Deoxyribonucleic Acid Integrity Research Articles

Improved semen parameters and sperm deoxyribonucleic acid fragmentation following microscopic varicocelectomy: short term results of a prospective study

Background: Assessment of sperm deoxyribonucleic acid (DNA) damage post varicocelectomy, has brought about new prospects in treating men with infertility. Improvements in spermatogenesis with decrease in sperm DNA damage has been demonstrated which has led to significant enhancement in fertility rates. Therefore, we evaluated the efficacy of microscopic varicocelectomy on the reduction in sperm DNA damage in concurrence to the conventional improvement of semen parameters. Methods: All patients with a diagnosis of varicocele leading to infertility and planned for microscopic varicocelectomy from January 2013 through September 2019, were included in the study. Sperm chromatin structure assay (SCSA) was done preoperatively and at 3 and 6 months postoperatively for sperm DNA integrity assessment and the results expressed as sperm percentage DFI were compared. A p value <0.05 was considered statistically significant. Results: 105 infertile men underwent microscopic varicocelectomy. Mean age and infertility period from the date of unprotected sex were 26.5±5.6 years and 30 months respectively. Baseline mean sperm concentration was 29 million/ml, mean progressive motility 24% and a percentage DFI ranged from 5–36% with a mean of 16%. Sperm DNA integrity improved significantly with a DFI decrease from 25±11% to 18±6%, 3 months post-surgery which was consistent at 6 months (13±5%). Sperm concentration and progressive motility increased with a mean sperm count improvement by 21.9 million/ml and mean motility by 20.3%. Conclusions: Microscopic varicocelectomy provides durable improvement in DNA integrity and semen parameters. Sperm DNA integrity assessment using SCSA is a useful tool to demonstrate change in the semen quality post treatment.

Inventory and quality control of biosample collection from pregnant women at different gestational ages to search for early biomarkers of pregnancy complications

Aim. To conduct an inventory and quality control of biosample collection from pregnant women at different gestational ages to search for early biomarkers of pregnancy complications.Material and methods. In this work, methods for assessing the sample preparation of biosamples were used, including the isolation of deoxyribonucleic acid (DNA)/ribonucleic acid from various biomaterials, polyacrylamide gel electrophoresis of protein, and database analysis.Results. Inventory and quality control of the collection (n=18390) was carried out, which confirmed the high safety of the biomaterial, regardless of storage period. The mean concentration of DNA was 69,96±6,56 ng/µl, extracellular DNA (ecDNA) — 0,20±0,02 ng/µl, ribonucleic acid — 38,16±5,69 ng/µl. DNA Integrity Number (DIN) >9, RNA integrity number (RIN) >7, A260/280 >1,8 were for all studied samples. Protein electrophoresis demonstrated no degradation of protein zones after longterm storage. The number of errors detected during the inventory was 84 (0,46% of all records in the database), while there were 64 donors with incomplete clinical information (15% of all donors in the collection).Conclusion. The necessity of mandatory implementation of standard operating procedures when creating and maintaining a collection, on the one hand, and periodic inventory with biosample quality assessment, on the other, has been demonstrated.

The effect of zinc supplementation on improving sperm parameters in infertile diabetic men.

Diabetes mellitus (DM) may have different adverse effects on the male reproductive system. Zinc (Zn) is one of the necessary elements in the human and mammalian diet that plays an important role in scavenging reactive oxygen species (ROS) by providing antioxidant and anti-apoptotic properties. The aim of this study was to determine the protective effects of zinc supplements on sperm chromatin and the evaluation of sperm deoxyribonucleic acid (DNA) integrity in diabetic men. In this interventional study, 43 infertile Iranian men in diabetic and non-diabetic groups were included. They were then randomly divided into two subgroups: normal saline intake and zinc sulfate intake (25 mg orally for 64 days each). Different indices of sperm analysis (number, morphology and motility) and testosterone levels were evaluated in four groups. Protamine deficiency and DNA fragmentation were assessed using chromomycin A3 (CMA3) and sperm chromatin dispersion (SCD) methods, respectively. Zinc supplementation reduced the deformity of neck and head of sperms (p < 0.05), as well as deformity of sperm tail in infertile diabetic men. Zinc administration ameliorated sperm motility types A, B and C (p < 0.05). Moreover, zinc administration reduced abnormal morphology and DNA fragmentation of sperms, which increased the SCD1 and SCD2 and reduced the SCD3 and SCD4 in both treated groups. Zinc supplementation, as a powerful complement, is able to balance the effect of diabetes on sperm parameters, sperm chromatin and DNA integrity. Consequently, zinc supplementation can probably be considered a supportive compound in the diet of diabetic infertile men.

