Interleukin-1ß and TNF-α systems in ovarian follicles and their roles during follicular development, oocyte maturation and ovulation.

Tumour necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) are cytokines that are involved in the development, proliferation and apoptosis of ovarian follicular cells in domestic mammals. The expression of these cytokines in various follicular compartments, depending on the stage of follicle development, demonstrates their involvement in the control of primordial follicle growth up to the preovulatory stage. The mechanism of action of these factors depends on the presence of their receptors that transduce their biological actions. This review shows the expression sites of TNF-α, IL-1ß and their receptors in ovarian follicles, and discusses the mechanism of action of these cytokines during follicle development, oocyte maturation and ovulation in domestic animals.

Cryopreservation of collared peccary (Pecari tajacu L., 1758) epididymal sperm using extenders based on Tris and powdered coconut water (ACP®-116c).

SummaryThe aim of this study was to establish a functional freezing-thawing protocol for epididymal sperm of collared peccaries (Pecari tajacu L., 1758) by comparing different extenders. The epididymal sperm from 12 sexually mature males was recovered by retrograde flushing using Tris-based or coconut water-based (ACP®-116c) extenders. After initial evaluation, samples were diluted and frozen with the same extenders to which 20% egg yolk and 6% glycerol were added. After 2 weeks, thawing was performed at 37°C/60 s and sperm motility, vigour, morphology, functional membrane integrity, sperm viability, sperm plasma membrane integrity, and a computer-assisted semen analysis (CASA) were assessed. In addition, to evaluate the survival of frozen-thawed sperm, a thermal resistance test (TRT) was executed. Samples preserved using Tris were in better condition compared with those preserved using ACP®, showing higher values for most assessments performed, including CASA and the TRT (P<0.05). After determining Tris to be the better of the two extenders, additional samples were thawed using different thawing rates (37°C/60 s, 55°C/7 s, 70°C/8 s). Sperm thawed at 37°C/60 s had the greatest preservation (P<0.05) of viability (54.1 ± 5.9%) and functional membrane integrity (43.2 ± 5.4%), and had higher values for various CASA parameters. In conclusion, we suggest the use of a Tris-based extender added to egg yolk and glycerol for the cryopreservation of epididymal sperm obtained from collared peccaries. In order to achieve better post-thawing sperm quality, we suggest that samples should be thawed at 37°C/60 s.

Characterisation and cryopreservation of the ovarian preantral follicle population from Spix's yellow-toothed cavies (Galea spixii Wagler, 1831).

The aim of the present study was to characterise the ovarian preantral follicle (PF) population and to establish a solid surface vitrification (SSV) process using dimethyl sulfoxide (DMSO) as a cryoprotectant for preservation of ovarian tissue from yellow-toothed cavies (Galea spixii). Ovaries were fixed for PF population analysis or were subjected to the SSV process. The mean (± s.e.m.) PF population per ovarian pair was estimated to be 416.0±342.8. There were 140.0±56.0 (63.4%) and 125.0±58.0 (64.0%) primary follicles on the right and left ovaries, respectively. The proportion of this follicle category was significantly greater than that of other follicle categories (P<0.05). The diameter of follicles (123.7±18.3µm), oocytes (50.1±5.0µm) and nuclei (14.27±2.01µm) was larger for secondary ones when compared with other PFs categories. Most PFs were morphologically normal (94.6%), with light microscopy identifying only a few atretic follicles (5.4%). After SSV, there was a reduction in the proportion of morphologically normal PFs compared with the non-vitrified group (69.5% vs 91.2%, respectively). Transmission electron microscopy revealed preservation of oocytes and granulosa cell membranes and the morphological aspect of follicles; the primary change observed in some vitrified PFs was the presence of vacuoles in the oocytes and granulosa cells cytoplasm and turgid mitochondria. In conclusion, the present study provides an estimative and characterization for the PF population in ovaries of G. spixii. Moreover, we report its PFs cryopreservation using an SSV process.

