Ejaculatory Abstinence Affects the Sperm Quality in Normozoospermic Men-How Does the Seminal Bacteriome Respond?

This study was designed to describe bacterial profiles of ejaculates collected following a long and short ejaculatory abstinence set in the context of changes in the conventional, oxidative, and immunological characteristics of semen. Two specimens were collected in succession from normozoospermic men (n = 51) following 2 days and 2 h, respectively. Semen samples were processed and analyzed according to the World Health Organization (WHO) 2021 guidelines. Afterwards, sperm DNA fragmentation, mitochondrial function, levels of reactive oxygen species (ROS), total antioxidant capacity, and oxidative damage to sperm lipids and proteins were evaluated in each specimen. Selected cytokine levels were quantified using the ELISA method. Bacterial identification by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry revealed that samples collected following two days of abstinence presented with a higher bacterial load and diversity, and a greater prevalence of potentially uropathogenic bacteria including Escherichia coli, Staphylococcus aureus and Enterococcus faecalis. Only staphylococci and Escherichia coli remained present in specimens obtained after 2 h of abstinence. Whilst all samples accomplished the criteria set by WHO, a significantly higher motility (p < 0.05), membrane integrity (p < 0.05), mitochondrial membrane potential (p < 0.05), and DNA integrity (p < 0.0001) were detected following 2 h of ejaculatory abstinence. On the other hand, significantly higher ROS levels (p < 0.001), protein oxidation (p < 0.001), and lipid peroxidation (p < 0.01) accompanied by significantly higher concentrations of tumor necrosis factor alpha (p < 0.05), interleukin-6 (p < 0.01), and interferon gamma (p < 0.05) were observed in specimens collected after two days of abstinence. It may be summarized that shorter ejaculatory abstinence does not compromise sperm quality in normozoospermic men, while it contributes to a decreased occurrence of bacteria in semen which is accompanied by a lower probability of damage to spermatozoa by ROS or pro-inflammatory cytokines.

Salvia sclarea Essential Oil Chemical Composition and Biological Activities.

Salvia sclarea essential oil (SSEO) has a long tradition in the food, cosmetic, and perfume industries. The present study aimed to analyze the chemical composition of SSEO, its antioxidant activity, antimicrobial activity in vitro and in situ, antibiofilm, and insecticidal activity. Besides that, in this study, we have evaluated the antimicrobial activity of SSEO constituent (E)-caryophyllene and standard antibiotic meropenem. Identification of volatile constituents was performed by using gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS) techniques. Results obtained indicated that the main constituents of SSEO were linalool acetate (49.1%) and linalool (20.6%), followed by (E)-caryophyllene (5.1%), p-cimene (4.9%), a-terpineol (4.9%), and geranyl acetate (4.4%). Antioxidant activity was determined as low by the means of neutralization of the DDPH radical and ABTS radical cation. The SSEO was able to neutralize the DPPH radical to an extent of 11.76 ± 1.34%, while its ability to decolorize the ABTS radical cation was determined at 29.70 ± 1.45%. Preliminary results of antimicrobial activity were obtained with the disc diffusion method, while further results were obtained by broth microdilution and the vapor phase method. Overall, the results of antimicrobial testing of SSEO, (E)-caryophyllene, and meropenem, were moderate. However, the lowest MIC values, determined in the range of 0.22-0.75 µg/mL for MIC50 and 0.39-0.89 µg/mL for MIC90, were observed for (E)-caryophyllene. The antimicrobial activity of the vapor phase of SSEO (towards microorganisms growing on potato) was significantly stronger than that of the contact application. Biofilm analysis using the MALDI TOF MS Biotyper showed changes in the protein profile of Pseudomonas fluorescens that showed the efficiency of SSEO in inhibiting biofilm formation on stainless-steel and plastic surfaces. The insecticidal potential of SSEO against Oxycarenus lavatera was also demonstrated, and results show that the highest concentration was the most effective, showing insecticidal activity of 66.66%. The results obtained in this study indicate the potential application of SSEO as a biofilm control agent, in the shelf-life extension and storage of potatoes, and as an insecticidal agent.

