Lung consolidation caused by Mycoplasma hyopneumoniae has a negative effect on productive performance and economic revenue in finishing pigs.

This study aimed to measure the impact of productivity and the consequent economic losses related to lung lesions caused by M. hyopneumoniae. Five-hundred 75 days-old pigs were selected and weighed at the beginning and at the end of the finishing phase to assess the average daily gain (ADG). These animals were evaluated at the slaughter, and samples were collected for laboratory analysis to confirm the presence of M. hyopneumoniae DNA. The lungs of each pig were examined and classified into groups based on the extension of macroscopic lung lesions. Four-hundred eighty-six lungs were examined and 68.5% (n = 333) had macroscopic lung lesions. All pigs with lesions were positive for M. hyopneumoniae in qPCR. Linear mixed regression models (proc Glimmix) were performed on SAS to estimate the effect of macroscopic lung lesion scores on the ADG of finishing pigs. All pairwise comparisons among lesion score groups were performed using p < 0.05. For each increase of one percent in the lesion area, there was a decrease of 1.8 g in the daily weight gain. All the groups had a numerically lower ADG when compared to Group 1 (no lesions). The economic analysis was performed by simulation on Excel to estimate and compare the financial performance of the different lung lesion score groups. The negative correlation found between the group with no lung lesions and the group with more than 15.1% of lesions, showed a statistical difference in ADG, which could mean an opportunity to gain up to $ 6.55 per pig at slaughter. The presence of lesions causes the animals to decrease their productive potential, causing financial loss and generating impacts on the production system.

Momento de aplicação do nitrogênio e algumas variáveis estruturais e bromatológicas do capim-massai / Time of nitrogen application and some structural and bromatologic variables of massaigrass

As plantas forrageiras tropicais apresentam alto potencial de produção de massa de forragem, condicionada às condições edafoclimáticas, principalmente à adubação nitrogenada. A magnitude da resposta depende do sincronismo entre a disponibilidade de nutrientes e as necessidades da planta forrageira na rebrotação, e os resultados encontrados na literatura são contrastantes. Assim, este trabalho foi realizado com o objetivo de avaliar características estruturais e bromatológicas de Panicum maximum Jacq cv. Massai submetido a quatro doses de nitrogênio (0, 40, 80 e 120mg/dm3) e três momentos de aplicação após o corte (um, três e sete dias), divididos em três cortes, em um esquema fatorial 4 x 3 e delineado inteiramente ao acaso, com quatro repetições. O experimento foi conduzido em casa de vegetação. O incremento na taxa de aparecimento de perfilhos, na densidade populacional de perfilhos, na produção de massa seca de folha total e nos teores de proteína bruta foi de 42%, 65%, 770% e 35%, respectivamente, para a dose de 120mg/dm³ em relação à ausência de adubação nitrogenada. As fibras em detergente neutro e em detergente ácido decresceram cerca de 6% e 9%, respectivamente, com a adição de nitrogênio. Apenas a massa seca de folha total respondeu às épocas de aplicação de nitrogênio após o corte, com valores pouco expressivos. A adubação nitrogenada exerce efeito positivo sobre as variáveis estudadas, porém novos estudos devem ser conduzidos para a definição da época de adubação de nitrogênio após o corte, para a melhor utilização do nutriente pelas plantas.(AU)

Tropical forage plants have a high potential for mass production of fodder, subject to soil and climatic conditions, especially with nitrogen fertilization. The magnitude of the response depends on the timing of availability of nutrients and forage plant needs in the regrowth. Thus, this work was carried out to evaluate structural and bromatologic characteristics of the Panicum maximum Jacq cv. Massai in response to four nitrogen doses (0, 40, 80 and 120mg/dm3) and with three application times after cutting (one, three and seven days) in a 4 x 3 factorial treatment combination, in a completely randomized design, with four replicates. The experiment was conducted in a greenhouse. The increase in the rate of tillering, tiller population density, dry mass total leaf production and crude protein levels was 42%, 65%, 770% and 35%, respectively for the dose of 120mg / dm³ regarding the absence of fertilization. The neutral detergent and acid detergent fibers showed a decrease of 6% and 9% with nitrogen addition. Dry mass total leaf production responded to times of nitrogen application after cutting, with little significant values. Nitrogen fertilization has a positive effect on the variables studied, but new studies should be conducted to define the timing of nitrogen fertilization after cutting, for better utilization of this nutrient by plants.(AU)

Human platelet antigen genotyping of platelet donors in southern Brazil.