Nanostructured Lipid Carriers as a Strategy to Improve the In Vitro Schistosomiasis Activity of Praziquantel.

Praziquantel (PZQ) is a pyrazinoisoquinoline anthelmintic that was discovered in 1972 by Bayer Germany. Currently, due to its efficacy, PZQ is the drug of choice against all species of Schistosoma. Although widely used, PZQ exhibits low and erratic bioavailability because of its poor water solubility. Nanostructured lipid carriers (NLC), second-generation solid lipid nanoparticles, were developed in the 1990s to improve the bioavailability of poorly water soluble drugs. The aim of this study was to investigate nanostructured lipid carriers as a strategy to improve the efficacy of PZQ in S. mansoni treatment. We prepared NLC2 and NLC4 by adding seventy percent glycerol monostearate (GMS) as the solid lipid, 30% oleic acid (OA) as the liquid lipid and two surfactant systems containing either soybean phosphatidylcholine/poloxamer (PC/P-407) or phosphatidylcholine/Tween 60 (PC/T60), respectively. The carriers were characterized by nuclear magnetic resonance, differential scanning calorimetry, thermogravimetric analysis and Fourier transform-infrared spectroscopy. The safety profile was evaluated using red cell hemolysis and in vitro cytotoxicity assays. The results showed that the encapsulation of PZQ in NLC2 or NLC4 improved the safety profile of the drug. Treatment efficacy was evaluated on the S. mansoni BH strain. PZQ-NLC2 and PZQ-NLC4 demonstrated an improved efficacy in comparison with free PZQ. The results showed that the intestinal transport of free PZQ and PZQ-NLC2 was similar. However, we observed that the concentration of PZQ absorbed was smaller when PZQ was loaded in NLC4. The difference between the amounts of absorbed PZQ could indicate that the presence of T60 in the nanoparticles (NLC4) increased the rigid lipid matrix, prolonging release of the drug. Both systems showed considerable in vitro activity against S. mansoni, suggesting that these systems may be a promising platform for the administration of PZQ for treating schistosomiasis.

The Caries Assessment Spectrum and Treatment (CAST) instrument: construct validation.

The Caries Assessment Spectrum and Treatment (CAST) is a newly developed epidemiological instrument. The aim of this study was to investigate its construct validity. Four calibrated examiners, using CAST codes 0-6, visually examined 109 surfaces of extracted and exfoliated teeth. These teeth were then hemisectioned, photographed, and assessed histologically by two calibrated examiners using the Downer criteria. Twenty-eight of the 109 teeth were scanned using micro-computed tomography (micro-CT) and assessed by the same examiners using the same criteria. Validation was determined through calculation of the sensitivity, specificity, and Youden index for two categories of carious lesions examined visually, with histology and micro-CT as gold standards. Interexaminer consistency was κ = 0.76: SE ± 0.05 between visual and histological assessments of teeth and was κ = 0.89: SE ± 0.08 between visual and micro-CT assessments. For the category 'healthy' vs. 'diseased' (CAST codes 0-2 vs. CAST codes 3-6), sensitivity, specificity, and Youden index values of 100%, 92.9%, and 93%, respectively, for micro-CT, and 96.6%, 86%, and 83%, respectively, for histology, were obtained. For the category 'dentine' vs. 'non-dentine lesions' (CAST codes 0-3 vs. CAST codes 4-6) sensitivity, specificity, and Youden index values of 90%, 100%, and 90%, respectively, for micro-CT, and 81.4%, 100%, and 81%, respectively, for histology, were obtained. Construct validity of the CAST instrument was obtained.

Testicular and color variation in the kissing bug, Rhodnius brethesi, in Amazonas, Brazil.

