Steroidogenic activity and morphological characterization of prenatal testes and epididymis of guinea pig (Cavia porcellus).

The present study aims to establish the morphological, morphometric, and immunostaining patterns of the steroidogenic enzymes 17ß-HSD and 5α-reductase and androgen receptors (AR) during the prenatal development of the male gonad and epididymis of Cavia porcellus. Fetuses at 22, 25, 30, 40, 45, 50, and 60 days of gestation (DG) were used. Specimens were dissected and subjected to macroscopic, histological, histomorphometric, and immunohistochemical analyses. Genital and scrotal protrusions were identified in 22 DG embryos. Gonocytes were identified at 25 DG and the formation of primary testicular cords was observed at 30 DG. Through anatomical evaluation, we observed differentiation of the epididymis into the head, body, and tail at 45 DG. During development, there is a progressive decrease in the diameters of the testicular cords and epididymal ducts. 17ß-HSD enzyme immunostaining was observed in Leydig cells at all ages, while 5α-reductase was observed in Leydig cell cytoplasm and gonocytes at 40, 50, and 60 DG. AR shows gonocyte labeling at 30 DG. Thus, from the second trimester of pregnancy, it is possible to observe patterns of anatomical development, such as genital and scrotal prominence (22 DG), the appearance of gonocytes in the testicular cords at 25 DG, and the beginning of the organization of primary testicular cords at 30 DG, suggesting sexual differentiation. The 17ß-HSD, 5α-reductase, and ARs play an essential role in sexual development and differentiation, presenting immunostaining at different reproductive process times.

Definition of protocols for cryopreservation and three-dimensional in vitro culture of prepubertal goat testicular tissue after histomorphological, ultrastructural, and functional analysis.

This study aims to define the best method (slow freezing or vitrification) and fragment size (1, 5, or 9 mm³) for prepubertal goat testis cryopreservation, as well as to evaluate testicular morphological integrity after cryopreservation and in vitro culture (IVC). Initially (experiment I), 1, 5, or 9 mm³ testis fragments were cryopreserved by slow freezing using a Mr. Frosty container with 20% Dimethylsulfoxide (DMSO) or vitrified using the Ovarian Tissue Cryosystem (OTC) device, (Equilibration solution - ES: 10% DMSO and 10% ethylene glycol - EG; Vitrification solution - VS: 20% DMSO and 20% EG) and then subjected to morphological analysis, type I and III collagen quantification and gene expression (Oct4, C-kit, Bax, and Bcl-2). Subsequently, (experiment II), fresh or cryopreserved by slow freezing testis fragments were cultured in vitro and submitted to morphological analysis by scanning electron microscopy. The data from the experiment I revealed fewer morphological alterations in 1 and 5 mm³ fragments after vitrification and slow freezing, respectively. The percentage of type I collagen fibers in 5 and 9 mm³ frozen was higher than in fresh or vitrified fragments. For type III collagen, fresh or frozen fragments of 1 and 5 mm3 showed a higher percentage than fragments of 9 mm3. Gene expression for Oct4 and C-kit after slow freezing or vitrification in the 5 mm3 fragments was lower than that observed in the fresh fragments. The Bax:Bcl-2 ratio in the 1 and 9 mm³ fragments was lower than in the 5 mm³ fragments for fresh fragments or after freezing. In experiment II, fragments cultured in vitro, previously frozen or not, showed more morphological alterations than fresh or frozen fragments. We concluded that slow freezing of 5 mm³ fragments was the best protocol for cryopreserving prepubertal goat testis and although the results of IVC are encouraging, it still needs improvement to restore testicular function after cryopreservation.

Impact of ethanol and heat stress-dependent effect of ultra-diluted Arnica montana 6 cH on in vitro embryo production in cattle.

