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Журнал цитологии и гистологии

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Объем 5, Проблема 5 (2014)

Обзорная статья

Classical Neurotransmitters and Neuropeptides involved in Parkinson's Disease: A Multi-Neurotransmitter System

Felix-Martin Werner and Rafael Coveñas

Parkinson’s disease is a neurodegenerative disease with motor and non-motor symptoms. In Parkinson’s disease, a neurotransmitter imbalance occurs in the extrapyramidal system with a dopamine and GABA deficiency and an acetylcholine and glutamate surplus. Other classical neurotransmitters such as serotonin, the neuroactive substance adenosine and neuropeptides such as dynorphin and substance P are also involved in the pathophysiology of the disease. Here, we describe the alterations of the involved neuroactive substances and the relationships between them in the extrapyramidal system. From the findings previously reported in the literature, here a neural network is developed in the extrapyramidal system. Additional anti-Parkinsonian drugs and their actions in the neural network are also pointed out, since a multimodal pharmacotherapy of the disease might improve its outcome.

исследовательская статья

Preventive Action of Honey on Methotrexate Induced Intestinal Mucositis in Albino Rats (Immunohistochemical Study)

Ali Sultan Al-Refai

Background and Objectives: Gastrointestinal mucositis is a severe and debilitating side effect of chemotherapy. Methotrexate (MTX) represents one of the most potent classical anti-tumor drugs. The aim of this study was to determine the preventive effects of honey on the intestinal mucositis caused by methotrexate in rats.
Materials and Methods: In current study forty females Albino rats, weighing 250-300g were used in the study. For the induction of mucositis, 60 mg/kg of MTX was administered intraperitoneally to each animal in the study group on day 4. The control animals were intraperitoneally injected by normal saline in the same manner and dose like MTX. At the beginning of the experiment, the rats in each group were randomly divided into two groups: Distilled water treated group and honey treated group (10 animals each). A volume of distilled water equal to honey was given by intragastric gavage tube, while the other group was gavaged with honey at a dose of (2.5g/kg) two times daily. The body weight for each rat was measured and then the animals were sacrificed on day 8. In each experiment, one centimeter of prox¬imal jejunum was removed for histopathological, intestinal morphometry, and immunohistochemical analysis using Ki-67 and Bcl-2 immunolabeling.
Results: The MTX / honey treated group showed significantly (p<0.05) less body weight loss and less histopathological scores than that of MTX / water treated group. But there were non significant differences (p>0.05) were seen in the Ki-67 and Bcl-2 immune expression in comparison with the MTX/water treated group.
Conclusion: Honey at a dose of 2.5g/kg two times daily, produced a partial protection against methotrexate induced mucositis in rat by decreasing the histopathological scores.

исследовательская статья

The Role of Technical Quality Control in Histology Laboratories

Zuhair M Mohammedsaleh

The importance of laboratories cannot be gainsaid as far as the health of any country is concerned. This is especially considering that laboratories have a bearing on the accuracy of research carried out on varied ailments, as well as the effectiveness of any remedial substance. Of course, there are numerous categories of laboratories with histology laboratories coming as some of the most fundamental. Histology laboratories come with a number of distinctive characteristics that can only be comprehended through comparing the laboratories with other areas of medical laboratories. Nevertheless, they also have to ensure the efficacy and integrity of their results. This underlines the importance of quality control in histology laboratories. Quality control underlines a system pertaining to routine technical activities that are aimed at developing and controlling the quality and accuracy of inventory while it is being developed. This system comes up with consistent and routine checks that identify and address omissions and errors, safeguard the integrity, completeness and correctness of data, while also recording all quality control activities. This Review Article examines the role of quality control in histology laboratories. It examines the varied aspects of quality management including risk identification, risk analysis and risk elimination. On the same note, it examines the convergence between quality management and quality assurance while paying close attention to the role of technical quality control in histology laboratories.