Because of the morphological and morphometric variation of testicular follicles in different genera of the subfamily Triatominae, it was of interest to associate those parameters with the different medial pronotal band patterns (wide and narrow) found in Rhodnius brethesi (Matta) (Hemiptera: Reduviidae). This is a wild species often associated with Leopoldina piassaba Wallace (Arecales: Arecaceae) palm, with a geographical distribution restricted to the Amazon region. The specimens used were from the state of Amazonas, and were kept under conditions of 29 ± 1 °C, 80 ± 5% RH, 12:12 L:D photoperiod, and were fed weekly on blood from Swiss mice. Three-day-old fasting males were separated in accordance with the patterns of the medial pronotal band, dissected, and the testicles removed. After removal of the testicular membrane, the follicles were spread, drawn by camera lucida, and measured. The results showed that the testis of R. brethesi consists of seven follicles, divided into two groups by length; two long and five short. In specimens with a wide medial pronotal band, the long follicles were 5.4 mm in length, but in specimens with a narrow medial band, the long follicles were 5.64 mm in length. The difference was significant. The short follicles were not different in length, suggesting the presence of a possible complex "brethesi" in the Amazon region.

The Caries Assessment Spectrum and Treatment (CAST) index: face and content validation.

OBJECTIVES: The aim of this study was to establish the face and content validity of the Caries Assessment Spectrum and Treatment (CAST) index. This hierarchical epidemiological index consists of 10 codes covering the spectrum of carious lesion progression and describing conditions ranging from the absence of carious lesions to the presence of sealants and restorations, the presence of lesions in enamel and dentine, and the presence of advanced stages of carious lesion in pulpal tissue and tissue surrounding the tooth. METHODS: Using the RAND modified e-Delphi consensus method, a set of 17 statements related to the content, including the codes and descriptions used, and suitability of the CAST index were scored on a scale of 1-9 by 15 senior epidemiologists from 15 countries over three rounds of assessment. Agreement of ≥ 75% was required to indicate consensus on a statement. RESULTS: Consensus was reached on 14 statements in the first round, none in the second and on the remaining three statements in the third round. To obtain feedback on the initial validation of the index and to test its external validity, 41 epidemiologists from 24 countries were requested to assess the validated statements. Minor changes to 10 of the 17 statements' content and descriptions were suggested; this necessitated the resubmission of the modified CAST index to the original 15 epidemiologists. Consensus of ≥ 80% was reached on all 10 statements regarding codes and their descriptions. CONCLUSIONS: After a total of four rounds with the panel members and one round with the feedback group, the CAST index was approved for face and content validity. External validity was obtained. The participating epidemiologists found the RAND modified e-Delphi consensus method to be a suitable instrument for reaching consensus.

Effects of Aloe vera extract on growth, viability, ultrastructure and expression of mRNA for antioxidant enzymes in bovine secondary follicles cultured in vitro.

This study aimed to investigate the effects of Aloe vera extract on follicular growth, viability, ultrastructure, and mRNA levels for superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase 1 (GPX1) and peroxiredoxin 6 (PRDX6) in bovine secondary follicles cultured in vitro. To this end, secondary follicles were mechanically isolated from the ovarian cortex and cultured at 38.5 °C, with 5% CO2 in air, for 18 days in TCM-199+ alone or supplemented with 2.5%, 5.0%, 10.0% and 20.0% Aloe vera extract. Follicular growth, morphology and antrum formation were evaluated every 6 days, while ultrastructure was evaluated at the end of culture. Analysis of viability was performed by calcein-AM and ethidium homodimer-1, while mRNA levels for SOD, CAT, GPX1 and PRDX6 were evaluated by real-time PCR at the end of culture. The results show that follicles cultured with 2.5% Aloe vera had increased the rate of antrum formation, while 2.5% and 5.0% Aloe vera improved follicular viability rate. Follicles cultured with 2.5% and 10.0% Aloe vera increased the levels of mRNA for SOD and GPX1 respectively, but the levels of CAT were reduced in follicles cultured with 2.5%, 5.0%, 10.0% and 20.0%. Additionally, follicles cultured with 2.5% of Aloe vera had their ultrastructure well preserved, while those cultured with 5.0%, 10.0% and 20.0% exhibited increased oocyte vacuolization and damaged organelles. In conclusion, 2.5% Aloe vera increases antrum formation, viability and expression of mRNA for SOD in cultured secondary follicles, but higher concentrations of Aloe vera have negative effects on follicular ultrastructure.

