Percentage Of DNA Fragmentation Research Articles

Phase angle at bioelectric impedance analysis is associated with detrimental sperm quality in idiopathic male infertility: a preliminary clinical study.

In 2020, 38% of adults were affected by obesity, while infertility globally affected 1 in 6 people at some stage of their lives.Body mass index (BMI) provides an easy but occasionally inaccurate estimation of body composition. To achieve a more precise assessment, bioelectric impedance analysis serves as a validated tool that administers electrical energy through surface electrodes. Phase angle as a function of the relationship between tissues resistance and reactance, is a trustworthy predictor of body composition and cell membrane integrity. We aim to assess whether there is an association between phase angle and seminal parameters, as well as sperm DNA fragmentation percentage. Semen samples of 520 idiopathic infertile patients were analyzed according to 2021 World Health Organization guidelines and evaluated for sperm DNA fragmentation rate. Each participants underwent bioelectric impedance analysis. Median age was 40 years old, median BMI was 26.3 kg/m2, median phase angle was 6.2°. In the logistic regression analysis adjusted for age and total intracorporeal water, phase angle (continuous) was significantly associated with oligozoospermia (odds ratio [OR]:0.4; p<0.01) and sperm morphology (OR: 0.65; p=0.05) and slightly with sperm DNA fragmentation (OR: 0.98; p=0.07). In subgroup analysis, the logistic regression analysis adjusted for the mentioned parameters showed that a phase angle between 6.2 and 7 (°) (OR: 0.63; p=0.02) and >7 (°) (OR: 0.12; p<0.01) were associated with a reduced risk of oligozoospermia compared to values <6.2 (°). Similarly, a phase angle between 6.2 and 7 (°) (OR: 0.57; p< 0.01 and OR: 0.58; p= 0.01) and PA > 7 (°) (OR: 0.12; p= 0.03 and OR: 0.21; p< 0.01) were associated with a reduced risk of lower sperm concentration and lower total sperm count, respectively, compared to a phase angle < 6.2 (°). Our study suggests a negative association between phase angle and detrimental sperm parameters in male idiopathic infertility.

Zinc oxide nanoparticles preserve the quality and fertility potential of rooster sperm during the cryopreservation process.

Sperm cryopreservation is one of the main methods for preserving rooster sperm for artificial insemination (AI) in commercial flocks. Yet, rooster sperm is extremely susceptible to reactive oxygen species (ROS) produced during the freezing process. Oxidative stress could be prevented by using nanoparticles containing antioxidants. The present study was conducted to investigate the effect of zinc oxide nanoparticles (ZnONP) in rooster semen freezing extender on quality parameters and fertility potential. For this aim, semen samples were collected and diluted in Lake extenders as follows: control: Lake without ZnONP, ZnO100: Lake with 100-μg zinc oxide (ZnO), ZnONP50: Lake with 50-μg ZnONP, ZnONP100: Lake with 100-μg ZnONP and ZnONP200: Lake with 200-μg ZnONP. After freezing and thawing, sperm motility, viability, membrane integrity, morphology, mitochondrial activity, acrosome integrity, DNA fragmentation, lipid peroxidation and ROS, as well as fertility and hatchability were assessed. According to the current results, higher rates of motility, membrane integrity, mitochondrial activity, acrosome integrity and live cells were detected in the ZnO100, ZnONP50 and ZnONP100 groups compared to other groups (p ≤ .05). Yet, the percentage of dead cells, DNA fragmentation, lipid peroxidation and ROS levels were lower in the mentioned groups (p ≤ .05). Furthermore, a higher percentage of fertility was observed in the ZnO100 and ZnONP100 groups than in the control group (p ≤ .05). In conclusion, the use of 100-μg ZnO and 50- to 100-μg ZnONP represents a valuable and safe additive material that could be used to improve the quality and fertility potential of rooster sperm under cryopreservation conditions.