исследовательская статья

The Relationship between the Intercellular Adhesion Molecule-1 Expression and the Response to BCG Immunotherapy in Non Muscle Invasive Bladder Cancer

Faouzia Ajili, Sonia Abdelhak, Amel Nedri, Nadia Kourda, Afifa maaloul, Salma Karay, Mohamed Slim Salmi and Samir Boubaker

Background: Bladder cancer is the second most common malignancy of the urogenital region. Although Bacillus Calmette-Guerin (BCG) is considered as the adjuvant treatment of choice for non-muscle invasive bladder cancer (NMIBC), there is no consensus for a predictive factor to assess BCG success. The intercellular adhesion molecule-1 (ICAM1) has been reported to function in multiple malignancies, but its effect on NMIBC hasn't been discussed yet. This study attempted to evaluate if ICAM-1 could be useful predictive markers in BCG responses.
Materials and Methods: Thirty primary resected NMIBC patients were included in the study. All patients received adjuvant BCG instillations. ICAM1 expression was inspected by immunohistochemistry and correlated with clinicopathologic variables. Association between protein expression and BCG therapy response was evaluated by univariate and multivariate analysis.
Results: Univariate Cox regression analysis of baseline characteristics and ICAM-1 expression showed that no significant association was found with BCG immunotherapy response. In the other hand, multivariate Cox regression analysis showed that ICAM-1 protein is not an independent factor of tumor response after BCG immunotherapy.
Conclusions: this study demonstrates that ICAM1 could not be a useful prognostic marker for BCG treatment in NMIBC. We emphasize that this was a preliminary study and therefore further confirmation on a larger set of tissues is necessary.

Обзорная статья

Cancer Associated Cachexia; Etiopathogenesis, Molecular Mechanisms and Therapeutic Strategies

Mohamed Hassan, Abdelouahid El-khattouiti, Youssef Haikel and Mossaad Megahed

Cachexia is a multifunctional syndrome that is characterized by anorexia and extensive loss of adipose tissue and skeletal muscle, is common in many chronic and advanced diseases. Most of cancer patients show variable patterns of body loss that is known as cancer associated cachexia (CAC). Although CAC share common characteristics, the severity of this disease is variable, and seems to be tumor type, site and stage-dependent. The etiology of cachexia is attributed to abnormal metabolism that is thought to be mediated by tumor- and host-derived cytokines and factors. Although the role of cytokines in the etiology of CAC has been reported, the molecular mechanisms regulating its occurrence in cancer patients are not described in details. This review focuses on the etiopathogenesis of CAC and the underlying mechanisms.

исследовательская статья

Phenotype Changes of Circulating Monocytes in a Hypercholesterolemic Swine Model of Coronary Artery Disease

Silverio Sbrana, Gualtiero Pelosi, Maria Rita Puntoni, Federica Viglione, Maria Giovanna Trivella and Oberdan Parodi

Objective: Inflammation and immunity activation play a key role in atherosclerosis (ATS) onset and progression. Aim of this study was to investigate the relationships between phenotype of circulating monocytes and coronary artery disease (CAD) development in a histologically well-characterized swine model of ATS.
Methods: Blood samples were obtained from 6 animals at baseline and after 16 weeks high fat cholesterolenriched diet. Flow cytometry monocyte identification was performed (CD172a marker). Adhesion (CD18a, CD11a, CD11R3, CD49d, CD29), differentiation (CD14) and activation receptors (SLA-DR, CD16, CD163) were quantified as percentage of positivity (%) and Relative Fluorescence Intensity (RFI). Lipid parameters (LDL, oxLDL, HDL) and soluble endothelial ICAM-1 were measured and histologic quantitative assessment of coronary ATS was performed.
Results: Flow cytometry analysis demonstrated a significant post-diet decrease of CD14 RFI and an increment of % SLA-DR. Pre-diet values of ICAM-1 and % SLA-DR correlated reciprocally (P=0.0191) and with several CAD severity indexes (P≤0.02). Positive correlations between RFI changes of CD29 (P=0.0213) and CD18a (P=0.0341) and morphometric indexes of coronary ATS were found. Post-diet RFI values of CD29, CD18a and CD16 were also closely related to morphometric parameters (P<0.03). A cumulative post-diet tendency to increase of CD14low/ CD163high monocyte fraction (45.07 ± 2.27 vs. 40.14 ± 3.16) and a tight correlation between changes of this monocyte subset and corresponding HDL variations (P=0.0100) were also observed.
Conclusions: Blood monocyte orientation towards a macrophage-like phenotype, similar to a HDL-induced maturation, and a close association between markers changes and severity of diet induced coronary ATS could provide new insights into plaque growth and progression in CAD.