Human platelet antigens (HPA) are immunogenic structures that result from single nucleotide polymorphisms (SNPs) leading to single amino acid substitutions. This study sought to determine the allele and genotype frequencies of HPA-1, HPA-2, HPA-3, HPA-4, HPA-5 and HPA-15 in platelet donors from the state of Rio Grande do Sul (RS), Brazil, and compare their allele frequencies to those observed in other populations. HPA genotyping was performed by PCR-SSP method. The study sample comprised 201 platelet donors (167 Caucasians and 34 non-Caucasians). Allele 'a' was that most commonly found for HPA-1 to 5 in both groups. The HPA-15ab genotype predominated over homozygous genotypes of this system. Fisher's exact test revealed statistically significant differences for the HPA-5 system, with a greater prevalence of the HPA-5b allele in non-Caucasians. The neighbour-joining method and principal components analysis revealed genetic proximity between our Caucasian group and European populations. We conclude that the allele frequencies of HPA-1 to 5 and HPA-15 found in our Caucasian sample are similar to those reported for European populations. These findings corroborate the ethnic makeup of the population of RS. The higher frequency of the HPA-5b allele found in the non-Caucasian group of our sample suggests the possibility of allosensitization in patients who receive platelet transfusions from genetically incompatible donors.

Temporal auditory processing and phonological awareness in children with benign epilepsy with centrotemporal spikes.

The aim of this research was to analyze temporal auditory processing and phonological awareness in school-age children with benign childhood epilepsy with centrotemporal spikes (BECTS). Patient group (GI) consisted of 13 children diagnosed with BECTS. Control group (GII) consisted of 17 healthy children. After neurological and peripheral audiological assessment, children underwent a behavioral auditory evaluation and phonological awareness assessment. The procedures applied were: Gaps-in-Noise test (GIN), Duration Pattern test, and Phonological Awareness test (PCF). Results were compared between the groups and a correlation analysis was performed between temporal tasks and phonological awareness performance. GII performed significantly better than the children with BECTS (GI) in both GIN and Duration Pattern test (P < 0.001). GI performed significantly worse in all of the 4 categories of phonological awareness assessed: syllabic (P = 0.001), phonemic (P = 0.006), rhyme (P = 0.015) and alliteration (P = 0.010). Statistical analysis showed a significant positive correlation between the phonological awareness assessment and Duration Pattern test (P < 0.001). From the analysis of the results, it was concluded that children with BECTS may have difficulties in temporal resolution, temporal ordering, and phonological awareness skills. A correlation was observed between auditory temporal processing and phonological awareness in the suited sample.

Quality of life in pregnant women with epilepsy versus women with epilepsy / Qualidade de vida em gestantes com epilepsia versus mulheres com epilepsia

It is assumed that 25 percent of patients with epilepsy are women of fertile age and 0.3 percent to 0.6 percent of all children are born of mothers with epilepsy. The aim of this study was to evaluate the quality of life on pregnant with epilepsy and compare with non-pregnant women with epilepsy. We evaluated two groups (Experimental Group - 29 pregnant women with epilepsy and Control Group - 30 women with epilepsy); they were attended at the HC/UNICAMP. The patients had three meetings to carry out and implement the anamnesis and the application of QQV-65. There were no significant differences in the measurement of quality of life when comparing both groups. However, when we analyzed individually in the pre- and post-partum periods, we observed significant differences in health aspects (p=0.0495), physical (p=0.02868) and emotional (p=0.0253) dimensions in QQV-65. This study shows that pregnancy could be interpreted as a stressor. In late pregnancy when this stressor was removed, women with epilepsy had improvement in their quality of life.

