Comparison between two cryopreservation techniques of human ovarian cortex: morphological aspects and the heat shock response (HSR)

This study was tailored to compare the cryopreservation of the human ovarian cortex using closed metal container vitrification or the slow-freezing technique. Superficial ovarian cortical tissue biopsies were collected from 12 participants who underwent gynaecological videolaparoscopy. The fragmented samples were allocated to three experimental conditions: (a) fresh ovarian tissue, (b) slow-freezing, and (c) vitrification with a metal closed container. After thawing or rewarming, cellular morphological analyses were performed to determine tissue viability. The cellular response to thermal stress was measured by a putative increase in the immune quantification of the heat shock protein 70 kDa (heat shock protein 70 kDa response — HSR) after a heat challenge (2 h exposure at 42 °C). Both the total number of intact follicles and the frequency of primordial follicles were higher in fresh ovarian tissue than in the preserved samples, regardless of the technique employed. There was a trend towards an increase in the absolute number of intact follicles in the tissue preserved by vitrification. After cryopreservation, a higher HSR was obtained after slow-freezing. These results indicate that both cryopreservation techniques present advantages and may be used as alternatives to ovarian tissue cryopreservation.     

Evaluation of Strongyloides stercoralis infection in patients with HTLV-1

Introduction: Individuals infected with the human T-lymphotropic virus type 1 (HTLV-1) may present severe and disseminated forms of Strongyloides stercoralis infection with low therapeutic response.Objective: To investigate the S. stercoralis infection and the seroprevalence of IgG anti-S. stercoralis antibodies in individuals infected with HTLV-1 attending the Reference Center for HTLV-1 (CHTLV) in Salvador, Bahia, Brazil.Materials and methods: We conducted a cross-sectional study in 178 HTLV-1-infected individuals treated at the HTLV specialized center between January, 2014, and December, 2018. The parasitological diagnosis of S. stercoralis was performed using the Hoffman, Pons and Janer, agar plate culture, and Baermann-Morais methods. The IgG anti-S. stercoralis detection was performed using an in house enzyme-linked immunosorbent assay (ELISA). The HTLV-1 infection was diagnosed using a commercial ELISA and confirmed by Western blot.Results: The frequency of S. stercoralis infection was 3.4% (6/178). Individuals infected with S. stercoralis from rural areas (50.0%; 3/6) also showed S. stercoralis hyperinfection (>3,000 larvae/ gram of feces). The frequency of circulating anti-S. stercoralis IgG antibodies was 20.8% (37/178). Conclusions: HTLV-1-infected people living in precarious sanitary conditions are more prone to develop severe forms of S. stercoralis infection. Considering the high susceptibility and unfavorable outcome of the infection in these individuals, the serological diagnosis for S. stercoralis should be considered when providing treatment.     

Effects <Emphasis Type="Italic">per se</Emphasis> of Organic Solvents in the Cerebral Acetylcholinesterase of Rats

Acetylcholinesterase (AChE) was studied in different rat brain regions (cerebellum, hypothalamus, striatum, hippocampus and cortex) in the presence of different organic solvents normally used in the in vitro assay. The organic solvents used were acetone (C₃H₆O), acetonitrile (C₂H₃N), ethyl alcohol (C₂H₆O), isopropyl alcohol (C₃H₈O), methyl alcohol (CH₄O), tert-butyl alcohol (C₄H₁₀O) and dimethyl sulfoxide (DMSO, C₂H₆OS) ranging from 0.6 to 10%. Ethyl and methyl alcohol presented no effect on AChE activity at any of the concentrations and brain structures tested. In the hippocampus, isopropyl alcohol did not demonstrate a significant inhibitory effect, even at high concentrations. Tert-butyl alcohol presented an interesting result, increased AChE activity (P < .05) in the hypothalamus (1.8%), cortex (1.8 and 2.5) and striatum (1.2, 1.8 and 2.5%) and decreased activity at a concentration of 10% in the cortex (P < .05) and striatum (P < .01). Acetone and acetonitrile presented similar results, both significantly inhibiting AChE in all structures (5%, P < .05 and 10%, P < .01). DMSO exhibited a highly inhibitory effect at practically all concentrations tested (P < .01). In conclusion, for testing new compounds on AChE activity in vitro, methyl and ethyl alcohol may be the best organic solvent choice.     