Possible Implications of Bacteriospermia on the Sperm Quality, Oxidative Characteristics, and Seminal Cytokine Network in Normozoospermic Men.

This study focused on the identification of bacterial profiles of semen in normozoospermic men and their possible involvement in changes to the sperm structural integrity and functional activity. Furthermore, we studied possible fluctuations of selected cytokines, oxidative markers, and antibacterial proteins as a result of bacterial presence in the ejaculate. Sperm motility was assessed with computer-assisted sperm analysis, while sperm apoptosis, necrosis and acrosome integrity were examined with fluorescent methods. Reactive oxygen species (ROS) generation was quantified via luminometry, sperm DNA fragmentation was evaluated using the TUNEL protocol and chromatin-dispersion test, while the JC-1 assay was applied to evaluate the mitochondrial membrane potential. Cytokine levels were quantified with the biochip assay, whilst selected antibacterial proteins were quantified using the ELISA method. The predominant species identified by the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry were Staphylococcus hominis, Staphylococcus capitis and Micrococcus luteus. The results revealed that the sperm quality decreased proportionally to the increasing bacterial load and occurrence of conditionally pathogenic bacteria, including Enterococcus faecalis, Staphylococcus aureus and Escherichia coli. Antimicrobial susceptibility tests revealed a substantial resistance of randomly selected bacterial strains to ampicillin, vancomycin, tobramycin, and tetracycline. Furthermore, an increased bacterial quantity in semen was accompanied by elevated levels of pro-inflammatory cytokines, including interleukin-1, interleukin-2, interleukin-6, tumor necrosis factor alpha as well as ROS overproduction and lipid peroxidation of the sperm membranes. Our results suggest that semen quality may be notably affected by the bacterial quantity as well as quality. It seems that bacteriospermia may be associated with inflammatory processes, oxidative stress, sperm structural deterioration, and a subsequent risk for the development of subfertility, even in normozoospermic males.

Chemical Composition and Biological Activities of Eucalyptus globulus Essential Oil.

Eucalyptus globulus essential oil (EGEO) is considered as a potential source of bioactive compounds with significant biological activity. The aim of this study was to analyze the chemical composition of EGEO, in vitro and in situ antimicrobial activity, antibiofilm activity, antioxidant activity, and insecticidal activity. The chemical composition was identified using gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). The main components of EGEO were 1,8-cineole (63.1%), p-cimene (7.7%), a-pinene (7.3%), and a-limonene (6.9%). Up to 99.2% of monoterpenes were present. The antioxidant potential of essential oil and results indicate that 10 µL of this sample can neutralize 55.44 ± 0.99% of ABTS•+, which is equivalent to 3.22 ± 0.01 TEAC. Antimicrobial activity was determined via two methods: disk diffusion and minimum inhibitory concentration. The best antimicrobial activity was shown against C. albicans (14.00 ± 1.00 mm) and microscopic fungi (11.00 ± 0.00 mm-12.33 ± 0.58 mm). The minimum inhibitory concentration showed the best results against C. tropicalis (MIC 50 2.93 µL/mL, MIC 90 3.17 µL/mL). The antibiofilm activity of EGEO against biofilm-forming P. flourescens was also confirmed in this study. The antimicrobial activity in situ, i.e., in the vapor phase, was significantly stronger than in the contact application. Insecticidal activity was also tested and at concentrations of 100%, 50%, and 25%; the EGEO killed 100% of O. lavaterae individuals. EGEO was comprehensively investigated in this study and information regarding the biological activities and chemical composition of the essential oil of Eucalyptus globulus was expanded.

Biological Activity of Cupressus sempervirens Essential Oil.

