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Introduction: The histology of fetal kidney differs significantly from the adult kidney, because the nephrogenesis, the synthesis of new nephrons, lasts until the end of the fetal period. The kidney develops from two mesodermal primordia: the metanephric blastema forms from the intermediate mesoderm and the ureteric bud forms from the mesonephric duct. Materials and methods: We studied the histological structure of the fetal renal tissues in 18 post mortem fetuses from the Department of Anatomy and Embryology, University of Medicine and Pharmacy Tîrgu Mureş. Results: On HE stained sections we identified the nephrogenic zone and the developing nephrons in different stages of their development. We noted the increase of renal size, cortical thickness and the number of glomeruli. Discussion: Our results are similar with those in the literature data. In the nephrogenic zone of the cortex, which is formed by undifferentiated stell cells, newgenerations of nephrons appear during fetal life. The nephrogenic zone is composed of thick undifferentiated blastema cells. The newly formed nephrons are organized in layers: the oldest nephrons are located in the inner portion of the cortex, while the youngest nephrons are located in the outer layer of the cortex. Conclusion: The development of the nephron in humans follows the development of the nephron understood from the studies made on mice and rats. [ABSTRACT FROM AUTHOR]

Introduction: Anatomical variations of the arterial supply of the abdominal organs are relatively frequent. The most variations were reported regarding the celiac trunk (CT), followed by the superior mesenteric artery (SMA) and the inferior mesenteric artery (IMA), which seems to be extremely rare. In case of the middle colic artery (MCA), multiple variations of origin and number have been described. Purpose: The aim of this study is to present the anatomical variations of the MCA based on a case report. Material and methods: We have dissected a formalin fixed male cadaver from the demonstration material of the Anatomy and Embryology Department of UMF Târgu-Mureş. Results: We have found two middle colic arteries: a right-middle and left-middle colic arteries arising independently from the SMA. The right-middle colic artery supplied the hepatic colic flexure and the right part of the transverse colon. The left-middle colic artery supplied the splenic colic flexure and the left part of the transverse colon. The middle part of the transverse colon was supplied by an arterial connection between the two mentioned arteries. Conclusions: Detailed knowledge of anatomical variations of the colic arteries is important especially during surgical and interventional radio-oncological approaches of the pathology involving the large intestine to prevent any inadvertent complication. [ABSTRACT FROM AUTHOR]

The aim of this work is to show that the bones with marrow, treated as a quasi-porous media, can be successfully used to study the effects of ovariectomy-induced osteoporosis. Proton one-dimensional (1D) nuclear magnetic resonance (NMR) T -distribution and two-dimensional (2D) T - T exchange maps combined with histological images were used to measure the proximal part of the femoris, diaphysis and distal epiphysis of ovariectomized and non-ovariectomized Wistar albino rats. The 1D normalized T distributions showed four peaks which were associated with protons in four major pools: (1) the protons from bounded water to collagenous matrix; (2) fluids in osteocyte lacunae and canaliculi channels; (3) fluids in secondary pores like Haversian and transverse Volkmann canals and (4) soft matter like bone marrow and fluids in primary pores like trabecular bone cavities. The peak's association and hierarchical structure of pores in femoral bone were supported by a 2D T - T exchange map and by a series of dehydration experiments monitored by NMR measurements. The bone marrow narrows the T -distributions, increasing resolution, but will not influence significantly the peaks positions; therefore, the NMR relaxometry is a valuable tool to characterize the pore distributions and effects of induced osteoporosis in diverse bones sections. [ABSTRACT FROM AUTHOR]

Chronic myeloid leukemiais the first human neoplasm with elucidated genetic background: the translocation between chromosome 9 and 22 represents the hallmark event. The resulting BCRABL fusion gene encodes a tyrosine kinase with abnormal activity. Depending on the breakpoint location of BCR gene, three main fusion gene subtypes are described, M-BCR-ABL being the most common, with two variants: b2a2 and b3a2.In this study our aim was to investigate the frequency of the two b2a2 and b3a2 variants and their relationship with clinical parameters. Material and methods: We tested 33 chronic myeloid leukemia patients at the Molecular Biology laboratory of the Anatomy Department of UMF Tg-Mures.We performed real time quantitative, and single step PCR. Results: From the 33 enrolled patients 31 expressed one of the M-BCR-ABL variants: 15 patients (48.38%) the b2a2 (51.61%) and 16 patients the b3a2 variant. None of the patients presented other subtypes. We found statistically significant correlation regarding platelet number (p=0.0036) and hematocrit value (p= 0.048) between the two variant subgroups, while other tested parameters (age, spleen size, white blood cell count, clinical phase, M-BCR-ABL expression level) did not differ significantly. Conclusions:Our findings are partially concordant with other publications. However, due to the variety of reported data regarding the phenotype of each variant, we consider a large multicenter study is required. [ABSTRACT FROM AUTHOR]