This study evaluated the effect of adding ultra-diluted and dynamized Arnica montana 6 cH, and its vehicle (0.3% ethanol) to the in vitro maturation (IVM) medium, in the absence (experiment 1) or presence (experiment 2) of heat stress (HS), on bovine oocyte maturation and in vitro embryo production (IVEP). In experiment 1 (n = 902 cumulus oocyte complexes, COCs), the treatments were 1) IVM medium (Control treatment), 2) IVM medium + 0.3% ethanol, and 3) IVM medium + Arnica montana 6 cH. In experiment 2 (n = 1064 COCs), the treatments were 1) IVM medium without HS, 2) IVM medium under HS, 3) IVM medium + ethanol under HS, and 4) IVM medium + Arnica montana under HS. In the absence of HS (experiment 1), the addition of Arnica montana to the IVM medium had a deleterious effect on the IVEP (cleavage and blastocyst rates) and the total cell number/blastocysts. On the other hand, ethanol (0.3%) increased IVEP in relation to the Control and Arnica montana treatments. However, in the presence of HS during IVM (experiment 2), the addition of ethanol or Arnica montana increased IVEP when compared to the HS treatment alone, and the Arnica montana treatment resulted in greater total cell number/blastocysts compared to the other treatments. In conclusion, this study showed for the first time that the negative or positive effect of Arnica montana 6 cH on IVEP depends on the culture condition (i.e., absence or presence of HS during IVM). On the other hand, ethanol showed beneficial and consistent results on IVEP regardless of exposure to HS.

Foetal development of skeletal muscle in bovines as a function of maternal nutrition, foetal sex and gestational age.

To determine the effects of maternal nutrition on modifications of foetal development of the skeletal muscle and possible increase in the potential of skeletal muscle growth in cattle, gestating cows were either fed 190% NRC recommendations (overnourished; ON) or 100% NRC recommendation (control; CO). Interaction between maternal nutrition (MN) and the foetal sex (FS) was also investigated. Foetuses were necropsied at four different time points throughout gestation (139, 199, 241 and 268 days of gestation) to assess the mRNA expression of myogenic, adipogenic and fibrogenic markers in skeletal muscle. Phenotypic indicators of the development of skeletal muscle fibres, intramuscular lipogenesis and collagen development were also evaluated. Modifications in mRNA expression of skeletal muscle of foetuses were observed in function of MN and FS despite the lack of effect of MN and FS on foetal weight at necropsy. Maternal ON increased the mRNA expression of the myogenic marker Cadherin-associated protein, beta 1 (CTNNB1) and adipogenic markers Peroxissome proliferator-activated receptor gamma (PPARG) and Zinc finger protein 423 (ZNF423) at midgestation. However, no differences on foetal skeletal muscle development were observed between treatments at late gestation indicating that a compensatory development may have occurred on CO foetuses making the effect of MN on skeletal muscle development not significant at late gestation. Moreover, our data have shown an evidence of sexual dimorphism during foetal stage with a greater skeletal muscle development in male than in female foetuses. In conclusion, providing a higher nutritional level to pregnant cows changes the trajectory of the development of skeletal muscle during midgestation, but apparently does not change the potential of post-natal growth of muscle mass of the offspring, as no differences in skeletal muscle development were observed in late gestation.

Stressors in ICU: perception of the patient, relatives and health care team.

OBJECTIVE: To compare the evaluation of the stressors present in the intensive care unit (ICU) from the point of view of the patient, relatives and the multiprofessional team and to identify differences and similarities with regard to the perception of stressors in order to optimize patient care. DESIGN: Cross-sectional analytical survey. SETTING: General ICU of a private hospital. PATIENTS AND PARTICIPANTS: From April 1st to June 30th, 1996, 50 ICU patients during the first week of their ICU stay, 50 of their respective relatives and 50 members of the professional team directly involved in the care of these patients. MEASUREMENTS AND RESULTS: The Intensive Care Unit Environmental Stressor Scale (ICUESS) was administered to all patients. The relatives and health care professionals were asked to complete the ICUESS on the basis of their perception of the patient's stressors. Being in pain, having tubes in the nose or mouth, being restrained by tubes and being unable to sleep were considered by the patients, relatives and health care professionals as the main stressors. The professional team evaluated the intensity of the stressors higher than either the family or the patient. No statistical significance was detected between the intensity of the stressors as evaluated by the patient and the intensity evaluated by relatives and by the professional team. CONCLUSIONS: Being in pain, being unable to sleep and having tubes in the nose and/or mouth were pointed out as the major stressors by the three groups. There was no statistically significant correlation between the total stress scores of the patients and their relatives (r = 0.193), between the patients and the team (r = -0.002), or between the total scores of the team and the relatives (r = -0.185). The results suggest that the views of the relatives and the professional team concerning the stressors have some similar points compared to the evaluation made by the patient himself, although the intensity of the evaluation for each group corresponds to its own perception.

