Jornal de Citologia e Histologia

Growth differentiation factor 9 (GDF9) plays a key role in the fertility of most mammalian species. Ovine GDF9 gene is localized in the 5th chromosome. The gene’s length is 2.5 kb consisting of two exons separated by one intron (1126 bp) and encodes a pro-peptide comprising 453 amino acids. The mature peptide includes 135 amino acids. The purpose of this paper is to determine the GDF9 polymorphism in sheep of Salskaya and Romanov breeds in Rostov region of Russia. Polymorphism was identified by PCR- RFLP method at points G1 (G260A) and G4 (G721A). AG and GG genotypes were detected at points G1 and AA and AG genotypes were detected at points G4 in Salskaya sheep breed, frequency of A was 0.05 and G allele 0.95 (at point G1) and frequency of A was 0.95 and G allele 0.05 (at point G4). In Romanov breed AG and GG genotypes were detected at points G1 and AA and AG genotypes were detected at points G4, frequency of A was 0.20 and G allele 0.80 (at point G1) and frequency of A was 0.80 and G allele 0.20 (at point G4).

Oxidized cellulose is a polysaccharide substance formed by the oxidation of cellulose. Depending on the amount used, oxycellulose is completely absorbable and can be used as hemostatic agents in general and dental surgery for hemostatic purposes. Each procedure on the bone also carries the risk of bleeding, but still not much is known about the bone tissue reaction on this material. Due to this fact, the aim of this study was to evaluate the tissue reaction and osteogenic potential of the natural oxycellulose material RESORBA-CELL® standard by its implantation into rat cranial bone defects. The materials were evaluated using an established rat cranial defect model in 22 animals. One bone defect with a diameter of 5 mm was created per animal. The defects were filled with oxycellulose and left to heal for 4 weeks. Twelve samples (n=6 oxycellulose; n=6 untreated control lesions) were processed for histological evaluation. The remaining 10 samples were processed for mRNA expression analysis of genes coding for growth factors and osteogenic differentiation using quantitative RT-PCR. In oxycellulose treated bone lesions significant reduced mRNA levels of Runx2, Bglap and COL1A1 were found. The expression reached 74%, 73% and 47% of the mRNA quantity of the untreated bone, respectively. In the histological sections, the bone defects were completely filled with connective tissue / bone marrow with embedded foam cells after four weeks. In contrast to the controls, the tested material do not induced early bone healing. The residual bone seems to be rather absorbed. In conclusion, the study has shown that the RESORBA-CELL® oxidized cellulose has no osteogenic potential, but it still can be considered as a useful wound dressing in the dental surgery.

Interleukin (IL)-33 is a cytokine that has key roles in the induction and perpetuation of allergic inflammation. It is strongly expressed by airway epithelial cells, but there is controversy about whether immunoreactivity is localised only to the nucleus or is also present in the cytoplasm. We investigated IL-33 immunoreactivity in airway tissues in a mouse model of an acute exacerbation of chronic asthma, in human asthmatic airway tissue, and in airway tissue from patients with allergic bronchopulmonary aspergillosis. We demonstrated that cytoplasmic immunoreactivity in airway epithelial cells was dependent both upon the target species and the severity of the allergic inflammatory response. Furthermore, using double immunolabelling, we showed that immunoglobulin-synthesising plasma cells express high levels of IL-33, which could contribute to allergic inflammation.

Hypericum perforatum L. (HPL) has been used as a beneficial herb on menopause related syndromes for many years by inhibiting lipid oxidation. Hypericum perforatum L. extract contain flavonoids and phenolic acids, but it was not a method to separate and identify flavonoids which was based on design of experiment. Moreover, upper complicated HPLC method might cause undesirable effects on feasibility and repeatability in fingerprint technology. It was necessity to establish a generally and systematized way which could develop a reliable fingerprint technology for Chinese medicine. Plackett-Burman design method was a conventional tool for variables randomization aiming at optimization. Our study was based on this optimized design of experiment about ethanol extract from Hypericum perforatum L. and established the corresponding method for separation and identification of flavonoids. With this Design-Expert software, the results showed HCRF have significant effect on variables. From One-factor plots for main effect of HCRF, 254 nm wavelength measurement have a significant influenced on the HCRF, and have provided the optimized parameters.

