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Phosholipase A2 (PLA2) activity in the seminal plasma and in sperm heads is closely related to sperm motility and male fertility. Therefore, the purpose of this study was to investigate the possible involvement of different isoforms of phospholipase in asthenozoospermia. To accomplish this, cPLA2, phospho-cPLA2, iPLA2, and sPLA2 were evaluated by immunofluorescence and immunoblot analyses in spermatozoa obtained from 22 normozoospermic men and 28 asthenozoospermic patients. We found significant differences in cPLA2 and its phosphorylated/activated form, iPLA2, and sPLA2 content and distribution in normal and asthenozoospermic patients. cPLA2 was localized in heads, midpieces, and tails of all spermatozoa as constitutive enzyme, less expressed in the tail of spermatozoa with low progressive motility. While active phospho-cPLA2 distribution was homogeneous throughout the cell body of control-donor spermatozoa, lower levels were detected in the tails of asthenozoospermic patients, as opposed to its strong presence in heads. Low immunofluorescence signal for iPLA2 was found in astenozoospermic patients, whereas sPLA2 was significantly lower in the heads of asthenozoospermic patients. Spermatozoa with low progressive motility showed differences both in terms of total specific activity and of intracellular distribution. cPLA2, iPLA2, and sPLA2 specific activities correlated positively and in a significantly manner with sperm progressive motility both in normozoospermic men and asthenozoospermic patients. In conclusion, PLA2s are expressed in different areas of human spermatozoa. Spermatozoa with low motility showed differences in total specific activity and enzyme distributions. We speculated that PLA2 expression and/or different distribution could be potential biomarkers of asthenozoospermia, one of the major causes of male factor infertility. [ABSTRACT FROM AUTHOR]

Introduction: Cytokines are glycoproteins that modulate reproductive function through a series of various mechanisms (by both conditioning gonadal steroidogenesis and contributing to the preservation of an inflammatory microenvironment). Aim of the Study: To evaluate the impact of certain clinical variables (i.e., age, obesity, insulin resistance index, serum antithyroid antibodies serum levels) on the serum concentrations of cytokines TNF-alpha, IL-6, and IL-10 in the follicular fluid of women undergoing a medically assisted procreation (PMA) cycle. Materials and Methods: A total of 40 female patients undergoing an intracytoplasmic sperm injection (ICSI) in oocytes, following ovarian stimulation by purified FSH and hCG carried out after suppression of ovarian function. The follicular fluid, obtained by surgical ultrasonography-guided withdrawal, was stored at -30 degrees C. Subsequently the cytokines were assayed by ELISA technique. Results: Women suffering from class II obesity showed follicular levels of TNF-alpha significantly higher (p<0.05) than women with a normal body mass index (BMI). Significantly higher concentrations of TNF-alpha and IL-6 were found in women with HOMA index >2.5. Women clinically presenting with concomitant obesity and high serum levels of antithyroid antibodies were found to have higher follicular levels of TNF-alpha and IL-6 (p <0.05) in comparison with women suffering from obesity only or low antithyroid antibodies levels only, or from both these conditions. Conclusion: Obesity is a common clinical condition associated with a higher concentration of inflammatory substances in the follicular fluid of infertile women. It is not understood, as yet, the possible pejorative role exerted by the presence of other clinical conditions, such as insulin resistance and high levels of antithyroid antibodies, that are conditions frequently encountered in the clinical practice. [ABSTRACT FROM AUTHOR]

