Inducible nitric oxide synthase in duodenum of children with Giardia lamblia infection.

The investigation were performed on children with Giardia lamblia infection, diagnosed on the basis of positive stool tests for Giardia antigen (Elisa) or by microscopical detection of trophozoites in duodenal fluid. In duodenal biopsies morphological studies and immunohistochemical reaction for inducible nitric oxide synthase (iNOS) were performed. The control group was made up of duodenal tissue of children with excluded giardiasis and inflammation of the upper part of gastrointestinal tract. The duodenal biopsies from children without Giardia lamblia infection were found to have a high immunoreactivity for iNOS in enterocytes, the cells of intestinal crypts, endothelial cells of vessels and connective tissue cells of lamina propria. In children with giardiasis: in some biopsies the expression of iNOS was as high as in control group, in others was weaker detectable and the shortening of intestinal villi was seen. There were also duodenal biopsies with the lack of immunoreactivity for iNOS, with shorter villi and a large amount of mucus in the intestinal epithelium. Beside of goblet cells, also enterocytes were loaded with mucus. The pathological changes may cause malabsorption and also may have a negative influence on the defense of the intestinal wall against Giardia lamblia infection. The different morphological and immunohistochemical results in the duodenum of children with giardiasis can elucidate a variety of clinical symptoms from asymptomatic to severe infection

Bone marrow morphology during haematopoietic stem cell mobilisation with cyclophosphamide in mice

The aim of the study was to examine the morphology of the bone marrow of mice after stimulation with cyclophosphamide (Cy). The experimental mice were given a single intraperitoneal injection with 250 mg/kg bw cyclophosphamide. After 2, 4 and 6 days of experiment the femurs were obtained for morphological study. On the 2nd day after the mobilisation of the mice with Cy destruction of the bone marrow was observed with a decrease in the haematopoietic compartment and an increase in the area occupied by sinusoids filled with erythrocytes. Erythrocytes were located among the haematopoietic cells, which indicated that the endothelial barrier had been disrupted. On the 4th day after treating the mice with Cy, repair processes in the bone marrow were conducted, including macrophages. The cells filled with haemosiderin migrated from the extravascular compartment of the bone marrow into the lumen of the sinusoids. There were proliferating cells among the haematopoietic cells. On the 6th day the morphology of the bone marrow was similar to the morphology of that in the control mice. However, more haematopoietic cells were visible compared to the control bone marrow. The presence of an increased number of leucocytes in the sinusoid lumen in comparison with the control suggested that at that time the migration of haematopoietic cells from the bone marrow had been initiated

DHT deficiency perturbs the integrity of the rat seminiferous epithelium by disrupting tight and adherens junctions

In rats with a DHT deficiency induced by finasteride, morphological changes in the seminiferous epithelium were observed. The structural alterations were manifested by the premature germ cells sloughing into the lumen of seminiferous tubules. The etiology of this disorder could be connected with intercellular junctions disintegration. We showed in the immunohistochemical study the changes in expression of some proteins building tight and adherens junctions. The depression of N-cadherin, β-catenin and occludin immunoexpressions could be the reason for the release of immature germ cells from the seminiferous epithelium. However, the observed increase of the immunohistochemical reaction intensity of vinculin, one of the cadherin/catenin complex regulators, could be insufficient to maintain the proper function of adherens junctions. The hormonal imbalance appears to influence the pattern of expression of junctional proteins in the seminiferous epithelium. It could lead to untimely germ cells sloughing, and ultimately could impair fertility. (Folia Histochemica et Cytobiologica 2011, Vol. 49, No. 1, 62–71

Possible involvement of microtubules and microfilaments of the epididymal epithelial cells in 17beta-estradiol synthesis.

The rat epididymal epithelial cells revealed features of steroidogenic cells and released 17beta-estradiol (E2) into the culture medium. In steroidogenic cells, elements of the cytoskeleton due to their influence on organelle distribution are implicated in the regulation of steroidogenesis. In the present study, the morphology of cultured epididymal epithelial cells in light, scanning and transmission electron microscopes was evaluated. The organization of microtubules and microfilaments revealed by fluorescence microscopy, and the concentration of E2 in cultured medium were also studied. The epididymal epithelial cells were cultured in different conditions: in the medium with or without exogenous testosterone (T) and in the co-culture with Leydig cells as a source of androgens. The cells in co-culture located close to Leydig cells were rich in glycogen, PAS-positive substances and lipid droplets, in higher amount than the cells cultured with addition of exogenous testosterone. Stress fibers and microtubules of epididymal epithelial cells cultured with exogenous T and in co-culture with Leydig cells presented typical structure, and numerous granular protrusions appeared on the surface of the cells. Disorganization of microtubules and shortening of stress fibers as well as the smooth cell surface deprived of granular protrusions were observed in the epididymal epithelial cells cultured without T. Change of the cytoskeleton organization caused by the absence of androgen in culture medium resulted in an increased E2 secretion

