Distribution of extracellular matrix related proteins in normal and cryptorchid ziwuling black goat testes

The Ziwuling black goat is an indigenously in China, their offspring are frequently affected by congenital cryptorchidism. The extracellular matrix (ECM) contains cytokines and growth factors that regulate the development of the testis, and component changes often result in pathological changes. Cryptorchidism is closely related to structural changes in ECM. In this study, the histochemical staining, immunohistochemical, immunofluorescence and Western blot combined with semi-quantitative analysis was used to describe the distribution of the important ECM components Collagen type IV (Col IV), laminin (LN)and heparan sulfate proteoglycans (HSPG) in the normal and cryptorchid testes of Ziwuling black goats. Results showed that: The histochemical staining showed that the dysplasia of seminiferous tubules and decreased number of Sertoli cells in cryptorchidism, as well as sparse collagen fiber. Meanwhile, the distribution of reticular fibers is relatively rich. Furthermore, the PAS and AB staining in the interstitial vessels and lamina propria of seminiferous tubules is weak. The immunohistochemical and immunofluorescence revealed that Col IV, LN was strongly expressed in Leydig, Sertoli cells of normal testes and moderately positive in the spermatogonia and spermatids, but HSPG was not expressed in the spermatogonia. However, cryptorchidism, the expression of Col IV, LN and HPSG in Leydig, Sertoli cells significantly decreased, as well as the expression of Col IV and LN in capillary endothelial cells, but HSPG was moderately expressed in spermatogonia. Based on these data, the underdevelopment of spermatogenic epithelium, decreased synthesis function of collagen fibers and Leydig cells develop usually in the cryptorchidism were shown to be closely related to the abnormal metabolism of Col IV and LN. The positive expressed of HSPG in the spermatogonia of cryptorchid testes is related to the compensatory development of spermatogonia.     

Potential role of cyanidin 3-glucoside (C3G) in diabetic cardiomyopathy in diabetic rats: An in vivo approach

The present study aimed to evaluate the importance of cyanidin 3-glucoside (C3G) of diabetic cardiomyopathy in diabetic rats. The rats were induced with diabetic using streptozotocin and total triglyceride (TG) and total cholesterol (TC) were determined. The range of myocardial enzymes such as aspartate aminotransferase (AST), creatine kinase (CK) and lactate dehydrogenase (LD) were also estimated, further, the Immuno histochemical analysis and western blot investigation were determined for the actual activity of C3G. Results indicated that the marker enzymes such as CK, LD and AST were significantly (P < 0.05) increased in STZ administered rats (DM group), while the levels of these elevated marker enzymes of cardiac injury significantly (P < 0.05) declined in the DM + C3G group, as compared to the diabetic group of rats. Additionally, a decrease in the level of TNF-alpha and interleukin-6, was noticed in the C3G treated group as compared to diabetic group. Finally, blotting analysis clearly confirmed that theC3G treatment resulted to higher level response of Bcl-2 and lower level response of caspase-3 and BAX. In conclusion, C3G a natural antioxidant may prevent cardiovascular complications by ameliorating oxidative damage, inflammation, metabolic dysfunctions and apoptosis pathways in type 2 diabetes.     

Anatomical substrate for neurotensin-acetylcholine interactions in the rat basal forebrain

We have previously shown by combined radioautography and acetylcholinesterase histochemistry that the distribution of ¹²⁵I-neurotensin (NT) binding sites was in register with that of cholinergic neurons in the rat nucleus basalis magnocellularis (NBM). The present study utilized three experimental approaches to elaborate on the type and cellular localization of NT binding sites in the NBM. Competition studies using levocabastine, a selective blocker of the low affinity NT binding component, revealed that most of the ¹²⁵I-NT binding sites labeled in the NBM are of the levocabastine-insensitive high affinity type, known to correspond to the physiologically active receptor. Ibotenic acid-induced lesions of the NBM produced a marked reduction in both cholinesterase reactivity and cellular ¹²⁵I-NT binding suggesting that most of the labeled sites are associated with the cholinergic neurons themselves rather than with an afferent input to those cells. Finally, examination of the high resolution radioautographic distribution of ¹²⁵I-NT binding sites in semithin sections revealed that a proportion of ¹²⁵I-NT-labeled receptors is associated with the plasma membrane of magnocellular perikarya and proximal processes, thereby providing an anatomical substrate for a local action of NT in the NBM.     