Phosholipase A2 (PLA2) activity in the seminal plasma and in sperm heads is closely related to sperm motility and male fertility. Therefore, the purpose of this study was to investigate the possible involvement of different isoforms of phospholipase in asthenozoospermia. To accomplish this, cPLA2, phospho-cPLA2, iPLA2, and sPLA2 were evaluated by immunofluorescence and immunoblot analyses in spermatozoa obtained from 22 normozoospermic men and 28 asthenozoospermic patients. We found significant differences in cPLA2 and its phosphorylated/activated form, iPLA2, and sPLA2 content and distribution in normal and asthenozoospermic patients. cPLA2 was localized in heads, midpieces, and tails of all spermatozoa as constitutive enzyme, less expressed in the tail of spermatozoa with low progressive motility. While active phospho-cPLA2 distribution was homogeneous throughout the cell body of control-donor spermatozoa, lower levels were detected in the tails of asthenozoospermic patients, as opposed to its strong presence in heads. Low immunofluorescence signal for iPLA2 was found in astenozoospermic patients, whereas sPLA2 was significantly lower in the heads of asthenozoospermic patients. Spermatozoa with low progressive motility showed differences both in terms of total specific activity and of intracellular distribution. cPLA2, iPLA2, and sPLA2 specific activities correlated positively and in a significantly manner with sperm progressive motility both in normozoospermic men and asthenozoospermic patients. In conclusion, PLA2s are expressed in different areas of human spermatozoa. Spermatozoa with low motility showed differences in total specific activity and enzyme distributions. We speculated that PLA2 expression and/or different distribution could be potential biomarkers of asthenozoospermia, one of the major causes of male factor infertility. [ABSTRACT FROM AUTHOR]

Airborne transmission is predicted to be a prevalent route of human exposure with SARS-CoV-2. Aside from African green monkeys, nonhuman primate models that replicate airborne transmission of SARS-CoV-2 have not been investigated. A comparative evaluation of COVID-19 in African green monkeys, rhesus macaques, and cynomolgus macaques following airborne exposure to SARS-CoV-2 was performed to determine critical disease parameters associated with disease progression, and establish correlations between primate and human COVID-19. Respiratory abnormalities and viral shedding were noted for all animals, indicating successful infection. Cynomolgus macaques developed fever, and thrombocytopenia was measured for African green monkeys and rhesus macaques. Type II pneumocyte hyperplasia and alveolar fibrosis were more frequently observed in lung tissue from cynomolgus macaques and African green monkeys. The data indicate that, in addition to African green monkeys, macaques can be successfully infected by airborne SARS-CoV-2, providing viable macaque natural transmission models for medical countermeasure evaluation., Competing Interests: The authors have declared that no competing interests exist.

Air pollution has been suggested to affect semen quality, but the evidence is still contradictory. To assess whether any differences occur in conventional sperm parameters of men life-long resident in low, middle-low, middle, and high industrial density zones in the province of Messina. We retrospectively analyzed the conventional sperm parameters of patients to whom the sperm analysis was requested during their female partner counseling for infertility in an assisted reproductive technique (ART) center. A total of 184 men were enrolled. Total sperm count was higher in patients living in low and middle-low industrial density areas compared with that of men living in middle and high ones (123.5 ± 146.8 vs. 80.7 ± 92.7 mil/ejaculate, p < 0.05). No difference was found for sperm concentration (37.2 ± 49.7 vs. 30.5 ± 37.2 mil/mL), progressive motility (15.4 ± 19.8% vs. 14.2 ± 18.4%), total motility (62.3 ± 20.5 vs. 58.4 ± 19.9 mil/mL), and normal forms (2.7 ± 1.5 vs. 2.3 ± 3.0 mil/mL). These results add further evidence to findings from Sicilian population. Effective control of air pollution should be accomplished to prevent its negative impact on human reproductive health.

The chloroquine resistance transporter of the human malaria parasite Plasmodium falciparum , PfCRT, is an important determinant of resistance to several quinoline and quinoline-like antimalarial drugs. PfCRT also plays an essential role in the physiology of the parasite during development inside erythrocytes. However, the function of this transporter besides its role in drug resistance is still unclear. Using electrophysiological and flux experiments conducted on PfCRT-expressing Xenopus laevis oocytes, we show here that both wild-type PfCRT and a PfCRT variant associated with chloroquine resistance transport both ferrous and ferric iron, albeit with different kinetics. In particular, we found that the ability to transport ferrous iron is reduced by the specific polymorphisms acquired by the PfCRT variant as a result of chloroquine selection. We further show that iron and chloroquine transport via PfCRT is electrogenic. If these findings in the Xenopus model extend to P. falciparum in vivo , our data suggest that PfCRT might play a role in iron homeostasis, which is essential for the parasite's development in erythrocytes., (© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.)

