Fermented soy product supplemented with isoflavones affected fat depots in juvenile rats

Objectives: This study investigated the effects of soy product fermented by Enterococcus faecium and Lactobacillus jugurti supplemented with isoflavones on adipose tissue, blood lipid, and glucose levels on juvenile rats. Methods: Rats were fed a cholesterol-enriched diet for 3 wk as a preliminary treatment to create hypercholesterolemia. They were then fed a chow diet (HC), a chow diet plus fermented soy product supplemented with isoflavones (HCFI), a chow diet plus placebo (HCP), or a chow diet plus placebo supplemented with isoflavones (HCPI), respectively, for an additional 3 wk. Results: The beneficial effects of fermented soy product supplemented with isoflavones on epididymal (EPI) and retroperitoneal (RET) fat pads was likely due to isoflavones because adipocyte circumference (micrometers) in the HC group was significantly larger (EPI: 105.66 ± 13.36; RET: 134.95 ± 25.40) than that in the HCFI group (EPI: 93.17 ± 12.80; RET: 108.62 ± 15.50) and HCPI group (EPI: 93.06 ± 15.10; RET: 112.34 ± 18.21). The probiotic micro-organism accentuated the antilipogenic effect of isoflavones on RET (HCFI: 108.62 ± 15.50 micrometers versus HCPI: 112.34 ± 18.21 micrometers). Moreover, the fermented product increased glucose concentration similar to that in the chow group but did not change blood lipids. Conclusion: This product may offer new approaches to obesity prevention. © 2005 Elsevier Inc. All rights reserved.

Ultrastructural features of the lining epithelium of the vas deferens of Agouti paca

The epididymal epithelium of Agouti paca, a wild South American rodent, was basically formed by principal and basal cells. Principal cells were closely related to processes of adsorptive endocytosis, phasefluid endocytosis and also secretion originating from their cytoplasmic ultrastructural features. Principal cells were also characterized by the presence of vesicles of several shapes, sizes and internalized content occurring in smaller pits, pale small vesicles next to the apical brush border of microvillus, as well as coated vesicles, smooth surface vesicles and great vesicles. Multivesicular bodies, endosomes and lysosomes were mainly observed in supranuclear position. Moreover, presence of an apocrine secretory process was demonstrated by the occurrence of apical cytoplasmic expansions projecting into the vas deferens luminal compartment. Basal flattened cells without luminal surface contact occurred next to the basement membrane of the ductus, and did no exhibit special ultrastructural features.

Localização histoquímica de algumas hidrolases no epidídimo do gerbilo

This study mainly showed that alkaline phosphatase expression had been present in the proximal regions of the epididymidis ductus of the gerbil which comprised the initial segment and proximal caput. The reactivities of acid phosphatase and ATPase were strong in the proximal and distal regions of the epididymidis ductus at the level of the apical cytoplasm and epithelium, except at the corpus level, a very thin isthmus located between the caput and cauda epididymidis, and as a general rule a low enzymatic reactive region of the epididymis of gerbil. SDH revealed also low activities in all the regions and regional structures of the duct, except into the luminal content formed by storaged spermatozoa, prior on the cauda level. The enzymes presented in the epididymis were correlated to some histophysiological roles such as the enzymatic mediation of endocytosis, secretion, absorption and active transport concerning to phosphatases and ATPase and a possible mitochondrial role of SDH could occur at the spermatozoa level in which the middle pieces were formed by a great amount of mitochondria.

Estudo morfológico e histomorfométrico sobre variações sazonais do ducto deferente de codorna da variedade italiana

In the autumn the vas deferens of the Italian variety of domestic quail appeared as a single, thin, and straight duct along its total extension. Thus, transversal histologic sections of this duct showed a circular tubular shape. The pseudoestratified columnar epithelium that lined the vas deferens presented longitudinal folds which invaded the lumen frequently empty of spermatozoa. Although in the winter, spring and summer the usual morphological appearance of the quail's vas deferens was seen as a highly coiling duct. So, each transversal histologic section of the vas deferens, in all the segments, showed parallel cut sections of the duct being irregular in shape and variable in number and interconnected by the adventitial loose connective tissue. In these observations, the tubular lumen was totally performed by spermatozoa and seminal fluid. Consequently, with previous base on the testis kinetics of the Italian quail variety, it was concluded that the spermatozoa production, followed by emission, storage and ejaculation of spermatozoa through the vas deferens did not stop during the winter, spring and summer, but ceased in the autumn the quiescent phase of the circannual reproductive cycle in this bird.