Lipid Nanocarriers for Hyperproliferative Skin Diseases.

Hyperproliferative skin diseases (HSD) are a group of diseases that include cancers, pre-cancerous lesions and diseases of unknown etiology that present different skin manifestations in terms of the degree and distribution of the injuries. Anti-proliferative agents used to treat these diseases are so diverse, including 5-aminolevulinic acid, 5-fluorouracil, imiquimod, methotrexate, paclitaxel, podophyllotoxin, realgar, and corticosteroids in general. These drugs usually have low aqueous solubility, which consequently decreases skin permeation. Thus, their incorporation in lipid nanocarriers has been proposed with the main objective to increase the effectiveness of topical treatment and reduce side effects. This manuscript aims to describe the advantages of using lipid nanoparticles and liposomes that can be used to load diversity of chemically different drugs for the treatment of HSD.

Influence of chromium propionate dose and feeding regimen on growth performance and carcass composition of pigs housed in a commercial environment.

Although chromium (Cr) feeding study results have been variable, our hypothesis was feeding a regimen that changed dosage over time would result in a larger positive response in growth performance and carcass characteristics. In Exp. 1, a total of 1,206 pigs (PIC 337 × 1050, initial BW 28.7 kg) were used with 27 pigs per pen and 9 pens per treatment. Diets were corn-soybean meal-dried distillers grains with solubles based and were fed in a five-phase feeding program. Treatments were arranged as a 2 × 2 + 1 factorial with a control diet containing no added Cr propionate (Kemin Industries Inc., Des Moines, IA), or diets with either 100 or 200 µg/kg added Cr during the grower (dietary phases 1 and 2) and/or finisher (dietary phases 3, 4, and 5) periods. During the grower period, ADG and G:F were similar among pigs fed the control or 100 µg/kg added Cr diets, but decreased in pigs fed 200 µg/kg Cr (quadratic, P ≤ 0.001). During the finisher period, pigs supplemented with 200 µg/kg added Cr had the greatest ADG and G:F (quadratic, P ≤ 0.019). Overall, increasing Cr had no effect on ADG or ADFI; but G:F was greatest (quadratic, P = 0.020) when pigs were fed 100 µg/kg of added Cr throughout. Carcass characteristics were not influenced by Cr dosage or feeding regimen. In Exp. 2, a total of 1,206 pigs (PIC 359 × 1050, initial BW 48.9 kg) were used with 27 pigs per pen and 15 pens per treatment. Diets were corn-soybean meal, dried distillers grains with solubles based and were fed in four phases. There were three dietary treatments: a diet with no added Cr for both grower (dietary phase 1 and 2) and finisher (dietary phase 3 and 4) periods, a diet with 200 µg/kg added Cr during the grower and 100 µg/kg added Cr during the finisher periods, or a diet with 200 µg/kg added Cr for both periods. Addition of 200 µg/kg Cr in both periods marginally increased (P < 0.10) ADG compared with pigs fed no added Cr. There was no evidence (P ≥ 0.523) of added Cr influencing overall ADFI and G:F. Percentage carcass yield was reduced (P = 0.018) when Cr was added at 200 µg/kg for both periods, with no evidence of differences (P ≥ 0.206) in other carcass characteristics. In summary, overall G:F was improved in Exp. 1, and ADG in Exp. 2, by added Cr, but there was no evidence that different feeding regimens will consistently result in improved performance. However, these data are consistent with the literature in that added Cr in growing-finishing pigs diets improves, albeit small, ADG or G:F.