Introduction: Previous reports described two main olfactory systems and several subsystems. The main systems comprise the main olfactory epithelium and the vomeronazal organ. The septal organ is an olfactory subsystem described as an olfactory island surrounded by respiratory epithelium, located in the ventral area of the posterior part of the nasal septum. There are no reports confirming the existence of the human septal organ. Our objective was the identification of the septal organ in human fetuses using morphological methods. Material and method: We used a total of 8 human fetuses and performed the dissection of the nasal area to remove the nasal cavity as a whole. The obtained sections were stained using Alcian blue-van Gieson and PAS staining. To confirm the presence of neural elements we used the primary monoclonal antibodies neuron specific enolase and CD56. Results: Using the histological stains we identified an area of epithelium in the antero-inferior part of the nasal septum that displays the characteristics of the septal organ described in animals: absence of goblet cells, thicker than the surrounding respiratory epithelium, the lamina propria contains PAS+ and alcian-blue+ acini, capillaries, peripheral nerves, and the glands of the lamina propria open through the basement membrane of the epithelium. NSE immunohistochemistry revealed a weak positive signal throughout the septal and turbinate epithelium, and very strong expression in the epithelium of the superior wall, superior part of the septum and superior turbinate, and the vomeronasal organ. CD56 expression was limited to zones known to be chemosensory areas. Conclusions: Histological stains raise the possibility of a chemosensory epithelium that would correspond to the septal organ, but immunohistochemistry did not confirm this aspect. [ABSTRACT FROM AUTHOR]

Phosphatase and tensin homologue mutated on chromosome 10 (PTEN) is a tumor suppressor gene that encodes a protein inhibiting the PI3K/AKT pathway stimulated by EGFR. PTEN gene mutation or a decrease in PTEN expression plays an important role in colorectal carcinogenesis. The objective of this study was the correlation of PTEN expression with the main clinico-pathological factors of colon adenomas/polyps (A/P). We performed immunohistochemistry using the Ultravision Labeled Polymer System method, on colon A/P and carcinomas (CC) from the material of the Pathology Department of the County Emergency Clinical Hospital of Tîrgu-Mureş, Romania. PTEN expression decreases significantly in CCs versus A/P. In hyperplastic polyps and the majority of sessile serrated, and traditionally serrated adenomas the expression of PTEN was decreased. In tubular and tubulo-villous adenomas the expression of this marker changes mostly in case of high grade intraepithelial neoplasia. Our results confirm early changes of PTEN expression in colorectal carcinogenesis. [ABSTRACT FROM AUTHOR]

Introduction: Anatomical variations of the arterial supply of the upper limb are relatively frequent. In case of the radial artery, multiple variations of origin and aberrant course have been described. The objective of this work is the study of anatomical variations of this artery based on a case presentation. Material and methods: We have dissected a formalin fixed male cadaver from the demonstration material of the Anatomy and Embryology Department of UMF Târgu Mureş. Results: The left radial artery has been found to originate from the third segment of the axillary artery, between the roots of the median nerve, covered by the pectoralis maior muscle. In the arm it was accompanied by the median nerve, brachial artery and radial veins. In the elbow region it passed under the aponeurosis of the biceps muscle, and in the forearm it was positioned between the pronator teres muscle and the medial margin of the brachioradialis muscle. Conclusions: The anatomical course described in this case is different from those reported in published studies. Anatomical variations of the radial artery are important in medical practice because it is a frequently utilized graft for surgical myocardial re-vascularization. [ABSTRACT FROM AUTHOR]

The most commonly used hypolipemiant agents are statins and fibrates. Several studies have shown different pathways that relate these drugs with bone metabolism. As both dyslipidemia and osteoporosis are frequent in elderly patients, statins and fibrates might become a dual-purpose therapy. In this study we aimed to compare the effect of the two drugs on healthy and osteoporotic bone and also on the fracture healing process. For our experimental study we used Wistar female rats divided into 6 groups, half of them being ovariectomized and all suffered a mediodiaphysary fracture. Each group was treated with simvastatin, fenofibrate 10mg/kg daily or received no treatment respectively. Bone structure and fracture healing were evaluated at 2, 4, 6 and 8 weeks post fracture by histological means, with haematoxylin- -eosine staining. On bone structure, at week 6 and 8 statins reduced the bone trabeculas size on nonovariectomized rats, but increased them in osteoporotic rats, while fibrates showed no difference in comparison with the control groups. The fracture healing process was enhanced when simvastatin was administred, regardless if the bone was osteoporotic or not. At the same time, no obvious positive effect of fenofibrate on fracture healing process was observed, the outcome of the fracture being similar in fenofibrate treated rats and control group, both without osteoporosis. In this study, simvastatin had a better positive effect on bone than fenofibrate, which might recommend statins as a double-potential drug in dylipidemic and osteoporotic patients. [ABSTRACT FROM AUTHOR]

Objective: Despite various treatment utilised, vascular anomalies represent difficulty in many cases for physicians. The Bleomycin is a cytotoxic antitumoral antibiotic, that cose modifications of DNA. It has been also successfully used in intralesional injection treatment of cystic hygromas and haemangiomas, based specifically on a high sclerosing effect on vascular endothelium. We used this agent in the treatment of congenital haemangiomas and lymphangiomas, with various results. Therefore we tried to work out an experimental model, to better understand the mechanism of this substance, and the differences on the results. Method: We used the chick embryo chorioallantoic membrane (CAM) to study the effect of Bleomicine on the angiogenesis. Results: After three days of bleomycin administration, we noticed several histopatological changes on CAM treated with bleomycin: thickness decreases and stroma densification occurs; tendency of the figurative elements to extravasate in the stroma; large vessels, wich keeps the form of blood vessels, but the red blood cells are agglutinated, with stasis, thrombosis; the endothelium is no longer visible morphological. Conclusions: Histopathological changes observed on the CAM treated with bleomycin supports our teory, that this substance in addition to the specific sclerosing effect on vascular endothelium, also has an inhibitory effect on angiogenesis, wich would explain wy the treatment goes better on younger children. [ABSTRACT FROM AUTHOR]

