Dobrava virus carried by the yellow-necked field mouse Apodemus flavicollis, causing hemorrhagic fever with renal syndrome in Romania.

Hemorrhagic fever with renal syndrome (HFRS) has been confirmed by serological methods during recent years in Romania. In the present study, focus-reduction neutralization tests (FRNT) confirmed Dobrava hantavirus (DOBV) as the causative agent in some HFRS cases, but could not distinguish between DOBV and Saaremaa virus (SAAV) infections in other cases. DOBV was detected by a DOBV-specific TaqMan assay in sera of nine patients out of 22 tested. Partial sequences of the M genomic segment of DOBV were obtained from sera of three patients and revealed the circulation of two DOBV lineages in Romania. Investigation of rodents trapped in Romania found three DOBV-positive Apodemus flavicollis out of 83 rodents tested. Two different DOBV lineages were also detected in A. flavicollis as determined from partial sequences of the M and S genomic segments. Sequences of DOBV in A. flavicollis were either identical or closely related to the sequences obtained from the HFRS patients. The DOBV strains circulating in Romania clustered in two monophyletic groups, together with strains from Slovenia and the north of Greece. This is the first evidence for the circulation of DOBV in wild rodents and for a DOBV etiology of HFRS in Romania.

Nestin and caveolin-1 in the diagnosis of GISTs.

Gastrointestinal stromal tumors (GISTs) are the most common primary mesenchymal neoplasias of the gastrointestinal tract, typically expressing c-kit (CD117) and CD34. Recently, it was reported that nestin and caveolin-1 are also expressed in some human sarcomas, GISTs included. We performed a retrospective study on formalin fixed, paraffin embedded samples from 81 cases of confirmed GISTs, aiming to characterize their immunohistochemical profile, including nestin and caveolin-1 expressions. Tissue samples were evaluated immunohistochemically for CD117, CD34, nestin and caveolin-1. The patients (M:F 36:45), aged 46 to 84 years, had spindle cell type GISTs in 56.7% of cases, epithelioid in 30.8% and mixed pattern in 12.3%. Immunohistochemically, CD117 was positive in 88.9% of GISTs, CD34 in 85.1%, nestin in 77.7% and caveolin-1 in 71.6% of the tumors. Of nine c-kit negative GISTs, 66.7% expressed nestin, the same as caveolin-1 and 44.5% expressed both nestin and caveolin-1. Statistical analysis using Kendall's and Spearman's tests revealed significant correlations between nestin and caveolin-1 expressions (p=0.024). Our results suggest that nestin and caveolin-1 could be considered sensitive markers in GISTs, together with CD117 and CD34, for diagnostic purposes. Their significant expression in CD117 negative GISTs could represent an immunohistochemical alternative in establishing the diagnosis of these tumors.

Statistical correlations between peripheral blood lymphocyte subpopulations and tumor inflammatory infiltrate in stage I of skin melanoma.

The article presents statistical correlations of immune cell parameters investigated in patients diagnosed with skin melanoma stage I. Recent data indicate a suppressed immune response, probably sustained by immune-regulating molecules expressed or shed from the tumor. These molecules block an efficient immune response and thus the tumor develops. All the molecules that are part of the tumor escape mechanisms can be targets for immune-mediated anti-tumor agents. We try to find the significance of some immunohistochemical markers (UCHL1, CD4, CD8) in tumoral inflammatory infiltrate and to establish the statistical correlations between molecular markers and tumor grade and stage. The studied parameters were: CD3+, CD4+, CD8+, CD56+16+ and CD19+. The statistical results were performed with SPSS v. 15.0. We demonstrate that a CD4+ on-going immune response is elicited in the investigated patients. We found a possible compensatory mechanism between T-lymphocytes and NK-cells and also between the antibodies generating cells and the natural cytotoxic cells. We are confident that these statistical correlations between clinical, immunological and immunohistochemical data can be useful in the disease management and personalized immune-therapy.

The histopathology analysis of the diffuse sclerosing variant of the papillary carcinoma of the thyroid: a distinctive and rare form.

Diffuse sclerosing variant (DSV) is a rare variant of papillary thyroid carcinoma (PTC) and its features have not yet been fully characterized. The aim of this case report is to analyze the immunohistochemistry profile of this disease and to highlight this rare entity. We analyzed the histology and immunohistochemistry of a female patient admitted in the Surgery Department of the "Sf. Ioan" Emergency Hospital, Bucharest, in April 2008. We especially used a wide panel of antibodies (Thyroglobulin mono, Thyroglobulin poly, Ki-67, TTF 1, Cytokeratin 19, Cytokeratin 34betaE12, and p63) in order to point out the follicular origin of the cells and to investigate the extensive squamous metaplasia lesions. The immunohistochemistry (IHC) was performed on 3 mum thick sections from 10% formalin-fixed paraffin-embedded tissues, according to the indirect tristadial Avidin-Biotin-Complex method. Our case report reveals that the DSV of PTC has distinctive pathologic features and its diagnosis importance is suggested by the high incidence of recurrence after operation.