исследовательская статья

P53 Overexpression in Ductal Carcinoma in situ of the Breast

Kikuchi S, Osako T, Nishiyama Y, Nakano M, Tashima R, Fujisue M, Toyozumi Y, Arima N and Reiki Nishimura

Background: Ductal carcinoma in situ (DCIS) is the most common type of non-invasive breast cancer. DCIS is not life threatening but can increase the risk of invasive ductal carcinoma (IDC) development. However, the role of P53 expression in the progression to invasiveness in DCIS is unclear. In this retrospective study, the differences in biological markers, including protein 53 (P53) expression, between DCIS and IDC ≤5 mm in diameter (T1a) were evaluated.
Methods: Among 2,497 primary breast cancer patients, who underwent initial surgery between 2001 and 2010, 267 (10.4%) patients were diagnosed with DCIS and 86 (3.4%) patients were diagnosed with T1a. P53 expression was evaluated using immunohistochemical analysis, and the distribution (<10%, <50%, and ≥50%) and the association with other biological markers was investigated in DCIS and T1a.
Results: Overexpression (≥ 50%) of P53 was seen in 13% of patients with DCIS and in 24% of patients with T1a, constituting a significant difference (p=0.001). P53 overexpression was significantly associated with higher nuclear grade, lower rates of estrogen receptor (ER)/progesterone receptor (PgR) positivity, higher rates of epidermal growth factor receptor 2 (HER2) positivity, and higher values of nuclear protein Ki-67 in patients with DCIS. Recurrence rates did not differ significantly between DCIS and T1a. Logistic regression analysis of factors associated with invasiveness revealed that P53 overexpression was one of the significant factors, and ER negativity was found to be a significant independent factor in multivariate analysis.
Conclusion: P53 overexpression reflected the aggressiveness and correlated with the invasiveness in DCIS. These findings suggest that P53 may play an important role in invasion of DCIS in the breast.

Краткое сообщение

Does the Acupuncture Point Shaoshang (Lu11) in Humans Match with an Increased Density of Connexines?

Ines Pilz-Klement and Christian-Albrecht May

Acupuncture points are clinically described by their localization and sensation measuring skin resistance. As a morphological correlate, gap junctions and connexines (Cx) were postulated but up to now only few animal studies tried to identify these structures histologically. Immunohistochemical staining of acupuncture point Lu11 of 4 human donors (76-81 years of age) were stained with antibodies against Cx 26, 30, 32 and 43. A densitometric evaluation was performed using analySIS Software 3.2 (Olympus). In the region of the acupuncture point Lu11 the density of Cx 26 and Cx 43 was increased. This increase was not restricted to the epidermal layers but also included the dermis and subcutis. Our findings give evidence that there are persistent regions of increased Cx density in the human skin. They provide a morphological base for fast alterations in electrical skin resistance dependent on the functional stage of the Cxs by forming either gap junctions or hemi-channels.

исследовательская статья

A Comprehensive Analysis of Markers for Neuroendocrine Tumors of the Lungs Demonstrates Estrogen Receptor Beta to be a Prognostic Markers in SCLC Male Patients

Curioni-Fontecedro Alessandra, Soldini D, Seifert B, Eichmueller T, Korol D, Moch H, Weder W and Stahel RA

Knowledge about the biology and prognostic markers of neuroendocrine tumours (NET) of the lungs is scarce. As sex steroids contribute to cell proliferation in various human tissues, we aimed at characterising the expression of steroid receptors in a broad cohort of probes from NET of the lungs and evaluated their possible prognostic impact. A total of 192 tumour specimens of NET of the lungs were collected, with 58 surgical specimens of typical carcinoids, 42 of atypical carcinoids, 32 of large cell neuroendocrine carcinomas and 60 of small cell lung cancer (SCLC). Analysis by immunohistochemistry for the following antigens was performed: estrogen receptor β (ERβ), ERα, PR (progesterone receptor), AR (androgen receptor), Thyroid transcription factor 1 (TTF1), synaptophysin and for chromograninA. ERβ was found to be expressed in more than 60% of tumor probes. Survival data were available from 126 patients. In SCLC patients a subgroup analysis showed a survival benefit in the group of male patients whose tumours expressed ERβ (p=0.008) and all SCLC patients whose tumours expressed both ERβ and chromograninA (p=0.02). Here we could demonstrate that ERβ expression is frequent through all NET of the lungs. Moreover, our results suggest that ERβ represents a favourable prognostic factor for SCLC male patients and all SCLC patients whose tumors co-expressed chromogranin A.

