of life. To understand the origin of life, the interactions among inorganic compounds that existed in the Precambrian era must be elucidated. In this context, calcium, barium, or strontium silico-carbonates (usually called silica-biomorphs) have been simply named biomorphs that emulate the morphologies of organisms, such as flowers, leaves, stems, helices,...

Heat Inactivation of Coccidioides posadasii and Coccidioides immitis for Use in Lower Biosafety Containment.

The difficulty involved in obtaining sufficient intact genomic deoxyribonucleic acid (DNA) from Coccidioides spp for downstream applications using published protocols prompted the exploration of inactivating mycelia and arthroconidia using heat under biosafety level 3 containment. This was followed by optimizing DNA extraction from mycelia using various methods at lower containment. Various exposure times and temperatures were examined to identify an effective heat inactivation procedure for arthroconidia and mycelia from both C immitis and C posadasii. Heat inactivation of mycelia was followed by DNA extraction using 2 commercially available kits, as well as a phenol:chloroform-based extraction procedure to determine DNA integrity and quantity among extraction methods using both live and heat-inactivated mycelia. Ten-minute and 30-minute exposure times at 80°C were sufficient to inactivate Coccidioides spp arthroconidia and mycelia, respectively. DNA yield between live versus heat-inactivated mycelia was similar for each extraction procedure. However, DNA obtained using phenol:chloroform was of higher quantity and integrity compared with DNA obtained using the commercially available kits, which was highly fragmented. The ability to heat-inactivate Coccidioides cultures for processing at a lower level of containment greatly increased the efficiency of DNA extractions. Therefore, this is an ideal method for obtaining Coccidioides spp DNA and inactivated arthroconidia.