Admite-se que 25 por cento dos pacientes com epilepsia sejam mulheres em idade fértil e que 0,3 por cento a 0,6 por cento de todas as crianças nascidas sejam filhas de mães epilépticas. O objetivo do presente estudo foi avaliar a qualidade de vida em gestantes com epilepsia e compará-la com a de mulheres com epilepsia não grávidas. Foram avaliados dois grupos (Grupo Experimental - 29 gestantes com epilepsia e Grupo Controle - 30 mulheres com epilepsia) atendidos no HC/UNICAMP. As pacientes foram submetidas a três encontros para a realização de anamnese e a aplicação do QQV-65. Não encontramos diferenças significativas na avaliação de qualidade de vida ao comparar ambos os grupos. No entanto quando avaliados individualmente no período pré e pós-natal, observamos diferenças significativas nos aspectos: saúde (p=0,0495), físico (p=0,02868) e emocional (p=0,0253) no QQV-65. Este estudo mostrou que a gravidez pode ser interpretada como um estressor. No final da gravidez, quando este estressor foi removido, mulheres com epilepsia mostraram melhora na qualidade de vida.

Evaluation of nanoparticles loaded with benzopsoralen in rat peritoneal exudate cells.

Psoralens are widely used for the treatment of hyperproliferative skin disease. In this work, we prepared nanoparticles (NP) containing a benzopsoralen (3-ethoxy carbonyl-2H-benzofuro[3,2-f]-1-benzopiran-2-one) by the solvent evaporation technique. We evaluated important NP parameters such as particle size, drug encapsulation efficiency, effect of the encapsulation process over the drug's photochemistry, zeta potential, external morphology, and in vitro release behavior. We also investigated the nanoparticle as a drug delivery system (DDS), as well as its target delivery to the action site, which is a very important parameter to increase the therapeutic use of psoralens and to reduce their side effects. The uptake of benzopsoralen-loaded PLGA nanoparticles by different kinds of cells found in rat peritoneal exudates was also studied. The photodamage promoted by irradiation with UV light revealed morphological characteristics of cell damage such as cytoplasmic vesiculation, mitochondrial damage, and swelling of both the granular endoplasmatic reticulum and nuclear membrane. This encapsulation method maintained the drug's properties and improved drug delivery to the target cell.

HIV-1 viremia during the first 28 weeks of pregnancy is not associated with mother-to-child transmission

It is currently recommended that antiretroviral prophylaxis to prevent mother-to-child transmission (MTCT) of HIV be initiated at 14 weeks of gestation. However, the relevance of early-gestation HIV viral load level for intrauterine MTCT is unknown. The objective of this study was to determine the relationship between prenatal maternal viral load and intrauterine MTCT. Records of HIV-infected pregnant women in two centers in Brazil, from 1999 to 2004 were analyzed. Three pregnancy periods were considered: earlier than 14 weeks, 14 to 27 6/7 weeks, and 28 weeks of gestation or more. Peripartum HIV exposure was also computed. Maximum viral load in each period was the measure of HIV exposure. Four hundred fifty-seven HIV-infected pregnant women were evaluated, but 53 were excluded. The MTCT rate was 0.49 percent (2/404-95 percent confidence interval (CI95) = 0.14-1.79 percent). Newborns were not breast-fed. Median viral load for the earlier-than-14-week period was 9,900 copies/mL (P25-75 1,000-50,775 copies/mL), 8,350 copies/mL (P25-75 707-42,000 copies/mL) for the 14 to 27 6/7-week period, and 435 copies/mL (P25-75 90-7,775 copies/mL) after the 28-week period. The peripartum median viral load was 400 copies/mL (P25-75 80-500 copies/mL). MTCT in mothers with VL > 1,000 copies/mL during the first 14 weeks (0.67 percent, 2/298) was not different from those with VL =1,000 copies/mL (0.0 percent, 0/96, P=1). Analogously, in the 14 to 27 6/7-week period, MTCT was similar in groups with VL higher (0.68 percent, 2/292) or lower (0 percent, 0/106) than 1,000 copies/mL (P=1). Regarding VL >1,000 copies/mL at 28-weeks-or-later and at peripartum periods, MTCT rates were 1.15 percent (2/173, P = 0.18) and 2.8 percent (2/71, P = 0.03), respectively. Intrauterine transmission does not seem to be influenced by HIV viremia during the first 28 weeks of pregnancy.