Immune- and enzyme histochemical characterisation of leukocyte populations within lymphoid and mucosal tissues of Atlantic halibut (Hippoglossus hippoglossus)

Leukocyte populations within the kidney, spleen, posterior intestine and gills of Atlantic halibut were investigated using a panel of histological, enzyme- and immunohistochemical methods. In the kidney and spleen, a diverse population of leukocytes was associated with the extensive network of sinusoids and larger blood vessels present in these tissues. IgM+ cells (B-cells, plasma cells and IgM-bearing macrophages) and large mononuclear cells showing reactivity for non-specific esterase (NSE) and acid phosphatase (ACP), representing macrophage populations, were often associated with vessel walls that were also the site of trapping of fluorescent microspheres. In the kidney, trapping of 0.1 and 0.5 microm diameter microspheres occurred at these sites but in the spleen, the 0.1 microm diameter microspheres were retained in ellipsoids. The lymphoid tissues of the kidney and spleen possessed a spread population of 5'-nucleotidase+ (5'N+) cells but compartmentalisation of the splenic white pulp was suggested by an absence of these 5'N+ reticular cells in areas associated with melanomacrophage accumulations and in areas rich in IgM+ cells. A striking feature of the mucosal tissues was the diversity of leukocyte populations within the epithelium particularly of the posterior intestine, including IgM+ cells and NSE+, ACP+ and 5'N+ mononuclear cells. Although limited in numbers in the posterior intestine, IgM+ cells were more common in the epithelium than in the lamina propria. In the gills, leukocytes as detected by enzymatic reactivity were scarce, but IgM+ cells were very abundant in the stratified epithelium of the gill arch and filaments. The difference in distribution of these leukocyte populations between the intestines and gills suggested a compartmentalisation of the mucosal immune system and the need to assess the immunological competence of mucosal tissues in Atlantic halibut.     

Optimizing specimen processing for ancient soft tissue specimens

Abstract

Despite many reports concerning processing of ancient soft tissues, scant attention has been paid to optimizing procedures for processing soft tissues that have been altered by taphonomic processes. To determine the best procedures, we investigated the rehydration solution, time of exposure to the solutions, fixative solution and exposure to heat. Processes were evaluated based on the minimum section thickness, degree of tissue fragmentation, definition of tissue architecture and penetration of stains. We found that in desiccated samples, tissue architecture was optimized by using Ruffer's solution for rehydration and Schaffer's solution as fixative, because these tissues require water restoration within the tissues due to their compacted character. Heating enhanced penetration of dyes in these specimens, which improved diagnosis. Saponified tissues that had suffered extensive decomposition were more labile and required slow water uptake. The best histological sections were obtained using Sandison's solution followed by fixation with formaldehyde and avoiding heat. To obtain the best results with paleohistological specimens, the procedure must be determined by the condition of the sample and by accounting for the nature of its damage.     