Objective: We sought to assess the prevalence of asymptomatic venous thrombosis in infertile women with thrombophilic disorders (TDs)., Methods and Results: A total of 73 infertile women with TDs underwent duplex ultrasound scan to evaluate superficial and deep venous circulation of lower limbs. A control group of 35 infertile women without TDs was included. A single TD was found in 13 (17.8%) subjects, and 40 (54.8%) women presented a combined defect (more than three alterations). No residual mural thrombosis (RT) was noted in any deep veins. We found RT in 48 (65.8%) patients of TD group, while no RT was found in the control group (p < 0.0001). None of the clinical and prothrombotic factors were predictors of RT (all p > 0.20), and frequency of TD did not correlate with multi-vessel RT (p = 0.252)., Conclusions: No signs of deep vein thrombosis but high prevalence of superficial RT is present in infertile women with TDs. Further studies are needed to assess the prognostic value of our findings., (© The Author(s) 2014.)

Mutations in the "chloroquine resistance transporter" (PfCRT) are a major determinant of drug resistance in the malaria parasite Plasmodium falciparum. We have previously shown that mutant PfCRT transports the antimalarial drug chloroquine away from its target, whereas the wild-type form of PfCRT does not. However, little is understood about the transport of other drugs via PfCRT or the mechanism by which PfCRT recognizes different substrates. Here we show that mutant PfCRT also transports quinine, quinidine, and verapamil, indicating that the protein behaves as a multidrug resistance carrier. Detailed kinetic analyses revealed that chloroquine and quinine compete for transport via PfCRT in a manner that is consistent with mixed-type inhibition. Moreover, our analyses suggest that PfCRT accepts chloroquine and quinine at distinct but antagonistically interacting sites. We also found verapamil to be a partial mixed-type inhibitor of chloroquine transport via PfCRT, further supporting the idea that PfCRT possesses multiple substrate-binding sites. Our findings provide new mechanistic insights into the workings of PfCRT, which could be exploited to design potent inhibitors of this key mediator of drug resistance., (© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.)

Mutations in the chloroquine resistance transporter (PfCRT) are the primary determinant of chloroquine (CQ) resistance in the malaria parasite Plasmodium falciparum. A number of distinct PfCRT haplotypes, containing between 4 and 10 mutations, have given rise to CQ resistance in different parts of the world. Here we present a detailed molecular analysis of the number of mutations (and the order of addition) required to confer CQ transport activity upon the PfCRT as well as a kinetic characterization of diverse forms of PfCRT. We measured the ability of more than 100 variants of PfCRT to transport CQ when expressed at the surface of Xenopus laevis oocytes. Multiple mutational pathways led to saturable CQ transport via PfCRT, but these could be separated into two main lineages. Moreover, the attainment of full activity followed a rigid process in which mutations had to be added in a specific order to avoid reductions in CQ transport activity. A minimum of two mutations sufficed for (low) CQ transport activity, and as few as four conferred full activity. The finding that diverse PfCRT variants are all limited in their capacity to transport CQ suggests that resistance could be overcome by reoptimizing the CQ dosage.

No studies have evaluated the ultrasound features of the male sex accessory glands in infertile patients with bacterial male accessory gland infection (MAGI) according to the microbiological outcomes of bacterial cultures (absent, partial or complete) following antibiotic therapy administration. Therefore, the aim of this study was to evaluate the ultrasound characteristics of the prostate, seminal vesicles, and epididymal tracts after treatment with levofloxacin (a common quinolone antibiotic), in patients with infections caused by Escherichia coli (a Gram-negative bacterium) according to the Naber's classification, which includes the following categories: eradication, eradication with superinfection, persistence and persistence with superinfection. The study was conducted in 100 patients aged 25±8 years (range: 20-40 years) with bacterial MAGI and bacterial cultures positive only for E. coli (colony forming units ≥10(6) per ml). Retrospective analysis was conducted only on patients treated with oral levofloxacin (500 mg) administered once daily for 28 days who were recruited over the last 5 years. Following antibiotic treatment, patients with microbiological persistence or persistence with superinfection had a significantly higher percentage of ultrasound abnormalities suggestive of prostato-vesiculitis (PV) (30.2% and 36.0%, respectively) or prostato-vesiculo-epididymitis (PVE) (60.2% and 70.0%, respectively) compared with patients with microbiological eradication (PV=10.2% and PVE=8.2%, respectively) or eradication with superinfection (PV=18.8% and PVE=21.2%, respectively). In conclusion, patients with microbiological persistence or persistence plus superinfection showed the highest prevalence of complicated forms of MAGI (PV and PVE), compared with patients with microbiological eradication or eradication with superinfection.

