Distribution patterns of PCNA and ANP in perinatal stages of the developing rat heart

Distribution patterns of proliferating cell nuclear antigen (PCNA) and atrial natriuretic peptide (ANP) were determined immunohistochemically and morphometrically in atrium and ventriculum of the developing rat heart in different stages of the perinatal period. During the prenatal period, PCNA and ANP were localized in opposite patterns, particularly in trabecular myocytes. A distinct reduction in the percentage of PCNA-positive nuclei was detected starting at day 19 of the prenatal period, and these cells were rarely observed on postnatal days 30 and 60. In cardiomyocytes, a distinct increase in ANP positivity was found, whereas PCNA positivity was very low. It is concluded that PCNA expression gradually decreased from prenatal day 19 onwards, whereas ANP expression increased in atria throughout the prenatal and postnatal periods, except for a decrease in ANP expression in ventricles from prenatal day 21 onwards. The opposite expression patterns of PCNA and ANP in trabecular myocytes of ventricles indicate that ANP may have antimitogenic/antiproliferative effects in trabecular myocytes.     

Localization of NGF and nNOS in varicocele-induced rat testis

Nerve growth factor (NGF) is synthesized in male germ cells. The presence of neuronal nitric oxide synthase (nNOS) in Leydig cells is related to its role in the regulation of testosterone release. Varicocele is often characterized by abnormal sperm quality and influences the fertilizing capacity of the haploid gamete. We investigated the localization of NGF and nNOS in testes of adult Wistar rats with experimentally induced varicocele after 9, 11, and 13 weeks, as well as in sham-operated controls by immunohistochemistry and Western blot. In control testis, we detected NGF in nuclei of Sertoli cells and also as small vesicular-like structures in the cytoplasm of primary spermatocytes, and in round and elongating spermatids. Varicocele-induction revealed a slight decrease of NGF at 13 weeks, especially in Sertoli cells. In control tissue, nNOS protein was present mainly in Leydig cells and in Sertoli cell cytoplasm. Additionally, nNOS immunoreactivity was present in the heads of elongated spermatids. Western blot results revealed that the decrease of NGF was not significant in the13-week varicocele group, moreover, the amount of nNOS was not altered in any of the varicocele groups. In conclusion, NGF and nNOS have important roles for normal gametogenesis and our data for the first time indicates that varicocele induction does not necessarily affect the expression of NGF and nNOS. Thus, these two molecules do not appear to be related to varicocele induction.     

Hyaluronic acid binding ability of human sperm reflects cellular maturity and fertilizing potential: selection of sperm for intracytoplasmic sperm injection

PURPOSE OF REVIEW: The current concepts of sperm biochemical markers and the central role of the HspA2 chaperone protein, a measure of sperm cellular maturity and fertilizing potential, are reviewed. RECENT FINDINGS: Because HspA2 is a component of the synaptonemal complex, low HspA2 levels and increased frequency of chromosomal aneuploidies are related in diminished maturity sperm. We also suggest a relationship between HspA2 expression in elongating spermatids and events of late spermiogenesis, such as cytoplasmic extrusion and plasma membrane remodeling that aid the formation of the zona pellucida binding and hyaluronic acid binding sites. The presence of hyaluronic acid receptor on the plasma membrane of mature sperm, coupled with hyaluronic acid coated glass or plastic surfaces, facilitates testing of sperm function and selection of single mature sperm for intracytoplasmic sperm injection. The frequencies of sperm with chromosomal disomy are reduced approximately fourfold to fivefold in hyaluronic acid selected sperm compared with semen sperm, comparable to the increase in such abnormalities in intracytoplasmic sperm injection offspring. Hyaluronic acid binding also excludes immature sperm with cytoplasmic extrusion, persistent histones, and DNA chain breaks. SUMMARY: Hyaluronic acid mediated sperm selection is a novel technique that is comparable to sperm zona pellucida binding. Hyaluronic acid selected sperm will also alleviate the risks related to intracytoplasmic sperm injection fertilization with sperm of diminished maturity that currently cause worldwide concern.     

Immunohistochemical distribution patterns of collagen type II,chondroitin 4-sulfate,laminin and fibronectin in human nasal septal cartilage

Collagen type II, chondroitin 4-sulfate, laminin and fibronectin are major components of cartilage matrix. It is important to know their distribution patterns to evaluate relationships between cartilage cells and surrounding cartilage matrix. In the present study, we investigated localization patterns of these macromolecules in human nasal septal cartilage by immunohistochemical methods. Samples of human nasal septal cartilage were obtained from patients with nasal septum deviations who underwent septoplastic operation and were prepared for immunohistochemical examination. Distribution patterns of cartilage matrix macromolecules correlated with those found in other cartilage tissues. Diffuse staining of collagen type II was found in the cartilage matrix, chondroitin 4-sulfate immunostaining was present in the cytoplasm and like a pericellular ring around chondrocytes. Laminin immunostaining was found in the cytoplasm of chondrocytes, and fibronectin was localized in the pericellular matrix and in capsules of human nasal septal cartilage. Moreover, fibronectin was also detected at high levels in the interconnecting segments between adjacent chondrons. In conclusion, similar localisation patterns of the components investigated in human septal cartilage as in other tissues indicate that these macromolecules may play a role in both cell-matrix adhesion and matrix-matrix cohesion in the pericellular microenvironment surrounding nasal septal cartilage chondrocytes as in other cartilage tissues.     

Expression of CCM2 and CCM3 during mouse gonadogenesis

Purpose

Three cerebral cavernous malformation (CCM) proteins, CCM1, CCM2, and CCM3, regulate cell-cell adhesion, cell shape and polarity, and most likely cell adhesion to extracellular matrix. Recently, CCM2 and CCM3 are known to be expressed in control and varicocele-induced rat testes, but little is known about these proteins during gonadogenesis. This led us to study the CCM proteins during the mouse gonadogenesis.

Methods

Neonatal (PND 0), postnatal, and adult mice testes and ovaries were obtained from mice. CCM2 and CCM3 expression were analyzed during mouse testicular and ovarian development by immunohistochemistry and quantitative real-time PCR.

Results

The results showed that in both sexes, Ccm2 and Ccm3 mRNA and protein were first detectable after gonadogenesis when the gonads were well differentiated and remained present until the adult stage. In the testis, CCM2 and CCM3 expression were restricted to the nuclei of Sertoli cells, suggesting a conserved role in testicular differentiation. In the ovary, the CCM2 and CCM3 proteins were localized in the cytoplasm of oocytes, suggesting an unexpected role during oogenesis. Quantitative real-time PCR (qRT-PCR) results showed that expression of Ccm2 and Ccm3 genes could play a role in the regulation of mouse gonadogenesis translational activation upon testicular and ovarian development.

Conclusions

The localization of CCM2 and CCM3 proteins show their different functions for CCM2 and CCM3 which may have important roles in testicular and ovarian differentiation. In conclusion, CCM2 and CCM3 may be involved in establishing the differential expression pattern in developing mouse testis and ovary.     

A case of lupus vulgaris with unusual location

Tuberculosis (TB) is one of the oldest diseases of our planet; today, it still is a serious medicosocial problem in both developing and developed countries. Cutaneous tuberculosis is part of the small percentage of extrapulmonary forms of the disease and has considerable morphological variability. It is often confused with various cutaneous disorders and some other granulomatous processes of the skin. Here, we present a case of long-standing and atypically-located lupus vulgaris in an old man. The patient was successfully treated with triple antituberculous therapy.