Morphological changes in distant muscle fibers following thermal injury in Wistar rats

Purpose: Thermal injury causes catabolic processes as the body attempts to repair the damaged area. This study evaluated the effects of a scald injury on the morphology of muscle fibers belonging to a muscle distant from the lesion. Methods: Thirty Wistar rats were divided into control (C) and scalded (S) groups. Group S was scalded over 45% of the body surface, standardized by body weight. Rats in both groups were euthanized at four, seven and 14 days following the injury. The middle portions of the medial gastrocnemius muscles were sectioned, stained with hematoxylin and eosin and Picrosirius, and submitted to histological analysis. Results: Control group sections exhibited equidistantly distributed polygonal muscle fibers with peripheral nuclei, characteristic of normal muscle. The injured group sections did not consistently show these characteristics; many fibers in these sections exhibited a rounded contour, variable stain intensities, and greater interfiber distances. A substantially increased amount of connective tissue was also observed on the injured group sections. Conclusion: This experimental model found a morphological change in muscle distant from the site of thermal injury covering 45% of the body surface.

Seasonal metabolic depression, substrate utilisation and changes in scaling patterns during the first year cycle of tegu lizards (Tupinambis merianae)

The tegus increase in body mass after hatching until early autumn, when the energy intake becomes gradually reduced. Resting rates of oxygen consumption in winter drop to 20% of the values in the active season (Vo(2)=0.0636 ml g(-1) h(-1)) and are nearly temperature insensitive over the range of 17-25degreesC (Q(10)=1.55). During dormancy, plasma glucose levels are 60% lower than those in active animals, while total protein, total lipids and beta-hydroxybutyrate are elevated by 24%, 43% and 113%, respectively. In addition, a significant depletion of liver carbohydrate (50%) and of fat deposited in the visceral fat bodies (24%) and in the tail (25%) and a slight loss of skeletal muscle protein (14%) were measured halfway through the inactive period. Otherwise, glycogen content is increased 4-fold in the brain and 2.3-fold in the heart of dormant lizards, declining by the onset of arousal. During early arousal, the young tegus are still anorexic, although Vo(2) is significantly greater than winter rates. The fat deposits analysed are further reduced (62% and 45%, respectively) and there is a large decrease in tail muscle protein (50%) together with a significant increase in glycogen (2-3-fold) and an increase in plasma glucose (40%), which suggests a role for gluconeogenesis as a supplementary energy source in arousing animals. No change is detectable in citrate synthase activity, but beta-hydroxyacyl CoA dehydrogenase activities are strongly affected by season, reaching a Mold and 5-fold increase in the liver tissue of winter and arousing animals, respectively, and becoming reduced by half in skeletal muscle and heart of winter animals compared with late fall or spring active individuals. From hatching to late autumn, the increase of the fat body mass relatively to body mass is disproportionate (b=1.44), and the mass exponent changes significantly to close to 1.0 during the fasting period. The concomitant shift in the Vo(2) mass exponent in early autumn (b=0.75) to values significantly greater than 1.0 in late autumn and during winter dormancy indicates an allometric effect on the degree of metabolic depression related to the size of the fat stores and suggests greater energy conservation in the smaller young.

Structure of the posthepatic septum and its influence on visceral topology in the tegu lizard, Tupinambis merianae (Teiidae : Reptilia)

The posthepatic septum (PHS) divides the body cavity of Tupinambis merianae into two parts: the cranial one containing the lungs and liver and the caudal one containing the remaining viscera. The PHS is composed of layers of collagenous fibers and bundles of smooth muscle, neither of which show systematic orientation, as well as isolated blood vessels, lymphatic vessels, and nerves. Striated muscle of the abdominal wall does not invade the PHS. The contractions of the smooth muscles may stabilize the pleurohepatic cavity under conditions of elevated aerobic needs rather than supporting breathing on a breath-by-breath basis. Surgical removal of the PHS changes the anatomical arrangement of the viscera significantly, with stomach and intestine invading the former pleurohepatic cavity and reducing the space for the lungs, Thus, the PHS is essential to maintain the visceral topography in Tupitionibis. J. Morphol. 258:151-157, 2003. (C) 2003 Wiley-Liss. Inc.