Background: The effects of two lipidlowering drugs, simvastatin and fenofibrate on osteoporosis in the femurs of healthy and ovariectomized female rats were investigated quantitatively by histological images and 1H NMR relaxometry. Methodology: The intertrabecular cavities percentage was estimated from numerically cleaned histological images which initially present both the trabecular bone architecture and bone marrow filling the intertrabecular pores. The eight weeks evolution of intertrabecular cavities percentage measured from histological images of randomly selected sections in ovariectomized rats femoral diaphysis is similar with the evolution of transverse relaxation time centre of gravity T2,CG, a selected parameter to quantify the changes in intertrabecular cavities dimensions from onedimensional (1D) transverse relaxation time T2 distributions. Results: The analysis of histological images recorded for non-ovariectomized rats show that the treatment with simvastatin and fenofibrate has a negative effect on the dimension of the femoral diaphysis trabecular bone. The analysis of 1D T2 distributions show that the simvastatin and fenofibrate lipid-lowering drugs produce different effects on different particular sections of studied rat bones, i.e. proximal part of femoris, diaphysis or distal epiphysis. Conclusions: Finally, it is shown that the effects of the treatments are strongly dependent on the duration of treatment.

Abstract Background The immunohistochemical demonstration of Enhancer of zeste homologue 2 (EZH2) proved to be a useful marker in several tumor types. It has been described to distinguish reliably hepatocellular carcinomas from liver adenomas and other benign hepatocellular lesions. However, no other types of malignant liver tumors were studied so far. Methods To evaluate the diagnostic value of this protein in hepatic tumors we have investigated the presence of EZH2 by immunohistochemistry in hepatocellular carcinomas and other common hepatic tumors. EZH2 expression was examined in 44 hepatocellular carcinomas, 23 cholangiocarcinomas, 31 hepatoblastomas, 16 other childhood tumor types (rhabdomyosarcoma, neuroblastoma, Wilms’ tumor and rhabdoid tumor), 17 metastatic liver tumors 24 hepatocellular adenomas, 15 high grade dysplastic nodules, 3 biliary cystadenomas, 3 biliary hamartomas and 3 Caroli’s diseases. Results Most of the malignant liver tumors were positive for EZH2, but neither of the adenomas, cirrhotic/dysplastic nodules, reactive and hamartomatous biliary ductules stained positively. Conclusions Our immunostainings confirm that EZH2 is a sensitive marker of hepatocellular carcinoma, but its specificity is very low, since almost all the investigated malignant liver tumors were positive regardless of their histogenesis. Based on these results EZH2 is a sensitive marker of malignancy in hepatic tumors. In routine surgical pathology EZH2 could be most helpful to diagnose cholangiocarcinomas, because as far as we know this is the first marker to distinguish transformed and reactive biliary structures. Although hepatoblastomas also express EZH2, the diagnostic significance of this observation seems to be quite limited whereas, the structurally similar, other blastic childhood tumors are also positive. Virtual Slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1173195902735693

Background: The elucidation of the genetic background of the myeloproliferative neoplasms completely changed the management of these disorders: the presence of the Philadelphia chromosome and/or the BCR-ABL oncogene is pathognomonic for chronic myeloid leukemia and identification of JAK2 gene mutations are useful in polycytemia vera (PV), essential thrombocytemia (ET) and myelofibrosis (PMF). The aim of this study was to investigate the role of molecular biology tests in the management of myeloproliferative neoplasms. Materials and methods: We tested the blood samples of 117 patients between April 2008 and February 2013 at the Molecular Biology of UMF Târgu Mures using RQ-PCR (for M-BCR-ABL oncogene) and/or allele-specific PCR (for JAK2V617F mutation). Results: Thirty-two patients presented the M-BCR-ABL oncogene, 16 of them were regularly tested as a follow-up of the administered therapy: the majority of chronic phase patients presented decreasing or stable values, while in case of accelerated phase and blast phase the M-BCR-ABL values increased or remained at the same level. Twenty patients were identified with the JAK2V617F mutation: 8 patients with PV, 4 with ET, 3 with PMF, 4 with unclassifiable chronic myeloproliferative disease and 1 patient with chronic myelomonocytic leukemia. There was no case of concomitant occurance of both molecular markers. Conclusions: Molecular biology testing plays an important role in the management of myeloproliferative neoplasms: identification of the molecular markers confirms the final diagnosis, excluding secondary causes of abnormal blood count parameters. Regular monitoring of M- BCR-ABL expression level is useful in the follow-up of therapeutic efficiency. [ABSTRACT FROM AUTHOR]