Ascorbic acid and flavonoid-peroxidase reaction as a detoxifying system of H(2)O(2) in grapevine leaves

Biosynthesis of both ascorbic acid (AsA) and peroxidase activity were induced by light in cv. Sultana grapevine leaves. Induced peroxidase activity mainly involved basic isoenzymes of pI 9.8 and 9.6 and catalyzed the oxidation of flavonoids like quercetin and kaempferol and derivatives of hydroxycinnamic acids such as ferulic and p-coumaric acids, but not AsA. However, the peroxidase-dependent oxidation of ferulic acid and quercetin was temporarily suppressed by AsA as long as it remained in the reaction medium. Kinetics and spectroscopic results indicated that AsA was oxidized to dehydroascorbic acid only in the presence of phenols or flavonoids, and did not interfere with the catalytic activity of the peroxidase. Ascorbate peroxidase isoenzymes (APx), whose activities are widely considered central for detoxification of H(2)O(2) in most plant cells, were not detected in grape leaves extracts. The significance of light stimulus on peroxidase activity and leaf AsA content is discussed in terms of a flavonoid-redox cycle proposed as an alternative system to detoxify H(2)O(2) in grapevine leaves.     

Continuous gametogenesis in the neotropical freshwater teleost, Bryconops affinis (Pisces:Characidae)

The gametogenesis of Bryconops affinis was studied by light, transmission and scanning electron microscopy. The spermatogenesis is semi-cystic and spermatids are released into the lumen of seminiferous tubules, where spermiogenesis is completed. Spermatozoa have an ovoid head, a rudimentary middle piece with a small number of mitochondria and long flagellum (primitive spermatozoa). The Sertoli and Leydig cells show secretory activity during spermatogenesis. By the end of this phenomenon, the Sertoli cells phagocytize the residual spermatozoa, while the Leydig cells show involuted characteristics. With regard to the oogenesis process, the oocyte development was divided into four stages based on the cytological characteristics of the oocyte and its surrounding layers. Ultrastructural analysis revealed that the zona pellucida is formed during the previtellogenic stage. Specializations associated to the outer layer of the zona pellucida may be related to the egg's adherence to the substrata.     

Comparative analysis of gonadal morphology in six fish species of the Incertae Sedis genera in Characidae of occurrence in the São Francisco River Basin,Brazil

Martins, Y.S., Arantes, F.P., Sato, Y., dos Santos, J.E., Rizzo, E. and Bazzoli, N. 2012. Comparative analysis of gonadal morphology in six fish species of the Incertae Sedis genera in Characidae of occurrence in the São Francisco River Basin, Brazil. —Acta Zoologica (Stockholm) 93 : 48–56. This study presents a comparative analysis aiming to show the remarkable differences found in gonad morphology between six fish species from the genera Incertae Sedis in the Characidae family. Maturing ovaries of Astyanax fasciatus presented grayish color whilst the other species’ were yellowish in color. The cortical alveoli of the analyzed species are small (5.20 ± 1.44–8.28 ± 2.02 μm) and are formed by various discontinuous layers of vesicles except in the Bryconops affinis, where they are large (12.87 ± 2.41 μm) and form various continuous layers of vesicles. The pellucid zone is thin and varies from 2.44 ± 0.90 to 5.79 ± 1.90 μm and is thickest in B. affinis (10.57 ± 2.08 μm). Folds in the pellucid zone were observed in Astyanax bimaculatus, A. fasciatus, Hemigrammus marginatus, and Moenkhausia costae. The follicular cells of B. affinis are columnar (28.19 ± 13.71 μm), while they are squamous in other species (1.52 ± 0.64–2.12 ± 0.78 μm). This study showed that the morphologic and biometric characteristics of the B. affinis vitellogenic oocytes are statistically different than the analyzed other ones Incertae Sedis of the Characidae family. In advanced maturing testis of A. bimaculatus and A. fasciatus, the spermatozoa were embedded in acidophilic secretion which was gradually reabsorbed in the seminiferous tubules when males were partially spent.     

Fit-climbing test: a field test for indoor rock climbing

The aim of this study was to develop an indoor rock-climbing test on an artificial wall (Fit-climbing test). Thirteen climbers (elite group [EG] = 6; recreational group [RG] = 7) performed the following tests: (a) familiarization in the Fit-climbing test, (b) the Fit-climbing test, and (c) a retest to evaluate the Fit-climbing test's reliability. Gas exchange, blood lactate concentration, handgrip strength, and heart rate were measured during the test. Oxygen uptake during the Fit-climbing test was not different between groups (EG = 8.4 ± 1.1 L; RG = 7.9 ± 1.5 L, p > 0.05). The EG performance (120 ± 7 movements) was statistically higher than the RG climbers' performance (78 ± 13 movements) during the Fit-climbing test. Consequently, the oxygen cost per movement during the Fit-climbing test of the EG was significantly lower than that of the RG (p < 0.05). Handgrip strength was higher in the EG when compared with that in the RG in both pre-Fit- and post-Fit-climbing test (p < 0.05). There were no significant differences in any other variables analyzed during the Fit-climbing test (p > 0.05). Furthermore, the performance in the Fit-climbing test presented high reliability (intraclass correlation coefficient = 0.97). Therefore, the performance during the Fit-climbing test may be an alternative to evaluate rock climbers because of its specificity and relation to oxygen cost per movement during climbing.