Risk Factors and Symptoms of Meibomian Gland Loss in a Healthy Population

Purpose. The aim of this study was to investigate the relationships between MGL and ocular symptoms, several systemic conditions, and key markers of ocular surface health. Methods. We included into the study 91 healthy volunteers between the ages of 20 and 77 years. We analyzed meibomian gland morphology, function, and lid margin alterations. We correlated our findings with self-reported ocular symptoms, systemic medical history, lifestyle factors, and tear film abnormalities. Results. We observed that a high ocular surface disease index, a history of either chalazion or hordeolum, experience of puffy eyelids upon waking, and foreign body sensation all appeared to be predictors of an abnormal meiboscore after adjusting for age and sex (p=0.0007; p=0.001; p=0.02; p=0.001, resp.). Multivariate logistic regression model including age and sex showed that there were three independent predictors of abnormal meiboscore: older age (OR = 1.03, 95% CI = 1.01–1.04 per year, p=0.006), postmenopausal hormone therapy (OR = 4.98, 95% CI = 1.52–16.30, p=0.007), and the use of antiallergy drugs (OR = 5.85, 95% CI = 2.18–15.72, p=0.0004). Conclusion. Our findings extend current knowledge on the pathophysiology of MGL

Bone marrow morphology during haematopoietic stem cell mobilization with G-CSF in mice

The aim of the work was to examine the morphology of the bone marrow of mice during stimulation with G-CSF. Experimental Balb C mice were daily injected subcutaneously with 250 μg/kg b.w. G-CSF (Neupogen). After 2, 4 and 6 days of the experiment femurs were obtained for morphological study. On day 2 of the mobilization the amount of haematopoietic cells in the bone marrow increased and dilatation of the sinusoids was observed. Only single leukocytes were observed in the lumen of the vessels. There were numerous leukocytes in the lumen of the sinusoids on day 4 of the mobilization. The morphology of the bone marrow on day 6 was similar to that of the control. Mobilization of mice with G-CSF resulted in migration of haematopoietic cells from the bone marrow and the process is most pronounced on day 4

Effect of hCG on the morphology of rat epididymal epithelial cells in vitro

The epididymis is an androgen-dependent organ. The hormones regulate the morphology and secretory activity of the epididymal epithelial cells. The cells in vitro resume their function as in vivo and also reveal features of steroidogenic cells. It can be expected that, as with Leydig cells, the morphology and function of the cells can be regulated by LH/hCG. The aim of the study was to assess the morphology of epididymal epithelial cells in vitro after stimulation with hCG. The experiment was performed on cells isolated from sexually mature rats. The epididymal epithelial cells were cultured in a medium with the addition of dihydrotestosterone (DHT). Moreover, the cells were cultured in the medium with DHT and without DHT but enriched with hCG. The epididymal epithelial cells cultured with DHT formed a monolayer and accumulated glycogen, a PAS-positive substance and lipid droplets. The cells cultured without DHT were stellate in shape and low in glycogen and PASpositive substance but they contained lipid droplets. The morphology of epididymal epithelial cells cultured without DHT but after stimulation with hCG was similar to the morphology of the cells cultured with DHT. This was the first sign that the morphology of the cells can be influenced by hCG

Rat epididymal epithelial cells and 17beta-estradiol synthesis under hCG stimulation in vitro.