Introduction: The BCR-ABL fusion gene [t(9;22) (q34;q11)] occurs in 3-5% of pediatric acute lymphoblastic leukemia (ALL) and predicts a very poor prognosis. Material and methods: 2 ml samples of bone marrow (BM) and peripheral blood (PB) in EDTA tubes from 24 ALL patients were examined in the molecular biology laboratory of our university with quantitative real-time PCR (qRT-PCR) method for BCR-ABL gene expression. Prognostic factors, like age, leucocyte count, lymphoblast morphology and immunology, absolute lymphoblast count on day 8, remission status on day 33 as well as treatment results were recorded from every patient. Results: All 24 qRT-PCR analysis for major and minor BCR-ABL gene expression from BM and PB were negative. Immunophenotyping performed in 25 patients revealed common B ALL in 12 patients, T-cell immunology in 3 and pre-B immunophenotype with aberrant myeloid marker expression and/or CD10 negativity in 11 patients. L1 morphology appeared in 85.7% of the pre-B ALL cases, while other immunophenotypes were more likely associated with L2 cytology (62.5%) (p=0.033). Early cortisone response was favourable in 22 patients, all 26 patients achieved complete remission on day 33. Common B immunophenotype was associated with lower WBC (mean 37,770, median 8,200) than other immunophenotypes (WBC mean 63,783, median 50,680). Conclusions: A new method, the qRT-PCR test was introduced in the investigation of pediatric ALL in our university from 2010. We found a statistically significant correlation between L1 blast morphology and common B immunophenotype. Poor cortisone response was found more frequently in T-cell ALL and pre-B ALL with aberrant myeloid markers or CD10 negativity. All our patients achieved complete remission on day 33. Lower WBC count at presentation was associated with L1 morphology and pre-B immunophenotype. [ABSTRACT FROM AUTHOR]

Background: Hip joint dislocation and dysplasia are frequent congenital malformations with a multifactorial etiology with a major role played by intrauterine mechanical forces during development. Our aim was to define the surface of the femur and acetabulum, and to study the changes in geometrical parameters and the relationships between them during development. Material and method: In this study we dissected the hip joints of 10 post-mortem fetuses. Fetal age was determined using crown-rump length (CRL) (min. 8.5 cm, max. 30 cm). Then we performed morphological and geometrical measurements on the articular components. We calculated the area of the articular surfaces, then determined the femoral head coverage and compared the antetorsion (AT) of the femur with the AT of the acetabulum. Results: in the 3 months fetus the surface areas of the femur and acetabulum are almost identical, the femoral head coverage is maximal, and later with age progression the head coverage decreases, as its growth is more intense than that of the acetabulum. During development the increase of femoral AT determines the increase of the acetabular AT. Conclusions: The decrease in femoral head coverage during development results in a decrease in articular stability. The femoral head's position and compression plays an important role in the development of the acetabulum. [ABSTRACT FROM AUTHOR]

Background: Adenomatous polyposis coli (APC) gene is thought to play a key role in the majority of sporadic colorectal cancers developed through the chromosomal instability pathway. In 10-15% of CRC the defect of the mismatch repair genes (MLH1, MSH2), the microsatellite instability is the underlying mechanism for carcinogenesis. The aim of this study was the correlation of APC, MLH1 and MSH2 immunoexpression in different types of colon adenomas/polyps (A/P). Materials and methods: We processed biopsies and surgical pieces of colon A/P and carcinoma developed in adenoma (CC). The APC, MLH1, MSH2 expression were graded, and used for establishing different immune phenotypes that have been compared by statistical tests. Results: The majority of tubular and tubulovillous adenomas have the MLH1+/MSH2+/APC+ immune phenotype, and the ratio of MLH1-/ MSH2-/APC+ cases increases in case of hyperplastic polyps and serrated adenomas. A/P developing in the right colon and in patients below 40 years were more frequently MLH1-/MSH2-/APC+. Conclusions: APC immunoexpression decreases in adenomas/polyps with dysplasia, and MLH1 and MSH2 expression is altered especially in hyperplastic polyps and serrated adenomas. [ABSTRACT FROM AUTHOR]

Background: Prevention of colorectal carcinomas is based on early detection and removal of precancerous lesions. Our goal was to perform a comparison between clinicopathological features of adenomas/polyps developed in the presence and in the absence of a colon carcinoma. Materials and methods: We studied a total of 117 colon adenomas and polyps selected from the material of the Pathology Department of the Emergency County Hospital of Tîrgu Mure. We compared and correlated through statistic analysis the clinicopathological factors, gender, age of the patients, localization, histological type, grade of dysplasia. Results: 24.6% of A/P develop synchronously with a carcinoma and affect especially women and patients between 50-60 and 70-80 years; these are mainly tubular adenomas, localized more frequently in the left colon. Adenomas and polyps developed without a carcinoma are more frequent in males, in patients between 60-70 years, and are mainly tubulovillous adenomas. Conclusions: One out of 4 colon carcinomas are associated with one or more precancerous lesions. [ABSTRACT FROM AUTHOR]

Objective: Immunohistochemical study of the left ventricle-aortic root and valvar junction complex and its relevant dynamics. Material and method: Serial sections of cardiac samples have been performed using usual histological procedures and stained with trichrome technique in order to evaluate collagen and elastic fibers. The valvar interstitial cells were immunohistochemically marked using SMA and vimentin antibodies, visualized by UltraVision Labeled Polymer System. Results: The ventriculo-aortic junction is a musculo-fibrous circle surrounding the clinical annulus. The interposed dense connective bundles and interfoliate triangles anchor the aortic root to the outflow tract of the aortic ostium. The valvular leaflets have a fibro-spongious core turning into the middle layer of the aortic root. The structure of the fibrous layer support has a typical mechanical architecture. Conclusions: The bindings between the aortic root and valve reflect the valvular loads and tensile forces to the compliant aortic root. The valvar interstitial cells, mechanoreceptive myofibroblasts, are activated by transforming growth factor signals in the process of adaptation of fiber orientation during junction remodeling. [ABSTRACT FROM AUTHOR]