The protective efficacy of dual dietary rosemary plus cinnamon mix against lead nitrate-induced immune suppression, genotoxicity, and oxidant/antioxidant status in Nile tilapia fingerlings

AbstractLead (Pb) is a common heavy metal, which negatively influences the health and performance of aquatic organisms. The aquatic pollution with Pb is increasing day after day due to the increase in human activities, agricultural, and mining activities among others. Lead was regarded as oxidative stressor causing mutagenic and physiological disorders in aquatic animals. Hence, the current perspective assessed the promoting effects of dietary mixture of rosemary plus cinnamon (R + C) intervention on growth, hemato-biochemical, and hepatic oxidative biomarkers in Nile tilapia (Oreochromis niloticus) exposed to a sub-lethal dose of lead nitrate (7.94 mg Pb/L) for 60 days. Fish were distributed into four equal groups; T1 was kept as a control group, T2 was fed on 10 g R + C/kg feed, and T3 and T4 were fed on the control diet and 10 g R + C/kg feed, respectively, along with lead nitrate exposure. Findings of the present study indicated that, upon exposure to Pb (T3), growth, hemato-biochemical, and immune indices in Nile tilapia were notably reduced compared with that of the control group (T1). Meanwhile, stress (blood glucose, cortisol, total cholesterol, lactate dehydrogenase, alanine and aspartate aminotransferase, and alkaline phosphatase), hepatic oxidative stress (superoxide dismutase, catalase, and glutathione peroxidase) biomarkers, and DNA fragmentation percentage were significantly augmented in T3 vs T1. Additionally, exposing fish to lead nitrate caused immune suppression compared with the control group (T3 vs T1). On the other hand, the dietary supplementation of R + C mix to Nile tilapia (T2) enhanced its growth performance, antioxidant, and immune status. Feeding Pb-intoxicated Nile tilapia on R + C mix (T4) mitigated the oxidative damage and immune suppression induced by lead nitrate exposure compared with T3. Based on the present study outcomes, we opine that feeding Nile tilapia with rosemary plus cinnamon mixture (10 g/kg feed) has protective effects against lead nitrate-induced stress, physiological alterations, and oxidative damage.

Effect of Supplemented Alcoholic Extract of Annatto (Bixa orellara) Seeds and α-Tocotrienol to Soy-Bean Lecithin Extender on Some Biomarkers of Cryopreserved Semen of Holstein Bulls

The purpose of this research was to examine the impact of Soy-bean lecithin extender combined with different quantities of alcoholic extract of annatto (Bixa orellara) and - tocotrienol on sperm abnormalities and certain biomarkers of Holstein bulls semen after being frozen for varying lengths of time. In the experiment, bull semen was pooled, diluted with Soy-bean lecithin extender, and distributed randomly among five groups: Diluted semen was put through the semen cryopreservation technique; the control treatment (C) consisted solely of soy-bean lecithin extender; T1 included 150mg annatto extract/25 ml; T2 contained 200mg annatto extract/25 ml; T3 contained 3mM tocotreinol; and T4 contained 4mM tocotreinol. After 48 hours, 3 months, and 6 months, both chilled and frozen semen were evaluated for abnormalities, Malondialdehyde (MDA), Total antioxidant capacity (TAC) concentration, and DNA fragmentation percentage. As shown by the data, there were no significant differences between the treatments in terms of total abnormalities in sperm after any of the cryopreservation times, but there were significant differences in terms of MDA and DNA fragmentation. Some of the biomarkers in Holstein bulls’ sperm were found to be significantly improved when a combination of alcoholic extract of annatto(Bixa orellara) and tocotrienol was added to Soy-bean lecithin.

Effects of Benzo[a]pyrene on Human Sperm Functions: An In Vitro Study.

Benzo(a)pyrene (BaP) is considered one of the most dangerous air pollutants for adverse health effects, including reproductive toxicity. It is found both in male and female reproductive fluids likely affecting spermatozoa after the selection process through cervical mucus, a process mimicked in vitro with the swim-up procedure. In vitro effects of BaP (1, 5, 10 µM) were evaluated both in unselected and swim-up selected spermatozoa after 3 and 24 h of incubation. BaP reduced total, progressive and hyperactivated motility and migration in a viscous medium both in swim-up selected and unselected spermatozoa. Viability was not significantly affected in swim-up selected but was reduced in unselected spermatozoa. In swim-up selected spermatozoa, increases in the percentage of spontaneous acrosome reaction and DNA fragmentation were observed after 24 h of incubation, whereas no differences between the control and BaP-treated samples were observed in caspase-3 and -7 activity, indicating no effects on apoptotic pathways. ROS species, evaluated by staining with CellROX® Orange and Dihydroethidium, did not differ in viable spermatozoa after BaP treatment. Conversely, the percentage of unviable ROS-positive spermatozoa increased. Our study suggests that BaP present in male and female genital fluids may heavily affect reproductive functions of human spermatozoa.