Biological behavior of the gerbil ventral prostate in three phases of postnatal development

In this study, we characterized the gerbil's ventral prostate histology ultrastructurally and quantitatively throughout three phases of postnatal development (young, adult, and old) in order to comprehend its biological behavior and propensity to developing spontaneous lesions with aging. The gerbil prostate is composed of alveoli and ducts immersed in a stroma composed of smooth muscle, fibroblasts, collagen and elastic fibers and vessels. The prostate tissue components present morphological and quantitative aspects that vary according to age. Young animals have an immature gland with modest secretory activity. Synthetic activity remained stable in adult and old gerbil. However, prostatic morphology was altered in the aging, showing an increased epithelium and stromal fibrosis. The nuclei of the secretory cells increased with aging, whereas nucleoli presented few alterations during postnatal development. The epithelial proliferation and stromal remodeling noted in this study indicate that the gerbil prostate may respond to the androgen declines typical of senescence through epithelial proliferation and stromal remodeling.

Morphological and autoradiographic studies on the corneal and limbal epithelium of rabbits

The investigation was centered on the morphological features of the conjunctiva-cornea transition (limbus) of the rabbit eye and the proliferative behavior of its epithelium. The eyes were processed for examination with light and electron microscopy, as well as for autoradiography after intravitreal injection of [H-3]thymidine ([H-3]TdR). At the sites of extraocular muscle insertion, the vascularization of the stroma extended to the peripheral cornea, and the limbal epithelium was thin with its basal stratum made up by clear cuboidal cells. In between the muscle insertions, the cuboidal clear cells, as well as the stroma blood vessels; were scarce. At the light microscope level, the basement membrane was distinct in the cornea but not in the limbus or the conjunctiva. Autoradiographs demonstrated that, at the limbus, the basal cells migrated very quickly to the suprabasal region and remained there up to the 28-day interval. Labeled cells were identified in all epithelial layers of the cornea, including the basal one, at 21 and 28 days but not in the limbal basal clear cells. The rate of renewal of conjunctival epithelium was similar to that observed for the transition with scarce clear cells. The high-resolution autoradiographs demonstrated that the basal cuboidal clear limbal cells exhibit a quick renewal and that they are not label-retaining cells. These latter ones were detected all over the corneal epithelium and in the suprabasal layers of the limbus up to 28 days, in physiological conditions, without the need of stimulation by damage to the corneal epithelium.

Production and body composition traits of broilers in relation to breast weight evaluated by path analysis

O peso do peito possui grande importância econômica na indústria de frangos, podendo estar associado a outras variáveis passíveis de seleção. Estimaram-se correlações fenotípicas entre características de desempenho (peso vivo aos 7, 28 dias e ao abate e profundidade de músculo peitoral por ultra-sonografia), carcaça (peso eviscerado e de pernas) e composição corporal (peso do coração, do fígado e da gordura abdominal), em uma linhagem de frangos, e quantificou-se a influência direta e indireta destas variáveis sobre o peso do peito. Para tanto, utilizou-se a análise de trilha, desdobrando-se a matriz de correlações parciais em coeficientes que forneceram a influência direta de uma variável sobre a outra, independentemente das demais. A manutenção das variáveis peso vivo ao abate e peso eviscerado na matriz de correlações pode ser prejudicial à análise estatística que envolve os sistemas de equações normais, como a análise de trilha, devido à multicolinearidade observada. O peso vivo ao abate e a profundidade do músculo peitoral por ultra-sonografia apresentaram efeitos diretos importantes sobre o peso de peito e foram identificadas como as principais responsáveis pela magnitude dos coeficientes de correlação obtidos. Assim, uma pré-seleção individual para estas características pode favorecer um aumento no peso de peito nesta linhagem, se mantidas as condições ambientais de criação dos frangos, uma vez que o peso vivo ao abate e a profundidade de músculo peitoral por ultra-sonografia afetam fenotipicamente e diretamente o peso de peito.