Effect of non-ionizing radiation from mobile phones in different time periods exposure on DNA integrity of infertile men

Exposure to radiation of various forms is one of the many environmental, health, and lifestyle variables that have been linked in recent years to an increase in the rate of male infertility. According to data from several recently published studies, mobile phone use may negatively affect sperm parameters, which may lower male fertility .Ninety-seven samples of semen from fertile and infertile individuals were sent to the "High Institute for Infertility Diagnosis and Assisted Reproductive Technologies. Al-Nahrain University in Baghdad, between November 2022 and May 2023. The samples were split into four groups: normozoospermia (n = 40), asthenozoospermia (n = 25), oligozoospermia (n = 16), and teratozoospermia (n = 16). Each group was further divided into two groups based on the amount of time spent using a mobile phone each day: less than 4 hours per day and equal to or more than 4 hours per day. A routine seminal fluid analysis was carried out to measure the sperm parameters and test the deoxyribonucleic acid fragmentation rate using an acridine orange stain .Samples exposed to electromagnetic waves of cell phones showed a significant decrease in DNA normality. deoxyribonucleic acid fragmentation index (p &lt; 0.001) in normozoospermia patients exposed to more than 4 hours/day of non-ionizing electromagnetic waves. The results of deoxyribonucleic acid fragmentation index after treatment were shown a significant improvement compared to before the treatment in all fertile and infertile groups. Electromagnetic waves emitted from cell phones are associated with a decrease of deoxyribonucleic acid normality when exposure for &lt;4hours/day. The study recommend that men of reproductive age avoid keeping their cell phones in the hip pocket and front pants pocket which may negatively affect spermatozoa deoxyribonucleic acid integrity and impair male fertility

A Comparative Study of Three Nucleic Acid Integrity Assay Systems.

Background: The measurement of nucleic acid quality, especially the analysis of integrity, is a key step for many downstream experiments in biomedical research and quality control of biomaterials. General gel electrophoresis is a traditional method for nucleic acid integrity analysis. Currently, more electrophoresis techniques are becoming standardized and automated operations with higher precision. In this study, we have evaluated the comparability and bias of the outcomes from three commercial assay systems. Methods: Seventy-two deoxyribonucleic acid (DNA) and 67 ribonucleic acid (RNA) samples were selected for methodological comparison among different systems. The DNA Quality Number (DQN) and RNA Quality Number (RQN) of BIOptic Qsep400, DNA Quality Score (DQS) and RNA Quality Score (RQS) of PerkinElmer Labchip GX Touch HT were separately compared with the DNA Integrity Number (DIN) and RNA Integrity Number (RINe) of the Agilent 4200 TapeStation according to Clinical and Laboratory Standards Institute (CLSI) guideline (EP09-A3). Results: The biases of the mean estimated between DQN and DIN, DQS and DIN both exceeded the acceptance criteria. The Passing-Bablok regression analysis between DQN and DIN, and the Deming regression analysis between DQS and DIN, showed the biases were both within the acceptance criteria, and the bias between DQN and DIN was smaller. For the comparisons of RQN and RINe, RQS and RINe, the regression analyses revealed the biases were both within the acceptance criteria. The bias of the mean estimated between RQS and RINe was outside of the acceptance criteria. Conclusions: There was a good comparability in nucleic acid integrity detection between BIOptic Qsep400 and PerkinElmer Labchip GX Touch HT with the Agilent 4200 TapeStation. However, the bias and linear correlations require more attention between systems.

Sperm selection by the oviduct: perspectives for male fertility and assisted reproductive technologies†.

