Revista de citología e histología

Background: The orchid plants of Dendrobium, belong to the largest family of flowering plants orchidaceae. The genus Dendrobium reported to possess various therapeutic activities including anticancer activity. In the present scenario, cancer is one of the most leading causes of death worldwide. Therefore, the main objective of the present study was to demonstrate the cytotoxic and apoptotic inducing activity of Dendrobium crepidatum and Dendrobium chrysanthum ethanolic extracts in T-cell lymphoma. Methods: MTT assay was done to evaluate the cytotoxic effect of the extracts on lymphoma cells. The reactive oxygen species (ROS) generation by the ethanolic extracts was also examined by DCFH-DA. Fluorescence microscopy studies and DNA fragmentation assay were done to investigate the apoptotic cell death. Flow cytometry was done to check the effect of ethanolic extracts on cell cycle phase distribution. Results: The results revealed that both the extracts led to dose dependent cytotoxic effect in T-cell lymphoma along with increased generation of intracellular reactive oxygen species in a dose and time dependent manner. The fluorescent microscopic studies showed that the extracts induced cellular shrinkage, chromatin condensation and nuclear fragmentation, the major hallmarks of apoptosis, in dose dependent manner. The presence of DNA ladder further confirms the process of nuclear fragmentation, and the cell cycle analysis showed significant delay at G2/M phase of the cell cycle with both the extracts. Conclusion: The whole study suggests that Dendrobium crepidatum and Dendrobium chrysanthum ethanolic extracts induced substantial cytotoxic and apoptotic activity in T-cell lymphoma. Therefore, further studies are needed to establish the detailed mechanism of anticancer activity of both the ethanolic extracts.

Schizoaffective disorder, which is combined with schizophrenic and affective, i.e. depressive or manic or alternating depressive and manic symptoms, has a prevalance of 0.5%. Here, we describe the alterations of the most important classical neurotransmitters in the brain regions involved in schizophrenic and affective symptoms. Schizoaffective is undoubtedly an inheritable chronic psychiatric disease, whereby traumata can enhance schizophrenic and affective symptoms in one third of patients. Neural networks are described in the brainstem, hippocampus and ventral tegmental area and the mentioned neurotransmitter alterations are considered. Prophylactic treatment of schizoaffective patients consists of administering mostly second-generation antipsychotic drugs alone or in combination with mood-stabilizing drugs. The clinical importance of the antipsychotic drug clozapine for a pharmacotherapy of treatment-resistant forms of the disease is underlined.

Cartilages growth requires activation of perichondrial cells and chondrocyte multiplication. Activated perichondrial cells produce an outer cartilage matrix and in appositional growth and chondrocytes division produce new chondrogenic cells which participate in interstitial growth of cartilage. This study was performed to identify a third type of cartilage growth in skeletal elements of quail embryos at 6, 7, 8, 9, 10, 13, 16, 17 days of incubation. Paraffin sections of the whole embryos were stained using H&E and Safranin O stains. Immunohistochemical staining for C-KIT, matrix metallpoproteinase-9 (MMP-9) and type II collagen was performed. Other samples were processed for scanning and transmission electron microscopy. We found that cells extended form mesenchymal tissue or perichondrium penetrated the cartilage templates of different skeletal elements. The morphological and immunohistochemical characteristics of these cells were identical to mesenchymal cells; characterized by their small size, having multiple cell processes and staining positive for C-KIT. MMP-9 positive cells were detected in cartilage matrix. The mesenchymal cells further differentiated into chondrocytes which were positive for type II collagen immunostaining and secreted Safranin O positive cartilage matrix. In conclusion, our results suggest that C-KIT positive mesenchymal cells secrete MMP-9 to penetrate growing cartilage, produce type II collagen and proteoglycan and have a role in interstitial growth of embryonic cartilage templates in quail. (Graphical Abstract).