Determining the influence of chromium propionate and Yucca schidigera on growth performance and carcass composition of pigs housed in a commercial environment.

Two experiments were conducted to determine the effects of feeding chromium propionate (Cr; Kemin Industries, Inc., Des Moines, IA) and a Yucca schidigera-based extract (YS; Distributors Processing, Inc., Porterville, CA) on growth performance of finishing pigs housed in commercial conditions. In experiment 1, a total of 1,188 pigs (PIC 337 × 1050; initially 27.3 ± 0.48 kg body weight [BW]) with 27 pigs per pen and 11 pens per treatment were split by sex upon arrival at the facility and were randomly allotted to groups of four pens blocked by BW. Diets were corn-soybean meal-dried distillers grains with solubles-based and were fed in five phases. Treatments were arranged as a 2 × 2 factorial with main effects of Cr (0 vs. 200 µg/kg) or YS (0 vs. 62.5 mg/kg YS-based feed grade concentrate). Overall, adding Cr alone increased (P = 0.049) average daily feed intake (ADFI), and inclusion of YS resulted in a marginally significant increase (P = 0.077) in ADFI. Backfat depth was increased (P = 0.043) and lean percentage was decreased (P = 0.011) with added Cr. In experiment 2, a total of 2,430 pigs (PIC 359 × 1050; initially 29.3 ± 0.43 kg BW) were placed in balanced mixed-sex pens with 27 pigs per pen, blocked by average pen BW, and randomly assigned to one of six dietary treatments with 14 pens per treatment. Diets were corn-soybean meal-based and were formulated in five dietary phases. Treatments were arranged in a 2 × 3 factorial with main effects of Cr (0 vs. 200 µg/kg added Cr), and YS extract (0, 62.5, or 125 mg/kg YS-based feed grade concentrate). Overall, a marginally significant (linear, P = 0.072) Cr × YS interaction was observed for average daily gain (ADG) where there was insufficient evidence of a difference with increasing YS in diets not including added Cr (P ≥ 0.109); however, ADG increased (quadratic, P = 0.026) with YS addition in treatments fed 200 µg/kg added Cr. For overall ADFI, a marginally significant (linear, P = 0.071) Cr × YS interaction was observed where YS increased ADFI with 200 µg/kg added Cr (linear, P = 0.031), however, did not when diets contained no added Cr (P = 0.700). A marginally significant reduction in gain:feed ratio was observed when 62.5 mg/kg YS was included (quadratic, P = 0.053), and final BW and hot carcass weight were lowest with 62.5 mg/kg YS (quadratic, P = 0.012). In summary, adding Cr propionate along with YS led to modest changes in performance with the greatest benefit observed with 200 µg/kg Cr and 125 mg/kg YS-based feed grade concentrate.

Effect of cryoprotectant type and concentration on the vitrification of collared peccary (Pecari tajacu) ovarian tissue.

The aim of the present study was to establish a protocol for solid surface vitrification of peccary ovarian tissue by using different cryoprotectants. Ovarian pairs from five adult females were fragmented and two fragments (fresh control group) were immediately subjected to morphological evaluation using classical histology, transmission electron microscopy, and viability analysis using fluorescent probes. The remaining fragments (n = 18) were vitrified using a solid surface method with different concentrations (3 or 6 M) of ethylene glycol (EG), dimethyl sulfoxide (DMSO) or dimethyl formamide (DMF). After 2 weeks, samples were re-warmed and evaluated. A decrease in the percentage of morphologically normal preantral follicles (PFs) was verified for all the groups in comparison to the fresh control (92.0 ± 2.8%); however, if only the primordial follicles are considered, the most effective preservation (P < 0.05) was achieved with the use of EG at 3 M (74.2±7.3%) or DMSO at 6 M (75.0 ± 4.2%). Ultrastructural analysis indicated there were well-preserved PFs in all the groups evaluated, having well-defined membranes, a few vacuoles, and organelles that were uniformly distributed throughout the cytoplasm, mainly round and elongated mitochondria in close association with lipid droplets. Viability was preserved (P < 0.05) with the use of EG at 3 (97%) or 6 (97%) M, DMSO at 3 (100%), and DMF at 6 (97%) M. Solid surface vitrification, therefore, is an effective method for conservation of peccary female germplasm, especially with the use of EG at 3 M, which was highly effective for preservation of both the morphology and viability of PFs.

