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OBJECTIVE: This study aimed at comparing the depression and anxiety levels, and health self-perception during the coronavirus disease 2019 pandemic among subjects who practice aerobic, strength, and mixed (aerobic and strength) exercises and nonsports participants. MATERIALS AND METHODS: We included 304 Brazilians of both sexes in this cross-sectional study. All participants were recruited through online advertisement and completed a self-administered questionnaire regarding the personal information, level of restriction adopted, physical activity, and mood state screening (Patient Health Questionnaire-9 and General Anxiety Disorder-7). We divided the participants into four groups: strength sports group (CrossFit or strength training), aerobic/endurance sports groups (running, cycling, triathlon, or swimming), mixed sports groups (individuals who practice endurance and strength sports), and nonsports group. RESULTS: The Kruskal-Wallis test showed a significant effect of the group on the depression and anxiety levels. Meanwhile, the post-hoc comparisons showed a significantly lower depression level in the mixed and aerobic sports groups than in the strength sports and nonsports groups, and a significantly lower anxiety level in the mixed and aerobic sports groups than in the nonsports group. Furthermore, participants in the mixed, strength, and aerobic sports groups presented a better level of health self-assessment than the nonsports group, and those in the mixed sports group had a better level of health self-assessment than the strength or aerobic sports groups. CONCLUSIONS: Individuals practicing aerobic exercises present lower depression and anxiety levels than those practicing strength training and are inactive. However, individuals who practice strength exercises and aerobics have the best levels of health perception.

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Lipoblastoma is a rare and benign tumour arising from embryonal fat cells, predominantly diagnosed in children younger than 3 years old. The most frequent locations are the extremities and trunk, while the head and neck areas are more rarely affected (10-15% of total cases). Clinically, the most common presentation is a fast-growing painless mass. Ultrasound is the first-line imaging examination, but Magnetic Resonance Imaging (MRI) allows for better definition of the relationships with the adjacent vascular and muscular structures. It can help to identify the lipomatous components, and it is useful for preoperative planning. However, the definitive diagnosis is provided by histopathological examination. Complete surgical excision is the first-line treatment, with a good prognosis in case of total eradication. We report the case of a 7-month-old male child with a rapidly growing mass that had typical radiological features of lipoblastoma.

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The antenatal diagnosis of congenital toxoplasmosis represents today an important application of molecular diagnostic methods such as PCR. Done directly in amniotic fluid, the PCR for T. gondii is able to detect in utero infected foetus with high probability. Doing so, it precludes to obtain foetal blood and to use more cumbersome and lengthy procedures such as inoculation to cell lines or to mice. Although it is use today in many centres, the molecular diagnosis of T. gondii by PCR is neither commercialized nor standardised. The objective of this report is to present the methodology used in our institution and to establish its degree of validation.

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A molecule isolated from the peritoneal fluids of women undergoing laparoscopy for in-vitro fertilization techniques has been chemically characterized and identified as 1-palmitic-3-phosphorylcholine (lysophosphatidylcholine, LPC). This lipid is able, at physiological concentrations, to completely inhibit sperm motility in vitro in a dose-dependent way. Synthetic LPC induced rapid and complete arrest of sperm motility when added to sperm suspensions at physiological concentrations without any damage to cell membranes. Taken together, these results suggest that LPC may represent a previously unrecognized in-vivo modulator of human sperm motility.

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OBJECTIVES: A study was conducted to evaluate the extent of a Q-fever epidemic through active case finding in the area of Vicenza (north-eastern Italy), and to identify risk factors for Q-fever in this outbreak. METHODS: 1) Descriptive epidemiology; 2) Seroepidemiological survey; 3) Case-control study. 1) Epidemic curve and maps with the location of cases. Identification of the road followed by the flocks of sheep. 2) Cross-sectional study on humans and flocks of sheep tested for anti-Coxiella burnetii antibodies. 3) Cases were defined by the presence of fever > 38 degrees C plus serological confirmation. Controls were 94 apparently healthy individuals attending outpatient facilities for control visits or certification, group-matched by geographical area, age and gender. A standardized questionnaire was administered by trained interviewers. Odds ratio and 95% confidence intervals (CI) were used to evaluate risk factors for Q-fever. RESULTS: A total 58 cases were identified in a 5-month period. Male to female ratio was 2.8:1; mean age was 42 years (range: 20-65 years). Twenty-eight patients (48%) were hospitalized. Fever was accompanied by asthenia (81%), headache (76%), chills (72%), and myalgia and arthralgia (53%); cough was present in 47% of patients. Rx abnormalities were found in 81% of the patients undergoing chest X-ray. Among 111 apparently healthy family members who underwent serological testing, four (3.6%) had antibodies to Coxiella burnetii. Three flocks which passed through the outbreak area between late May and early June were shown to be infected, with prevalence of antibodies ranging between 45 and 53%. The case-control study showed a significant association with exposure to flocks of sheep (Odds ratio = 6.1; 95% CI 2.5, 16.3). Other potential risk factors were not more commonly reported by cases with respect to controls. CONCLUSIONS: Indirect exposure to flocks of sheep was a determinant of this outbreak of Q-fever. This finding suggests that transmission occurred through inhalation of contaminated airborne particles. The importance of control measures should be stressed in areas traversed by flocks of sheep.