Cytogenetical aspects of testicular cells in economically important species of coreidae family (Heteroptera)

Some cytogenetical aspects of spermatozoa formation were studied in 9 Coreidae Brazilian species: Anasa bellator, Athaumastus haematicus, Chariesterus armatus, Dallacoris obscura, Dallacoris pictus, Leptoglossus gonagra, Leptoglossus zonatus, Sphictyrtus fasciatus, and Zicca annulata. Similarly to the other species described to date, all the species studied herein showed cystic spermatogenesis, a reddish membrane covering the testes, a X0 sex determining system, a pair of m-chromosomes, intersticial chiasmata in most autosomes, and autosomes dividing reductionally at first meiotic division and equationally in the second 1 while sex chromosomes, divide equationally and reductionally at first and second meiotic division, respectively. In addition, it was observed that the sex chromosome is heteropycnotic at prophase and that heteropycnotic chromosomal material is found in the nuclei at spermiogenesis. In the species studied, the diploid chromosome number ranged from 19 to 25. It was 19 in S. fasciatus (16A+2m+X0); 21 in A. bellator, A. haematicus, D. obscura, D. pictus, L. gonagra, and L. zonatus (18A+2m+X0); 23 in Z. annulata (20A+2m+X0); and 25 in C. armatus (22A+2m+X0). © 2007 The Japan Mendel Society.

Localization of total proteins and lactate dehydrogenase in hamster epididymis

The concentration of total protein measured by photocolorimetric methodology and reported as units per 100 mg of tissue decreased from the initial segment to the cauda epididymidis of the Golden hamster, being significant the numeric difference observed between these two regions. This observation was related with an increased synthesis and secretion of proteins to the lumen in proximal segments of the epididymidis duct, mainly in initial segment, as proposed for other rodents. LDH activity was higher in initial segment and distal cauda than in the caput and corpus epididymidis, although no significant differences in mean values had been observed. The high LDH activity observed in initial segment and cauda epididymidis of hamster had been related to an expressive epithelium metabolic activity presented in these regions. This metabolic activity help to guarantee the survival of spermatozoa stored in cauda epididymidis. Furthermore, lower LDH activity noted in the caput and corpus epididymidis might be related with a progressive reduction of glycolysis in initial maturation step of spermatozoa mainly verified in corpus epididymidis. © 2007 Sociedad Chilena de Anatomía.

Decrease of the DNA damage levels after sperm preparation by layering method

The aim of this prospective study was to determine the DNA fragmentation levels before and after sperm preparation by layering method. A total of 78 patients submitted to assisted reproduction technology (ART) for infertility treatment were evaluated. Ejaculated spermatozoa were obtained by masturbation on the day of ART procedure. The evaluation of DNA fragmentation was performed in the fresh semen and after preparation by a layering method, respectively. After washing with PBS, the sperm pellets were smears and then processed for the terminal deoxyribonucleotidyl transferase (TdT)-mediated dUTP nick-end labelling (TUNEL) assay that was performed using a Cell Death Detection Kit with tetramethylrhodamine-labelled dUTP. For quantitative evaluation, 200 spermatozoa in randomly selected areas on microscope slides were evaluated and the percentage of TUNEL positive spermatozoa was determined. If ≥20% of selected sperm were TUNEL positive, the exam was considered abnormal. The mean percentage of DNA sperm fragmentation before sperm preparation was 17±8.3% and after 7.8±6.5% (p<0.0001). The exam was considered normal in 49 patients before preparation and in 73 patients after (p<0.0001). The sperm preparation with a layering method for the ART procedure is effective to select sperm with a significant decrease of the DNA damage.