海金沙卵发生的组织化学研究

采用组织化学方法对海金沙（Lygodium japonicum （Thunb.） Sw.）卵发生进行了研究。高碘酸-锡夫反应（PAS反应）结果显示：颈沟和腹沟细胞外产生的黏性物质为多糖类物质;在卵细胞发生早期颈卵器细胞内均含有质体,且淀粉粒含量丰富,随着颈卵器的发育,卵细胞以及颈沟和腹沟细胞内淀粉粒数量和体积均逐渐减少,最后退化消失。苏丹黑B反应结果显示：海金沙颈沟和腹沟细胞外产生的黏性物质也含有脂类物质,而卵细胞内并未有明显脂类物质产生;卵细胞质膜处也并没有糖类或脂类物质的积累。本研究从组织化学角度佐证了海金沙的特殊性,即既有原始蕨类的特征,又有进化蕨类的特征。     

中华蟾蜍消化道6种酶的组织化学定位

目的研究中华蟾蜍消化道酸性磷酸酶(ACP)、碱性磷酸酶(ALP)、腺苷三磷酸酶(ATPase)、非特异性酯酶(NSE)、过氧化物酶(POX)和琥珀酸脱氢酶(SDH)等6种酶的分布。方法在消化道的8个部位取材,采用冰冻切片技术、石蜡切片技术、酶的组织化学方法和光密度定量分析。结果 ACP主要分布于胃贲门中贲门腺部,十二指肠和回肠中酶反应呈弱阳性。ALP主要分布于食管、十二指肠至回肠的粘膜上皮,十二指肠酶活性最高。ATPase在消化道各部位均有分布,胃中胃腺部和回肠粘膜上皮酶活性显著较高(P0.05)。NSE和POX在整个消化道粘膜上皮和粘膜固有层均有分布,胃各部位酶活性显著较低(P0.05)。SDH除在食管和直肠酶活性显著较低外,其它部位均有大量分布,十二指肠和回肠酶活性显著较高(P0.05)。结论中华蟾蜍消化道粘膜6种酶的分布同其它动物有相似之处,也有其自身特点。6种酶在消化道中的分布与消化道各部位的生理机能密切相关。     

A polydisperse linear random coil model for the quaternary structure of pig colonic mucin

The distribution of molecular weights for polymeric colonic mucus glycoprotein or ``mucin' isolated and solubilised in the presence of protease inhibitors from pig colons is shown to be considerably greater than its ``subunit' (thiol reduction product) and papain digested forms using the technique of size-exclusion chromatography coupled to multi-angle laser light scattering, and confirmed by sedimentation equilibrium measurements. The conformation of this mucin is probed by examining the molecular weight – intrinsic viscosity relationship in terms of the Mark-Houwink-Kuhn-Sakurada analysis for its polymeric (or ``whole'), reduced and papain-digested forms: an exponent ``a' of (1.1±0.1) is obtained indicating a linear random coil conformation consistent with other mucins. Size-exclusion chromatography coupled to multi-angle laser light scattering is shown to provide a relatively simple complementary technique to sedimentation equilibrium for the molecular weight distribution analysis of polydisperse materials. Received: 29 November 1996 / Accepted: 2 December 1996     

Identification and Pattern of Primary Sensory Cells in the Body Wall Epithelium of the Tubificid Worm, Limnodrilus hoffmeisteri

This study compares the effectiveness of two examination methods suitable for morphological observations, scanning electron microscopy (SEM) and light microscopic NADH-diaphorase (NADP-d) histochemistry, to identify location and distribution pattern of primary sensory cells situated in the body wall epithelium of Limnodrilus hofmeisteri. SEM observations revealed that grouped sensory cells forming two distinct types of sensillas (or sense organs) are scattered over the body surface. The easily identifiable first type (ciliated sensilla) contains numerous penetrative ciliate sensory cells while no ciliate sensory cells were seen in the second type (sensory bud). NADH-d staining proved to be a suitable method to identify both structures further staining solitary sensory cells, and certain sensory fibres that could not be visualised by SEM preparations. Our results show that NADH-d staining, as a histochemical marker of primary sensory cells, is a suitable and effective method to reveal their morphology and distribution pattern in whole mount preparations in the smallest detail, suggesting that the application of this relatively simple and cheap method may contribute to understand the basic organisation and evolution of the peripheral sensory structures of oligochaetes.