Inflammatory, degenerative and vascular lesions in long-term dialysed patients.

UNLABELLED: No other medical field but nephrology showed so important achievements as a result of concerted efforts of doctors, biologists and technicians. Considering that in renal insufficiency, regardless of its aetiology, the common path is represented by the transitory or definite damage of the "renal filter", many attempts have been made in order to reproduce the process of blood cleaning by the kidney. AIM: The high prevalence of cardiovascular diseases in hemodialysed population suggested that the disease could begin before or during the stage of chronic renal insufficiency. We investigated the vascular lesions, especially the immunologic features of patients involved in hemodialysis programs. Our study reflects a general picture of the immunologic status of hemodialysed patients, helping to understand the special profile ofa hemodialysed patient. MATERIAL AND METHODS: Two groups have been selected and analyzed: one of 15 patients from the Hemodialysis Department in "Sfântul Ioan" Hospital, and another one composed of 30 patients with other diseases (the control group), from the Medical Department. The detection of specific antibodies against some HCV proteins corresponding to the most conserved regions of the viral genome has been done using the immune test LiaTEK HCV III. In the hemodialysed group, a blood sample has been drawn before and after the hemodialysis session, at 15-20 minutes, while in the other group the blood sample has been drawn together with the other tests. A flow cytometry examination was made at the Center of Immunology, in order to determine simultaneously several physical and chemical parameters. We analyzed the two groups of patients (HD/n=15; the reference group/n=30) regarding immunophenotyping, all types of lymphocytes and interleukin 2 (IL-2). RESULTS: The results have classified the HD patients into three subgroups, depending on the mean of the results from flow cytometry exam, referred to normal values. The assessment of the patients with or without HD to each group was made on the basis of the similar behavior of the markers investigated. The most affected age group in patients with HD was 31-40 years, followed by the age group 41-50 years (26%). The majority patients of the control group were of the same age 31-40 years old (40%), while 33% of them were between 41 and 50 years old. One subgroup (A) of HD patients showed the improvement of the total number of T lymphocytes (CD3+/CD19-) after the session, while the total number of B lymphocytes was stable. The number ofT lymphocytes with receptors for interleukin 2 (CD25) improved after hemodialysis. The second group (B) presented from the beginning a low number of total T lymphocytes (CD3+/CD19-). We found that the value of B lymphocytes (CD19+/CD3-) decreased after hemodialysis. Activated T lymphocytes (CD25), with receptors for interleukin 2, achieved greater values (3.66%), which cannot be found in the other groups. The third group (C) showed normal values for total T lymphocytes (CD3+/CD19-) before HD, which did not modify significantly after the session. The patients had the same decreased values for B lymphocytes, which have continued to decrease after the HD (7.98%). CONCLUSIONS: Post dialytic immunologic changes of the mononuclear cells represent the hallmark of the complexity of the immune response generally and especially too, in hemodialysed patients. We have noticed patients that presented an increase of the total number of T lymphocytes after the dialysis, but only T and NK lymphocytes and not B lymphocytes as well, suggesting the susceptibility to infections. The evaluation of the immune response using flow cytometry has confirmed the presence of high variations of the immune profile in hemodialysed patients, the decrease of T-cells activation but, it does not support the data regarding intrinsic functions of T and B cells. The high diversity of the immune response in hemodialysed patients is a consequence of the genetic individuality of each patient and also of the associated pathology or equipment used (viral infection, membrane type).

The usefulness of immunohistochemistry in sporadic colorectal cancer.

BACKGROUND: Colorectal cancer is an important disease with a large morbidity and mortality and also with increasing health care costs because of widespread of the multi-modal therapy and of the new drugs that continue to appear. There are 678.000 colorectal cancer cases and 400.000 deaths from the disease worldwide. It is the second commonest cause of cancer death in the European Union but, unlike the commonest cause of cancer related death that is lung cancer, the basis of the disease initiation is currently not understood. At the same time, the incidence increases with age, the carcinomas being rare before the age of 40 years, excepting individuals with genetic predisposition or predisposing conditions such as inflammatory bowel diseases. The early detection of colorectal cancer is potential associated with an important decrease of the cancer related mortality. AIM: Our study proposes to find out the significance of some immunohistochemical markers (VEGF, p53, CK20 and CEA) in sporadic colorectal carcinoma cases and to establish the statistical correlations between molecular markers and tumor grade and stage. MATERIAL AND METHODS: We investigated histopathological 40 inpatients (19 female and 21 males) who undergone surgery for colorectal carcinomas in "Sf. Ioan" Emergency Hospital, Bucharest, between September 2005-September 2006. We proceeded the histopathological examination to establish the grade, stage and the main features of the tumors, and then we analyzed using ABC method for immunohistochemistry the following markers for 20 selected cases: vascular endothelial growth factor (VEGF), carcinoembrionic antigen (CEA), cytokeratin 20 (CK20), and p53 oncoprotein. Finally, we analyzed statistical the results using t-Student test. RESULTS: The distribution of colorectal cancer cases (n = 40) regarding the age has showed the preponderance of patients older than 70 years (22/55%) and a small percentage of younger adults (2/5%). The repartition of colorectal tumors of sex ratio outlines a small difference between males (21/52.5%), and females (19/47.5%). The histopathological analysis of tumor grade in the 40 cases has revealed a high percent of moderate grade tumors (23/57.5%), in comparison with the poor differentiated tumors (11/27.5%) and the well-differentiated cancers (6/15%). The neovascularity within the stroma, the main features of tumor growth, has been noticed in 15 cases (3.75%), and also an important inflammatory lymphocyte infiltrate in nine cases (22.5%). We have noticed positive correlation between VEGF1 and CK20 (r = 0.4, p = 0.05), and between VEGF1 and CEA (r = 0.88, p = 0.001). In addition, our results demonstrate a positive correlation between tumor grade and CEA (r = 0.43, p = 0.009), and no relation among the other markers. CONCLUSIONS: Our present study shows that CK20 and CEA are positive immunostaining markers no matter the stage (100%). The oncoprotein p53 has been negative in T1 and T2 stages, but in advanced stages has been positive in a half of cases (50%). Regarding the location, p53 and VEGF showed positively results whatever the topography. We have noticed a direct proportional relation in VEGF expression and CEA, and CEA and tumor grade (r = 0.88, p<0.001).