Orthodontic attachments interfere with oral hygiene maintenance leading to plaque accumulation and ultimately enamel demineralization caused by oral microflora. Orthodontists must have a clear understanding of the nature and the type of microflora present to combat with the above stated problem. This study was done to isolate and identify the microflora from orthodontic attachments and to study the quality of oral microflora during fixed mechanotherapy at regular intervals. In this prospective clinical study, ten patients were selected for arch wire ligation with commonly used elastic modules. One elastic module was removed from each patient at regular intervals of 24 hrs, 7, 14 and 21 days for isolation and qualitative assessment of the microbial growth. Aerobic inoculation was done using nichrome inoculating wire. Further subspeciation was done using certain biochemical reactions like fermentation of sugar. For anaerobic growths Mac-Intosh jar was used. The study revealed that there was minimal or no growth of aerobes and anerobes after 24 hours. However, with increased duration of time there was progressive colonization of microbes (both commensals and pathogenic) on the orthodontic attachments. On examination of orthodontic attachments over a scheduled period of time anaerobic microorganisms were isolated which indicates that patients undergoing fixed mechanotherapy should exercise maximum oral hygiene with oral hygiene adjunctive aids other than routine brushing and should also undergo periodic oral prophylaxis so as to avoid any iatrogenic damage and alteration of the oral environment.

A novel human inhibitor of apoptosis protein family member, termed Livin, was demonstrated in the pathogenesis of different human malignancies, and also it was studied as a potential treatment target in malignancies. However, there is no report on the Livin expression profile in Hodgkin Lymphoma. In this study, we evaluated the Livin expression in 78 paraffin embedded blocks including 39 staged cases of Hodgkin Lymphoma and 39 control subjects (normal and reactive hyperplasia lymph nodes). Tissue Microarray-based Semi-quantitative Immuno-flourecent Staining was applied for protein expression profiling in both infiltrating non-neoplastic cells (morphologically typical Lymphocytes) and neoplastic cells (Hodgkin and Reed-Sternberg) of cases and control samples. Our results demonstrated that the mean ratio of Livin/GAPDH expression was significantly increased between infiltrating background cells in Hodgkin Lymphomas and control cases (0.54596 vs. 0.50827, P<0.001). Also, a significant difference was found in the mean ratio of Livin/GAPDH expression between neoplastic cells and major background cells in the tumor microenvironment (0.59024 vs. 0.54596, p<0.001). Furthermore, this study confirmed a significant increase of Livin expression from early-stage to advanced-stage of Hodgkin Lymphoma (0.52888 vs. 0.580146, P<0.01).These findings suggest that Livin may play a critical role in the pathogenesis of Hodgkin Lymphoma. It also can be a novel prognostic marker and a potential therapeutic target in this type of lymphoma.

Alzheimer’s and Parkinson’s diseases are neurodegenerative diseases which cannot be cured so far. In both diseases, a GABAergic-glutaminergic neurotransmitter imbalance might occur. Neural networks are suggested for Parkinson’s disease in the extrapyramidal system and for Alzheimer’s disease in the hippocampus and the temporal cortex. Combined GABAA agonists and NMDA antagonists might have therapeutic and maybe prophylactic properties in Alzheimer’s and Parkinson’s disease. It is important to examine the neural networks in the brain areas involved in both diseases. This would offer the possibility to try an early beginning pharmacotherapy of these, up to now, incurable diseases.

Although rare, osteosarcoma is the most frequent post-radiation second malignancy of the bone. We present a case of an osteoblastic osteosarcoma that occurred in a woman with a history of two neoplasms, as an example of multiple treatment-associated neoplasms. In fact, prior to the development of this osteosarcoma, our patient had been diagnosed with a breast carcinoma 24 years earlier, which had been treated by quadrantectomy, followed by radiotherapy, but it had recurred after 7 years. Thus, the patient had undergone mastectomy, followed by 4 years of tamoxifen. Then, 14 years later, she developed the second tumor, an endometrial adenocarcinoma, which was treated by surgery and adjuvant radiotherapy. Afterwards, a neoplasm developed in the pelvic bones, which histologically proved to be an osteosarcoma. Athough considered low, there is some risk of developing an endometrial cancer after tamoxifen protracted therapy. Since germline mutation analyses for p53 and RB1 resulted negative in our case, Thus, the treatment could have played a prominent role in our patient.