Addition of Equex STM to Extender Improves Post-Thawing Longevity of Collared Peccaries' Sperm.

The effect of Equex STM® paste supplementation on the Tris-extender for collared peccaries' semen cryopreservation was assessed. Semen from 12 mature individuals was obtained by electroejaculation and evaluated for morphology, membrane integrity, osmotic response, and sperm kinetic metrics. Samples were diluted in Tris plus 20% egg yolk and divided into three aliquots. The first aliquot was without any supplementation, the second and third contained 0.5 and 1.0% Equex STM, respectively. The samples were added with 3% glycerol, frozen in liquid nitrogen, thawed, and assessed for the same parameters after a thermal resistance test (TRT) for 120 minutes. Similar values were detected for the different treatments immediately after thawing, except for the amplitude lateral head that was reduced in samples containing Equex (p < 0.05). During TRT, samples containing Equex were more efficient in preserving the sperm motility (at 0.5%: 25.5% ± 4.4%; at 1%: 33.3% ± 6.3%) at 30 minutes, in comparison with the control group (16.6% ± 6.0%), in which sperm motility decreased at 15 minutes (p < 0.05). Moreover, Equex, especially at 0.5% concentration, was able to maintain plasma membrane integrity and sperm motility in all the samples after incubation for 60 minutes. In conclusion, we recommend the addition of Equex STM at 0.5% to the Tris-extender to improve post-thawing sperm longevity in collared peccaries.

Composition of collared peccary seminal plasma and sperm motility kinetics in semen obtained during dry and rainy periods in a semiarid biome.

The aim of this study was to evaluate environmental effects in a semiarid region on collared peccary seminal plasma content and sperm motility. Ejaculates from 12 mature males were obtained during the peak of rainy and dry periods of the Caatinga biome. Samples were evaluated for semen volume, pH, as well as sperm concentration, morphology, osmotic response, membrane integrity, chromatin condensation, and kinetic motility. Seminal plasma was evaluated for ions and organic compounds. The values for chloride, iron, magnesium, phosphorus, citric acid, cholesterol, triglycerides, total proteins, albumin, and fructosamine were similar during the dry and rainy periods; however, concentrations of fructose (849.2 mg/dL compared with 119.4 mg/dL) and calcium (32.3 mg/dL compared with 15.6 mg/dL) were greater during the rainy compared with dry period (P < 0.05). There were correlations (P < 0.05) among values for semen variables and biochemical contents, particularly between fructose and sperm velocity average pathway (r = 0.65), velocity straight line (r = 0.78), velocity curvilinear (r = 0.57), amplitude lateral head (r = 0.62), linearity (r = 0.41), and subpopulation with a medium velocity (r = -0.75). Furthermore, values for relative humidity were positively correlated with concentrations of fructose (r = 0.49), while air temperature (r = -0.43) and wind velocity values (r = 0.66) were negatively affected by concentration of fructose (P < 0.05). There were novel results regarding collared peccary seminal plasma biochemistry indicating there are important correlations with values for semen variables that are affected by the environment in a semiarid climate.

Epididymal sperm from Spix's yellow-toothed cavies sperm successfully cryopreserved in Tris extender with 6% glycerol and 20% egg yolk.