The contribution of sperm to embryogenesis is gaining attention with up to 50% of infertility cases being attributed to a paternal factor. The traditional methods used in assisted reproductive technologies for selecting and assessing sperm quality are mainly based on motility and viability parameters. However, other sperm characteristics, including deoxyribonucleic acid integrity, have major consequences for successful live birth. In natural reproduction, sperm navigate the male and female reproductive tract to reach and fertilize the egg. During transport, sperm encounter many obstacles that dramatically reduce the number arriving at the fertilization site. In humans, the number of sperm is reduced from tens of millions in the ejaculate to hundreds in the Fallopian tube (oviduct). Whether this sperm population has higher fertilization potential is not fully understood, but several studies in animals indicate that many defective sperm do not advance to the site of fertilization. Moreover, the oviduct plays a key role in fertility by modulating sperm transport, viability, and maturation, providing sperm that are ready to fertilize at the appropriate time. Here we present evidence of sperm selection by the oviduct with emphasis on the mechanisms of selection and the sperm characteristics selected. Considering the sperm parameters that are essential for healthy embryonic development, we discuss the use of novel in vitro sperm selection methods that mimic physiological conditions. We propose that insight gained from understanding how the oviduct selects sperm can be translated to assisted reproductive technologies to yield high fertilization, embryonic development, and pregnancy rates.

An internal pilot study of a novel rectal mucocellular sampling device to allow next-generation sequencing for colorectal disease.

BackgroundThe ORI-EGI-02 study was designed to test the hypothesis that rectal mucus collected using a novel rectal sampling device (OriCol™), contains sufficient human deoxyribonucleic acid (DNA) of the required quality for Next Generation Sequencing (NGS), for colorectal disease genetic signature discovery.MethodsUsing National Institute for Health and Care Research methodology, an internal pilot study was performed in January 2020–May 2021, at four sites in the United Kingdom, to assess the process of recruitment, consent, specimen acquisition and viability for analysis. Following an OriCol™ test, the sample was stabilized with a buffer solution to preserve the material, which was posted to the laboratory. Samples were processed using QIAamp® DNA Blood Midi kit to extract DNA and Quant-iT™ PicoGreen® dsDNA Reagent to quantify the retrieved DNA. DNA integrity was measured by Agilent TapeStation system. 25 ng of human amplifiable DNA was prepared for Next Generation Sequencing (NGS), which was performed on an Illumina NextSeq550 sequencer using the 300-cycle high output kit v2.5.ResultsThis study assessed the first 300 patients enrolled to the ORI-EGI-02 Study (n = 800). 290/300 (96.67%) were eligible to undergo OriCol™ sampling procedure and 285/290 (98.27%) had a successful OriCol™ sample taken. After transportation, extraction and quantification of DNA, 96.20% (279/290) of the samples had NGS successfully performed for bioinformatic analysis.ConclusionsOur internal pilot study demonstrated that the OriCol™ sampling device can capture rectal mucus from unprepared bowel in subjects who could undergo a digital rectal examination. The technique could be applied irrespective of age, frailty, or co-morbidity. Completion of the study to 800 patients and analysis of NGS data for colorectal cancer mutations will now proceed.

The Correlation Between Sperm DNA Integrity and Conventional Semen Parameters

The integrity of sperm deoxyribonucleic acid (DNA) and chromatin is very important for genetic material transmission into offspring. Therefore, the aim of the present study was to investigate the correlation between sperm deoxyribonucleic acid integrity and sperm parameters. The current study was conducted on 96 semen samples. This study was done at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University for the period between November 2020 and May 2021. The samples were collected, and seminal fluid analysis was performed according to World Health Organization guidelines. Sperm Chromatin Dispersion test was performed. The results showed that the deoxyribonucleic acid fragmentation of sperms in infertile men in this study was higher than reported in fertile men in previous studies indicating that deoxyribonucleic acid fragmentation (DNA) is actively contributing at least partially to male infertility. In terms of enrolled men, sperm deoxyribonucleic acid fragmentation (SDF) showed significant positive correlation with round cell count.

The Correlation Between Sperm DNA Integrity and Conventional Semen Parameters

The integrity of sperm deoxyribonucleic acid (DNA) and chromatin is very important for genetic material transmission into offspring. Therefore, the aim of the present study was to investigate the correlation between sperm deoxyribonucleic acid integrity and sperm parameters. The current study was conducted on 96 semen samples. This study was done at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University for the period between November 2020 and May 2021. The samples were collected, and seminal fluid analysis was performed according to World Health Organization guidelines. Sperm Chromatin Dispersion test was performed. The results showed that the deoxyribonucleic acid fragmentation of sperms in infertile men in this study was higher than reported in fertile men in previous studies indicating that deoxyribonucleic acid fragmentation (DNA) is actively contributing at least partially to male infertility. In terms of enrolled men, sperm deoxyribonucleic acid fragmentation (SDF) showed significant positive correlation with round cell count

DNA Supramolecular Assembly on Micro/Nanointerfaces for Bioanalysis.