Introduction: Dyslipidemias represent a major public health problem. Statins are inhibitors for 3-hydroxy-3-methylglutaril coenzyme A (HMG CoA) reductase, which plays an important role in the second step of cholesterol biosyntesis, but also in bone metabolism. The purpose of this study has been to determine the effect of statins on bone and to correlate the statistical and clinical results in order to establish recommendations regarding the treatment of dyslipidemias. Material and method: In the clinical part we studied the incidence of dyslipidemias in 20 family medicine practices randomly chosen: 10 from the rural environement and 10 from the urban area, during the National Assessment Program of Public Health. In the experimental part, we compared histologically the process of callus formation at ovariectomised rats. We administered simvastatine and we examined with hematoxylin-eosin staining the femoral fracture, prelevated on the 2nd, 4th, 6th and 8th postoperatory week. Results: In the rural environement, 14.35% of patients were diagnosed with dyslepidemia while in the urban area the percentage was of 12.8%. Out of the newly diagnosed diseases, 55.6% were dyslipidemias. Eight weeks after fracture, the control group (non-ovariectomised, no treatment) presented a late ossification, while in rats treated with simvastatine (ovariectomised and non-ovariectomised) the callus was bony. The non-treated ovariectomised group revealed a chondro-hyaline callus. Conclusions: In our experiments, the treatment with statins enhanced callus formation. As both young and old patients are exposed to traumatic or osteoporotic fractures, it is important to establish a series of treatment protocols for dyslipidemia, that should not only reduce the cardiovascular risk but also the development of osteoporosis and its complications. We recommend the treatment with simvastatine in cases which associate osteoporosis and dyslipidemia. [ABSTRACT FROM AUTHOR]

Introduction: Breast cancer is the leading type of cancer affecting women, with an increasing trend of mortality, and a lifetime risk of more than 10%. There are cases of inherited predisposition, which is mostly caused by mutations of the tumor suppressor genes BRCA1 and BRCA2. The objective of this retrospective study is to identify presumptive candidates for BRCA1 mutation carriers using available clinical, histopathological and immunohistochemical data. Material and method: We processed the available recorded data of 1334 patients diagnosed with breast cancer between the years 2005 and 2010 from the archives of the Pathology Department of the Clinical County Hospital Tg.-Mures. We considered age under 45, histopathological characterization and available immunohistochemical information. Results: The main selection criteria (age below or equal to 45 years) excluded the majority from further analysis, while 188 cases (12.35%) remained. After performing distribution into histopathological type groups, we looked at the immunohistochemical data. We obtained 19 ER-/ PR- negative cases without Her2/neu information, and only 12 cases could be proven to be triple negative, and thus to be considered as primary candidates for BRCA mutation screening. Conclusions: We identified a very small number of presumptive BRCA mutation carrier candidates. This is due mostly to the fact that the information available comprised macroscopic and classical histopathological data. Immunohistochemical characterization is not widespread, but this shortcoming will be circumvented by prospective reprocessing of the archived biological materials, in order to characterize the BRCA mutation patterns of our population. [ABSTRACT FROM AUTHOR]

Introduction: The kidney develops from two mesodermal primordia. Aquaporin 1 (AQP1) is a membrane protein characteristic to epithelial and endothelial cell of the human body. The Pax family of genes encodes transcription factors with important role in intrauterine development. Connexins are transmembrane proteins found in gap junctions. We monitored the changes in the expression of AQP1, paired box gene 2 (PAX2), paired box gene 8 (PAX8), connexin 36 (Cx36) and connexin 43 (Cx43) proteins in fetal renal tissue., Materials and Methods: We studied 34 post mortem fetuses of 9 to 24 weeks from the Laboratory of Pathology, Emergency County Hospital of Târgu Mureş, Romania, using immunohistochemistry., Results: AQP1 expression appeared in the apical and basolateral parts of cells, lining the proximal convoluted tubules and the descending limb of Henle's loop, then in the tubule pole of Bowman's capsule also. Nuclear expression of PAX2 was observed in structures developed both from the ureteric bud and the metanephric mesenchyme, and of PAX8 was observed in the proximal convoluted tubule's epithelium, Henle's loop, and collecting ducts. Cytoplasmic expression of Cx36 was localized to nephrons in different developmental stages, glomerular vessels and collecting ducts, and of Cx43 was localized to the endothelium of glomerular and peritubular vessels, as well as to the epithelium of the proximal tubules., Discussions and Conclusions: Nephrogenesis begins in the embryonic period, and continues into the fetal period as well. It is regulated by a wide array of markers. The current study supplements literature data regarding immunoexpression of these markers during renal development in the fetal period.

Unlabelled: The study of epidermal growth factor receptor (EGFR) gene mutations in lung adenocarcinoma patients has a special clinical significance in the selection of patients for tyrosine-kinase inhibitor therapy. The aim of this study was to identify patients with EGFR mutations using allele-specific polymerase chain reaction (PCR), from formalin-fixed paraffin-embedded (FFPE) tissue and fresh tissue (FT)., Materials and Methods: We performed a retrospective study using 13 cases of FFPE lung adenocarcinoma, and a prospective study using seven fresh samples of lung carcinomas (FT), collected by intraoperative dissection of the tumors. Using the DNA extracted from the FFPE tissue and FT, we attempted to identify deletions of exon 19 and point mutations of exon 21, according to the allele-specific PCR method described by Dahse et al. (2008)., Results: In all seven cases of FT (three adenocarcinomas, three squamous carcinomas, one large-cell carcinoma), we identified the wild type allele and the internal control in case of exon 19, and the wild type allele for exon 21, but not the mutated alleles. Considering that no standard method for formalin fixation and paraffin embedding has been implemented at the Laboratory of Pathology, the DNA extracted from these samples became fragmented and damaged, which compromised the results of PCR testing aimed at the detection of EGFR mutations., Conclusions: The presented method can be implemented at our laboratory to identify these mutations from fresh tissue collected during surgical resection. Additionally, standardization of formalin fixation and paraffin embedding of surgical samples is required, in order the enable subsequent processing using molecular biology methods.