Analyzing and Examining Smoking Effects and Oxidative Stress Markers in Seminal Plasma in Iraqi Infertility Men

The oxidative stress (OS) and sperm DNA fragmentation have been associated with sub-infertility in males and infertility due to a lack of means of protection and defense (antioxidants) for spermatozoa and an increase in reactive oxygen spices (ROS) related to wrong lifestyle (cigarettes and hookah) or what also known as Shisha. The study included (90) men between the ages of (23-40) years, (60), infertile men who smoked cigarettes and hookahs. They were divided into two groups, the first oligozoospermia and the second asthenozoospermia, according to the guidelines of the World Health Organization for seminal fluid analysis (SFA), 5 th edition, WHO(2010). Each group included (30) smoking infertile males, with a control group included (30) non-smoking fertile males. The study aimed to estimate the concentration level of Ascorbic Acid (AA) in the seminal plasma of two groups of infertile men smoking cigarettes and hookahs using enzyme-linked immunosorbent assay (ELISA) method, with calculating the concentration of Malondialdehyde (MDA) as well as estimating the percentage of sperm DNA fragmentation (SDF) using sperm chromatin dispersion (SCD), Halo view. Compared with healthy fertile men of non-smokers of cigarettes and hookahs as one of the biomarkers of oxidative stress in infertile men in Nineveh Governorate-Iraq. The results indicated that there were significant differences at the probability level (p&lt;0.01) for the studied parameters (AA, MDA, SDF) compared to the control group (fertile men). The results showed an increase in the percentage of sperm DNA fragmentation with increasing the concentration of MDA in seminal plasma and decreasing the concentration of AA. Cigarette and hookah smoking increased the generation of ROS with a decrease in the water-soluble non-enzymatic antioxidant Ascorbic acid (AA) and thus increased the oxidative processes of the plasma membranes of spermatozoa and the occurrence of oxidative stress and impaired fertility and caused infertility. Keywords: male infertility, oxidative stress, sperm DNA fragmentation, Ascorbic acid, MDA. Bionatura http://dx.doi.org/10.21931/RB/CSS/2023

Design, synthesis, in vitro anticancer, molecular docking and SAR studies of new series of pyrrolo[2,3-d]pyrimidine derivatives

The current study involves the design and synthesis of a newly synthesized pyrrolo[2,3-d]pyrimidine derivatives to contain chlorine atoms in positions 4 and 6 and trichloromethyl group in position 2 using microwave technique as a new and robust approach for preparation of this type of pyrrolo[2,3-d]pyrimidine derivatives. The chemical structure of the synthesized pyrrolo[2,3-d]pyrimidine derivatives 3–19 was well-characterized using spectral and elemental analyses as well as single-crystal X-ray diffraction. All compounds were tested in vitro against seven selected human cancer cell lines, namely, MCF7, A549, HCT116, PC3, HePG2, PACA2 and BJ1 using MTT assay. It was found that compounds 14a, 16b and 18b were the most active toward MCF7 with IC50 (1.7, 5.7, and 3.4 μg/ml, respectively) relative to doxorubicin (Dox.) (26.1 μg/ml). Additionally, compound 17 exerted promising cytotoxic effects against HePG2 and PACA2 with IC50 (8.7 and 6.4 μg/ml, respectively) relative to Dox. (21.6 and 28.3 μg/ml, respectively). The molecular docking study confirmed our ELISA result which showed the promising binding affinities of compounds 14a and 17 against Bcl2 anti-apoptotic protein. At the gene expression level, P53, BAX, DR4 and DR5 were up-regulated, while Bcl2, Il-8, and CDK4 were down-regulated in 14a, 14b and 18b treated MCF7 cells. At the protein level, compound 14b increased the activity of Caspase 8 and BAX (18.263 and 14.25 pg/ml) relative to Dox. (3.99 and 4.92 pg/ml, respectively), while the activity of Bcl2 was greatly decreased in 14a treated MCF7 (2.4 pg/ml) compared with Dox. (14.37 pg/ml). Compounds 14a and 14b caused cell cycle arrest at the G1/S phase in MCF7. Compounds 16b and 18b induced the apoptotic death of MCF7 cells. In addition, the percentage of fragmented DNA was increased significantly in 14a treated MCF7 cells.