Comparison of the contractile performance of the hypertrophied myocardium from spontaneous hypertensive rats and normotensive infarcted rats.

The sarcoplasmic reticulum (SR) exerts a key role on the excitation-contraction coupling process in the myocardium. Since the relation between the volume of cellular organelles, such as SR, and the sarcolemmal area of myocytes is not uniform in myocardial hypertrophy of different etiologies, we compared the contractile performances of hypertrophied left ventricular papillary muscles from rats with pressure overload and with volume overload. Hemodynamically compensated spontaneous hypertensive rats (SHR, 3 months old, systolic blood pressure = 189 +/- 4 mmHg, n = 8) and Wistar rats with healed (30 days) myocardial infarction (MI, n = 7) produced by ligation of the left coronary artery were used. Results were compared with age-matched Wistar control (CON) rats (n = 13). Force (F), corrected to muscle cross-sectional area (g/mm2), and dF/dt were recorded in muscles contracting isometrically and stretched to Lmax. The inotropic response to increasing extracellular Ca2+ concentrations (1.25 to 5.0 mM) was compared in twitches (0.5 Hz) and during tetanic stimulation (5 Hz, 30 s) in the muscles treated with 1 microM ryanodine. F recorded in basal conditions (Ca = 1.25 mM, 0.5 Hz) in the CON group (1.34 +/- 0.20 g/mm2) was higher (p < 0.05) than in the MI (0.73 +/- 0.13 g/mm2) and lower (p < 0.05) than in the SHR group (2.08 +/- 0.25 g/mm2). Similar differences between groups were also observed in relation to +dF/dt. Increasing extracellular Ca produced a parallel increase of F and +dF/dt in the three groups of muscles. Ryanodine treatment reduced F and +dF/dt in all groups and completely inhibited the development of force in post-rest contractions, indicating SR inhibition. SHR muscles were more sensitive to ryanodine than CON and MI (F decrease = 64 +/- 7, 51 +/- 5, and 22 +/- 5%, respectively, p < 0.05). The tetanic tension (Ca = 1.25 mM) was similar in SHR and CON (0.82 +/- 0.19 and 0.92 +/- 0.18 g/mm2; p > 0.05) and depressed in the MI group (0.35 +/- 0.12 g/mm2). These data suggest an increased participation of SR as source of activator Ca in the hypertrophied muscle of SHR. This adaptation likely contributes to maintain the normal cardiac function in hemodynamically compensated SHR, despite increasing afterload levels. This adaptation seems not to occur after MI, which may contribute to the depressed contractile performance of the left ventricular muscle surviving to infarction.

Stressors in ICU: patients' evaluation.

OBJECTIVE: To study the physical and psychological stressors in the intensive care unit (ICU) and to correlate stressors with different demographic variables. DESIGN: Cross-sectional analytical survey. SETTING: Intensive care unit of a private hospital. PATIENTS AND PARTICIPANTS: 50 randomly selected ICU patients during the first week of their ICU stay. MEASUREMENTS AND RESULTS: The Intensive Care Unit Environmental Stressor Scale was administered to 50 patients. Pain and the impossibility of sleeping due to noise and having tubes in the nose and mouth were considered the most important physical stressors. Loss of self control and lack of understanding about the attitudes and procedures were the main psychological stressors. CONCLUSIONS: Interventions should be aimed at relieving the patient's pain and at controlling the level of noise to make sleep possible. From the psychological standpoint, the independence of the patient should be encouraged, thus stimulating the recovery of self-control. The team should also inform the patient about the procedures which will be carried out.

Contractility changes of the right and ventricular muscle after chronic myocardial infarction.