Neuroanatomical approaches of the tectum-reticular pathways and immunohistochemical evidence for serotonin-positive perikarya on neuronal substrates of the superior colliculus and periaqueductal gray matter involved in the elaboration of the defensive behavior and fear-induced analgesia.

Deep layers of the superior colliculus, the dorsal periaqueductal gray matter and the inferior colliculus are midbrain structures involved in the generation of defensive behavior and fear-induced anti-nociception. Local injections of the GABA(A) antagonist bicuculline into these structures have been used to produce this defense reaction. Serotonin is thought to be the main neurotransmitter to modulate such defense reaction in mammals. This study is the first attempt to employ immunohistochemical techniques to locate serotonergic cells in the same midbrain sites from where defense reaction is evoked by chemical stimulation with bicuculline. The blockade of GABA(A) receptors in the neural substrates of the dorsal mesencephalon was followed by vigorous defensive reactions and increased nociceptive thresholds. Light microscopy immunocytochemistry with streptavidin method was used for the localization of the putative cells of defensive behavior with antibodies to serotonin in the rat's midbrain. Neurons positive to serotonin were found in the midbrain sites where defensive reactions were evoked by microinjection of bicuculline. Serotonin was localized to somata and projections of the neural networks of the mesencephalic tectum. Immunohistochemical studies showed that the sites in which neuronal perikarya positive to serotonin were identified in intermediate and deep layers of the superior colliculus, and in the dorsal and ventral columns of the periaqueductal gray matter are the same which were activated during the generation of defense behaviors, such as alertness, freezing, and escape reactions, induced by bicuculline. These findings support the contention that serotonin and GABAergic neurons may act in concert in the modulation of defense reaction in the midbrain tectum. Our neuroanatomical findings indicate a direct neural pathway connecting the dorsal midbrain and monoaminergic nuclei of the descending pain inhibitory system, with profuse synaptic terminals mainly in the pontine reticular formation, gigantocellularis nucleus, and nucleus raphe magnus. The midbrain tectum-gigantocellularis complex and midbrain tectum-nucleus raphe magnus neural pathways may provide an alternative output allowing the organization of the fear-induced anti-nociception by mesencephalic networks.

HIV-1 viremia during the first 28 weeks of pregnancy is not associated with mother-to-child transmission.

It is currently recommended that antiretroviral prophylaxis to prevent mother-to-child transmission (MTCT) of HIV be initiated at 14 weeks of gestation. However, the relevance of early-gestation HIV viral load level for intrauterine MTCT is unknown. The objective of this study was to determine the relationship between prenatal maternal viral load and intrauterine MTCT. Records of HIV-infected pregnant women in two centers in Brazil, from 1999 to 2004 were analyzed. Three pregnancy periods were considered: earlier than 14 weeks, 14 to 27 6/7 weeks, and 28 weeks of gestation or more. Peripartum HIV exposure was also computed. Maximum viral load in each period was the measure of HIV exposure. Four hundred fifty-seven HIV-infected pregnant women were evaluated, but 53 were excluded. The MTCT rate was 0.49% (2/404-95% confidence interval (CI95) = 0.14-1.79%). Newborns were not breast-fed. Median viral load for the earlier-than-14-week period was 9,900 copies/mL (P25-75 1,000-50,775 copies/mL), 8,350 copies/mL (P25-75 707-42,000 copies/mL) for the 14 to 27 6/7-week period, and 435 copies/mL (P25-75 90-7,775 copies/mL) after the 28-week period. The peripartum median viral load was 400 copies/mL (P25-75 80-500 copies/mL). MTCT in mothers with VL > 1,000 copies/mL during the first 14 weeks (0.67%, 2/298) was not different from those with VL =1,000 copies/mL (0.0%, 0/96, P=1). Analogously, in the 14 to 27 6/7-week period, MTCT was similar in groups with VL higher (0.68%, 2/292) or lower (0%, 0/106) than 1,000 copies/mL (P=1). Regarding VL >1,000 copies/mL at 28-weeks-or-later and at peripartum periods, MTCT rates were 1.15% (2/173, P = 0.18) and 2.8% (2/71, P = 0.03), respectively. Intrauterine transmission does not seem to be influenced by HIV viremia during the first 28 weeks of pregnancy.