P-040 The Role of ACE2 and DNA fragmentation index on covid-19 post infected, healthy and infertile men

Abstract Study question to investigate the relationship between ACE2 and DNA fragmentation index and their impact on men’s reproductive system in covid-19 post-infected patients. Summary answer the drastic effect of covid-19 on the male reproductive system might be due to elevated DFI and ACE2 levels. What is known already In the last two years, many studies had been devoted to uncovering the depraved impact of covid-19 by measuring ACE2 concentrations or sperm DNA fragmentation but no study was applied to measure both parameters, this study performed on both parameters to figure out the role of angiotensin-converting enzyme (ACE2) with seminal fluid parameters and DNA fragmentation index in covid-19 post infected men and infertile patient in comparison to healthy fertile men. Study design, size, duration This prospective cohort study was conducted from November 2021 to June 2022. one hundred and nine men who participated in this study. Participants/materials, setting, methods The participants were divided into four groups: post-infected men with Covid-19 who recovered from Covid-19 infection with a recovery interval of no more than three months from the time of symptoms onset and all were fertile married men, healthy fertile men who haven't been infected with Covid-19 in the last three months obtain normal seminal fluid parameters as control, The third group is composed of oligospermia, while the fourth group is constituted of azoospermia volunteers Main results and the role of chance In this study, the result showed that the seminal fluid parameters for post-infected men with covid-19; were demotion in sperm concentration, total count, motility, and morphology with a p-value &amp;lt; 0.001 when compared with healthy men. DNA fragmentation in post-infected men was 4 times higher than in non-infected and it was found that post-infected men had 10 times more ACE2 concentrations in comparison with healthy men. The study showed that in oligospermia and azoospermia volunteers own a high ACE2 level compared to healthy volunteers and that the oligospermia subjects have a significantly high sperm DNA fragmentation index. In conclusion, the drastic effect of covid-19 on the male reproductive system might be due to elevated DNA fragmentation and ACE2 levels, and there is a statistically significant positive correlation between the DNA fragmentation index and ACE2 levels. Limitations, reasons for caution number of post-infected men with covid-19 Wider implications of the findings 1- A positive relationship exists between DNA fragmentation index percentage (DFI) and angiotensin-converting enzyme level (ACE 2). 2- Covid-19 infection alters sperm parameters such as concentrations, total sperm count, motility, and morphology. 3- The percentage of DNA fragmentation rose significantly after covid-19 infection. Trial registration number self funding

Eulophia gracilis pseudobulb extract mitigates cyclophosphamide-induced genotoxicity and oxidative stress on murine hepatic tissue

The reports over the years on chemotherapeutic regimen involving cyclophosphamide (CYP), a bifunctional alkylating agent, demonstrated hepatotoxic side effect. Eulophia gracilis (EG) is a medicinal plant with folkloric utility in the treatment of liver damage and blood related diseases. However, there is a knowledge gap on the impact of E. gracilis effectiveness on CYP-associated hepatic toxicity in the literature. We investigated on potency of aqueous methanolic extract of E. gracilis (AMEG) and CYP-mediated hepatic toxicity in rats. Experimental rats were administered with CYP (2 mg/kg) or co-treated with AMEG (200 or 400 mg/kg) for 7 days consecutively. The result showed that co-treatment with AMEG significantly reduces alanine aminotransferase, aspartate aminotransferase, gamma glutamyl transferase and lactate dehydrogenase activities compared to the CYP group. Moreover, AMEG abated CYP-induced decreases in superoxide dismutase, catalase and glutathione peroxidase enzymes in the liver homogenate. AMEG alleviated CYP-facilitated surges of hepatic concentration of advanced oxidized protein product (AOPPs) and lipid peroxidation in rats. Additionally, AMEG reduced pathological lesions in the liver of co-treated rats and elicited anti-genotoxic effect by mitigating CYP-mediated increases of frequency of formation of polychromatic erythrocyte in the bone marrow and hepatic percentage DNA fragmentation in CYP-exposed rats. Overall, AMEG protective effect improved liver dysfunction occasioned by CYP-mediated toxicities in rats by abating oxidative stress and alleviating genotoxic responses.

Lactobacillus plantarum secretions may exert a cryoprotective effect on human sperm motility: a prospective in vitro study.