The aim of this study was to evaluate the antioxidant, antibiofilm, antimicrobial (in situ and in vitro), insecticidal, and antiproliferative activity of Cupressus sempervirens essential oil (CSEO) obtained from the plant leaf. The identification of the constituents contained in CSEO was also intended by using GC and GC/MS analysis. The chemical composition revealed that this sample was dominated by monoterpene hydrocarbons α-pinene, and δ-3-carene. Free radical scavenging ability, performed by using DPPH and ABTS assays, was evaluated as strong. Higher antibacterial efficacy was demonstrated for the agar diffusion method compared to the disk diffusion method. The antifungal activity of CSEO was moderate. When the minimum inhibitory concentrations of filamentous microscopic fungi were determined, we observed the efficacy depending on the concentration used, except for B. cinerea where the efficacy of lower concentration was more pronounced. The vapor phase effect was more pronounced at lower concentrations in most cases. Antibiofilm effect against Salmonella enterica was demonstrated. The relatively strong insecticidal activity was demonstrated with an LC50 value of 21.07% and an LC90 value of 78.21%, making CSEO potentially adequate in the control of agricultural insect pests. Results of cell viability testing showed no effects on the normal MRC-5 cell line, and antiproliferative effects towards MDA-MB-231, HCT-116, JEG-3, and K562 cells, whereas K562 cells were the most sensitive. Based on our results, CSEO could be a suitable alternative against different types of microorganisms as well as suitable for the control of biofilms. Due to its insecticidal properties, it could be used in the control of agricultural insect pests.

Thymus zygis, Valuable Antimicrobial (In Vitro and In Situ) and Antibiofilm Agent with Potential Antiproliferative Effects.

With the growing issues of food spoilage, microbial resistance, and high mortality caused by cancer, the aim of this study was to evaluate T. zygis essential oil (TZEO) as a potential solution for these challenges. Here, we first performed GC/MS analysis which showed that the tested TZEO belongs to the linalool chemotype since the abundance of linalool was found to be 38.0%. Antioxidant activity assays showed the superiority of TZEO in neutralizing the ABTS radical cation compared to the DPPH radical. The TZEO was able to neutralize 50% of ABTS•+ at the concentration of 53.03 ± 1.34 µg/mL. Antimicrobial assessment performed by employing disc diffusion and minimal inhibitory concentration assays revealed TZEO as a potent antimicrobial agent with the highest inhibition activity towards tested gram-negative strains. The most sensitive on the treatment with TZEO was Enterobacter aerogenes showing an MIC 50 value of 0.147 ± 0.006 mg/mL and a MIC 90 value of 0.158 ± 0.024 mg/mL. Additionally, an in situ analysis showed great effects of TZEO in inhibiting gram-negative E. coli, P. putida, and E. aerogenes growing on bananas and cucumbers. Treatment with the TZEO vapor phase in the concentration of 500 µg/mL was able to reduce the growth of these bacteria on the food models to the extent > 90%, except for E. coli growth on the cucumber, which was reduced to the extent of 83.87 ± 4.76%. Furthermore, a test on the antibiofilm activity of the tested essential oil revealed its biofilm prevention effects against Salmonella enterica which forms biofilms on plastic and stainless-steel surfaces. Performed tests on the TZEO effects towards cell viability showed no effects on the normal MRC-5 cell line. However, the results of MTT assay of TZEO effects on three cancer cell lines (MDA-MB-231, HCT-116, and K562) suggest that TZEO exerted the strongest effects on the inhibition of the viability of MDA-MB-231 cells, especially after long-term treatment in the highest concentration applied with reducing the viability of the cells to 57%. Additionally, results of NBT and Griess assays suggest that TZEO could be a convenient candidate for future testing for developing novel antitumor therapies.

An In-Depth Study on the Chemical Composition and Biological Effects of Pelargonium graveolens Essential Oil.