Breast metastases from gynecologic cancer are infrequent and are mainly from the ovary. They usually appear in advanced stages and have poor survival rates after diagnosis. Two cases of breast metastasis of serous ovarian carcinoma are presented. In both cases fine-needle aspiration (FNA) study of breast revealed a lesion with morphologic characteristics of serous carcinoma, while one of them was with immunocytochemical study. Origin was confirmed by histological examination of this lesion.

We report a case of a primary lung tumor characterized by complex gland-like spaces lined by simple cuboidal to columnar epithelium surrounded by a hyalinized spindle-cell fibroblastic proliferation reminiscent of adenofibroma of the female genital tract. The lesion occurred in a 56 year-old male with a history of left thoracic pain and hemoptysis. The tumor presented clinically as 1.5 cm solitary lesion and was discovered on chest X-rays. The lesion was treated by lobectomy. The main importance lies in distinguishing this lesion histologically from other types of pulmonary hamartoma, pulmonary blastoma, intrapulmonary solitary fibrous tumor and metastases from soft tissues and visceral sarcomas.

Oxidative stress has been recommended as a contributory factor in complication and progression of diabetes. The aim of the study was to evaluate the effects of pomegranate on body weight and histopathology of the steptozotocin–nicotiamide (STZ-NA) generated oxidative stress induced diabetic rats. Thirty-two male Sprague- Dawley rats were randomly divided into non-diabetic, diabetic untreated and diabetic treated with pomegranate and Glibenclamide. Experimental diabetes was induced by a single intraperitoneal (i.p) administration of STZ 60 mg/kg body weight (b.w), 15 minutes after the i.p injection of NA (120 mg/kg b.w). Histopathological analyses were conducted on the liver and kidney tissues. While the body weight significantly decreased in the diabetic control group, however it was a significant increase in the diabetic treated group (P<0.05). Destruction of the liver architecture of the hepatocytes in the diabetic group showed the signs of necrosis, degeneration, dilatation, and inflammation in the central vein and blood vessels. In the kidney, shrinkage and lesion in Bowman’s capsule were observed. Pomegranate acted as an antioxidant thereby preventing oxidative damage in the diabetic liver and kidney.

The present study was conducted to assess the histopathological and cytotoxic effect of lead exposure on male reproductive function. Adult male rats were divided in 2 groups. The first as a control group was given distilled water as drinking, while the second was exposed to 2 g/l of lead acetate in distilled water during 35 days. The structural damage on tests parenchyma was investigated by histological study and supplemented by biochemical assay of testosterone level. DNA fragmentation in germinal cells was determined using terminal desoxynucleotidyl transferase mediated dUTP nick end-labeling (TUNEL) assay. The histological study showed that lead can induce pronounced alterations on germ cells. The testosterone level was significantly decreased in treated rats compared to controls. TUNEL assay revealed a significant DNA fragmentation index in rats exposed to lead. This study showed TUNEL assay can be used to determinate DNA fragmentation in germinal cells. Moreover, we conclude that lead acetate may be considered as strong cytotoxic and genotoxic agent on male reproductive function.

Background and objectives: Phylloides tumors are categorized as benign, borderline and malignant on the basis of histological features which are subjected to significant inter-observer variability. Our aim was to study the role of immunohistochemical expression of CD10 in discriminating benign, borderline and malignant categories of phylloides tumor.
Methods: The expression of CD10 was studied in 85 phylloides tumors (27 benign, 25 borderline, and 33 malignant) using immunohistochemistry to see whether the expression differed between these histologic categories. Chi-square test was applied to determine the significance of difference in CD10 expression among tumor categories. Significance was established at p<0.05.
Results: 5/27 (18.5%) benign, 16/25 (64%) borderline and 26/33 (78.8%) malignant cases expressed significant (2+ or 3+) staining. This expression of CD10 significantly varied among histological categories (p<0.001). Histological features such as stromal atypia, stromal cellularity, tumor margins, mitotic activity and tumor size correlated significantly with tumor categories as well as with CD10 expression.
Conclusion: From these highly significant results, we believe that there is a strong correlation between CD10 expression and tumor grade and it can be used as an adjunct diagnostic tool for categorizing Phylloides tumors along with other histological features.