Sustained release of lidocaine from Poloxamer 407 gels.

In this work, we show that alteration of P407 gel content can affect drug release rates. The inorganic salts and PEG 400 commonly included in the formulation of P407 gels can also change the rate at which a drug is released. Lidocaine was selected as a model drug because, although widely used in the treatment of pain, its use is limited by short duration of its effects. The use of P407 gels prolongs the residence time of the lidocaine at the injection site, sustains drug release and increases therapeutic efficacy. Release studies were performed in a diffusion system. During release, data followed the Higuchi square root law time kinetic (r>0.98). Increased polymer concentration in the gel increases viscosity and reduces lidocaine release rates and diffusion coefficients via extended gel dissolution time and prolonged drug diffusion through the gel matrix. Lidocaine release rates and diffusion coefficients increased in gels composed of NaCl or PEG 400 aqueous solution. Because these additives are hydrophilic, they reduce gel dissolution time, thereby accelerating drug diffusion. Poloxamer is biocompatible and the results support the possibility of using Poloxamer gel as a sustained release injectable formulation.

Morphology and histoenzymology of eosinophilic granulocytes in the circulating blood of the turtle (Chrysemys dorbignih).

The studies on the characterization of eosinophils and neutrophils/heterophils of turtles are contradictory. Some authors have pointed out the existence of two distinct cell types: eosinophils and heterophils. Other authors have proposed that eosinophils and heterophils may be the same cells in different stages of maturation. These interpretations are based only on a morphological analysis. In the blood of the turtle (Chrysemys dorbignih), a South American freshwater species, there are two types of granulocytes with eosinophilic staining pattern: the first with round cytoplasmic granules and the second with ellipsoidal cytoplasmic granules. In the present study by using histoenzymological methods for the analyses of enzymological cellular content, we found that the cells with round cytoplasmic granules were positive for nonspecific esterase and the cells with ellipsoidal granules were positives for acid phosphatase, alkaline phosphatase, nonspecific esterase and peroxidase. The results show that these cells are distinct cells and that the cells with ellipsoidal cytoplasmic granules have the same histoenzymological characteristics as the neutrophils/heterophils of mammalians and other vertebrates.

The thrombocyte aggregation process in the turtle Phrynopys hilarii (Chelonia). An ultrastructural study.

This study investigates the thrombocyte aggregation process in the South American fresh water turtle (Phrynopys hilarii) using electron microscopy. Blood was taken from surgically exposed lateral neck vessels of ten turtles Phrynopys hilarii during the spring and summer seasons, when the mean temperature is 37 degrees C. Blood samples were fixed with Karnovsky solution for processing by transmission electron microscopy. The turtle thrombocytes were spindle-shaped with lobulated nuclei. Prominent vesicles and canaliculi were found throughout the cytoplasm. The cytoplasm organelles showed an agranular endoplasmatic reticulum, Golgi complex near the centrioles and scattered free ribosomes. These cells are similar to bird thrombocytes but distinct from fish and frog thrombocytes. Blood clotting time was 5 min +/- 30 sec measured by the Lee and White method. Structural alterations resulting from the aggregation process occurred after activation. Thrombocytes developed numerous filopodial projections, an increased number of vacuoles and changed from spindle to spherical shape. P. hilarii thrombocytes have different morphologic characteristics compared to other non-mammalian vertebrate cells. These cells can participate in the aggregation process, as observed in birds.

European sea bass (Dicentrarchus labrax L.) cytochrome P450arom: cDNA cloning, expression and genomic organization.