Semen cryopreservation is a widely used procedure for fertility preservation, despite some level of cryodamage that may occur in spermatozoa after thawing. However, there is some evidence that lactobacilli, one of the bacteria found in semen, might benefit spermatozoa quality. This study aims to determine whether the addition of Lactobacillus plantarum (L. plantarum) secretions to a sperm freezing medium has an impact on sperm motility, morphology, and DNA fragmentation. This is a prospective auto-controlled study. It was conducted on 30 raw semen samples from 30 infertile men attending a fertility center for semen analysis. Before freezing, all the samples were analyzed for motility, morphology, and DNA fragmentation percentages. Each sample was then divided equally into three aliquots. Cryopreservation was performed on each aliquot using one of the following three media: without L. plantarum secretions (control group), with 107 CFU/ml, or with 108 CFU/ml L. plantarum secretions. Sperm motility, morphology, and DNA integrity were evaluated after the cryopreservation media were added and after semen thawing. The results of this study indicated that after thawing, no statistically significant decrease in progressive motility and non-progressive percentages were detected in the sperm freezing medium supplemented with 108 CFU/ml L. plantarum secretions compared to the fresh raw semen. Moreover, multivariate linear regression model analyses showed that the progressive motility (p = 0.02), non-progressive motility (p = 0.016), and non-motile spermatozoa (p = 0.012) percentages were significantly decreased in the freezing medium (without L. plantarum secretions) compared to the fresh raw semen. To the great of our knowledge, this is the first study showing that L. plantarum secretions had a cryoprotective effect on sperm motility when added to the sperm freezing medium. Furthermore, L. plantarum secretions were found to protect sperm DNA integrity more effectively than the freezing medium without L.plantarum secretions in non-normozoospermia group. Cryopreservation procedures must therefore be optimized to minimize any iatrogenically-induced sperm DNA damage, given the correlation between sperm DNA damage and increased mutation loads in progeny. This article is protected by copyright. All rights reserved.

Molecular and physiochemical evaluation of buck semen cryopreserved with antioxidants.

The current study evaluated the physiochemical quality and gene expression profile of post-thawed buck semen after supplementation with antioxidants [melatonin (M), L-carnitine (LC), cysteine (Cys), LC + M, M + Cys, LC + Cys, LC + Cys + M] in comparison with the non-treated control group. Physical and biochemical characteristics of semen were evaluated following freezing and thawing. Transcript abundance of six selected candidate genes was profile using quantitative real-time PCR. The data demonstrated significant enhancement of post-freezing total motility, progressive motility, percentage of live sperm, CASA parameters, plasma membrane and acrosome integrity in all groups supplemented with Cys, LC, M + Cys and LC + Cys compared with the control group. The biochemical analysis of semen indicated that semen groups supplemented with LC and LC + Cys recorded increased levels of GPX and SOD that were coupled with up-regulation of antioxidant genes (SOD1, GPX1 and NRF2) and mitochondrial transcripts (CPT2 and ATP5F1A). Moreover, H2O2 level and DNA fragmentation percentage were reduced compared with other groups. In conclusion, supplementation of Cys alone or in combination with LC positively improved the post-thaw physiochemical properties of rabbit semen through activation of bioenergetics-related mitochondrial genes and cellular antioxidant defence mechanism.

Association between The SIRT1 and SIRT3 Levels and Gene Polymorphisms with Infertility in War Zones of Kermanshah Province, Iran: A Case-Control Study.

War toxin, mustard gas, alkylating agent results in male infertility via inducing reactive oxygen species (ROS) production and DNA mutagenesis. SIRT1 and SIRT3 are multifunctional enzymes that involve in the DNA repair, oxidative stress responses. This study aim is to assess the correlation between serum levels of SIRT1, SIRT3 and both rs3758391T>C and rs185277566C>G gene polymorphisms with infertility in the war zones of Kermanshah province, Iran. In this case-control study based on the semen analysis, samples were divided into two groups infertile (n=100) and fertile (n=100). High-performance liquid chromatography (HPLC) method was used to determine the malondialdehyde level, and also a sperm chromatin dispersion (SCD) test was used to evaluate the DNA fragmentation rate. Using the colorimetric assays, superoxide dismutase (SOD) activity was measured. SIRT1 and SIRT3 protein levels were determined by using ELISA. The genetic variants of SIRT1 rs3758391T>C, and SIRT3 rs185277566C>G, were detected by polymerase chain reaction-restriction fragment length (PCR-RFLP) technique. Malondialdehyde (MDA) level and the percentage of DNA fragmentation were higher in infertile samples, but serum levels of SIRT1 and SIRT3, and SOD activity was lower in infertile compared to fertile samples (P<0.001). The TC+CC genotypes and the C allele from SIRT1 rs3758391T>C polymorphism, and CG+GG genotypes and the G allele from SIRT3 rs185277566C>G polymorphism could increase risk of infertility (P<0.05). The results of this study suggest that war toxins through the impact on genotypes, decreasing levels of SIRT1 and SIRT3 and increasing levels of oxidative stress, lead to defects in the concentration, motility and morphology of sperms and thus, infertility in men.