Facing increasing demand for precision medicine, materials chemistry systems for bioanalysis with accurate molecular design, controllable structure, and adjustable biological activity are required. As a genetic biomacromolecule, deoxyribonucleic acid (DNA) is created via precise, efficient, and mild processes in life systems and can in turn precisely regulate life activities. From the perspective of materials chemistry, DNA possesses the characteristics of sequence programmability and can be endowed with customized functions by the rational design of sequences. In recent years, DNA has been considered to be a potential biomaterial for analysis and has been applied in the fields of bioseparation, biosensing, and detection imaging. To further improve the precision of bioanalysis, the supramolecular assembly of DNA on micro/nanointerfaces is an effective strategy to concentrate functional DNA modules, and thus the functions of DNA molecules for bioanalysis can be enriched and enhanced. Moreover, the new modes of DNA supramolecular assembly on micro/nanointerfaces enable the integration of DNA with the introduced components, breaking the restriction of limited functions of DNA materials and achieving more precise regulation and manipulation in bioanalysis. In this Account, we summarize our recent work on DNA supramolecular assembly on micro/nanointerfaces for bioanalysis from two main aspects. In the first part, we describe DNA supramolecular assembly on the interfaces of microscale living cells. The synthesis strategy of DNA is based on rolling-circle amplification (RCA), which generates ultralong DNA strands according to circular DNA templates. The templates can be designed with complementary sequences of functional modules such as aptamers, which allow DNA to specifically bind with cellular interfaces and achieve efficient cell separation. In the second part, we describe DNA supramolecular assembly on the interfaces of nanoscale particles. DNA sequences are designed with functional modules such as targeting, drug loading, and gene expression and then are assembled on interfaces of particles including upconversion nanoparticles (UCNPs), gold nanoparticles (AuNPs), and magnetic nanoparticle (MNPs). The integration of DNA with these functional particles achieves cell manipulation, targeted tumor imaging, and cellular regulation. The processes of interfacial assembly are well controlled, and the functions of the obtained bioanalytical materials can be flexibly regulated. We envision that the work on DNA supramolecular assembly on micro/nanointerfaces will be a typical paradigm for the construction of more bioanalytical materials, which we hope will facilitate the development of precision medicine.

Mitigation of Toxic Effects of 2 Naphthalene Sulfonate after its Treatment with Microbial Consortia

Present study aimed to treat 2 Napthalene Sulfonate (2NS), a dye intermediate with microbial consortia and assess its potential toxicological impact before and after treatment on the integrity of Deoxyribo-Nucleic Acid (DNA) in blood cells of Channa punctatus. Symbolic elevation in DNA damage with untreated 2NS administered fishes was observed as revealed by comet assay and micronucleus test. However, 2NS after being degraded using mixed bacterial population showed significant reduction in toxicological effects of 2NS. Thus, this study not only illustrated the adverse impact of such toxic contaminant of industrial waste but also suggested a highly efficient and eco-friendly way to remove the harmful xenobiotics from the environment which may help to reduce the exposure of aquatic fauna and flora to such lethal toxicants.

Use of Diverse Extraction Protocols to Decide the Integrity of Deoxyribonucleic Acid (DNA) Samples Extracted from Bovine Bone Samples of Different Ages obtained from an Abattoir in Ikorodu, Lagos State, Nigeria

Techniques for the identification human and non-human biological samples are developing at very high rates with the advent of different DNA extraction methods and polymerase chain reaction (PCR) based assays. This study aimed at using different extraction protocols to determine the integrity of DNA samples extracted from bovine bone samples of different ages collected from abattoir in Ikorodu, Lagos State, Nigeria. The DNA was extracted using CTAB, PCI protocols and a DNA kit (Quick DNA MiniPrep Plus Kit). Bovine mtDNA fragment containing the gene encoding ATPase 8 was amplified via Polymerase Chain Reaction (PCR). The PCR products were analysed on 1.8% agarose gel. It was observed that the DNA samples extracted using the PCI method at 48 h incubation time had the highest purity (1.68) and concentration (336 ng/µl) compared to other extraction methods employed in the study. However, DNA kit extracted samples had mean ± SE purity (1.52 ± 0-05) and concentration (192.25 ± 31.41 ng/µl) values that were higher than CTAB protocol values but lower than PCI protocol values. All isolated DNA samples were PCR-worthy and thus yielded PCR products which were within the expected amplicon size of 126-bp. All DNA extraction protocols employed in this study are stable and efficient for use in the identification of aged non- human bones. This study also revealed that these protocols can be used to isolate PCR amplifiable DNA from old bones.