Background: Reproducible structural analysis was made on cirrhotic human liver samples in order to reveal potential connections between morphological and laboratory parameters., Material and Methods: Large histological samples were taken from segment VII of 56 cirrhotic livers removed in connection with liver transplantation. Picro Sirius red and immunohistochemically (smooth muscle actin [SMA], cytokeratin 7 [CK7], Ki-67) stained sections were digitalized and morphometric evaluation was performed., Results: The Picro Sirius-stained fibrotic area correlated with the average thickness of the three broadest septa, extent of SMA positivity, alkaline phosphatase (ALP) values and it was lower in the viral hepatitis related cirrhoses than in samples with non-viral etiology. The extent of SMA staining increased with the CK7-positive ductular reaction. The proliferative activity of the hepatocytes correlated positively with the Ki-67 labeling of the ductular cells and inversely with the septum thickness. These data support the potential functional connection among different structural components, for example, myofibroblasts, ductular reaction and fibrogenesis but challenges the widely proposed role of ductular cells in regeneration., Conclusion: Unbiased morphological characterization of cirrhotic livers can provide valuable, clinically relevant information. Similar evaluation of routine core biopsies may increase the significance of this 'Gold Standard' examination.

Unlabelled: The Philadelphia chromosome and the resulting BCR-ABL fusion gene represent the hallmark event in chronic myeloid leukemia (CML) and their discoveries radically changed the management of these patients. Currently Wilms tumor 1 gene (WT1) is intensively investigated as high WT1 expression levels have been demonstrated in case of multiple solid tumors and malignant hematological syndromes (acute myeloid and lymphoid leukemia, myelodysplastic syndromes and chronic myeloid leukemia). The aim of our study was to investigate the WT1 expression in CML patients and its possible contribution to disease evolution., Patients and Methods: In the Laboratory of Molecular Biology, University of Medicine and Pharmacy of Tirgu Mures, Romania, we regularly determined the M-BCR-ABL and WT1 expression levels by RQ-PCR (real-time quantitative polymerase chain reaction) testing in case of 19 CML patients: six patients monitorized from the diagnosis and 13 patients first tested during therapy., Results: Eight CML (four advanced stage and four CP) patients showed high WT1 expression level, and in case of 11 patients the WT1 expression levels were undetectable or lower than 0.02%. The only significant difference between the high and low WT1 expression groups was represented by the clinical stage. In the majority of pretreated patients (10 out of 13 patients), the WT1 expression levels were low or undetectable., Conclusions: High WT1 expression in CML patients is detected especially in the advanced stages of the disease. Efficient Imatinib therapy may contribute to low WT1 levels in CP patients.

Introduction: HER2, EGFR, p53 and PTEN are important in organization of the germ layers, in embryonic development and morphogenesis, in the development and differentiation of certain organ systems and in embryonic morphogenesis. Our goal is the comparative examination of the expression of these markers in the digestive tract of 9-24-week-old fetuses., Materials and Methods: We studied using immunohistochemical techniques esophagus, stomach, small and large intestine tissue samples collected from 18 post mortem fetuses of 9-24 weeks., Results: HER2 and PTEN expression appears as early as the 9-12 weeks period in the digestive tract, but HER2 expression decreases in the 21-24 weeks period and then disappears. EGFR expression appears only during the 13-16 weeks period. The expression of p53 is strong until week 21, and then it is restricted to the deeper layers of the epithelium., Conclusions: Our findings suggest that these markers have role also in the fetal period and complete the scarce data found in literature about the expression of the studied markers in the development of the digestive tract.

The effects of two wonder drugs, simvastatins and fenofibrates on the proximal part of the femoris of a series of ovariectomized and non-ovariectomized Wistar albino rats was estimated qualitatively and semi-quantitatively by the modern method of 1D 1H-NMR T2-distribution. The 72 rats subjected to this study were divided in six groups and were sacrificed at two, four, six and eight weeks after ovariectomy and the proximal part of femoris was harvested. The CPMG (Carr-Purcell-Meiboom-Gill) echoes train curves were measured for the bones fully saturated with water during two months after two months of natural drying. These decays were analyzed by Laplace inversion and an average of normalized T2-distributions was considered for all rat's groups. The 1D averaged T2-distributions present four peaks, which were associated with protons in four major environments, from which the free water protons are used as spy molecules to explore the boundaries of cavities. In the approximation of spherical pores, the averaged T2-distributions were transformed in distributions of pores diameters. These were found in the range from 2 μm up to 2 mm. The relative amplitudes, widths and position of deconvoluted distributions of small, medium and large cavities are used for a qualitatively analysis of the effect of our lipid-lowering drugs. For a semi-quantitatively analysis, we chose the diameter d of proximal part of femoris' trabecular cavities. We show that the positive or negative effects of treatments with simvastatins and fenofibrates are strongly dependent on the duration of treatment. Moreover, the treatment of healthy bone is generally counter-indicated.