Epithelial cells of human and animal epididymis display features of steroidogenic cells. Rat epididymal epithelial cells in vitro produce androgens which are converted to 17beta-estradiol, and released into the medium. The regulation of the epididymal steroidogenesis is not fully understood but it could be expected that it remains under LH influence. In previous study we observed that the morphology of rat epididymal epithelial cells in vitro was affected by hCG and the increase of amount of lipid droplets, glycogen and PAS-positive substances was observed. The present studies show the organelles which take part in synthesis of steroids in rat epididymal epithelial cells in vitro and the effect of hCG on E2 synthesis. The cells were cultured in the medium with/without DHT and without DHT in supplementation with hCG. After hCG stimulation the amount of an active mitochondria were increased when compared to the amount of mitochondria in the epididymal epithelial cells cultured without DHT. Ultrastructure of the cells was similar to the cells cultured with DHT, while the cytoplasm of the cells cultured without DHT was disorganized. The synthesis of 17beta-estradiol was stimulated by hCG, that exerted its effect through LH/hCG receptors, localized in the epididymal epithelial cells

The symptomatology of aneurysmal bone cyst : the value of diagnostic imaging

Background: An aneurysmal bone cyst (ABC) is a benign lesion occurring mainly in children. Generally it is found in the metaphyses of long bones and in flat bones. Osteolysis, bone expansion, thinning of the cortex, osseous septa and fluid levels belong to the typical imaging findings in ABC cases. The purpose of the study was to determine the value of imaging methods: plain x-ray films, US, CT and MRI for the diagnosis of aneurysmal bone cyst and to work out the diagnostic algorithm for an ABC suspected on plain radiograms. Material/Methods: The material consists of 72 patients (44 boys, 28 girls) aged 2-23 years (mean age 12 yrs) with bone lesions diagnosed as aneurysmal bone cysts on pathologic examination. In all patients, plain radiographs were performed, in 26 - sonography (US), in 41 - computed tomography (CT), in 8 - magnetic resonance imaging (MRI). All the examinations were analyzed according to own system of evaluation of typical presentation in the particular methods. The histopathological diagnosis was established after biopsy or surgery. Results: The aneurysmal bone cysts were most frequently located in the long bones of the lower limbs (43%), in the long bones of the upper limbs (26.4%) and in flat bones (25%). The localization was mainly in the bone methaphyses (56.6%). On pathological examination, primary ABC was diagnosed in 65.3% of cases, secondary ABC in 23.6%, ABC and concomitant osteitis in 7%, ABC and giant cell tumor in 2.7%, ABC and chondrosarcoma in 1.4%. The application of four methods resulted in 72.2% consistency between diagnostic imaging and pathological examination. In case of only one method used the consistency was 75% for MRI, 63% for CT, 55.4% for plain films and 50% for US. In 100% of misdiagnosed cases there were no septa within osteolytic lesions on plain films, whereas in 43.8% features of malignancy were found. In 53% of misdiagnosed cases no fluid levels were observed on CT and in 69% on US. Conclusions: 1. The diagnosis of aneurismal bone cyst based on plain radiography is possible in cases with typical radiological signs. 2. Application of other methods significantly increases the percentage of correct diagnoses. 3. Lack of fluid levels does not exclude the diagnosis of ABC. 4. Imaging features of malignancy do not exclude the diagnosis of ABC

Androgen levels and apoptosis in the testis during postnatal development of finasteride-treated male rat offspring

Introduction. The hormone-dependent events that occur throughout the first wave of spermatogenesis, such as the establishment of the number of Sertoli cells (SCs) and spermatogonial stem cells (SSCs) within the seminiferous cords and the setting up of the blood-testis barrier, are important for adult male fertility. Any changes in the T/DHT ratio can result in male subfertility or even infertility. In this study we aimed to evaluate effects of paternal exposure to 5-alpha reductase type 2 inhibitor, finasteride on litter size, androgen levels and germ cell apoptosis in male offspring during postnatal development. Material and methods. The subjects of the study were 7, 14, 21/22, 28, and 90-day-old Wistar male rats (F1:Fin) born from females fertilized by finasteride-treated rats. Offspring born from untreated parental animals were used as a control group (F1:Control). Animals and the collected testes were weighed, blood and intratesticular levels of T and DHT were measured by ELISA, and the apoptotic index of testicular cells was evaluated by TUNEL technique. Results. We observed difficulties in obtaining male newborns from female rats fertilized by finasteride-treated male rats. In the F1:Fin rats, changes in the body and testes weights occurred, and a lower number of apoptotic cells was found during postnatal maturation of the seminiferous epithelium. Changes in androgen concentrations during the first spermatogenesis wave and adult life were also evident. Conclusion. Finasteride treatment of male adult rats may not only cause a decrease in the fertility of parental rats, but also could lead to incorrect, androgen-sensitive course of spermatogenesis in their offspring