Detection of Sperm DNA Integrity and Some Immunological Aspects in Infertile Males

Immunoinfertility caused by anti-sperm antibodies (ASAs) represents about 10–20% of infertility among couples, which interfere with sperm motility and ability to penetrate cervical mucus, sperm-oocyte binding, fertilization, and embryo development. In addition, deoxyribonucleic acid (DNA) damages are increasingly found with infertile cases affecting male reproduction potency and progeny. This study aims to assess the semen, presence of ASAs, and DNA fragmentation index in normozoospermic patients. A total number of 116 cases with an average age of 20–51 years old, and duration of infertility at 4.70 ± 2.77 are classified into 77 and 39 primary and secondary types of infertility, respectively. Mixed agglutination reaction test was used to estimate the ASAs in semen (direct method) and in seminal plasma and blood serum (indirect method), for both immunoglobulins IgG and IgA. Acridine orange test was used to detect DNA fragmentation index. The results showed a significant difference (P &gt; 0.05) for those with a secondary type of infertility at means 24.37 and 31.48 for IgG, and 14.46 ± 1.76 and 6.86 ± 0.39 for IgA by both direct and indirect methods, respectively. The direct method showed a significant difference only for the sperm tail, while that for indirect method was in sperm mid-piece. The mean of DFI for all cases was 38.25 ± 2.08, at 41.61 ± 2.19 and 31.63 ± 4.29, for both primary and secondary cases, respectively. The percentage of ASAs revealed no significant difference with DFI, except in some parts of sperm.

P-074 Sperm deoxyribonucleic acid integrity decreases with age and exhibits a rapid decline beyond the age of 35: a retrospective evaluation of 3446 semen samples

Abstract Study question Does advancing paternal age correlate with sperm DNA fragmentation index (DFI) and is there a cut-off age beyond which sperm DFI increases significantly? Summary answer In infertile men, DFI correlate with advancing paternal age and should be routinely screened starting 35 years of age. What is known already In recent decades, birth rates have substantially increased for men older than 30 years because of advanced age of marriage, rising life expectancy at birth, modern societal norms, and accessibility to assisted reproductive technology (ART). Advanced paternal age has been associated with a decline in conventional semen parameters (volume, concentration, motility, DFI), as well as, reduced fertility, increased risk of miscarriage, structural chromosomal aberrations and complex epigenetic disorders. Several studies have recommended testing sperm DFI in infertile men with advanced age (≥40 years) as it may provide prognostic information for couple attempting natural and assisted reproduction. Study design, size, duration This is a retrospective study of 3446 semen samples from patients under investigation for infertility between April 2016 and January 2022. Semen samples were obtained after 2-3 days of sexual abstinence. Patients were stratified into seven groups based on their age: patients ≤ 29 years (n = 127; 3.7%), 30-35 years (n = 868; 25.2%), 36-39 years (n = 863; 25.0%), 40-45 years (n = 1017, 29.5%), 46-49 years (n = 321; 9.3%), 50-55 years (n = 179, 5.2%) and ≥ 56 years (n = 71, 2.1%). Participants/materials, setting, methods Conventional semen parameters were assessed according to the WHO criteria and DFI was evaluated by TUNEL assay using the APODirect Kit run on BDAccuriC6 flow cytometer. Pearson’s r was used for correlation analysis between sperm concentration, DFI and paternal age. DFI results for each stratified patient group were evaluated by one-way ANOVA, followed by Tukey pos-hoc multiple comparison test. Results are presented as the mean±standard error and a P-value of &amp;lt; 0.05 was considered statistically significant. Main results and the role of chance In this cohort of men with a mean age of 39.5 years ± 0.1 (range 23-76 years), sperm deoxyribonucleic acid (DNA) fragmentation (21.1% ± 0.2) was positively correlated with age (r = 0.23, p˂0.001). In contrast, the correlation between sperm concentration and age was non-significant (r = 0.03, p = 0.07). Mean DFI in patients segregated into seven age groups were: ≤29 years (15.7% ± 0.8), 30-35 years (17.7% ± 0.4), 36-39 years (19.7% ± 0.4), 40-45 years (22.6% ± 0.4), 46-49 years (26.2% ± 0.9), 50-55 years (26.7% ± 1.2) and ≥ 56 years (31.1% ± 2.0). Mean %DFI level in the 26-29 and 30-35 age groups were non-significantly different (p = 0.65). However, mean %DFI level in the 36-39 age group was significantly higher than in the 26-29 and 30-35 age groups (p = 0.02 and p = 0.03, respectively). Mean %DFI level in the older age groups (40-45, 46-49, 50-55 and ≥ 56 years) were all significantly higher than in the 26-29 or 30-35 age groups (p˂0.001). Using a %DFI threshold level of 16.9%, 46.0% of patients ˂36, 52.2% of men aged 36-39, 60.2% of men aged 40-45, 67.3% of men aged 46-49, 72.6% of men aged 50-55 and 74.7% of men aged ≥56 years had an elevated DFI. Limitations, reasons for caution This is a retrospective analysis that did not account for confounding variables (e.g., clinical diagnosis, gonadotoxin exposure, febrile illness) that may affect conventional sperm parameters and DFI. Wider implications of the findings Our results underline the relationship between paternal age and sperm DFI and demonstrate a significant decline in sperm DNA fragmentation in men over the age of 35 years. The data suggest that we may want to reconsider the age cut-off we traditionally use to define advanced paternal age. Trial registration number No trial registration