Caloric restriction and resveratrol intake have beneficial effects on health, especially in the prevention and improvement of diabetes. Based on this evidence, this study has been conducted in order to examine the normal architecture of pancreatic islets and the long term effects on endocrine pancreas of standard diet, caloric restriction, or resveratrol in 19 adult grey mouse lemurs. Immunohistochemistry, with anti-insulin and anti-glucagon antibodies, and morphometric analysis were performed. The normal pancreatic islet was composed of an outer circle of α cells and a large inner body of β cells, the percentages of each cell types being 10.3 ± 3.3% and 69.7 ± 1.8%, respectively. A marked increase of the pancreatic islet mass was detected in 6 animals: 2 animals under standard diet,1 animal under calorie restricted diet, and 3 animals under standard diet supplemented with resveratrol. This lesion consisted mainly of an increase of β-cell mass in 4/6 animals, and seemed to appear in middle-aged adult primates (5.8 ± 0.8 years old). To our knowledge, there are no other studies which aim at describing the changes of the pancreatic islet mass in grey mouse lemurs.

Introduction: In the context of axon regeneration, certain molecules containing the basal lamina, particularly fibronectin, laminin and heparin sulfate promote axonal elongation in vitro and in vivo. This structure could lead the axons to distal stump in nerve defect repair. Material and Methods: In twenty male Wistar rats, a critical defect of 15 mm was created in the sciatic nerve, which was bridged by an acellular muscle graft, which were obtained from the gluteus medius muscle of two donor rats, and chemically treated to cellular extraction by protocol consisted of 7 h in distilled water changed three times every 2-3 h, one night in Triton detergent, and 24 h in deoxycholate. After 90 days, the explanted pieces of sciatic nerve with grafts were carved and cut into seven pieces equally in all animals; pieces were processed in resin (toluidine blue staining) and also for paraffin embedding: hematoxylin-eosin and Masson trichrome. Results: Microscopically, proximal ends appeared as a normal nerve, with proper, regular and orderly fascicular distribution, perfectly defined by its three wraps, with little connective tissue. At the graft, regenerated axons used the muscle as a bridge, with many small nerve fascicles separated by host tissue and plenty of blood vessels. Axons followed their basal lamina, defining small fascicles with large vessels and abundant connective tissue. At the distal end, there was no a clear division in fascicles in some animals. There was little connective tissue between the fibers and the epineurium was quite developed. Conclusions: The selected biological method for chemical treatment of skeletal muscle achieves acellular grafts. We successfully could repair a critical lesion of the sciatic nerve in rats using this acellular muscle graft. Acellular muscle that we employed as a graft provides a bridge tool for the advancement of the regenerating axons, perhaps due to the structural proximity of laminin to nerve sheaths.

Two common types of cells involved in cartilage growth; the undifferentiated perichondrial cells and chondrocytes. The perichondrial cells provide new external layers of cartilage matrix and are responsible for appositional growth. Chondrocytes proliferate to produce chondrogenic cells which generate a new interstitial cartilage matrix (interstitial cartilage growth). We previously described a third type of cartilage growth which is a mesenchymal cell-dependent type in the cartilage of the air breathing dendritic organ of catfish. The current study aimed to investigate this phenomenon using histochemical techniques, semi-thin sectioning, TEM and immunohistochemical staining. Typical mesenchymal cells penetrated the cartilage of the air breathing organ. Differentiating chondrogenic cells were detected adjacent to mesenchymal cells. Mesenchymal cells were continued with either the perichondrium or the external mesenchyme. Different stages of chondrogenic cells were identified adjacent to mesenchymal cells. Mesenchymal, chondrogenic and mature chondrocytes expressed type II collagen. Chondrogenic cells secrete elastic and safranin O positive cartilage matrix. Cartilage remnants and broken lacunae were observed at the site of mesenchymal cells. Mesenchymal cells, perichondrial cells, and chondrocytes had strong immunoreactivity for MMP-9. Mature chondrocytes undergo death leaving empty lacunae which were penetrated by mesenchymal cells. In conclusion, mesenchymal cells were responsible for cartilage matrix degradation and formation of the new matrix. They play a role in growth and maintenance of cartilage.