исследовательская статья

NAD(P)H:Quinone Oxidoreductase 1 Expression in Human Primary Melanotic Melanomas of the Skin

Francesco Zappa, Timothy Ward, Ennio Pedrinis, AIan Mc Gown and Nicholas Thatcher

NAD(P)H:quinone oxidoreductase 1 (NQ01; DT-diaphorase; DTD) is a two-electron reducing enzyme which is often over-expressed in cancers and can activate quinones to form cytotoxic species. This over-expression is used as target in the design of novel anti-cancer drugs. DTD is also expressed in normal tissues, but little is known about its “body mapping”. Because of the dearth of information on the cell-specific expression of DTD in pigmentproducing cells of the skin, we analysed its expression in normal and tumoral samples. Twenty skin biopsies of primary malignant melanotic melanomas (ten with vertical invasion, ten with superficial spreading) were analysed by immunohistochemistry. In normal skin DTD expression was weak in all epidermal layers, including normal melanocytes. Malignant melanomas showed a very strong DTD expression in tumoral cells of all samples and in endothelial lining of peri-tumoral vessels. These results suggest that DT-diaphorase may be a new marker that could be useful in determining the extent of a malignant melanocytic lesion, and also tend to support the potential of malignant melanomas as target for DTDdirected antitumour agents.

История болезни

Myxofibrosarcoma of the Thyroid: Second Case in Africa

Asmaa Salama, Nesreen Hafez, Eman Abu-Sinna, Ashraf Hassouna and Ayman A. Amin

Myxofibrosarcoma is a common sarcoma in the extremities, but is rare in the head and neck region. Here, we report a case of 76 year old female patient in whom myxofibrosarcoma generated from the thyroid. The tumor was characterized by spindle cell proliferation with low to moderate cellular density in myxoid stroma. Immunohistochemically, the tumor cells showed positive reactivity for vimentin and actin, but negative staining for S-100, CK and desmin. Tumor cells showed low proliferative activity as demonstrates by low Ki-67 labelling index (5%). The tumor was diagnosed as a low-grade myxofibrosarcoma. Despite the suboptimal general conditions of the patient, yet, surgery was the sole treatment of her and was successfully done. Adjuvant radiotherapy was indicated due to close circumferential margin. Myxofibrosarcoma is a very rare tumor in the head and neck region, and morphology beside immunohistochemistry is powerful tools to establish the diagnosis. Surgery is the main line of treatment followed by adjuvant radiotherapy to improve patient's survival.

исследовательская статья

Hepatotoxic Effects of Lead Acetate in Rats: Histopathological and Cytotoxic Studies

Zohra Haouas, Sallem A, Zidi I, Hichri H, Mzali I and Mehdi M

Lead toxicity is probably the most common form of heavy metal intoxication. The present study was conducted to assess the histopathological and cytotoxic effects of lead exposure on rat liver. Adult male Wistar rats were randomly divided in 2 groups. The first was exposed to 2 g of lead acetate in distilled water during 35 days, while the second served as a control group and was given distilled water. The structural damage in the liver was investigated by histological study and supplemented by biochemical assay of liver enzyme levels. DNA fragmentation in somatic cells was determined using terminal desoxynucleotidyl transferase mediated dUTP nick end-labeling (TUNEL) assay. The results obtained show increase in liver enzyme levels in treated rats compared to controls. The histological study showed that lead can induce several alterations such as hypertrophy of hepatocytes, portal space and central vein dilatation, vacuolation and lymphocytic infiltration. Tunel assay revealed significant DNA fragmentation in rats exposed to lead. This study showed that TUNEL assay can be used to determinate DNA fragmentation in somatic cells of rat liver. Moreover, we conclude that lead acetate may be considered as a strong hepatotoxic and genotoxic agent.

