Proteomic profile of vitrified in vitro-produced bovine embryos (Bos taurus indicus).

BACKGROUND: The proteomic profile of cryopreserved in vitro produced bovine embryos is little known but can provide insights on the successful application of cryo procedures in support of animal breeding. OBJECTIVE: To identify embryonic proteins and biomarkers related to improved cryotolerance of vitrified in vitro produced bovine embryos. MATERIALS AND METHODS: Proteins were isolated from embryo pools (n = 25 embryos per replicate) and analyzed using the nanoLC - MS/MS system. Further, the UniProtKB database (Uniprot -http://www.uniprot.org/) was used for protein identification. Proteins were classified based on their molecular mass, isoelectric point, and enzymatic activity. Post-translational modification predictions and functional gene ontology analysis were performed as well. Finally, a protein-protein interaction network was created to shed light on the embryo interactome. RESULTS: Based on the MS/MS approach, 66 proteins were identified from vitrified Bos taurus embryos. The retrieved proteins were presumably annotated, which allowed a description of the qualitative and functional aspects of the embryo proteome after the vitrification process. CONCLUSION: These findings allowed us to conclude that in vitro-produced vitrified embryos expressed proteins that underlie biological processes related to reproduction, stress and lipid metabolic process, which are essential to maintain embryo viability. doi.org/10.54680/fr22410110512.

Cryopreservation of Bovine Semen Using Extract of Bertholletia excelsa (Brazil Nuts).

BACKGROUND: Semen cryopreservation is essential in animal breeding programs for improving the availability of genetic resources from animals with high breeding value. OBJECTIVE: To evaluate the addition of Brazil nut extract as a replacement for egg yolk in bovine semen cryopreservation. MATERIALS AND METHODS: Semen was collected from five Nelore bulls and cryopreserved with the addition (treatments) of 0, 25, 50, 75, or 100% Brazil nut extract in the cryoprotectant medium. After thawing, spermatic cells were evaluated for morphology, plasma membrane integrity, spermatic kinetics, and in vitro fertilization. The experimental design was in randomized blocks, and the data were submitted to regression analysis. RESULTS: The minor-type and total defects, and plasma membrane integrity were affected (P < 0.05) as a function of egg yolk substitution with Brazil nut extract. There was a significant effect (P < 0.05) of Brazil nut extract addition on the spermatic kinetics and cleavage rate. CONCLUSION: The addition of Brazil nut extract in the cryoprotective medium as a substitute of egg yolk for freezing bovine semen negatively affects sperm quality and fertility.

The Addition of AçaÍ Palm Fruit Extract in the Cryopreservation Diluent of Bull's Semen with Low Performance on Freezing.

BACKGROUND: Semen freezing is of great importance for animal production because it allows the use and the rapid diffusion of the genetic material from economically important animals. OBJECTIVE: To evaluate the effect of açai (Euterpe oleracea; Arecaceae family) extract addition to the semen cryopreservation diluent on the morphology, sperm motility parameters, and plasma membrane integrity of spermatozoa. MATERIALS AND METHODS: The ejaculates, obtained from five bulls with low performance on semen freezing, were fractionated and distributed according to the experimental group. The control samples did not have açaí extract added, whereas to the treated groups were added 5, 10, 15 or 20 mg ml-1 of açaí extract into the semen diluent. The sperm morphology was evaluated with a formalin-saline-buffered solution. The plasma membrane integrity was evaluated by the epifluorescent test, while the cellular kinetics was assessed by an automated analysis of the spermatic movement. RESULTS: The sperm defects showed a linearly decreasing effect (P < 0.05) with the addition of different concentrations of açaí extract. The plasma membrane integrity was higher (P < 0.05) after the açaí addition to the cryopreservation diluent. There was no significant effect (P > 0.05) of the açaí extract on the kinetics of spermatozoa. CONCLUSION: The addition of açaí extract to the cryopreservation diluent provided better preservation of the structural integrity of the sperm plasma membrane in the bull's semen with low tolerance to the cryopreservation process.