Phytochemical Analysis, Antimutagenic and Antiviral Activity of Moringa oleifera L. Leaf Infusion: In Vitro and In Silico Studies.

Moringa oleifera (M. oleifera) leaves are rich in nutrients and antioxidant compounds that can be consumed to prevent and overcome malnutrition. The water infusion of its leaf is the easiest way to prepare the herbal drink. So far, no information is available on the antioxidant, antimutagenic, and antivirus capacities of this infusion. This study aimed to determine the composition of the bioactive compounds in M. oleifera leaf infusion, measuring for antioxidant and antimutagenic activity, and evaluating any ability to inhibit the SARS-CoV-2 main protease (Mpro). The first two objectives were carried out in vitro. The third objective was carried out in silico. The phytochemical analysis of M. oleifera leaf infusion was carried out using liquid chromatography-mass spectrometry (LC-MS). Antioxidant activity was measured as a factor of the presence of the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). The antimutagenicity of M. oleifera leaf powder infusion was measured using the plasmid pBR322 (treated free radical). The interaction between bioactive compounds and Mpro of SARS-CoV-2 was analyzed via molecular docking. The totals of phenolic compound and flavonoid compound from M. oleifera leaf infusion were 1.780 ± 5.00 µg gallic acid equivalent/g (µg GAE/g) and 322.91 ± 0.98 µg quercetin equivalent/g (µg QE/g), respectively. The five main bioactive compounds involved in the infusion were detected by LC-MS. Three of these were flavonoid glucosides, namely quercetin 3-O-glucoside, kaempferol 3-O-neohesperidoside, and kaempferol 3-α-L-dirhamnosyl-(1→4)-β-D-glucopyranoside. The other two compounds were undulatoside A, which belongs to chromone-derived flavonoids, and gentiatibetine, which belongs to alkaloids. The antioxidant activity of M. oleifera leaf infusion was IC50 8.19 ± 0.005 µg/mL, which is stronger than the standard butylated hydroxytoluene (BHT) IC50 11.60 ± 0.30 µg/mL. The infusion has an antimutagenic effect and therefore protects against deoxyribonucleic acid (DNA) damage. In silico studies showed that the five main bioactive compounds have an antiviral capacity. There were strong energy bonds between Mpro molecules and gentiatibetine, quercetin, undulatoside A, kaempferol 3-o-neohesperidoside, and quercetin 3-O-glucoside. Their binding energy values are −5.1, −7.5, −7.7, −5.7, and −8.2 kcal/mol, respectively. Their antioxidant activity, ability to maintain DNA integrity, and antimutagenic properties were more potent than the positive controls. It can be concluded that leaf infusion of M. oleifera does provide a promising herbal drink with good antioxidant, antimutagenic, and antivirus capacities.