Unlabelled: The balance between apoptosis and proliferation is tipped towards a decrease of apoptosis as the colonocyte progresses in the adenoma to carcinoma sequence of colon carcinogenesis. According to literature data, proteins like p53, Ki-67, APAF-1, Ets-1, PTEN contribute to inhibition of apoptosis and stimulation of proliferation., Aim: Considering the complex interference among colorectal carcinogenetic mechanisms, our aim was to study the markers Ets-1 and APAF-1 relative to p53, Ki-67 and PTEN expression in colon adenomas/polyps (A/P)., Materials and Methods: We performed immunohistochemistry on 99 colon A/P cases from the material of the Department of Pathology, Emergency County Hospital of Tirgu Mures, Romania. Secondary EnVision Flex/HRP (Horseradish peroxidase) (20 minutes) was used for signal amplification., Results: The majority of A/P show increased Ki-67, p53, Ets-1 expression, decreased APAF-1 expression and preserved PTEN expression. p53, Ki-67, Ets-1 and APAF-1 demonstrated statistically significant correlations with histological type and grade of dysplasia. We also observed that expression of these proteins in the intestinal crypts has a typical distribution according to histological type and grade of dysplasia., Conclusions: In case of hyperplastic polyps APAF-1 expression decreases as p53 and Ki-67 expression increases, followed by a decrease in PTEN expression in serrated adenomas, and an increase of Ets-1 expression in conventional adenomas.

The hepatic stem cells reside periportally forming the canals of Hering in normal liver. They can be identified by their unique immunophenotype in rat. The oval cells, the progenies of stem cells invade deep the liver parenchyma after activation and differentiate into focally arranged small-and eventually trabecularly ordered regular hepatocytes. We have observed that upon the completion of intense oval cell reactions narrow ductular structures are present in the parenchyma, we propose to call them parenchymal ductules. These parenchymal ductules have the same immunophenotype [cytokeratin (CK)7-/CK19+/alpha-fetoprotein (AFP)-/delta-like protein (DLK)-] as the resting stem cells of the canals of Hering, but different from them reside scattered in the parenchyma. In our present experiments, we have investigated in an in vivo functional assay if the presence of these parenchymal ductules has any impact on a progenitor cell driven regeneration process. Parenchymal ductules were induced either by an established model of oval cell induction consisting of the administration of necrogenic dose of carbontetrachloride to 2-acetaminofluorene pretreated rats (AAF/CCl4) or a large necrogenic dose of diethylnitrosamine (DEN). The oval cells expanded faster and the foci evolved earlier after repeated injury in the livers with preexistent parenchymal ductules. When the animals were left to survive for one more year increased liver tumor formation was observed exclusively in the DEN treated rats. Thus, repeated oval cell reactions are not necessarily carcinogenic. We conclude that the expansion of hepatic stem cell compartment conceptually can be used to facilitate liver regeneration without an increased risk of tumorigenesis.

Aim of the Study: Establishment of Ki67, p53 and CD34 expression in human tooth buds of different stages of odontogenetic development., Materials and Methods: Tissue samples containing tooth buds were removed from the incisor areas of human fetuses in different stages of development (weeks 9-10, 12-13, 13-16, 21-24), and from the canine and molar areas of 21-24 weeks fetuses. The tissue fragments were fixed using formalin and were processed using common histological techniques with paraffin embedding. Immunostaining for Ki67, p53 and CD34 has been performed using the dextran method and moist heat antigen retrieval (except for CD34). The resulting slides were photographed and quantitatively evaluated., Results: Ki67 immunoexpression decreases with advancement of the developmental stage of the tooth bud: in the inner enamel epithelium, between weeks 9 and 16 (IEE), in the preameloblasts (PB) between weeks 13 and 16, in the ameloblasts (AB) between weeks 21 and 24; outer enamel epithelium (OEE); stratum intermedium (SI); in the dental papilla: between weeks 9 and 10 in the dental papilla (DP), between weeks 13 and 16 in the outer layer of the dental papilla (DP1) and in the central layer of the dental papilla (DP2). Likewise, we noted Ki67 expression in the odontoblast layer (O) and pulp (P), between weeks 21 and 24. Concerning CD34 expression, we observed a decrease from weeks 9-10 until weeks 13-16, followed by an increase until weeks 21-24 of intrauterine life. From weeks 9-10, we observed a constant decrease of expression until weeks 13-16, followed by an increase during weeks 21-24., Conclusions: All Ki67, p53 and CD34 have been identified in the tooth bud. Ki67 expression gradually decreases with the embryonic development of the tooth, while p53 and CD34 expression decreases from weeks 9-10 to weeks 13-16 of intrauterine life, followed by an increase until weeks 21-24.

Lymphangiomas are uncommun congenital malformations of the lymphatic system, that involve the skin and subcutaneous tissues. Of the several types of treatment, surgical excision has been the preferred. There is a high recurrence rate because lymphangiomas tend to infiltrate the surrounding tissues. The bleomycin is a cytotoxic antitumoral antibiotic, that causes modifications of DNA. It has been also successfully used in intralesional injection treatment of cystic hygromas and haemangiomas, based specifically on a high sclerosing effect on vascular endothelium. We report the cases of five patients, with congenital lymphangioma, localized on the leg, in cervical and latero-thoracal region, treated with repeated intralesional bleomycin injections. The treatment indication was given by the location of this lesions and the infiltration of the surrounding vital tissues, that made the complete surgical excision impossible. Intralesional injection of bleomycin into the lymphangiomas was given at a dose, not exceeding 0,5 mg/kg of body weight, at intervals of 4 weeks. Complete resolution (n = 4) or significant improvement (n = 1) occurred in all patients treated. No other treatment was needed. We didn't notice local or general adverse effects. With this method we set the purpose to treat effectively this congenital malformations, obviating the need for invasive primary surgery or systemic treatment regimens. Toward other methods, intralesional bleomycin injections have a minimal risk of side effects (ulceration, pulmonary fibrosis).