Development and chemical characterization of biodegradable polymeric implants containing sirolimus for the treatment of malignant solid tumors.

The use of sirolimus and its analogs has been evaluated in studies aimed at combating several types of cancer; however, because of the limited bioavailability of the drug, the search for new forms of administration is required. Biodegradable polymeric implants containing sirolimus were developed and assessed as an alternative method of drug administration. Implants containing 25 % (w/w) sirolimus were prepared employing the polymer matrices chitosan, polycaprolactone and poly(lactic-co-glycolic acid) (PLGA) in two proportions: PLGA 50:50 and PLGA 75:25. Thermal analysis techniques such as thermogravimetry and differential scanning calorimetry, combined with x-ray diffraction were used to characterize and evaluate the compatibility of the constituents of the formulation. No incompatibilities were found between the components, but drug amorphization was observed in all samples. Implants made from the polymers chitosan and PCL may accelerate the degradation of SRL when these polymers are dissolved in methanol at 50 °C. HPLC analysis showed that the implant prepared with PLGA 75:25 did not present degradation products and maintained its appropriate drug content, even when dissolved in methanol and heated to 50 °C. Therefore, it represents the most suitable biodegradable polymer for use in implants developed for the treatment of malignant solid tumors. However, it is still necessary to further study the drug effects after amorphization of the crystal and also to perform stability and solubility analysis.

Temporal expression of pluripotency-associated transcription factors in sheep and cattle preimplantation embryos.

SummaryPluripotency-associated transcription factors (PATFs) modulate gene expression during early mammalian embryogenesis. Despite a strong understanding of PATFs during mouse embryogenesis, limited progress has been made in ruminants. This work aimed to describe the temporal expression of eight PATFs during both sheep and cattle preimplantation development. Transcript availability of PATFs was evaluated by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) in eggs, cleavage-stage embryos, morulae, and blastocysts. Transcripts of five genes were detected in all developmental stages of both species (KLF5, OCT4, RONIN, ZFP281, and ZFX). Furthermore, CMYC was detected in all cattle samples but was found from cleavage-stage onwards in sheep. In contrast, NR0B1 was detected in all sheep samples but was not detected in cattle morulae. GLIS1 displayed the most significant variation in temporal expression between species, as this PATF was only detected in cattle eggs and sheep cleavage-stage embryos and blastocysts. In silico analysis suggested that cattle and sheep PATFs share similar size, isometric point and molecular weight. A phenetic analysis showed two patterns of PATF clustering between cattle and sheep, among several mammalian species. In conclusion, the temporal expression of pluripotency-associated transcription factors differs between sheep and cattle, suggesting species-specific regulation during preimplantation development.

Reproductive ecology of phorid parasitoids in relation to the head size of leaf-cutting ants Atta sexdens Forel.

The leaf-cutting ant Atta sexdens Forel (Hymenoptera: Formicidae) is one of the most damaging agricultural pests in the Neotropics. Management strategies predominantly rely on the use of general insecticides. What is needed are more species-specific and environmentally friendly options. Parasitioids such as phorid flies (Diptera: Phoridae) may be one such option, but a greater understanding of the ecology of the flies and their ant hosts is essential to devise biological control strategies. Here we report parasitism rates, ant host size, parasitoid abundance per host and resultant sex ratios of two phorid species Apocephalus attophilus Borgmeier and Eibesfeldtphora tonhascai Brown parasitizing A.sexdens. The two species achieved parasitism rates of 1.48 and 1.46%, respectively and the pupal period was 14.7 ± 1.1 days and 22.1 ± 2.8 days, respectively. There was no significant difference between the head capsule width of ants parasitized by either A. attophilus or E. tonhascai. Likewise, there was no significant effect between the head capsule width of parasitized and unparasitized ants for both species. A significant positive correlation was found between the head capsule width of the parasitized ants and the number of adult parasitoids A. attophilus emerged. Ants parasitized by E. tonhascai survived significantly longer than those parasitized by A. attophilus. There was no significant effect of ant head width on the sex ratio of the offspring of either parasitoid species and no significant difference in the sex ratio (male: female) of their offspring. In summary, these data addressed here are important steps when considering natural enemies for biological control. Studying survival of the parasitized ants, parasitoid offspring sex ratio and host size preference allows for a better understanding of ant natural biological control in the field and can help in rearing of A. attophilus and E. tonhascai in laboratory.