The effect of natural products combination on MCF-7 cells exceeds tamoxifen therapeutic dose effects in vitro

Cancer remains to be one of the most severe sicknesses globally. Cases have kept rising over the years. Breast cancer (BC), which is among the leading types of cancers and predominantly affects women, is the second leading cause of cancer mortality. Researchers have developed interventions over the years; however, the BC survival rate has not improved since the 1980s. This has created the need for novel drug interventions that would manage and treat BC more effectively. This study focused on using a combination of natural product extracts such as phytoestrogen (Ziziphus jujube) and Tannin nanoparticles (NP99) together, which we have referred to as (Z.NP99) and tamoxifen (Tam) as one of the leading BC drugs since the 70s, in treating BC. The effectiveness of Tam if used alone in the treatment and if combined with Z.NP99 was evaluated using MCF-7 cells in vitro. The findings showed that the combination treatment of Z.NP99 affected the proliferation and viability of MCF-7 cells more than Tam at a 10 μg/mL dose. Moreover, Z.NP99 with Tam stimulated the maximum reduction of MCF-7 proliferation and viability in a time-dependent manner. Furthermore, Tam and Z.NP99 augmented the DNA fragmentation percentage combined with the upregulation of the apoptotic genes. Additionally, the results showed that the apoptotic impact of Z.NP99 and Tam on MCF-7 cells may be intermediated by down-regulating some genes such as Claudin-1 followed by down-regulating mRNA expression of MMP-9, VEGF, and BCL-2 genes of treated cells. Combining Tam with Z.NP99 considerably enhanced the effectiveness of conventional therapy. As a result, this study suggested that the Z.NP99 was ideal for developing effective natural treatments that would improve BC outcomes.

Study of Synthesis, Characterization, DFT, and In Vitro Biological Activity of Cu(II), Co(II), and Fe(II) Metal Complexes Based on Heterocyclic Azo Pyrazole Dye Ligand

A new azo dye ligand was synthesized by coupling 5-amino-3-methyl-1-phenylpyrazole with a diazonium ion of 2-amino-1-naphthalene sulfonic acid. Its Co(II), Fe(II), and Cu(II) complexes were also synthesized. This ligand and its complexes were characterized based on elemental analysis, IR, 1H NMR, UV-vis spectra, and thermogravimetric analysis. The data showed that the proposed complexes have an octahedral geometry around the central metal ion. The nonelectrolytic nature of the complexes was confirmed by molar conductance measurement. X-ray powder diffraction indicates that Co(II) and Fe(II) complexes are amorphous in nature, whereas the Cu(II) complex has crystalline and amorphous phases. The bond strength and molecular stability between the interaction of metal ions and the ligand were investigated by density functional theory (DFT). The in vitro antibacterial activity of the ligand and complexes was tested against two Gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus) and two Gram-negative bacteria (Escherichia coli and Neisseria gonorrhoeae) and a fungus (Candida albicans). The Cu(II) complex exhibited a high antimicrobial effect against all tested microorganisms. The DNA fragmentation percentage and cytotoxic activity against human liver cancer cells (HepG2) were evaluated. All tested compounds showed potential anticancer activity, and the Co(II) complex showed better activity results than the other tested compounds, with IC50 = 19.55 μg/mL.