Cytochrome P450arom, a key enzyme in the hormonal steroidogenic pathway, mediates the conversion of androgens to estrogens. This work describes the molecular cloning of the cDNA encoding the European sea bass (Dicentrarchus labrax L.) cytochrome P450arom by means of reverse transcriptase and polymerase chain reaction (RT-PCR) and 5' and 3'-rapid amplification of cDNA ends (RACE) analyses. The cDNA is 1822bp in length and encodes a putative protein of 517 amino acids. Northern blot analysis revealed that the ovary expressed a transcript of about 2.2kb in size. Analysis of the deduced amino acid sequence indicated 62-86% identity with ovarian P450arom of other teleost fish, the highest identity being found with the Japanese flounder, Paralichthys olivaceous. Identity was lower (56-65%) with the P450arom forms first reported in teleost brain. Only 52% identity was observed with the corresponding fragment of the cartilaginous fish, Dasyatis sabina. RT-PCR revealed that the sea bass P450arom mRNA was also expressed, at low levels, in testis and brain. Between the 5' and 3'-untranslated terminal regions (UTR), the sea bass CYP19 gene contains eight introns. All introns conform to the GT/AG rule for RNA splicing and are inserted in exactly the same positions as those found in Oryzias latipes and the human CYP19 gene.

Localization of peroxidase activity in blood mononuclear phagocytes in pacu fish (Piaractus mesopotamicus).

The localization of peroxidase activity in different cell regions is used as a criterion for classifying the stage of maturity of mammalian mononuclear phagocytes, with a positive peroxidase reaction indicating the presence of monoblasts, promonocytes, monocytes, and macrophages. Peroxidase activity was observed ultrastructurally in the circulating blood of pacu fish (Piaractus mesopotamicus), identifying monoblasts, promonocytes, monocytes, and macrophages. These observations suggest that differentiation of mononuclear phagocytes occurs in the blood circulation of fish, whereas in mammals, monoblasts and promonocytes are detected in bone marrow, with only monocytes detected in circulating blood and differentiation into macrophages occurring in other body compartments.

Effects of castration and testosterone replacement on peritoneal histamine concentration and lung histamine concentration in pubertal male rats.

Mast cells, which are the main source of histamine, are significantly affected by sex steroids. The present study was undertaken to determine the effects of bilateral castration and testosterone replacement on peritoneal histamine concentration and lung histamine concentration in pubertal male rats (Wistar strain). Three groups of animals were used in this study: (1) untreated castrated animals, (2) castrated animals subjected to androgen replacement by injection of propionate of testosterone, and (3) intact males as a control group. Castration alone produced a dramatic reduction in peritoneal histamine concentration. In addition, androgen replacement was effective in restoring the histamine concentration to the normal value detected in the control males (P<0.05, Kruskal-Wallis test). On the other hand, there was no significant variation in the lung histamine concentration between control males, untreated castrated males and castrated males that received androgen replacement (P<0.05, Kruskal-Wallis test). These results demonstrate for the first time that castration markedly reduces the peritoneum histamine concentration in pubertal male rats, and testosterone replacement prevents the decrease. Further, these procedures do not affect lung histamine concentration, demonstrating that mast cells from different tissues may respond differently to the same biological factors.

Endocytic activity in the thrombocytes of the turtle Phrynopys hilarii (freshwater South American species).

The phagocytic process in cells depends on lysosomal enzymes, high-energy metabolism and cellular recognition. In this paper, we investigated the presence of energy and recognition factors in thrombocytes of turtle Phrynopys hilarii (a freshwater South American species). Turtle thrombocytes (P. hilarii) present glycogen - possibly beta particles - dispersed in their cytoplasm and glycoproteins in the cell surface, as well as a large number of enzymes involved in the endocytic process (Pellizzon, 1996). The activity of these enzymes depends on high-energy metabolism and on cellular recognition provided by specific glycoconjugates (Alberts et al., 1994). This metabolic characterization is demonstrated by the large amount of glycogen particles observed in the cytoplasm by Thiéry's method. Glycogen labeling was also observed when concanavalin A-peroxidase was used as a marker for thrombocytes and for endocyted charcoal particles. Our results show that these cells have phagocytic ability, suggesting that their function in blood circulation is not limited to aggregation but may also involve a great potential for phagocytosis.

Immunocytochemical localization of chymase to cytoplasmic vesicles after rat peritoneal mast cell stimulation by compound 48/80.