In this study, we investigated the prevalence of Ureaplasma urealyticum and Mycoplasma hominis infection among 250 unselected infertile men, the presence of urogenital symptoms in infected men and the effects of these microorganisms on the conventional sperm parameters. Urethral samples were obtained using a swab inserted 3-4 cm into the urethral meatus. Ureaplasma urealyticum and Mycoplasma hominis were detected by the kit Mycofast R evolution 3 Elitech Microbiology (Elitech Microbiology, Signes, France). Ureaplasma urealyticum was detected in 15.6% of the cases and Mycoplasma hominis in 3.6%. One patients had a co-infection with both pathogens. About 41% of the infertile patients with mycoplasma infection had urogenital symptoms. A lower number of patients with mycoplasma infection had normal sperm parameters compared with non-infected infertile men, but this frequency showed only a trend compared to non-infected patients (Chi-square=3.61; P=0.057), and a significantly higher percentage of patients with oligo-astheno-teratozoospermia (Chi-square=127.3; P<0.0001), or asthenozoospermia alone (Chi-square=5.74; P<0.05) compared to non-infected infertile patients. In conclusion, this study showed an elevated prevalence of Ureaplasma urealyticum and Mycoplasma hominis infection in unselected men attending an infertility outpatient clinic and that the presence of these microorganisms is associated with a higher percentage of patients with abnormal sperm parameters.

Chlamydia (C.) trachomatis, an obligate intracellular bacterium, is responsible for the most common sexual transmitted disease and infertility. The purpose of this study was to evaluate: a) the frequency of chlamydial infection in unselected infertile couples and b) whether chlamydial infection could be identified in the semen sample as effectively as in the urethral swab of infertile patients. To accomplish this, 73 unselected, consecutive infertile couples were enrolled. Both male and female partners underwent a complete work-up to identify the cause of their infertility. A PCR method was used to detect C. trachomatis in urethral swabs and the semen samples of the male partners and in the cervical swabs of the female partners. C. trachomatis infection was found in 6 couples (8.2%). Three couples had both partners infected, 2 couples had only the male partner infected, and 1 only the female partner. C. trachomatis infection was found in the urethral swab of all 5 men infected, whereas the bacterial DNA was found in the semen sample of 2 of them. These findings suggest that C. trachomatis infection is present in about 8% of unselected infertile couples and that the bacterium should be searched in the male partner urethral swab which has a higher sensitivity.

Studies suggest Candida albicans infection has a negative effect on sperm function, including fertilizing ability. Assisted reproduction treatment using spermatozoa from a patient with unrecognized C. albicans infection did not result in fertilization. Preliminary evidence suggested an effect on sperm motility and apoptosis. This study was undertaken to evaluate the effects of experimentally induced C. albicans infection on motility, membrane mitochondrial potential (MMP), chromatin packaging and apoptosis [membrane phosphatidylserine (PS) externalization and DNA fragmentation] of spermatozoa isolated from normozoospermic healthy men. Motile spermatozoa were isolated by swim-up from 13 normal volunteers and exposed to increasing concentrations (0, 1000, 10,000, and 100,000 cfu/ml) of the fungus for 3 and 24 h. C. albicans was isolated from vaginal swabs, after identification, freshly prepared for experiments. Following incubation, sperm motility decreased significantly (P < 0.05 from 10,000 cfu/ml) and spermatozoa with reduced MMP or PS externalization, an early sign of apoptosis, increased in a time- and concentration-dependent manner. Sperm DNA fragmentation and chromatin integrity increased slightly after exposure to C. albicans, but the increase did not reach statistical significance. This study showed that C. albicans infection may decrease the functional competence of spermatozoa by reducing motility and MMP and by promoting molecular apoptosis mechanisms.