DISTRIBUTION OF DIFFERENT FIBER TYPES OF M-EXTENSOR CARPI RADIALIS LONGUS OF THE RAT

As revealed by the NADH-diaphorase and myosine ATPase, the M. extensor carpi radialis longus of the rat possesses at least 3 main kinds of fibres, with different distribution on the superficial and deep portions of the muscle. The superficial portion revealed that 67.68 % are FG (fast-twitch-glycolytic) fibres, 14.72 % are FOG (fast-twitch-oxidative) fibres and 17.60 % are SO (slow-twitch-glycolytic) fibres. Already the deep portion revealed that 71.29 % are SO (slow-twitch-glycolytic) fibres, 17.46 % are FOG (fast-twitch-oxidative-glycolytic) fibres and 11.25 % are FG (fast-twitch-glycolytic) fibres. The miosine ATPase reaction was used to demonstrate contracting characteristics. These findings suggest that the movements of fast contraction of the M. extensor carpi radialis longus are predominant.

Cytoarchitecture and musculotopic organization of the facial motor nucleus in Cebus apella monkey

The architecture and musculotopic organization of the facial motor nucleus in the Cebus apella monkey (a New World primate) were investigated using histological techniques and a multiple labelling strategy, in which horseradish peroxidase-conjugated neuroanatomical tracers (CTB-HRP and WGA-HRP) and fluorescent tracers were injected into individual facial muscles. The facial motor nucleus was formed by multipolar motoneurons and had an ovoid shape, with its rostrocaudal axis measuring on average 1875 mum. We divided the nucleus into four different subnuclei: medial, intermediate, dorsal and lateral. Retrograde labelling patterns revealed that individual muscles were innervated by longitudinal functional columns of motoneurons. The columns of the orbicularis oculi, zygomaticus, orbicularis oris, auricularis superior, buccinator and platysma muscles were located in the dorsal, intermediate, lateral, medial, lateral and intermediate subnuclei, respectively. However, the motoneuron columns of the levator labii superioris alaeque nasi muscle and frontalis muscle could not be associated with a specific subnucleus. The present results confirm previous studies regarding the musculotopic organization of the facial motor nucleus. However, we observed some particularities in terms of the relative size of each column in C. apella, which might be related to the functional and behavioral importance of each muscle in the particular context of this primate.

Elastic system of the rat ventral prostate and its modifications following orchiectomy

BACKGROUND. The extracellular matrix (ECM) has important roles in prostatic development, and marked stromal changes take place in the rat ventral prostate (VP) after androgen deprivation. However, little knowledge exists about individual ECM components.METHODS. The distribution of elastic fibers (EF) and elastic-related fibers (ERF) in the VP of castrated and control rats was investigated, using histochemistry and transmission electron microscopy (TEM).RESULTS. EF are barely detected in the prostatic stroma, but ERF are relatively abundant. Castration results in a relative increase in the number and thickness of ERF. TEM showed an open network of ECM microfibrils throughout the stroma and thin and short EF, which increase in number and thickness after orchiectomy.CONCLUSIONS. The presence of elastic system components in the rat VP warrants the deformability required for the secretion exclusion under the action of smooth muscle cells, and the castration-induced modification may be related to the contraction of the tissue and maintenance of peculiar arrangements of other ECM components. (C) 1997 Wiley-Liss, Inc.

Analysis of the inflammatory reaction induced by the catfish (Cathorops spixii) venoms