The cerebral cortex ensures an optimal interaction of mammals, including humans, with their environment, by encoding, storing and combining information about the surrounding world and the internal milieu. Probably the simplest and the most popular region for studying the cortical network is the hippocampal CA1 area, because it has the least heterogeneous neuronal population, the somata and dendrites of principal neurons (pyramidal cells) are arranged into well defined layers and the extrinsic and intrinsic inputs are segregated. The relatively homogeneous pyramidal cell population is supported by a very heterogeneous GABAergic interneuron population, which provides not only general inhibition, but also regulates the precise timing of pyramidal cell activity. Interneurons usually innervate distinct domains of the surface of their target cell. The strategic placement of inhibitory synapses, indicate that GABAergic interneurons belonging to different classes serve distinct functions in the hippocampal network. Neuron types are usually defined according to various morphological, molecular and physiological features. Under typical experimental conditions only some of these parameters are available, therefore an important scientific question is: which partial measures are sufficient for correct recognition of a class of cell. By immunohistochemistry it is possible to stain all neurochemically identical neurons in a given brain region, therefore it is the most widely used method for identifying neuron classes. This review presents the neuron types identified so far in the area CA1 of the rat hippocampus with special emphasis on the immunocytochemical characterization of these cells.

Inhibition in the central nervous system is largely mediated by local-circuit neurons that release GABA (gamma-amino-butyric acid). GABAA-receptors play a major role in virtually all brain physiological functions and serve as targets for numerous classes of drugs, used both in clinical practice and as research tools. These receptors are heteropentamers, alpha1 being the most widely occurring subunit; therefore it is the best candidate to be studied in pathological conditions where the inhibitory system might be altered (e.g. epilepsy). We compared quantitatively the regional distribution of GABAA-receptor alpha1-subunit (GABAAR-alpha1) expression in three brain areas: neocortex, hippocampus and cerebellum by RT-qPCR. TaqMan probe was used in order to avoid detection of non-specific amplification products and synaptophysin as internal control. This substance was chosen because it has a stable expression restricted to neurons, and contrary to GAPDH, the most commonly used reference gene for expression analysis, synaptophysin expression is not modified in animal models of epilepsy. Expression of synaptophysin was higher than expression of GABAAR-alpha1 in all samples from the central nervous system. The latter was significantly different among the studied brain areas. It was the smallest in the hippocampus, intermediate in the neocortex and the highest in the cerebellum. Interanimal differences were small for any brain region under study. These results indicate that combination of TaqMan real-time PCR method with synaptophysin as internal control can reliably measure the relative expression of GABAAR-alpha1 mRNA, and are suitable for investigating the modifications that appear under pathological conditions and/or diverse experimental paradigms.

It is thought that dysregulation of E-cadherin, syndecan-1 (CD138) and Ets-1 is involved in carcinoma development. E-cadherin is an important epithelial cell adhesion molecule; syndecan-1 (CD138) is a regulatory proteoglycan in both cell-cell and cell-matrix adhesion and Ets-1 is a proto-oncogene and transcription factor, which takes part in extracellular matrix remodeling. Our goal was to study the changes in the expression of these molecules during colon carcinoma development and progression. We tested 117 colon adenomas and 149 de novo and ex adenoma carcinomas of the colon, using the Ultravision Polymer system. The positive reaction rate was 100% for E-cadherin, 98.3% for syndecan-1 and 22.4% for Ets-1 in adenomas, while in carcinomas it was 88.5%, 62.4% and 56.3% respectively. We found decreasing expression of E-cadherin and syndecan-1 throughout colon carcinoma progression and an opposite regulation for the Ets-1 protein. Decrease in expression of syndecan-1 is more pronounced in carcinomas compared to E-cadherin. De novo carcinomas have lower E-cadherin and syndecan-1 expression, and higher Ets-1 expression compared to ex adenoma carcinomas. These findings support the hypothesis that there are differences in the carcinogenesis of these tumors.

The aim of this study is to present the diagnostic and differential diagnostic criteria of the bone marrow specimen involved by lymphomas based on the histomorphological immunophenotype features and clonality of the tumor cells, patterns of lymphoproliferation and diagnostic pitfalls. BMB material obtained from the right posterior iliac crest was represented from 87 untreated and treated patients with BM involving malignant lymphoma, stained with Hematoxylin-Eosin, Giemsa, Periodic Acid Schiff and Gömöri's Silver. In order to perform immunohistochemistry examination we used a large antibody panel. B-cell clonality was determined in six cases. We found eight reactive lymphoproliferative responses and 79 lymphoid neoplasms of which 45 were diagnosed as de novo lymphoma, the rest of 34 samples being examined for staging. The predominant lymphoma was CLL (30 cases), over followed by DLBCL (18 cases). The most frequent patterns of involvement were the interstitial (29%) and mixed (15%) ones. In eight cases, we found reactive lymphoid aggregates. The B-cell clonality test showed four monoclonal, one oligoclonal and one polyclonal diseases form. Diagnosis of lymphoma versus reactive aggregate has been based on the combination of a lot of antibodies and involvement pattern. Although investigation of gene rearrangement was necessary for the establishment of the correct diagnosis in only 6.9% of cases, it should be emphasized that it is of great importance in disease monitoring.