The essential oil of Pelargonium graveolens (PGEO) is identified in the literature as a rich source of bioactive compounds with a high level of biological activity. This study aimed to examine the chemical profile of PGEO as well as its antioxidant, antibacterial, antibiofilm, and insecticidal properties. Its chemical composition was analyzed using gas chromatography-mass spectrometry (GC-MS), achieving comprehensive identification of 99.2% of volatile compounds. The predominant identified compounds were ß-citronellol (29.7%) and geraniol (14.6%). PGEO's antioxidant potential was determined by means of DPPH radical and ABTS radical cation neutralization. The results indicate a higher capacity of PGEO to neutralize the ABTS radical cation, with an IC50 value of 0.26 ± 0.02 mg/mL. Two techniques were used to assess antimicrobial activity: minimum inhibitory concentration (MIC) and disk diffusion. Antimicrobial evaluation using the disk diffusion method revealed that Salmonella enterica (14.33 ± 0.58 mm), which forms biofilms, and Priestia megaterium (14.67 ± 0.58 mm) were most susceptible to exposure to PGEO. The MIC assay demonstrated the highest performance of this EO against biofilm-forming S. enterica (MIC 50 0.57 ± 0.006; MIC 90 0.169 ± 0.08 mg/mL). In contrast to contact application, the assessment of the in situ vapor phase antibacterial activity of PGEO revealed significantly more potent effects. An analysis of antibiofilm activity using MALDI-TOF MS demonstrated PGEO's capacity to disrupt the biofilm homeostasis of S. enterica growing on plastic and stainless steel. Additionally, insecticidal evaluations indicated that treatment with PGEO at doses of 100% and 50% resulted in the complete mortality of all Harmonia axyridis individuals.

Phytonutrients of Bitter Apricot Seeds Modulate Human Lipid Profile and LDL Subfractions in Adults with Elevated Cholesterol Levels.

The objective of the present study was to evaluate the effect of short-term consumption of bitter apricot seeds phytonutrients on cardiovascular risk factors with a special focus on LDL cholesterol subfractions using the Lipoprint system. A group of 34 adult volunteers (21 female/13 male) consumed 60 mg kg-1 of body weight of bitter apricot seeds daily for 42 days. Subjects were divided into two groups: one with normal cholesterol levels (NTC) and one with elevated total cholesterol levels (ETC). Blood serum levels of total cholesterol (T-C), low-density cholesterol (LDL-C), high-density cholesterol (HDL-C), and triglycerides (TG) did not change significantly (p > 0.05) in NTC group. However, there were significant decreasing of T-C (p ˂ 0.05) and LDL-C (p < 0.01) in ETC group. The LDL1, LDL2, and atherogenic LDL3-7 subfractions progressively decreased after 42 days of apricot seeds consumption in ETC group (p < 0.05). Apricot seeds consumption was associated with a significant increase in the mean LDL particle size especially in ETC group (p ˂ 0.01). The results of the present study support the hypothesis that daily consumption of bitter apricot seeds for 42 days positively modified the lipoprotein profile in the group with elevated total cholesterol.

Cornelian cherry fruit improves glycaemia and manifestations of diabetes in obese Zucker diabetic fatty rats.

Cornelian cherry (Cornus mas L.) was in the past frequently used in Slovak Republic; meanwhile fell into oblivion despite the fact that it is known as antidiabetic supplement. However, there is no research investigated its effect on animal model of Diabetes mellitus (DM) 2 type as it is Zucker diabetic fatty (ZDF) rats. Thus, the aim of this study was to determine the effect of C. mas fruit given orally on the development of DM symptoms in ZDF rats. In the experiment male ZDF rats (fa/fa) and their age-matched non-diabetic lean controls (fa/+) were used aged 12 weeks. Male ZDF rats were administered C. mas in two doses (500 and 1000 mg/kg body weight) using a gastric gavage for 10 weeks. One group of diabetic animals served as positive control and received only distilled water. We found significant decrease of glucose level after oral administration of C. mas in dose of 1000 mg/kg bw in pre-diabetic state of animals (until 7th week of the experiment) and significant restriction of water intake in both C. mas groups against the diabetic control. We presume that the higher dose of Cornelian cherry could be beneficial and helpful in prevention of diabetic symptoms when consumed regularly in young animals.

The effects of 12-week progressive strength training on strength, functional capacity, metabolic biomarkers, and serum hormone concentrations in healthy older women: morning versus evening training.