Cathorops spixii is one of the most abundant venomous fish of the southeastern coast of the State of São Paulo, and consequently causes a great part of the accidents seen there. The accidents affect mainly fishermen, swimmers and tourists and are characterized by punctiform or wide wounds, erythema, edema, pain, sudoresis, indisposition, fever, nausea, vomiting and secondary infection. The objective of this work was to characterize the inflammatory response induced in mice by both venoms (mucus and sting) of the catfish C spixii. Our results demonstrated that both venoms induced a great number of rolling and adherent leukocytes in the post-capillary venules of cremaster muscle of mice, and an increase in the vascular permeability in peritoneal cavity. Mucus induced the recruitment of neutrophils immediately after injection followed later by macrophage infiltration. In contrast, the cellular infiltration elicited by sting venom was rapidly resolved. The peritonitis reaction provoked by venoms was characterized by cytokine (IL-6), chemokines (MCP-1 and KC) or lipid mediator (LTB4) production in the peritoneal cavity. The macrophages from 7-day mucus venom-induced exudates upon in vitro mucus venom stimulation, expressed CD1 Ic x MHC class II and release bioactive IL-12p70. on the other hand, sting venom-elicited peritoneal macrophages lost the ability to differentiate into dendritic cells, following re-stimulation in vitro with sting venom, they do not express CD11c, nor do they exhibit sufficient levels of MHC class II. In conclusion, both types of venoms (mucus or sting) promote inflammatory reaction with different profiles, and the inflammatory reaction induced by the first was characterized by antigen persistence in peritoneal cavity that allowed the activation of phagocytic cells with capacity of antigenic presentation. (C) 2007 Elsevier Ltd. All rights reserved.

Automated NMR structure determination and disulfide bond identification of the myotoxin crotamine from Crotalus durissus terrificus

Crotamine is one of four major components of the venom of the South American rattlesnake Crotalus durissus terrificus. Similar to its counterparts in the family of the myotoxins, it induces myonecrosis of skeletal muscle cells. This paper describes a new NMR structure determination of crotamine in aqueous solution at pH 5.8 and 20 degrees C, using standard homonuclear (1)H NMR spectroscopy at 900 MHz and the automated structure calculation software ATNOS/CANDID/DYANA. The automatic NOESY spectral analysis included the identification of a most likely combination of the six cysteines into three disulfide bonds, i.e. Cys4-Cys36, Cys11-Cys30 and Cys18-Cys37; thereby a generally applicable new computational protocol is introduced to determine unknown disulfide bond connectivities in globular proteins. A previous NMR structure determination was thus confirmed and the structure refined. Crotamine contains an alpha-helix with residues 1-7 and a two-stranded anti-parallel beta-sheet with residues 9-13 and 34-38 as the only regular secondary structures. These are connected with each other and the remainder of the polypeptide chain by the three disulfide bonds, which also form part of a central hydrophobic core. A single conformation was observed, with Pro13 and Pro21 in the trans and Pro20 in the cis-form. The global fold and the cysteine-pairing pattern of crotamine are similar to the beta-defensin fold, although the two proteins have low sequence homology, and display different biological activities. (c) 2005 Elsevier Ltd. All rights reserved.

CD34 EXPRESSION IN STROMAL TUMORS OF THE GASTROINTESTINAL-TRACT

Gastrointestinal stromal tumors (GISTs) are mesenchymal tumors that may exhibit varied morphologic appearances (spindle, epithelioid) and biologic potentials. Given the continuing controversy regarding the type of cell differentiation present in these tumors (muscle versus nerve sheath versus null), we evaluated a set of GISTs, most of which had been previously examined for the presence of smooth muscle differentiation, for expression of CD34, a 115 kDa cell-surface progenitor cell marker also recently identified in a subset of mesenchymal tumors. Using antibody My 10 in deparaffinized, formalin-fixed tissue after pretreatment with microwave energy, we found that 46 of 57, or 81%, of GISTs were CD34+; this fraction of CD34+ tumors exceeded the fraction of these same GISTs found to show muscle actin (72%) expression. In addition, a consistently higher fraction of the tumor cell population was CD34+ than was muscle actin positive. These findings suggest that CD34 is a very sensitive marker for the identification of GISTs. CD34 is normally expressed by endothelial as well as perivascular cells, perhaps related to, but distinct from, vascular smooth muscle cells. While the nature of these latter cells is uncertain, the expression of CD34 in such a large fraction of GISTs may provide evidence of a unique differentiation pathway in these tumors.