Targeting endoplasmic reticulum stress, Nrf-2/HO-1, and NF-κB by myristicin and its role in attenuation of ulcerative colitis in rats

AimsUlcerative colitis (UC) is characterized by the up-regulation of pro-inflammatory mediators, apoptotic signals, and oxidative stress that can lead to an increased risk of colorectal cancer. The present study aims to investigate the possible role of myristicin in modulating endoplasmic reticulum stress (ERS) and risk-associated conditions in acetic acid (AA)-induced UC. Materials and methodsAdult male rats were treated with 150 mg/kg body weight of myristicin or mesalazine orally either as pre/post treatment or post-treatment only. The gene expression of glucose-related protein 78 (GRP78), CCAAT/enhancer-binding protein homologous protein (CHOP), and nuclear factor kappa B (NF-κB), percentage of DNA fragmentation, and serum levels of some oxidative and inflammatory markers were measured. Key findingsThe results indicated the potential upregulation of ERS, pro-apoptotic, lipid peroxidation, and pro-inflammatory cascades by induction of UC in rats. However, myristicin could effectively reverse the deteriorated effects of ulceration in colonic mucosa. It was mediated through downregulation of the ERS markers GRP78 and CHOP genes expression, reduction of NF-κB mRNA expression, DNA fragmentation, reduced lipid peroxidation, myeloperoxidase (MPO) activity and pro-inflammatory markers (Tumor necrosis factor-α (TNF-α), Interleukin-1β (IL-1β) and cyclo‑oxygenase (COX-2) activity). Accompanied by elevated levels of IL-10, colonic Nuclear erythroid factor (Nrf-2) and Heme oxygenase (HO-1) activity as well as blood antioxidant enzymes activity. Results of docking might confirm the biological results of our study. SignificanceMyristicin could effectively modulate important stress, and inflammatory effectors and protect mucosal DNA from oxidative damage which can serve as a promising candidate for the treatment of ulcerative colitis.

Male reproductive health after 3 months from SARS-CoV-2 infection: a multicentric study

PurposeWhile SARS-CoV-2 infection appears not to be clinically evident in the testes, indirect inflammatory effects and fever may impair testicular function. To date, few long-term data of semen parameters impairment after recovery and comprehensive andrological evaluation of recovered patients has been published. The purpose of this study was to investigate whether SARS-CoV-2 infection affect male reproductive health.MethodsEighty patients were recruited three months after COVID-19 recovery. They performed physical examination, testicular ultrasound, semen analysis, sperm DNA integrity evaluation (TUNEL), anti-sperm antibodies (ASA) testing, sex hormone profile evaluation (Total testosterone, LH, FSH). In addition, all patients were administered International Index of Erectile Function questionnaire (IIEF-15). Sperm parameters were compared with two age-matched healthy pre-COVID-19 control groups of normozoospermic (CTR1) and primary infertile (CTR2) subjects.ResultsMedian values of semen parameters from recovered SARS-CoV-2 subjects were within WHO 2010 fifth percentile. Mean percentage of sperm DNA fragmentation (%SDF) was 14.1 ± 7.0%. Gelatin Agglutination Test (GAT) was positive in 3.9% of blood serum samples, but no positive semen plasma sample was found. Only five subjects (6.2%) had total testosterone levels below the laboratory reference range. Mean bilateral testicular volume was 31.5 ± 9.6 ml. Erectile dysfunction was detected in 30% of subjects.ConclusionOur data remark that COVID-19 does not seem to cause direct damage to the testicular function, while indirect damage appears to be transient. It is possible to counsel infertile couples to postpone the research of parenthood or ART procedures around three months after recovery from the infection.

Effect of Adding α-tocopherol on Fertility Parameters in Bovine Semen Cryopreservation

Background: Cryopreservation of bovine semen affects survival of spermatozoa. It is necessary to improve semen quality to increase fertility. The aim was to determine the effect of α-tocopherol added to the bovine semen diluent at different time of freezing, upon sperm quality post thawing. Methods: Semen was collected from Charolais breed bulls. The semen was divided into two portions and one portion was added with α-tocopherol and the other was used as control. These portions were further divided into three parts and frozen for 8, 11 and 15 minutes, respectively. After 60 days of cryopreservation, the progressive motility, non-progressive motility, absence of motility, vitality and DNA fragmentation were evaluated. Result: The lower percentages of DNA fragmentation were treated with antioxidants (15.52, 14.83 and 14.71%). The highest vitality and progressive motility were found in the sample with α-tocopherol and frozen for 11 min with 72.91 and 58.61%, respectively. The addition of α-tocopherol to the semen diluent decreased damages to spermatozoa and enhanced the semen quality.