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The scope of this study was to evaluate the accuracy, precision and specificity of the sperm concentration measurements by the Strömberg-Mika Cell Motion Analyser (SM-CMA). Our data show that the instrument generally underscores the sperm concentration and therefore the uncorrected measurements must be corrected by the operator using the 'mouse'-driven option. In terms of precision, the system appears to have an excellent internal precision whereas its repeatability is influenced by the sperm concentration, the sample's homogeneity and the correction of the raw data. In order to increase the system's repeatability, we suggest that sperm counts should be carried out in various fields of the counting chamber, and the mean of the corrected values be taken as representative of the sperm concentration in the ejaculate if the various measurements show a homogeneous (poissonian) distribution. The correction of the raw data with the 'mouse'-driven correction option was also shown to improve the system's reproducibility. Concerning specificity, our data evidenced that, without technical correction, the instrument failed to correctly classify certain spermatozoa as such, thereby grossly underscoring sperm counts. This finding was more evident at low sperm counts. Overall, the SM-CMA requires additional laboratory time but the corrected sperm counts are comparable to manual counts and semi-automated counts with the added option that it provides the andrologists with various motility characteristics not possible with the latter methodologies.

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Aspergillus fumigatus secretes a serine alkaline protease (ALP) and a metalloprotease (MEP) when the fungus is cultivated in the presence of collagen as sole nitrogen and carbon source. The gene encoding ALP was isolated and characterized previously. We report here the cloning and the sequencing of the gene encoding MEP. Genomic and cDNA clones were isolated from A. fumigatus libraries using synthetic oligonucleotides as probes. Stretches of the deduced amino acid sequence were found to be in agreement with the N-terminal amino acid sequence of MEP and with internal peptide sequences. The amino acid sequence of the enzyme contains a putative active-site sequence HEYTH homologous to the active site of other bacterial and eukaryotic zinc metalloproteases. Sequence analysis reveals that MEP has a pre-proregion consisting of 245 amino acid residues preceding the 388 amino acid residues of the mature region (molecular mass of 42 kDa). An alp mep mutant, deficient in proteolytic activity at neutral pH in vitro, was constructed and tested for pathogenicity in a murine model. No difference in pathogenicity was observed between the wild-type strain and the alp mep double mutant, suggesting that ALP and MEP are not essential for the invasion of the lung tissues by A. fumigatus.

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The secreted aspartic proteinases (SAP) of Candida sp. are presumed to be potential virulence factors. In the opportunistic pathogen Candida albicans the proteinase genes identified to date, SAP1, SAP2, SAP3 and SAP4, constitute a multigene family. Before addressing the possible role of each proteinase in virulence, we sought to isolate all the members of this multigene family by screening a genomic library with a SAP1 probe for additional C. albicans SAP genes using low-stringency hybridization conditions. Three putative new members, SAP5, SAP6 and SAP7 were isolated and sequenced. The N-terminal segments of the deduced amino acid sequences of SAP5 and SAP6 contained secretion signal sequences similar to those of other Candida SAPs. Upon comparison and alignment with the other reported SAP amino acid sequences, SAP7 is not only the most divergent protein but also exhibits a much longer putative pro-sequence with a single Lys-Lys putative processing site. Using SAP1 to SAP7 as probes, the overall number of SAP genes in C. albicans was tentatively estimated by low-stringency hybridization to EcoRI-digested genomic DNA. While each isolated SAP gene could be assigned to distinct EcoRI bands, the existence of two additional genes not isolated after screening of the C. albicans gene library was inferred. Furthermore, evidence was obtained for the existence of SAP multigene families in other Candida species such as C. tropicalis, C. parapsilosis and C. guillermondii.

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Since progesterone has been claimed to induce acrosomal reaction and hyperactivated motility of human spermatozoa, the present study was undertaken to determine if its presence at concentrations similar to those of peri-ovulatory follicular fluid could influence the effect of peritoneal fluid on sperm motility in vitro. To this end, 11 sperm samples were incubated at 37 degrees C with five peritoneal fluids with/without exogenous progesterone, and sperm motility was assessed using a computer-assisted analyser at time (t) = 0, 2.5, 5 and 24 h. Overall there was no observable constant trend for enhancement or inhibition of sperm motility. Progesterone generally induced a negative effect on those sperm samples with high velocities in the native peritoneal fluids and a positive effect on those sperm samples demonstrating low motility in the native peritoneal fluids. The incorporation of progesterone into the incubation medium seemed to result in a 'tuning' of sperm velocity to around 30-50 micron/s. However, a given sperm sample reacted differently when incubated with various peritoneal fluids and, reciprocally, different semen samples incubated with the same peritoneal fluid showed very variable motility patterns. The greater variability of the effects exerted by progesterone on sperm motility could arise from the fact that each sperm sample may contain subpopulations of gametes with different sensitivity to progesterone.