Tissue tolerance of diclofenac sodium encapsulated in liposomes after intramuscular administration

In this work the effect of the encapsulation of diclofenac sodium within liposomes on the reduction of the myotoxicity after intramuscular administration in rats was studied. Diclofenac sodium was encapsulated in small unilamellar liposomes obtained from phosphatidylcholine, cholesterol, and a-tocopherol (40:10:0.04 mM), and administered by intramuscular injection in the quadriceps femoral muscle of male Wistar rats. After a single dose of 0.2 mg diclofenac formulations the local tissue damage was assessed by plasma creatine kinase (CPK) activity and histological analysis. It was demonstrated that formulations containing free diclofenac produced a higher increase in CPK activity, while those encapsulated in liposomes exhibited CPK activity similar to the control groups. Histopathological analysis of local muscle tissue performed on the third and seventh days following the injection showed intense cellular damage when free drug solution was used, while encapsulation in liposome protected the tissue against the local tissue inflammation.

Treatment of urinary stress incontinence by intravaginal electrical stimulation and pelvic floor physiotherapy

Treatment of urinary stress incontinence (USI) by intravaginal electrical stimulation (IES) and pelvic floor physiotherapy represents an alternative to other therapies. The purpose of this work was to evaluate the effectiveness of this treatment inpatients with urinary incontinence. From January 1998 to May 2000, 30 women (mean age 54 years) were studied. All patients had USI and 70% urge incontinence; average follow-up was 7 months. Selection criteria were based on clinical history, objective evaluation of perineal musculature by perineometry, and urodynamics. The treatment protocol consisted of three sessions of IES per week for 14 weeks using INNOVA equipment. Physiotherapy was initiated in the fifth week of IES. A significant decrease in the number of micturitions and urgency was observed after treatment (P<0.01). The pad test showed a reduction in urinary leakage from 13.9 to 5.9 g after treatment (P<0.01). Objective evaluation of perineal muscle strength showed a significant improvement in all patients after treatment (P<0.01). A positive correlation was observed between maximum flow rate (Q(max)) and all three variables: urethral pressure profile at rest and on straining (stop test), and abdominal leak-point pressure (ALPP). A positive correlation was also observed between ALPP and the stop test. Over 100 different surgical and conservative treatments have been tried to manage USI. The majority of these procedures reveal that despite progress already made in this area, there is no ideal treatment. Satisfactory results can be achieved with this method, especially with patients who are reluctant to undergo surgery because of personal or clinical problems.

Histochemical and SEM evaluation of the neuromuscular junctions from alcoholic rats

In the present study morphological changes occurring in the neuromuscular junctions (NMJ) of the extensor digitorum longus (EDL) and soleus muscles from albino rats (Rattus norvegicus) submitted to experimental chronic alcoholism were evaluated. Seventy two male animals aged 4 months and weighing on average 400 g were divided into three groups: control, alcoholic and isocaloric. Six rats from each group were anesthetized and sacrificed after 5, 10, 15 and 18 months. The NMJ did not show detectable morphological changes in either muscle after treatment when examined by light microscopy. With respect to the dimensions, statistical analysis demonstrated a tendency to a statistically significant treatment x time interaction for the length of soleus muscle NMJ. The ultrastructural study, however, revealed that the NMJ of the soleus muscle of animals submitted to 18 months of experimental alcoholism presented important morphological alterations. Characteristically, the NMJ of these muscles is located on an elevation on the surface of the muscle fiber, presenting a regular round, oval or elliptical shape and continuous and not very deep synaptic grooves. Approximately 30% of the NMJ of alcoholic rats are irregular in shape, with the sarcolemmal elevations typical of the synapse region being flattened on at least one side, with discontinuous synaptic grooves, and deep and punctiform contacts of the synaptic buds. These data suggest that, although skeletal muscle has a greater natural resistance against the direct or indirect effects of alcohol, some submicroscopic morphological alterations are detectable in the NMJ, especially in muscles with oxidative metabolism (soleus) following long periods of ingestion of alcohol. (C) 2002 Elsevier B.V. Ltd. All rights reserved.