The effects of male age on sperm DNA damage in an infertile population

The objective was to investigate the influence of age on sperm DNA damage. Semen samples were collected from 508 men in an unselected group of couples attending infertility investigation and treatment. DNA fragmentation in spermatozoa was measured by TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick-end labelling (TUNEL) assay; at least 200 spermatozoa in randomly selected areas of microscope slides were evaluated using a fluorescent microscope and the percentage of TUNEL positive spermatozoa was determined. The number of cells with red fluorescence (TUNEL positive) was expressed as a percentage of the total sample [DNA fragmentation index (DFI)]. Age was treated as a continuous variable for regression and correlation analysis. The following male age groups were used: Group I: ≤35 years, Group II: 36-39 years, and Group III: ≥40 years. DFI was significantly lower in Group I than in Group II (P = 0.034) or III (P = 0.022). There was no difference in DFI between Groups II and III. In addition, regression analysis demonstrated a significant increase in sperm DFI with age (P = 0.02). TUNEL assay clearly demonstrates an increase in sperm DNA damage with age. © 2007 Published by Reproductive Healthcare Ltd.

Efeito de um quelante de cálcio, um detergente e da lecitina de soja sobre a qualidade do sêmen caprino congelado-descongelado

The aim of this paper was verifying the effect of the Equex STM Paste and EDTA addition to a Tris-egg yolk extender, on the postthaw goat sperm viability. Nine semen samples of two adult goats were collected by artificial vagina and cryopreserved. It was also objective of this study, to evaluate the utilization of a soybean lecithin based commercial extender (Bioexcell® - IMV, L'Aigle, French) for the goat semen freezing. They were formed five experimental groups: TRIS; TRIS+EDTA; TRIS+EQUEX; TRIS+EDTA+EQUEX e Bioexcell. After evaluation, the semen was diluted in the five extenders and packed in 0.25mL straws with 100 million of motile spermatozoa. The samples were cooled at 0,46°C/min to 5°C, submitted at 75min of equilibration time and frozen in liquid nitrogen vapour. The thawing was accomplished in 37°C water bath for 50s. There were no differences (P>0,05) on the means of post-thaw total and progressive sperm motility among the groups TRIS, TRIS+EQUEX and TRIS+EQUEX+EDTA. The Bioexcell group obtained the least (P<0,05) percentage of post-thawing total and progressive sperm motility. After the thermotolerance test, it was observed the greatest (P<0,05) rates of total and progressive sperm motility in the Equex STM groups (TRIS+EQUEX and TRIS+EQUEX+EDTA). Thus, it can be affirmed that the Equex addition promotes better maintenance rates in the pos-thaw sperm viability, when compared with the extenders that did not contain it.

Semen variables of sheep (Ovis aries) experimentally infected with Toxoplasma gondii

The influence of Toxoplasma gondii on semen variables and sperm morphology of sheep was evaluated in eight reproductive males distributed into three experimental groups: GI, three sheep inoculated with 2.0 × 105 of P strain oocytes; GII, three sheep infected with 1.0 × 106 of RH strain tachyzoites and; GIII two control sheep. Clinical (rectal temperature, cardiac and respiratory frequencies), parasite and serology exams (IIF) were realized. Sperm variables (volume, motility, vigor and concentration) and semen morphology for each sheep were also evaluated. Thus, semen and blood collections were assessed on post-inoculation days (PIDs)-1,3,5,7,11,14 and weekly thereafter up to PID 70. Clinical alterations were observed (hypothermia and anorexia) in infected sheep from groups GI and GII. Parasitic outbreaks were detected in five sheep. All the infected sheep produced antibodies against T. gondii from PID 5 onwards, reaching a peak of 4096 and 8192 for group GI and GII sheep, respectively. Differences (P < 0.05) were observed regarding the ejaculate volume between the inoculated groups (oocytes and tachyzoites) and control. Even though experimental toxoplasmic infection resulted in clinical symptomology in the inoculated sheep, the minimal alterations in sperm pathologies could not be directly attributed to T. gondii. © 2008 Elsevier B.V. All rights reserved.