Vulvar and gastric involvement in plasmablastic lymphoma.

Plasmablastic lymphoma is highly malignant and invasive. It is most commonly found in gastrointestinal tract and is strongly asso- ciated with human immunodeficiency virus (HIV). The authors report a case of a 37-year-old HIV-positive African American woman with a vulvar mass diagnosed as plasmablastic lymphoma. Involvement of the upper gastrointestinal tract had been detected by endoscopic biopsy, suggesting that any patient diagnosed with plasmablastic lymphoma in unusual locations should be investigated for primary lesions in the gastrointestinal tract.

Influence of negative dental experiences in childhood on the development of dental fear in adulthood: a case-control study.

The aim of this study was to evaluate the factors associated with high dental fear among Brazilian university students, especially the effect of a negative dental experience in childhood. This paired case-control study was conducted at the Universidade Federal de Minas Gerais in Brazil. Dental, psychology and mathematics students were divided into cases (high fear) and controls (low fear), defined by cluster analysis, according to the items of the Dental Fear Survey (DFS). Cases (n = 65) and controls (n = 260) participants were paired (1:4) by gender, undergraduate course and social vulnerability. The students self-reported the DFS and a questionnaire about oral health. Descriptive analysis, bivariate and multivariate conditional logistic regression were used as statistical tests with a significance level of 5%. The multivariate model showed that students who reported negative dental experiences in childhood (OR = 2·97; 95% CI: 1·44-6·14), toothache in the last 12 months (OR = 11·31; 95% CI: 4·79-26·68), discomfort during dental treatment (OR = 5·36; 95% CI: 2·53-11·36) and poor self-evaluation of oral health (OR = 3·82; 95% CI: 1·61-8·11) were more likely to have high dental fear. Negative dental experiences in childhood influence dental fear in adulthood. Oral health education should be addressed among university students to reduce dental fear.

Predicting Pt-195 NMR chemical shift using new relativistic all-electron basis set.

Predicting NMR properties is a valuable tool to assist the experimentalists in the characterization of molecular structure. For heavy metals, such as Pt-195, only a few computational protocols are available. In the present contribution, all-electron Gaussian basis sets, suitable to calculate the Pt-195 NMR chemical shift, are presented for Pt and all elements commonly found as Pt-ligands. The new basis sets identified as NMR-DKH were partially contracted as a triple-zeta doubly polarized scheme with all coefficients obtained from a Douglas-Kroll-Hess (DKH) second-order scalar relativistic calculation. The Pt-195 chemical shift was predicted through empirical models fitted to reproduce experimental data for a set of 183 Pt(II) complexes which NMR sign ranges from -1000 to -6000 ppm. Furthermore, the models were validated using a new set of 75 Pt(II) complexes, not included in the descriptive set. The models were constructed using non-relativistic Hamiltonian at density functional theory (DFT-PBEPBE) level with NMR-DKH basis set for all atoms. For the best model, the mean absolute deviation (MAD) and the mean relative deviation (MRD) were 150 ppm and 6%, respectively, for the validation set (75 Pt-complexes) and 168 ppm (MAD) and 5% (MRD) for all 258 Pt(II) complexes. These results were comparable with relativistic DFT calculation, 200 ppm (MAD) and 6% (MRD). © 2016 Wiley Periodicals, Inc.

Mixed inheritance in the genetic control of ramulosis (Colletotrichum gossypii var. cephalosporioides) resistance in cotton.