The association between age-related infertility and deoxyribonucleic acid (DNA) integrity parameters of granulosa cells and lymphocytes

This study aimed to investigate the association between deoxyribonucleic acid (DNA) integrity parameters and advanced maternal age (AMA)-related infertility. The granulosa cells and the lymphocytes obtained from 119 infertile women were recruited. Patients were divided into two groups: the AMA group (≥35 years, n = 26) and the non-AMA group (<35 years, n = 93). The tail length, tail moment and tail DNA percentage were evaluated as the DNA integrity parameters using comet assay. Infertility duration (p=.001), luteinising hormone (p=.01) and progesterone levels (p<.0001) were higher and smoking was more prevalent in the AMA group (p=.001). AMA group was stimulated with higher gonadotropin doses (p=.04) and had decreased anti-mullerian hormone levels (p<.0001). All of DNA integrity parameters were distributed homogenously between the groups; however, the tail length of lymphocytes was higher (p=.02) in the AMA group. Fertilisation was lower (p=.02), oocyte quality was tended to be poor (p=.03) and blastocyst transfer was lower in the AMA group (p=.03). Embryo quality was distributed homogenously between the groups. Implantation, clinical pregnancy and live birth rates were similar between the groups. Impact Statement What is already known on this subject? Advanced maternal age (AMA)-related infertility is associated with diminished ovarian reserve and alteration in follicular environment resulting in poor oocyte quality; however, the exact pathophysiologic mechanism is not clear. What do the results of this study add? Tail length, tail deoxyribonucleic acid (DNA) percentage, tail moment of granulosa cells were nonsignificantly higher in the AMA group compared to younger patients. All of the DNA integrity parameters of lymphocytes were nonsignificantly higher; however, only tail length of lymphocytes was statistically higher in the AMA group than the non-AMA group. A positive correlation was observed between DNA integrity parameters of lymphocytes and body mass index. There were no correlations between DNA integrity parameters of granulosa cells and lymphocyte and infertility duration, gonadotropin dose, duration of ovarian stimulation, oocyte score, embryo score, basal hormone levels and anti-mullerian hormone levels. What are the implications of these findings for clinical practice and/or further research? Our findings offer new insight for further understanding the role of granulosa cells in mediating the poor reproductive outcome of ageing patients. Understanding the mechanisms of ovarian ageing and poor oocyte quality in women with AMA may help to identify specific targets for improving oocyte quality with ageing.

The Integrity of Deoxyribonucleic Acid (DNA) Extracted from Whole Teeth Samples Burnt with Different Accelerants Using Two Extraction Protocols for Forensic Sex Determination

Sex determination is usually the first step in forensic identification of victim(s) in disasters, jungle justice and arsonscenarios for many medical and legal reasons. The use of DNA in forensic analysis offers a good method in sexdetermination and the quality of extracted DNA is very important for downstream PCR. The aim of this study is toinvestigate the viability of DNA obtained from burnt teeth for forensic sex-determination. Two DNA extractionmethods consisting of silica based commercial kit and phenol-chloroform organic method, followed by polymerasechain amplification of amelogenin gene for sex determination were employed. Based on the DNA yield and opticaldensity values, the kit extraction method performed better than the phenol-chloroform method with 100% and85.71% success respectively. A Mann-Whitney U test of 260/280 absorbance showed no statistically significantdifference in the median absorbance for aviation fuel (median = 1.34) and gasoline (median =1.32) burnt samples.Amplification of AMEL genes using the commercial kit and phenol-chloroform method were 52.38% and 22.22%positive respectively compared to pre-extraction sex determination. This study shows that the use of the silica basedcommercial kit technique yielded higher DNA quality and quantity from whole teeth burnt with gasoline andaviation fuel as accelerants for downstream PCR amplification of AMEL genes compared to organic phenolchloroform method.

PARP Inhibitors: An Innovative Approach to the Treatment of Inflammation and Metabolic Disorders in Sepsis.