Organização celular dos testículos em Hylidae e leptodactylidae, no Pantanal (Estado do Mato Grosso do Sul, Brasil)

The objective of this work was to make a comparative analysis of germ cell organization at different stages of cellular differentiation in adult males of Dendropsophus nanus (Boulenger, 1889), Pseudis limellum (Cope, 1862), P. paradoxa (Linnaeus, 1758), and Scinax acuminatus (Cope, 1862), belonging to the family Hylidae; and Leptodactylus chaquensis (Cei, 1950) and L. podicipinus (Cope, 1862), belonging to the family Leptodactylidae, collected in the Pantanal and in Serra da Bodoquena, Mato Grosso do Sul State, Brazil. The testes were removed and fixed, dehydrated in a graded series of ethanol and embedded in methacrylate glycol resin (Historesin Leica®). The sections were stained by 1% toluidine blue and observed under light microscope. It was detected that all individual of the Hylidae family show, throughout the year, the presence of all germ cell types of spermatogenesis. However, all Leptodactylidae family individuals only show the presence of all germ cell types during the rainy season. The variations of characteristics in seminiferous epithelium organization, as well as the evident difference in the amount of spermatozoa inside the tubules, are evidence that the anurans in this work show different forms of spermatogenesis development throughout the year: the cycle is continuous for the Hylidae family, and discontinuous with explosive release of spermatozoa for the Leptodactylidae family.

Seleção de espermatozoides para ICSI

The application of assisted reproduction techniques has provided help to many men seeking to father a child, although the current success of these procedures remains suboptimal. Today some protocols allow sperm to be selected according to their ultrastructural morphology or surface molecular characteristics. On the other hand, successful human reproduction relies partly on the inherent integrity of sperm DNA. Therefore, it is now necessary to improve the safety of the sperm selection method. It is urgent to optimize procedures to isolate spermatozoa for ICSI with low risk of DNA damage. In recent years, two technologies have attracted the attention of specialists as methods capable of identifying a spermatozoon with low risk of DNA damage: Ultrastructural morphology sperm selection at high magnification and sperm head birefringence selection. This review analyses these two technologies. © Todos os direitos reservados a SBRA - Sociedade Brasileira de Reprodução Assistida.

Influência da raça do touro (Bos indicus x Bos taurus) na tolerância ao estresse térmico calórico de embriões bovinos produzidos in vitro

To better understand the differences related to HS resistance between Bos indicus and Bos taurus, we aim to verify if the HS tolerance is due mostly to the genetic contribution from the oocyte, spermatozoa or both. Oocytes from Nelore and crossbreed Holstein cows (cHST) were collected, matured and fertilized with semen from Nelore (N), Angus (An), Brahman (Bra) and Gir (Gir) bulls. Nine six hours post insemination (hpi), ≥ 16 cells embryos were separated in two groups: control and HS. In control group, embryos were cultured at 39°C, whereas in the HS group, embryos were subjected to 41°C for 12 h, and then returned to 39°C. There was no effect of HS on blastocyst and hatched blastocyst rates in all breeds analyzed. The percentage of oocytes that cleaved and reached morula stage was significantly lower (p < 0.05) in cHST x Gir as compared to the other breeds. Additionally, blastocyst rates was higher in cHST x N than in cHST x An and cHST x Gir (p < 0.05). It was concluded that the oocyte is more important than the spermatozoa for the development of thermotolerance, since the breed of the bull did not influence embryo development after HS.

Análise espermática de perdizes (Rhynchotus rufescens) criadas em cativeiro e suplementadas com selênio

Due to the commercial importance of the red-winged tinamou (Rhynchotus rufescens), for the past few years, the employment of reproductive biotechnologies has been attempted. Thirty animals were randomly assigned into two groups: control group (no selenium) and treatment group (supplemented with 0,2 a 0,8 mg selenium/ 1000 kg ration). Animals were allocated at the FCAV - UNESP/Jaboticabal (2007-2008). Semen collections were performed by digital manipulation and divided in pools of at least 150 μL. After the immediate evaluation of motility, vigour, concentration and morphology, an aliquot of 20 μL was diluted in 300 μL of physiologic solution in order to test acrosome and membrane integrities, which were performed by counting 200 cells for each test. Cells were evaluated as follows: 1) Intact acrosome: lilac acrosome; Non-intact acrosome: pink acrosome; 2) Live cells: non stained; Dead: stained. Data was statistically analysed using the SAS System for Windows. No differences were found between treatment and control groups for volume, motility, vigour, mean number of spermatozoa per animal, concentration, Intact acrosome, Intact membrane. The difference found on midpiece sperm defect (Se = 1,33 ± 0,53 and control = 3,78 ± 0,69, p = 0.0107) may be due to the damages caused by the selenium deficiency to the architecture of the midpiece, which compromises sperm mobility and fertilization capacity.