Ramulosis is one of the most aggressive diseases in cotton, and understanding the genetic control of its resistance is imperative for selecting superior cotton genotypes in breeding programs. This study analyzed the inheritance pattern of this resistance using chi-square goodness-of-fit tests to determine the phenotypic proportions of the F2 generation, and a mixed inheritance approach to jointly model major gene and polygenes effects. F1, F2, Rc1, and Rc2 generations were obtained by crossing resistant (BRS Facual, CNPA 2984, or CNPA 2043) and susceptible (Delta Opal, CNPA 999, or CNPA 2161) genotypes, and were assessed under field conditions with artificial inoculation of the pathogen (Colletotrichum gossypii var. cephalosporioides). Genetic control of the trait varied among the crossings. For Delta Opal x BRS Facual and CNPA 2161 x BRS Facual, phenotypic segregations in the F2 generation did not differ from the expected proportions for the hypothesis of duplicate genes (15:1). For Delta Opal x CNPA 2043, the segregation did not differ from the expected proportions for dominant recessive epistasis (13:3). The hypothesis of genetic control by one major gene was supported only for the Delta Opal x CNPA 2043 crossing. Three other crossings showed evidence of polygenes in the inheritance of the trait. In conclusion, major genes and polygenes are likely involved in the genetic control of ramulosis resistance in cotton.

Epicardial fat accumulation, cardiometabolic profile and cardiovascular events in patients with stages 3-5 chronic kidney disease.

BACKGROUND: It has been hypothesized that epicardial adipose tissue (EAT) exerts pathogenic effects on cardiac structures. We analysed the associations between EAT and both cardiovascular (CV) disease risk factors and CV events in patients with chronic kidney disease (CKD). PATIENTS AND METHODS: We included 277 nondialysed patients [median age 61, interquartile range (IQR) 53-68 years; 63% men] with stages 3-5 CKD in this cross-sectional evaluation. EAT and abdominal visceral adipose tissue (VAT) were assessed by computed tomography. Patients were followed for median 32 (IQR 20-39) months, and the composite of fatal and nonfatal CV events was recorded. RESULTS: With increasing EAT quartiles, patients were older, had higher glomerular filtration rate, body mass index, waist, VAT and coronary calcification, higher levels of haemoglobin, triglycerides, albumin, C-reactive protein and leptin and higher prevalence of left ventricular hypertrophy and myocardial ischaemia; total and high-density lipoprotein cholesterol, 25-hydroxy-vitamin D and 1, 25-dihydroxy-vitamin D progressively decreased. Associations between EAT and cardiac alterations were not independent of VAT. During follow-up, 58 CV events occurred. A 1-SD higher EAT volume was associated with an increased risk of CV events in crude [hazard ratio (HR) 1.41, 95% confidence interval (CI) (1.12-1.78) and adjusted (HR 1.55, 95% CI 1.21-1.99) Cox models. However, adding EAT to a standard CV disease risk prediction model did not result in a clinically relevant improvement in prediction. CONCLUSION: Epicardial adipose tissue accumulation in patients with CKD increases the risk of CV events independent of general adiposity. This is consistent with the notion of a local pathogenic effect of EAT on the heart or heart vessels, or both. However, EAT adds negligible explanatory power to standard CV disease risk factors.

Leishmania braziliensis amastigotes stimulate production of IL-1ß, IL-6, IL-10 and TGF-ß by peripheral blood mononuclear cells from nonendemic area healthy residents.