Impact of Escherichia coli Outer Membrane Vesicles on Sperm Function.

Reproductive tract infections account for approximately 15% of male infertility cases. Escherichia coli (E. coli) represents the most frequently isolated bacterial strain in the semen of infertile men. All Gram-negative bacteria constitutively produce outer membrane vesicles (OMVs). The present study proved, for the first time, the involvement of OMVs in human sperm function. E. coli OMVs were isolated by ultracentrifugation and characterized via sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis. Human sperm was exposed to OMVs (8 µg/mL) for different times (30, 45, 60 and 90 min). The vitality, motility, morphology, ROS level and DNA fragmentation of spermatozoa were evaluated. OMVs reduced the progressive motility and increased the immobile spermatozoa amount after 30 min of treatment. In addition, a significant increase in the percentage of intracellular ROS and sperm DNA fragmentation was recorded for each vesicular exposure time. These preliminary findings prove that OMVs contribute to altering human sperm function via two mechanisms: (i) impaired motility and (ii) DNA fragmentation.

Selenium nanoparticles impart robust neuroprotection against deltamethrin-induced neurotoxicity in male rats by reversing behavioral alterations, oxidative damage, apoptosis, and neuronal loss

This study investigated the neuroprotective role of selenium nanoparticles (SeNPs) on deltamethrin-induced neurotoxicity in rats. A total of 32 adult male Wister rats were allocated into the following four groups: 1) control, 2) deltamethrin (0.6 mg/kg), 3) SeNPs (0.5 mg/kg), and 4) deltamethrin + SeNPs. All agents were administered orally three times per week for 2 months. Locomotor behavior, anxiety-like behavior, biochemical parameters, including brain oxidative damage biomarkers (Malondialdehyde (MDA) and reduced glutathione (GSH)), brain acetylcholinesterase (AChE), and brain genotoxicity were evaluated. The gene expression levels of IGF-1 and Bcl2 were also determined. Moreover, a brain histopathological examination associated with the immunohistochemical determination of Bax in brain tissue was performed. Deltamethrin-intoxicated rats showed a reduction in the locomotor activity associated with a highly anxious state. They also displayed a disturbance in the brain redox state with a decrease in the brain AChE levels and a high DNA fragmentation percentage. Furthermore, they showed a decrement in the immunohistochemical GFAP levels as well as IGF-1 and Bcl2 gene expression levels with an increase in the immunohistochemical Bax levels. All these changes were confirmed by brain histopathology. Interestingly, SeNPs ameliorated all these changes and restored the normal brain architecture. In conclusion. SeNPs possess a potent medicinal activity due to their antioxidant and anti-inflammatory activity. Therefore, SeNPs can be a potential agent in ameliorating deltamethrin-induced neurotoxicity.

Absolute quantification of plasma mitochondrial DNA by droplet digital PCR marks COVID-19 severity over time during intensive care unit admissions.

Increased plasma mitochondrial DNA concentrations are associated with poor outcomes in multiple critical illnesses, including COVID-19. However, current methods of cell-free mitochondrial DNA quantification in plasma are time-consuming and lack reproducibility. Here, we used next-generation sequencing to characterize the size and genome location of circulating mitochondrial DNA in critically ill subjects with COVID-19 to develop a facile and optimal method of quantification by droplet digital PCR. Sequencing revealed a large percentage of small mitochondrial DNA fragments in plasma with wide variability in coverage by genome location. We identified probes for the mitochondrial DNA genes, cytochrome B and NADH dehydrogenase 1, in regions of relatively high coverage that target small sequences potentially missed by other methods. Serial assessments of absolute mitochondrial DNA concentrations were then determined in plasma from 20 critically ill subjects with COVID-19 without a DNA isolation step. Mitochondrial DNA concentrations on the day of enrollment were increased significantly in patients with moderate or severe acute respiratory distress syndrome (ARDS) compared with those with no or mild ARDS. Comparisons of mitochondrial DNA concentrations over time between patients with no/mild ARDS who survived, patients with moderate/severe ARDS who survived, and nonsurvivors showed the highest concentrations in patients with more severe disease. Absolute mitochondrial DNA quantification by droplet digital PCR is time-efficient and reproducible; thus, we provide a valuable tool and rationale for future studies evaluating mitochondrial DNA as a real-time biomarker to guide clinical decision-making in critically ill subjects with COVID-19.