As a non-threatened hystricognath rodent species, Spix's yellow-toothed cavies can be used as a model for the development of assisted reproductive techniques for the conservation of closely related species. The objective was to establish a functional protocol for cryopreservation of epididymal sperm from these cavies. Twelve sexually mature males, ∼2 y old and weighing ∼300 g, were euthanized. Sperm were recovered by retrograde flushing of the vas deferens and cauda epididymis with Tris extender. Thereafter, sperm were extended in Tris plus 20% egg yolk, with 3%, 6% or 9% glycerol or dimethyl sulfoxide (DMSO), placed in 0.25 mL straws and cryopreserved in liquid nitrogen. Sperm concentration, motility (using computer-assisted sperm analysis; CASA), plasma membrane integrity, osmotic response, morphology and sperm binding-ability were determined in fresh and frozen-thawed sperm. For most sperm endpoints, glycerol was a more desirable cryoprotectant than DMSO. Data (mean ±â€¯SEM) were similar with use of 3%, 6%, and 9% glycerol (P > 0.05) in osmotic response (40.66 ±â€¯6.3%, 42.5 ±â€¯7.1%, and 39.5 ±â€¯5.0% respectably), and membrane integrity (55.17 ±â€¯5.5%, 68.4 ±â€¯4.1%, and 59.1 ±â€¯4.9% respectably). Among concentrations assessed, the use of 6% glycerol resulted in the greatest (P < 0.05) post-thaw values for total motility (60.9 ±â€¯4.4%), rapid subpopulation motility (27.7 ±â€¯3.1%) and sperm-binding capability (227.0 ±â€¯20.2). In conclusion, epididymal sperm from the Spix's yellow-toothed cavies (G. spixii) are optimally cryopreserved in Tris extender with 6% glycerol and 20% egg yolk.

Day/night melatonin content in cerebral palsy.

Changes in the sleep-wake cycle are frequent and may impair quality of life in individuals with cerebral palsy (CP). To investigate if a lack of a day/night variation of melatonin content could be related with sleep disorders (SD), the SD were evaluated with a Sleep Questionnaire and the melatonin content using ELISA in 33 individuals with CP and 24 controls. The indicative of SD were present in 47% of CP group, and the most frequent was the indicative of sleep breathing disorder. The CP group showed higher diurnal and lower nocturnal melatonin content than controls. Individuals with CP that had indicative of SD showed lower nocturnal content of melatonin than those without SD. These results showed that the lack of the day/night variation of melatonin was related to SD in individuals with CP.

Environmental Factors Related to a Semiarid Climate Influence the Freezability of Sperm from Collared Peccaries (Pecari tajacu Linnaeus, 1758).

The influence of environmental factors in a semiarid climate on characteristics of fresh and frozen/thawed sperm collected from collared peccaries (Pecari tajacu) was assessed. Semen from 11 male collared peccaries was collected by electroejaculation during the peaks of the dry and rainy periods while rainfall indices, air temperatures, relative humidity levels, and wind speeds were measured. The number, motility, morphology, osmotic response, and membrane integrity of sperm in the collected ejaculates were assessed. Samples were then frozen in liquid nitrogen, thawed, and reassessed. The rainfall index of the rainy period (73.2 mm) was significantly higher than that of the dry period (13.6 mm) and the relative humidity was significantly higher during the rainy period (74.6%) than it was during the dry period (66.8%). Air temperature and wind speed did not differ between the two periods. Characteristics of sperm in the fresh samples were not affected by environmental parameters. In contrast, computerized analysis revealed that sperm in samples frozen during the rainy period exhibited better post-thaw membrane integrity (28.6 ± 6%), motility (29.5 ± 7.7%), and rapid sperm population (13.7 ± 6.2%) than did sperm in samples frozen during the dry period (23.4 ± 3% membrane integrity, 14.6 ± 4.1% motility, and 4.1 ± 1.2% rapid sperm; p < 0.05). Other characteristics of the frozen/thawed sperm did not differ depending on the period in which they were collected. We demonstrated that environmental parameters did not affect the quality of fresh sperm, but could influence the freezability of sperm collected from collared peccaries raised under a semiarid climate.