The subcellular events responsible for release of mediators by mast cells may help to clarify roles for mast cells in health and disease. In this study we show that the granule-associated protease chymase is also within cytoplasmic vesicles in appropriately stimulated rat peritoneal mast cells. Rat peritoneal mast cells were recovered before or 1-10 sec after exposure to the secretogogue compound 48/80 (10 micrograms/ml) and then were examined by radioimmunoassay to quantify histamine release or were processed, using routine methods for postembedding immunoelectron microscopy, to identify the subcellular localization of chymase. In comparison to unstimulated cells, compound 48/80 stimulated cells in two independent experiments showed an increase (15%, 28%) in the surface area of the cell and a decrease (12%, 6%) in the surface area of the total granule compartment before degranulation channel formation. These global cellular changes occurred in a background of transient but significant (p < 0.01) increases in the area and number of chymase-immunoreactive vesicles per microns2 cytoplasm. These changes were detectable at 5 or 7 sec after stimulation with compound 48/80 but returned to near prestimulation levels by 9 or 10 sec after addition of compound 48/80 (total cumulative histamine release was 28% by 8 sec and 47% by 14 sec). These observations suggest that vesicles participate in the early stages of regulated secretion of chymase from rat peritoneal mast cells.

Male gonadal denervation by guanethidine at pre-puberty: different doses, different results. The intervention of the pineal deafferentation.

Since gonadal denervation and pineal deafferentation by cervical superior ganglionectomy affect sexual development, this study was performed to evaluate testicular steroidogenesis, spermatogenesis and the cervical superior ganglion (CSG) histology in rats treated with guanethidine (GD). The treatment was performed by GD s.c. injections for 3 weeks, from the 21st day of age to the 41st day of age (pre-puberty), when the animals were sacrificed. Different doses were used: group A = 10 mg/kg/day, group B = 50 mg/kg/day and saline (control group). Testicular denervation was confirmed by HPLC for catecholamines in testicular tissue. Testicular concentrations (TC) of progesterone (P4) and testosterone (T) were measured by RIA. Significantly higher TC of P4 and lower TC of T were observed only in group A in comparison with group B and the control group. No alteration of sperm production was observed in either treated group. Histological analysis of CSG showed only few neuronal alterations in group A rats, while in group B the nervous cells were practically destroyed. This suggests that 10 mg/kg/day GD treatment probably produces a specific blockade of 17 alpha-hydroxylase/17,20 desmolase at pre-puberty leading to a decrease of the androgen production. However, in the 50 mg/kg/day group no differences were observed concerning the steroid profiles, this result being attributed to the extensive damage to the CSG observed only in group B. The CSG destruction causes deafferentation of the pineal gland producing abolishment of the inhibition of the 17 alpha-hydroxylase/17,20 desmolase promoted by melatonin or by an out of phase production of androgen.

Ovarian granulosa and theca interstitial cells: a morphological and physiological analysis in guanethidine denervated rats at pre-puberty.

Since ovary denervation causes delayed puberty, we investigated the relative importance of ovary innervation on the morphology and physiology of theca interstitial cells (TIC) and granulosa cells (GC) in female rats at pre-puberty. Elimination of the sympathetic innervation was performed by long term post natal treatment with guanethidine (GD), an adrenergic blocking agent. The sympathectomized rats exhibited: reductions in follicular volume (40%), granulosa cells area (43%) and theca interstitial cell volume (50%). Ovarian concentrations of pregnenolone (P5) and progesterone (P4) were decreased whereas no differences were observed in androstenedione (A) and estradiol (E2). The intensity of the immunocytochemical reaction for 3 beta hydroxysteroid dehydrogenase (3 beta-HSD) detected only in interstitial cells, did not show any difference. These in vivo results include the TIC in the bulk of ovarian structures affected by GD denervation at pre-puberty as it was already observed for GC. The reduced area/volume occupied by these cells in the GD treated ovary is associated to a blockade of the initial steps of the steroidogenic pathway, probably at the level of the cholesterol side chain cleavage enzyme (P450 s.c.c.), previously to P5 synthesis, since P5 is reduced. Similar intra ovarian concentrations of androgens are discussed in terms of possible pineal deafferentation promoted by GD at high doses.