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The relationship between the ability to secrete a specific acid proteinase (ACP) by Candida tropicalis in the presence of bovine serum albumin as a nitrogen source and virulence for mice was studied using two stable proteinase-positive and proteinase-negative strains (DSY68 and DSY65), which were constructed from the wild-type pathogenic yeast C. tropicalis ATCC 750. The inactivation of the gene encoding the secreted acid proteinase was produced by targeted gene disruption. Mortality rate was slightly lower in groups of mice infected with the proteinase-negative mutant. All other parameters analysed were similar for two strains of yeast. Our results therefore conclude that the ACP secreted by C. tropicalis did not contribute significantly to fungal virulence in systemic infections.

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Candida parapsilosis secretes an inducible acid protease (ACP) when cultivated in the presence of bovine serum albumin as the sole nitrogen source. In order to clone the ACP gene (ACP) of C. parapsilosis, a genomic library was screened with C. tropicalis ACP as the probe. Two different ORFs, ACPR and ACPL, were found to hybridize with the C. tropicalis ACP. ACPR contained a DNA sequence in agreement with the N-terminal amino acid sequence of C. parapsilosis ACP isolated from culture supernatants. ACPR was shown to be expressed and functional in a C. tropicalis acid protease mutant (acp) and with SDS-PAGE the protein product showed the same mobility as the ACP secreted by C. parapsilosis. These results imply that ACPR encodes the C. parapsilosis ACP. The deduced amino acid sequence of ACPR is similar to the amino acid sequence of proteases of the pepsin family. As in the case of the C. tropicalis and C. albicans ACP, the 5' extremity of ACPR revealed a propeptide containing two Lys-Arg amino acid pairs that have been identified as peptidase processing sites in several yeast-secreted peptides and protein precursors. As judged from the deduced amino acid sequences, the ACPL product would be similar to that of ACPR; however, a protein corresponding to ACPL was not found in supernatants from C. parapsilosis liquid cultures. In addition, ACPL did not complement the C. tropicalis acp mutant. We conclude that ACPL is a pseudogene or serves an as yet unidentified function.

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The gene for the secreted acid protease (ACP), a potential virulence factor of Candida species, was inactivated in Candida tropicalis by gene disruption. The disruption was performed by cotransformation of an ade2 C. tropicalis mutant with a linear DNA fragment carrying a deletion in ACP, and the replicative vector pMK16 which carries a selectable ADE2 gene marker. Few of the transformants exhibited lower protease secretion levels and were shown to have one deleted and one unaffected ACP copy, since C. tropicalis is a diploid yeast. These transformants were rendered homozygotic for this deletion by mild UV-treatment. One of the homozygotic acp deletion mutants obtained was completely devoid of extracellular protease activity and grew poorly on bovine serum albumin-containing medium. This mutant could be complemented by an ACP fragment inserted in pMK16, but also by an acid protease gene isolated from C. parapsilosis.

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The extracellular acid protease of Candida tropicalis was purified from the supernatant fraction of culture medium containing bovine serum albumin as nitrogen source and the NH2-terminal amino acid (aa) sequence of the protein was determined. The gene for the acid protease (ACP) was isolated using a pool of synthetic oligonucleotides as a probe and a segment of the deduced aa sequence was found to be in agreement with the NH2-terminal aa sequence of the protein. The deduced aa sequence of ACP is similar to the aa sequence of proteases of the pepsin family. The nucleotide sequence of the 5' portion of this gene revealed a coding sequence for a 60 residue propeptide containing two Lys-Arg amino acid pairs that have been identified as sites for peptidase processing of several exported peptides and proteins. The final Lys-Arg site occurs at the junction with the mature extracellular form of the acid protease.

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Aspergillus fumigatus secreted an inducible alkaline protease (AlPase) when cultivated in the presence of collagen (200 micrograms/ml) as sole nitrogen and carbon source. Proteolytic activity was maximum at pH 9.0 with azocollagen as substrate. The enzyme, which was the major protein found in the supernate of a liquid culture, was purified by ammonium sulphate precipitation and gel filtration. The Mr was determined to be 33 Kda by gel filtration and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The isoelectric point was estimated to be pH 8.2. Divalent cations strongly inhibited enzyme activity, whereas non-ionic detergents and reducing agents had no effect. A. fumigatus AlPase was totally inhibited by phenylmethanesulphonyl fluoride, antipain, chymostatin and alpha-2-macroglobulin. A. fumigatus AlPase is closely related to the A. oryzae AlPase, a serine protease of the subtilisin family, as attested by the antigen pattern seen by immunoblotting. The high collagenic activity and the ability of A. fumigatus AlPase to digest elastin could play a role in the invasion of the tissues by the fungus.

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