Sleep findings in Brazilian children with congenital Zika syndrome.

Study Objectives: Zika virus infection during pregnancy may result in congenital Zika syndrome (CZS), whose characteristics are being described. Methods: The present study aimed to investigate the sleep characteristics of 136 infants/toddlers (88 with CZS and 48 with typical development (TD), age and gender matched, 60% girls and 40% boys in both groups) using the Brief Infant Sleep Questionnaire. The ages of children in both groups ranged from 5 to 24 months (CZS 15.9 ± 0.4 vs. TD 15.8 ± 1.0 months, P= 0.90). Results: The results show that 34.1% of CZS and 2% of TD children were defined as poor sleepers, 15% of CZS and 2% of TD children remained awake at night for a period longer than 1 hour, and 24% of CZS and 2% of TD children slept less than 9 hours. The CZS group showed shorter total sleep time (CZS 11.24 ± 2.6 vs. TD 12.02 ± 1.9 hours, P= 0.03) and shorter nocturnal sleep duration than the TD group (CZS 8.2 ± 0.2 vs. TD 9.4 ± 0.2 hours, P= 0.0002). In contrast to the control group (P= 0.02, r= -0.34), in the CZS group, no correlation was found between age and nocturnal wakefulness. Future studies should explore these data in relation to the development and maturation of the central nervous system of these children. Conclusions: Considering the well-known consequences of poor sleep quality on health in several populations, the presence of sleep disorders should be considered in CZS using multidisciplinary treatments.

Neisseria meningitidis disease-associated clones in Amazonas State, Brazil.

BACKGROUND: The aim of this study is to describe the molecular epidemiology of Neisseria meningitidis invasive disease before the introduction of serogroup C conjugate vaccine in Amazonas State in 2010. METHODS: Meningococcal disease reported cases were investigated in Amazonas State during the period 2000-2010. N. meningitidis isolates (n = 196) recovered from patients were genotyped by multilocus sequence typing (MLST) and sequencing of porB, porA, fetA, fHbp and penA. Antimicrobial susceptibility was determined using E-test. RESULTS: In the study period, 948 cases were reported; the incidence was 2.8 for the entire state and 4.8 per 100,000 in the capital of Manaus. Most meningococcal disease was caused by N. meningitidis belonging to ST-32 (72%; 141/196) or ST-103 (21%; 41/196) clonal complexes. Capsular switching (B→C) was suggested within clonal complex (cc) 32. There were 6 (3%; 6/196) strains with intermediate susceptibility to penicillin and a single strain was resistant to rifampicin. Since 2007, serogroup C strains belonging to the cc103 have predominated and case-fatality has increased. CONCLUSION: We demonstrate a high rate of meningococcal disease in Amazonas State, where, like other parts of Brazil, serogroup C replaced serogroup B during 2000s. These data serve as a baseline to measure impact of serogroup C conjugate vaccine introduction in 2010. This study emphasizes the need for enhanced surveillance to monitor changes in meningococcal disease trends following the introduction of meningococcal vaccines.

Ophthalmic patterns of captive brown brocket deer (Mazama gouazoubira).

Captive brown brocket deer (Mazama gouazoubira) were manually restrained to assess tear production by the Schirmer tear test I to measure intraocular pressure by applanation tonometry, to examine ocular conjunctival epithelial cells via cytologic and histologic samples, and to survey ocular conjunctival microflora by microbiologic culture. The mean value for the Schirmer tear test I was 8.9 +/- 1.8 mm/min, and the mean intraocular pressure was 15.3 +/- 3.1 mm Hg. Conjunctival epithelium contained stratified pavimentous layers of cells, and the microflora consisted of predominantly gram-positive bacteria.