The relationship between motor maps and cytoarchitectonic subdivisions in rat frontal cortex is not well understood. We use cytoarchitectonic analysis of microstimulation sites and intracellular stimulation of identified cells to develop a cell-based partitioning scheme of rat vibrissa motor cortex and adjacent areas. The results suggest that rat primary motor cortex (M1) is composed of three cytoarchitectonic areas, the agranular medial field (AGm), the agranular lateral field (AG1), and the cingulate area 1 (Cg1), each of which represents movements of different body parts. Vibrissa motor cortex corresponds entirely and for the most part exclusively to AGm. In area AG1 body/head movements can be evoked. In posterior area Cg1 periocular/eye movements and in anterior area Cg1 nose movements can be evoked. In all of these areas stimulation thresholds are very low, and together they form a complete representation of the rat's body surface. A strong myelinization and an expanded layer 5 characterize area AGm. We suggest that both the strong myelinization and the expanded layer 5 of area AGm may represent cytoarchitectonic specializations related to control of high-speed whisking behavior.

The authors used multiple ovulation induction by CC+hMG+hCG and intrauterine insemination with capacitized sperm in 63 women, aged between 22 and 43 (mean age 32.474 +/- 5.364) with primary or secondary infertility of various origins (tubal problems being excluded) to enhance the chances of pregnancy and evaluate the pregnancy rate. Partners were aged between 26 and 47 (mean age 36.571 +/- 6.709). The following parameters were considered in all patients: mean age of the partners of the couple, parity, seminal fluid characteristics, duration of infertility, dose of each drug administered, mean number of ampuls per patient and mean 17 beta-estradiol level at the time of administration of hCG. There were 189 cycles induced and 26 suspended (13.75%) including 7 (3.70%) because of protocol drop-out, 13 (6.87%) because of the development of slight hyperstimulation and 6 (3.17%) because of marked hyperstimulation. There were 27 pregnancies (42.85%), 4 of which ended in miscarriage between 7 and 10 weeks. There were 4 twin pregnancies, i.e. 14.81%. Of the 27 pregnancies, 13 (48.14%) were obtained during the first cycle, 9 (33.33%) during the second cycle and 5 (18.51%) during the 3rd cycle. The rate of pregnancies per cycle was 14.89%. With regard to 17 beta-estradiol levels at the time of administration of hCG, 18 pregnancies (69.23%) were obtained in patients with a 17 beta-estradiol level > or = to 1300, 7 (25.92%) in patients with a 17 beta-estradiol level > or = to 1000 and < or = to 1300 pg/ml and 3 (11.11%) in those with a 17 beta-estradiol level < or = to 900 pg/ml.

The authors have compared the results obtained using four main drugs (calcitonin, ipriflavone, transdermal estrogens, fluorine-calcium) actually employed for the treatment of post-menopausal osteoporosis, administered to four groups of fast loser patients (442) in natural (months mean 19.97 +/- 5.99) and surgical (months mean 16.94 +/- 4.29) menopause. After six months of treatment, the efficacy of therapy has been evaluated on the basis of BD (bone density) and osteoarticular pain changes. The BD results have been compared with those of 100 non-treated patients, in the same clinical conditions. The authors have noticed an increase in bone mass (from +0.47% to +1.59%) and a great improvement in osteoarticular pain with all therapeutical protocols used while in the control group there was a progressive decrease of BMC (-1.23%) and a worsening pain. Comparing the results obtained with different therapies, the difference of mean mineralometric gain is not particularly significant among several treatments; but this difference is very significant between treated and non-treated patients who have continued to lose bone mass. The collateral effects, observed during administration of different drugs, have been minimal and the suspension of therapy has been always associated to their disappearance. In the opinion of the authors the good results, achieved with different therapies, depend on the precocity of the treatment, but also on the fact that, in peri-menopausal period, their effect has been increased by the estrogens. Being osteoporosis a multifactorial pathology, a careful control of the risk factors is appropriate and needs to be enforced in order to carry out a precocious treatment with specific drugs on bone metabolism and try to balance the natural turnover with the loss of bone mass.