Leishmania (Viannia) braziliensis causes cutaneous and mucosal leishmaniasis in several countries in Latin America. In mammals, the parasites live as amastigotes, interacting with host immune cells and stimulating cytokine production that will drive the type of the specific immune responses. Generation of Th17 lymphocytes is associated with tissue destruction and depends on IL-1ß, IL-6, TGF-ß and IL-23 production, whereas IL-10 and TGF-ß are associated with tissue protection. Here, we evaluate whether amastigotes stimulate peripheral blood mononuclear cells (PBMCs) from healthy donors to produce the major cytokines responsible for the generation of Th17. Seven L. (V.) braziliensis isolates from patients with different clinical forms of leishmaniasis were expanded in interferon-γ knockout mice to obtain amastigotes and in culture to get promastigotes. The parasites were used to stimulate PBMCs from healthy donors, and cytokine production was evaluated by ELISA or qPCR. Amastigotes and promastigotes induced IL-10 production in PBMCs; however, only amastigotes induced IL-1ß, IL-6 and TGF-ß. These data demonstrate for the first time that L. (V.) braziliensis amastigotes directly stimulate production of a unique pattern of cytokines that could contribute to the generation of Th17.

Neutrophils have a protective role during early stages of Leishmania amazonensis infection in BALB/c mice.

Neutrophils are involved in the early stages of immune responses to pathogens. Here, we investigated the role of neutrophils during the establishment of Leishmania amazonensis infection in BALB/c and C57BL/6 mice. First, we showed an accumulation of neutrophils between 6 and 24 h post-infection, followed by a reduction in neutrophil numbers after 72 h. Next, we depleted neutrophils prior to infection using RB6-8C5 or 1A8 mAb. Neutrophil depletion led to faster lesion development, increased parasite numbers and higher arginase activity during the first week of infection in BALB/c mice, but not in C57BL/6 mice. Increased susceptibility was accompanied by augmented levels of anti-L. amazonensis IgG and increased production of IL-10 and IL-17. Because IL-10 is a mediator of susceptibility to Leishmania infection, we blocked IL-10 signalling in neutrophil-depleted mice using anti-IL-10R. Interestingly, inhibition of IL-10 signalling abrogated the increase in parasite loads observed in neutrophil-depleted mice, suggesting that parasite proliferation is at least partially mediated by IL-10. Additionally, we tested the effect of IL-17 in inflammatory macrophages and observed that IL-17 increased arginase activity and favoured parasite growth. Taken together, our data indicate that neutrophils control parasite numbers and limit lesion development during the first week of infection in BALB/c mice.

Systematic screening of plant extracts from the Brazilian Pantanal with antimicrobial activity against bacteria with cariogenic relevance.

This study proposes a bioprospection methodology regarding the antimicrobial potential of plant extracts against bacteria with cariogenic relevance. Sixty extracts were obtained from ten plants--(1) Jatropha weddelliana, (2) Attalea phalerata, (3) Buchenavia tomentosa, (4) Croton doctoris, (5) Mouriri elliptica, (6) Mascagnia benthamiana, (7) Senna aculeata, (8) Unonopsis guatterioides, (9) Allagoptera leucocalyx and (10) Bactris glaucescens--using different extraction methods - (A) 70° ethanol 72 h/25°C, (B) water 5 min/100°C, (C) water 1 h/55°C, (D) water 72 h/25°C, (E) hexane 72 h/25°C and (F) 90° ethanol 72 h/25°C. The plants were screened for antibacterial activity at 50 mg/ml using the agar well diffusion test against Actinomyces naeslundii ATCC 19039, Lactobacillus acidophilus ATCC 4356, Streptococcus gordonii ATCC 10558, Streptococcus mutans ATCC 35688, Streptococcus sanguinis ATCC 10556, Streptococcus sobrinus ATCC 33478 and Streptococcus mitis ATCC 9811. The active extracts were tested to determine their minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), cytotoxicity and chemical characterization. Forty-seven extracts (78%) were active against at least one microorganism. Extract 4A demonstrated the lowest MIC and MBC for all microorganisms except S. gordonii and the extract at MIC concentration was non-cytotoxic. The concentrated extracts were slightly cytotoxic. Electrospray ionization with tandem mass spectrometry analyses demonstrated that the extract constituents coincided with the mass of the terpenoids and phenolics. Overall, the best results were obtained for extraction methods A, B and C. The present work proved the antimicrobial activity of several plants. Particularly, extracts from C. doctoris were the most active against bacteria involved in dental caries disease.