Previous findings suggest that performing strength training (ST) in the evening may provide greater benefit for young individuals. However, this may not be optimal for the older population. The purpose of this study was to compare the effects of a 12-week ST program performed in the morning vs. evening on strength, functional capacity, metabolic biomarker and basal hormone concentrations in older women. Thirty-one healthy older women (66 ± 4 years, 162 ± 4 cm, 75 ± 13 kg) completed the study. Participants trained in the morning (M) (07:30, n = 10), in the evening (E) (18:00, n = 10), or acted as a non-training control group (C) (n = 11). Both intervention groups performed whole-body strength training with 3 sets of 10-12 repetitions with 2-3 minutes rest between sets. All groups were measured before and after the 12-week period with; dynamic leg press and seated-row 6-repetition maximum (6-RM) and functional capacity tests (30-second chair stands and arm curl test, Timed Up and Go), as well as whole-body skeletal muscle mass (SMM) (kg) and fat mass (FM-kg, FM%) assessed by bioelectrical impedance (BIA). Basal blood samples (in the intervention groups only) taken before and after the intervention assessed low-density lipoprotein (LDL-C), high-density lipoprotein (HDL-C), blood glucose (GLU), triglycerides (TG), high-sensitive C-reactive protein (hsCRP) concentrations and total antioxidant status (TAS) after a 12 h fast. Hormone analysis included prolactin (PRL), progesterone (P) estradiol (ESTR), testosterone (T), follicle stimulating hormone (FSH), and luteinizing hormone (LH). While C showed no changes in any variable, both M and E significantly improved leg press (+ 46 ± 22% and + 21 ± 12%, respectively; p < 0.001) and seated-row (+ 48 ± 21% and + 42 ± 18%, respectively; p < 0.001) 6-RM, as well as all functional capacity outcomes (p < 0.01) due to training. M were the only group to increase muscle mass (+ 3 ± 2%, p < 0.01). Both M and E group significantly (p < 0.05) decreased GLU (-4 ± 6% and -8 ± 10%, respectively), whereas significantly greater decrease was observed in the E compared to the M group (p < 0.05). Only E group significantly decreased TG (-17 ± 25%, p < 0.01), whereas M group increased (+ 15%, p < 0.01). The difference in TG between the groups favored E compared to M group (p < 0.01). These results suggest that short-term "hypertrophic" ST alone mainly improves strength and functional capacity performance, but it influences metabolic and hormonal profile of healthy older women to a lesser extent. In this group of previously untrained older women, time-of-day did not have a major effect on outcome variables, but some evidence suggests that training in the morning may be more beneficial for muscle hypertrophy (i.e. only M significantly increased muscle mass and had larger effect size (M: g = 2 vs. E: g = 0.5).

Intake of bilberries (Vaccinium myrtillus L.) reduced risk factors for cardiovascular disease by inducing favorable changes in lipoprotein profiles.

Cardiovascular diseases (CVDs) are the leading causes of death, and lifestyle modification, including dietary changes, is recommended to improve this condition. In this study, regular consumption of bilberries was hypothesized to have beneficial effects on CVD risk reduction, by changes in human health indicators such as decreasing low-density lipoprotein cholesterol (LDL-C) and triglycerides (TGs) and increasing high-density lipoprotein cholesterol (HDL-C). The research involved women (n=25) and men (n=11) who consumed 150 g of frozen stored bilberries 3 times a week for 6 weeks. Anthropometric parameters, blood pressure, lipid profile, glucose, liver enzymes, creatinine, albumin, magnesium, and antiradical activity were measured. Except for the body mass index of women (P=.019), no significant changes were found for anthropometric indicators. The consumption of bilberries led to a decrease in the following parameters: total cholesterol (P=.017), LDL-C (P=.0347), TG (P=.001), glucose (P=.005), albumin (P=.001), γ-glutamyltransferase (P=.046), and a positive increase in HDL-C (P=.044). In men, additionally, favorable changes were observed in total cholesterol (P=.004), glucose (P=.015), albumin (P=.028), aspartate aminotransferase (P=.012), γ-glutamyltransferase (P=.013), and HDL-C (P=.009; in this group, LDL-C increased [P=.007]). Changes in other parameters were not significant, for both women and men. Thus, the regular intake of bilberries can be important to reduce CVDs risk, by decreasing LDL-C/TG and increasing HDL-C.