Contractility changes and adaptive responses resulting from acute left ventricular (LV) myocardial infarction are not restricted to the LV myocardium. The reduced LV function increased the right ventricular (RV) pressure load and neurohumoral factors, activated by the infarction

Contractility changes of the right and left ventricular muscle after chronic myocardial infarction

Contractility changes and adaptive responses resulting from acute left ventricular (LV) myocardial infarction are not restricted to the LV myocardium. The reduced LV function increases the right ventricular (RV) pressure load and neurohumoral factors, activated by the infarction episode, might have pan-cardiac effects. In the present study we investigated the mechanical activity of RV and LV isolated papillary muscles from 30-day infarcted male Wistar, 3-4-months old rat hearts. LV myocardial infarction was produced by ligature of the descending anterior branches of the left coronary artery (INF group). Control animals were submitted to sham surgery (SO group). Both groups were studied 30 days after the infarction procedure. Post-infarction hypertrophy was evaluated by measuring the cell diameters in the nuclear region. Contractility changes were analyzed by determining the isometric force (F) and the rate of force development (dF/dt) of papillary muscles from LV and RV. The effects of variations in extracellular Ca2+ concentrations (0.6, 1.25, 2.5 and 3.75 mM)were determined on twitches and tetanic contractures obtained during caffeine perfusion (2.5 mM) and were used to assess changes at the contractile protein level. The activity of the sarcoplasmic reticulum was evaluated by using the post-rest potentiation phenomenon. Hypertrophy occurred in both ventricles after infarction, with the RV chamber showing a pressure overload pattern while LV myocytes developed a volume overload pattern. F and dF/dt of LV papillary muscles decreased after infarction but did not change in the RV preparations. Positive inotropic changes obtained with increasing Ca2+ concentrations and the development of tetanic tension were reduced after infarction only in LV papillary muscles. The relative potentiation of post-rest contractions was only affected in the LV myocardium showing a decrease after infarction. These results suggest that different adaptive changes occur in the LV and RV myocardium after infarction. While the RV myocardium maintains its contractility the LV myocardium displays a depressed mechanical activity problably due to changes at the contractile mechinery level and to alterations in the Ca2+ handling process.

[Intensive Care Unit stay. Variables that affect emotional reactions]. / Internação em UTI. Variáveis que interferem na resposta emocional.

PURPOSE: To evaluate the anxiety and depression levels during five days of stay in a Clinical Cardiological Intensive Care Unit (ICU) and verify the relation to different variables. METHODS: Forty seven patients were submitted to IDATE scale for anxiety and an avaliation for depression. The scores were compared to medical diagnoses, age, local of stay before the ICU, number of hospitalizations. RESULTS: Anxiety and depression scores decrease significant from first to fifth day of ICU stay. No significant correlation was found between age, number of hospitalizations, medical diagnoses and the anxiety and depression scores. Patients admitted from the emergency unit presented a correlation between anxiety and depression scores. CONCLUSION: Anxiety and depression decrease during the five days of ICU stay. Emotional reactions are not influenced by medical diagnoses, age and number of hospitalizations. The emergency unit stay affects emotionally the patient in the first days of ICU admission.

Introduçäo à informática nas áreas de saúde e biológicas na UFPE / Introduction to informatic in health and biological areas in UFPE

Neste trabalho é apresentado o desenvolvimento de um programa de ensino e pesquisa para consolidação de um grupo em bioinformática na Universidade Federal de Pernambuco. O grupo vem desenvolvendo projetos e organizando cursos de informática aplicada às ciências da saúde e biológicas para alunos de graduação e pós-graduação, bem como para professores visando à inserção da informática no programa de suas disciplinas.

Edubiosoft: um software auxiliar no ensino de bioquímica / Edubiosoft: auxiliary software in biochemistry lectures

Com a finalidade de aprimorar o ensino de buioquímica, através de novas técnicas de lecionamento, foi desenvolvido nos Setores de Biotecnologia e de Bioinformática do LIKA, o programa EDUBIOSOFT, que através da utilização integrada de textos e imagens fornece ao aluno uma ferramenta de apoio ao aprendizado e avaliação do conhecimento de temas da bioquímica.