Immunostimulating activity of Uncaria tomentosa in RAW 264.7 macrophages.

Uncaria tomentosa is a plant native to the Amazon that has immunomodulatory and antitumor properties due to the alkaloids found in the plant, being able to modify the immune response by potentiating or suspending the action of cytokines secreted by macrophages that induce the immune response, either by the classical route (M1) or through the alternative route (M2). Macrophages activated by M1 convert L-arginine into L-citrulline and nitric oxide (NO), whereas macrophages activated by the M2 pathway use the enzymatic activity of arginase to convert the same substrate into L-ornithine and urea. The aim of this work was to evaluate the immunostimulating activity of the crude hydroalcoholic extract from the bark of the U. tomentosa stem in RAW 264.7 macrophages. Concentrations of 0.2, 0.1 and 0.05 mg/mL of U. tomentosa extract associated with LPS, INF-γ and IL-4 inducers were tested by determining NO production and arginase enzyme activity. Nitric oxide production was enhanced by the extract when associated with LPS and LPS + INF-γ inducers. In the activity of the arginase enzyme, the extract decreased the stimulation of IL-4 on the enzyme, mainly at 0.2 mg/mL concentration. Therefore, it is concluded that the crude hydroalcoholic extract of the stem bark of U. tomentosa in RAW 264.7 cells, at a concentration of 0.2 mg/mL, showed considerable pro-inflammatory activity.

Polyvinylidene fluoride - Hydroxyapatite 0-3 biocomposite filaments processed by twin-screw extrusion.

Polyvinylidene fluoride - hydroxyapatite composite filaments were processed by twin-screw extrusion at different processing angular velocities and characterized by scanning electron and atomic force microscopies, differential scanning calorimetry and tensile tests. Polymer-ceramic composites with a 0-3 connectivity were successfully obtained. Regardless of the used processing parameters, all composite filaments present very similar melting (∼152°C) and solidification (∼139°C) points and elastic moduli (∼1.0 GPa) for hydroxyapatite as dispersed phase in the composite with concentrations up to 25 wt%, indicating that they are adequate for twin-screw extrusion and 3D printing. However, the yield strength (∼29 MPa), ultimate tensile strength (∼36 MPa) and tensile point (∼29 MPa) parameters are similar only for hydroxyapatite concentrations up to 15 wt%, once higher concentrations of hydroxyapatite as dispersed phase result in fragile samples (∼50% lower for each studied property).

Evaluation of miRNAs regulation of BDNF and IGF1 genes in T2DM insulin resistance in experimental models: bioinformatics based approach.

microRNAs (miRNAs) are recognized as diabetes mellitus type 2 (T2DM) biomarkers useful for disease metabolism comprehension and have great potential as therapeutics targets. BDNF and IGF1 increased expression are highly involved in the benefits of insulin and glucose paths, however, they are down-regulated in insulin resistance conditions, while their expression increase is correlated to the improvement of glucose and insulin metabolism. Studies suggest the microRNA regulation of these genes in several different contexts, providing a novel investigation approach for comprehending T2DM metabolism and revealing potential therapeutic targets. In the present study, we investigate in different animal models (human, rat, and mouse) miRNAs that target BDNF and IGF1 in skeletal muscle tissue with T2DM physiological conditions. Bioinformatics tools and databases were used to miRNA prediction, molecular homology, experimental validation of interactions, expression in the studied physiological condition, and network interaction. The findings showed three miRNAs candidates for IGF1(miR-29a, miR-29b, and miR-29c) and one for BDNF (miR-206). The experimental evaluations and the search for the expression in skeletal muscle from T2DM subjects confirmed the predicted interaction between miRNA-mRNA for miR-29b and miR-206 through human, rat, and mouse models. This interaction was reaffirmed in multiple network analyses. In conclusion, our results show the regulation relationship between miR-29b and miR-206 with the investigated genes, in several tissues, suggesting an inhibition pattern. Nevertheless, these data show a large number of possible interaction physiological processes, for future biotechnological prospects.