Sepsis is not only a threat to the health of individual patients but also presents a serious epidemiological problem. Despite intensive research, modern sepsis therapy remains based primarily on antimicrobial treatment and supporting the functions of failing organs. Finding a cure for sepsis represents a great and as yet unfulfilled need in modern medicine. Research results indicate that the activity of poly (adenosine diphosphate (ADP)-ribose) polymerase (PARP) may play an important role in the inflammatory response and the cellular metabolic disorders found in sepsis. Mechanisms by which PARP-1 may contribute to inflammation and metabolic disorders include effects on the regulation of gene expression, impaired metabolism, cell death, and the release of alarmins. These findings suggest that inhibition of this enzyme may be a promising solution for the treatment of sepsis. In studies using experimental sepsis models, inhibition of PARP-1 has been shown to ameliorate the inflammatory response and increase survival. This action was described, among others, for olaparib, a PARP-1 inhibitor approved for use in oncology. While the results of current research are promising, the use of PARP inhibitors in non-oncological diseases raises some concerns, mainly related to the enzyme’s role in deoxyribonucleic acid (DNA) repair. However, the results of studies on experimental models indicate the effectiveness of even short-term PARP-1 inhibition and do not confirm concerns regarding its impact on the integrity of nuclear DNA. Current research presents PARP inhibition as a potential solution for the treatment of sepsis and indicates the need for further research.

Detection of human papillomavirus subtypes harbored in the foreskin of asymptomatic boys: Controlled study

Human papillomavirus infection (HPV) is one of the most common sexually transmitted infections and can cause penile and anal cancer in men, and invasive cervical cancer in women. To evaulate the colonization of 32 HPV subtypes in the foreskin of boys. A prospective analysis was made of the data of 62 healthy boys who had undergone standard circumcision. Deoxyribonucleic acid (DNA) was isolated from the foreskin tissues, and the integrity of DNA was tested. The DNA of each patient was further assessed with real-time polymerase chain reaction (PCR) and the presence of 32 subtypes of HPV was explored. To confirm the results, melting curve analysis and agarose gel electrophoresis (AGE) were performed for all samples. Further analysis was made using LCD-array on six randomly selected samples to confirm the results together with negative and positive controls. The mean age of the boys was 6.8±2years at the time of surgery. All positive controls and samples were positive, all negative controls were negative in the first HPV amplification assay. All positive controls had typical melting curve peaks, whereas all sample amplifications had non-specific, atypical melting curves not fitting with those of the positive controls. Two bands of expected sizes (124 and 405bp) were only observed in positive controls, but not in negative controls or samples on AGE. The same results were observed on the 6 randomly selected samples using LCD-array. Consequently, all the foreskin samples were evaluated as negative for the 32 HPV types investigated in the study. Literature shows a high prevalence of genital HPV in newborns, in early infancy, late adolescence and adulthood. However there is a lack of data in literature on the prevalence in early and late childhood. The negative results of HPV colonization on the foreskin in the current study may be attributed to the conservative and mostly monogamous nature of most family structures in Turkey. The results of the present study have shown that foreskin tissue is not a natural reservoir for HPV and subclinical HPV infection is not likely in the absence of suspected sexual contact.

Multi-scale classification of single-cell gel electrophoresis assay using deep learning algorithm

Structural and functional integrity of deoxyribonucleic acid (DNA) is crucial for the maintenance of hereditary information. However, by-products of cellular metabolism and physical or chemical factors may cause spontaneous DNA damage. The alkaline single-cell gel electrophoresis or comet assay analysis is an easy and reliable method for the determination genotoxic effects of chemical and physical factors. Simply, it is the electrophoretic analysis of intact/damaged DNA of a single cell on in a thin layer of agarose gel. The quantitative analysis of the comet assay images is performed manually by an expert researcher. In visual scoring, DNA nuclei are scored as 0, 1, 2, 3, and 4; and the correct scoring is crucial for the determination of the DNA damage. However, visual scoring depends on the professional experience of the researcher and it is a time consuming and exhausting task. Therefore, this evaluation is inevitable to have subjective results. To avoid this subjectivity and to show the effectiveness of deep learning algorithm on cell images, a Convolution Neural Network (CNN) based deep learning method is proposed to classify comet assay images. According to the results, CNN is trained and tested with high accuracy. The results show that CNN algorithm can successfully classify five different scores of comet assay images, and these results can also reduce the subjectivity.