Dermasoft: um sistema de instruçäo assistido por computador (CAI) para o ensino de dermatologia / Dermasoft: computer assisted instruction system (CAI) for dermatology lectures

Com o intuito de desenvolvimento novas técnicas de divulgação e ensino em dermatologia, os Setores de Biotecnologia e de Bioinformática do Laboratório de Imunopatologia Keizo Asami (LIKA) juntamente com o Departamento de Dermatologia da Universidade federal de Pernambuco (UFPE), está desenvolvendo através da tecnologia Computer Assisted Instuction (CAI), o DERMASOFT. Este software consiste na utilização integrada de bases de textos, imagens e casos clínicos no ambiente Windows com o objetivo de ser um instrumento de apoio ao ensino e de atualização para dermatologia e profissionais interessados na área.

Propranolol reduces myocardial hypertrophy in the right cardiac chambers after infarction in rats.

The effects of chronic propranolol (Prop) therapy on the postinfarction myocardial hypertrophy of infarcted rats were studied by histological techniques. Male albino rats were submitted to left coronary artery ligation to produce infarction or to sham surgery (Con, N = 6). Infarcted rats (Inf) were divided into 2 groups receiving Prop (2.5 mg/kg, twice a day, N = 6) or saline (N = 6) for one month, respectively. Myocyte diameters were measured in longitudinally oriented sections in the four heart chambers (60 cells/chamber). Inf produced a significant increase in mean diameter of myocytes from the right atrium and ventricle and from the left atrium. In the right ventricle, myocyte diameter increased from 8.9 +/- 0.5 microns in the Con group to 12.5 +/- 0.6 microns in the Inf group (P < 0.05). Under Prop, myocyte diameter was reduced (P < 0.05) to 9.8 +/- 0.9 microns. Similar values were observed in the right atrium. In the left atrium, Prop produced only a partial reversion of the postinfarction hypertrophy. In the left ventricle, myocyte diameter was not significantly changed after Inf or Prop therapy. These data show that beta blockers reduce the myocardial hypertrophy in the right heart chambers after experimental infarcts in rats. This effect can be secondary to reduction of pulmonary hypertension or to blockade of direct effects of catecholamines on myocardial fibers or both.

Propanolol reduces myocardial hypertrophy in the right cardiac chambers after infarction in rats

The effects of chronic propanolol (Prop) therapy on the post infarction myocardial hypertrophy of infarction rats were studied by histological techniques. male albino rats were submitted to left coronary artery ligation to produce infarction or to sham surgery (Con, N=6)., Infarction rats (Inf) were divided into 2 groups receiving Prop (2.5 mg/kg, twice a day, N+6) or saline (N=6) for one month, respectively. Myocyte diameters were measured in longitudinally oriented sections in the four heart chambers (60 cells/chamber). inf produced a significant increase in mean diameter of myocytes from the right atrium and ventricle and from the left atrium. In the right ventricle, myocyte diameter increased from 8.9 ñ 0.5 um in the Con group to 12.5 ñ 0.6 um in the the Inf group (P<0.05). Under Prop, myocyte diameter was reduced (P<0.05) to 9.8 ñ 0.9 um. Similar values were observed in the right atrium. In the left atrium, Prop produced only a partial reversion of the postinfarction hypertrophy. In the left ventricle, myocyte diameter was not significantly changed after Inf or Prop therapy. These data show that beta blockers reduce the myocardial hypertrophy in the right heart chambers after experimental infarcts in rats. This effect can be secondary to reduction of pulmonary hypertension or to blockade of direct effects of catecholamines on myocardial fibers or both

Demonstration of actin in neurosecretory cells by treatment with nitrobenzoxadiazole-phallacidin.

Serial sections containing neurosecretory cells from chicken hypothalamus were cut after fixation in formaldehyde and embedding in paraffin. Sections were exposed to NBD-Ph (nitrobenzoxadiazole-phallacidin) and showed evidence of containing actin. By using a medium with sodium borohydride, non-specific fluorescence could be excluded.