Preventive oral kefir supplementation protects mice from ovariectomy-induced exacerbated allergic airway inflammation.

Asthma is an inflammatory lung disease that affects more women than men in adulthood. Clinical evidence shows that hormonal fluctuation during the menstrual cycle and menopause are related to increased asthma severity in women. Considering that life expectancy has increased and that most women now undergo menopause, strategies to prevent the worsening of asthma symptoms are particularly important. A recent study from our group showed that re-exposure of ovariectomised allergic mice to antigen (ovalbumin) leads to an exacerbation of lung inflammation that is similar to clinical conditions. However, little is known about the role of probiotics in the prevention of asthma exacerbations during the menstrual cycle or menopause. Thus, our objective was to evaluate the effects of supplementation with kefir, a popular fermented dairy beverage, as a preventive strategy for modulating allergic disease. The results show that the preventive kefir administration decreases the influx of inflammatory cells in the airways and exacerbates the production of mucus and the interleukin 13 cytokine. Additionally, kefir changes macrophage polarisation by decreasing the number of M2 macrophages, as shown by RT-PCR assay. Thus, kefir is a functional food that potentially prevents allergic airway inflammation exacerbations in ovariectomised mice.

Is the wearing of elastic stockings for half a day as effective as wearing them for the entire day?

Background There is good evidence for the use of compression for some clinical indications but little is known about dosimetry in compression. Objective The aim of this work was to evaluate whether or not the use of compression stockings during part of the day would help in the reduction of evening oedema in patients with clinical, epidemiological, anatomical and physiopathological (CEAP) classifications C0 and C1. Methods The effects of elastic compression stockings on volumetric variations during the working day were evaluated for the legs of two men and 18 women (40 legs). The inclusion criterion was classification as C0 (10 legs) or C1 (30 legs) according to the CEAP criteria. Participants used three-quarter-length elastic compression stockings (20-30 mmHg) on three consecutive days for the entire day or only for the morning or they did not use the stockings at all. Volumetry using the water displacement technique was performed in the morning and in the evening. When the patients wore the stockings only during the morning, volumetry was also performed at 13:00 h. Results Significant increases in volume were observed for both legs when stockings were not used compared with the use of stockings in the morning only. After removing the stockings, both legs had significant increases in volume in the afternoon. However, use for half the day was better than not using the stockings at all. Conclusions The use of elastic compression stockings can reduce volumetric variations during working hours, with the use of stockings for the entire day being better than for just half the day.

Reliability of ultrasound for early sexing of goat fetuses derived from natural mating and from fresh, frozen and vitrified embryo transfer.

The aim of the present study was to identify the migration period of the genital tubercle and its later differentiation into external genital structures in fetuses derived from natural mating and fetuses from fresh, frozen and vitrified embryo transfer. A transrectal ultrasound with a double-frequency linear transducer (6.0 and 8.0 MHz) was used to monitor 123 goat fetuses, which were allocated to one of four groups: fetuses originating from controlled natural mating (G1, n = 32) and fetuses derived from fresh (G2, n = 34), frozen (G3, n = 30) and vitrified (G4, n = 27) embryo transfer. The transferable embryos were collected 7 days after mating by laparoscopy. Migration of the genital tubercle occurred significantly earlier (P < 0.05) in G1 than in G2, G3 and G4. The visualisation of the scrotum, prepuce and vulva occurred significantly earlier (P < 0.05) in G1 than in G2, G3 and G4. Our results show that fetal sexing is feasible after 55 days for fetuses from natural mating and after 60 days in fetuses from fresh and cryopreserved embryos. Thus, real-time ultrasonography is a reliable tool for fetal sex determination in goats after Day 50 of pregnancy, taking into account both the location of the genital tubercle and the identification of external genital structures.