исследовательская статья

Epithelial Dysplasia in Oral Mucosa and its Association with Betel Quid Chewing

Waris S and Nagi AH

Objectives: Current study was performed to determine the frequency of epithelial dysplasia in pan chewers and to calculate association between the quantity and quality of pan chewing and epithelial dysplasia and its grades.
Methods: This was a cross-sectional, analytical study. Cytological smears from 300 subjects were collected. After fixation with alcohol, smears were then air dried and stained with Haematoxylin and Eosin, Giemsa and Pap’s stain. The material was then examined under an optical microscope. Epithelial dysplasia was identified and graded using the method of Speight.
Results: Epithelial dysplasia was observed in 57.7% of the habitual pan chewers which was significantly associated with number of pans consumed per day as well as duration of pan consumption. These were variables were positively correlated with grading of epithelial dysplasia.
Conclusions: The study concludes that epithelial dysplasia was common among chronic pan chewers in Lahore city. Significant association and correlation was found between epithelial dysplasia and number of pans consumed per day as well as duration of pan consumption.

исследовательская статья

Matricellular Protein Expression and Cell Ultrastructure as Parameters to Test In Vitro Cytotoxicity of a Biomimetic Scaffold

Sandra Durante, Gabriella Teti, Viviana Salvatore, Stefano Focaroli, Monica Mattioli-Belmonte, Concetta Ferretti, Adriana Bigi, Roberto Giardino and Mirella Falconi

Following scaffold implantation, cell sufferance, in-vivo encapsulation, foreign body reaction and inflammatory response has been reported and the up- regulation of matricellular proteins is often connected with this condition. Cytotoxicity of biomaterials is generally tested according to ISO standard 10993-5 based mainly on viability tests. Additional assays, based on improved cytotoxicity knowledge, are suggested in order to better analyze the biocompatibility of implant materials. The purpose of the study was to evaluate the matricellular protein expression as biomarker for in vitro-testing the biocompatibility of implant materials. Tenascin-C, osteocalcin and osteopontin belong to the matricellular protein family and were chosen as cytotoxicity markers. Mesenchymal stem cells were seeded on collagen/hydroxyapatite scaffold and on carboxymethyl cellulose based hydrogel in order to evaluate gene/protein expression by cell viability test, Real Time PCR and western blot. Electron microscopy was carried out to evaluate the morphological changes induced by cell/scaffold interactions. A low expression of tenascin-c, osteonectin and osteopontin was demonstrated in collagen/hydroxyapatite scaffold compared to the cells cultured on tissue flasks and on hydrogel scaffold. Based on our results, we propose matricellular protein expression as parameter for testing in vitro biocompatibility of implant materials.

исследовательская статья

Effect of Methotrexate on the Jejunum of Adult Albino Rat and the Possible Protective Role of Vitamin A: Histological and Immunohistochemical Study

Mohamed BM Ali, Maha H Ashmawy, Ehab A A El-shaarawy, Inas A Mohamed and Tarek I Abd El-Galil

Background: Methotrexate (MTX) is an antineoplastic drug that is widely used in the treatment of neoplasms. Patients undergoing MTX therapy have a variety of side effects affecting many important organs.
Aim of the work: The aim of the present study was to detect the histological and the immunohistochemical adverse alterations that might occur in the jejunum of the adult albino rats following methotrexate therapy and to determine the possible protective role of vitamin A. Material and
Methods: Seventy two adult male albino rats were divided into six equal groups. In all groups the animals were sacrificed 24 hours after the last injection. The jejunal specimens were examined using H& E and Masson’s trichrome stains and immunohistochemical examination to study the alkaline phosphatase activity using light microscopy. Morphometric study measuring villous height, crypt depth, number of goblet cells, percentage area of fibrosis and the optical density of alkaline phosphatase activity was done in all groups.
Results: The methotrexate treated groups’ revealed different changes in the jejunum of rats. Distortion and cystic dilatation of the crypts, cellular shedding and dilated blood vessels were found. Morphometric study showed decreased villus height and goblet cells, increased the crypts depth. Immunohistochemical examination of the jejunal sections of the methotrexate treated groups showed weak reaction of alkaline phosphatase enzyme when compared with that of the control group.
Conclusion: It can be concluded that the toxic effect of methotrexate on the jejunum of rats can be partially improved with the concomitant use of vitamin A.

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