We report a case of idiopathic calcinosis cutis diagnosed by fine needle aspiration cytology in a 70 year old female who presented with subcutaneous swellings near the bilateral iliac crests. Cytological finding of amorphous calcium salts with histiocytes and the appropriate clinical background led to the cytodiagnosis of idiopathic calcinosis cutis as subsequently confirmed on histopathology. Pitfalls in the diagnosis of calcinosis cutis on cytology smears are also discussed

The present study describes for the first time histology, histochemistry, and ultrastructure of all rodlet stages. By ultrastructure, we identified rodlet progenitors in the stroma of the olfactory organ in Red-Fin shark. Rodlet progenitors were mesenchymal-like cells synthesis the primitive rodlet granules. Rodlet progenitors differentiated to vesicular rodlet cells, which was rich in vesicles and vacuoles, and accumulate intracellular fibrillar-like components. Granular rodlet cells were observed in the basal epithelia, contained premature rodlet granules and began to organize the fibrillar components of the rodlet capsule. Transitional rodlet cells continued to deposit the fibrils of the capsule and synthesis of rodlet granules. Mature rodlet cells were polarized bear-shape and had typical rodlet granules. Histochemistry showed rodlet cells had a wide range of staining affinities including carbohydrate, lipid, and protein staining. All stages of rodlet cells revealed the presence of PAS-positive granules in the cytoplasm of rodlet cells where, While the ruptured ones stained strongly with alcian blue, sliver stain and Sudan black B. Rodlet granule was positive for bromophenol blue Rodlet cells were positively immunostained against Matrix Metalloproteinase-9 (MPP-9) in all stages indicate invasion properties of rodlet cells in tissue. Conclusion rodlet cells originated from the stroma of the olfactory organ.

Fine needle aspiration (FNA) cytology of two cases of breast pure squamous cell carcinoma (SCC) and one invasive carcinoma mixed with SCC is reported. Cytopathologic findings of breast cancer vary greatly because mammary carcinomas are composed of many subtypes. Squamous cells, both malignant and benign, are occasionally seen in the breast FNA smears. Many differential diagnoses including various neoplastic and non-neoplastic diseases should be raised when squamous cells are observed in the mammary FNA smears. Squamous cells endowed with severe grade atypia obviously indicate malignancy. However, apparently benignlooking squamous cells are seen in the FNA specimens of both malignant and benign breast disorders. Accurate interpretation of cytologic findings is essential for discriminating malignant diseases from benign conditions. As prognosis and therapeutic options depend on tumour subtypes, deducing tumour histologic subtypes from cytologic specimens is useful for the appropriate treatment planning. FNA samples of pure SCC and invasive carcinoma mixed with SCC were examined in detail in an attempt to clearly define the cytopathologic characteristics of pure breast SCC. Careful review of the FNA slides of these cases helped us to recognize cytomorphologic findings useful in the differential diagnoses and predicting of histologic pictures of mammary disorders containing squamous cells.

Epithelioid osteosarcoma is an uncommon subtype of conventional osteosarcoma that comprised of both typical osteosarcoma component and epithelioid cells. It often occurs in adolescent and children. In here, we report a case of epithelioid osteosarcoma of a 12-year-old Chinese boy. Microscopically, the tumor contains large epithelioid cell with abundant cytoplasm and osteoid forming malignant cells, and partially, the tumor cells arranged in pseudoglandular structures. To our knowledge, it is the first report of epithelioid osteosarcoma with pseudo-glandular structure. The epithelioid cells expressed vimentin and epithelial membrane antigen but did not expressed cytokeratin.

This study concluded that diabetes type I resulted in pronounced oxidative stress that affected renal function and structure and reduced testosterone level. Sildenafil administration for 8 weeks proved to have a potential renal protective capacity at both morphological and functional levels. Thus, Sildenafil might have a potential novel in restoration of kidney function and structure in diabetic male patient especially those with low testosterone level.