Clinical factors and biomarkers in ovarian tumors development.

Ovarian cancer is a disease difficult to detect in early stages due to nonspecific symptoms and has a rapid progression with frequent relapses after radical surgical procedure. For these reasons, ovarian cancer generally represents the fourth cause of death through cancer in females, while in our country it is surpassed only by cervix cancer. The reduced survival is associated with the absence of symptoms, especially in early stages. Therefore, the diagnosis is delayed, when the metastases are already present and the prognosis is poor. While the etiology of the ovarian cancer is less understood, the histopathological studies and experiments regarding ovarian cancer development suggest that the majority of the tumors refined to the surface epithelium, a cuboidal layer that lays the ovary. It is still unclear if the molecular changes in this layer generate a neoplastic precursor that can be used for establishing an early diagnosis. None of the changes of the involved genes (p53, k-Ras, Her-2/neu, c-Myc, etc.) does seem to follow certain steps. We analyzed histological and immunohistochemical a group of 60 female patients admitted during January 2004 and January 2005 in Surgery Clinic of "Sfantul Ioan" Emergency Hospital, Bucharest. Our study reveals that a high percent (68.33%) of females had a correct diagnosis at admission, only five patients (8.33%) being diagnosed with other diseases. In 86.66% of cases, total hysterectomy with bilateral anexectomy has been made, in two cases (3.33%) tumor resection was the only needed therapy and in 19 cases (31.66%) peritoneal implants were found. More than 75% were serous tumors, 20% mucinous carcinoma and 5% borderline ovarian tumors. We found three cases of borderline tumors (5%) that histopathological proved to be serous tumors. The analysis of hormone receptors showed estrogen receptors in 32 cases (71.1%) of serous ovarian adenocarcinoma, in seven cases (58.33%) of mucinous adenocarcinoma, all three cases (100%) of borderline tumors and in four cases (21.05%) of the 19 with peritoneal implants. Progesterone receptors were found in 27 cases (60%) of serous carcinoma, five cases (41.66%) of mucinous carcinoma, one case (33.33%) of borderline tumors and five cases (26.31%) with peritoneal metastases. Immunohistochemical study of CerbB-2 showed positively only in 37 cases (82.22%) of serous carcinomas, five cases (41.66%) of mucinous carcinomas, one case (33.33%) of borderline tumor and eight cases (42.10%) with peritoneal metastases. All tumor types presented positives for CA125 (91.1% in serous tumors, 83.33% in mucinous tumors, 33.33% in borderline tumors and 73.68% in tumors with peritoneal implants. The investigated proliferative factors p53 and Ki-67 demonstrated correlation with tumor aggressiveness. Lack of positivity in borderline tumors and strong positivity in serous and mucinous carcinomas shows correlations with literature. This study outlines that an immunohistochemical analysis of certain antibodies cannot offer useful data regarding the prognosis or the screening for ovarian cancer.

Molecular changes in superficial bladder cancer.

Urinary bladder cancers represent a spectrum of diseases that can be grouped into three general categories: superficial, invasive and metastasis. Each differs in clinical behavior, prognosis and primary management. For superficial tumors, the aim is to prevent recurrences and progression to an incurable stage, recognizing that surgical removal of the bladder (over treatment for most tumors) is curative up to a point. For more invasive disease, the issue becomes how to determine which tumors can be cured with a single therapy such as surgery, and which, by virtue of a high metastatic potential, requires an integrated systemic approach to achieve cure. For metastatic disease, combination chemotherapy is the standard yet, despite responses in more than 50% of cases, overall cure rates remain low, and progression has been minimal over the past few years. We analyzed histopathological and immunohistochemical 70 patients with bladder carcinomas searching the stage, the grade and other associated lesions. The results showed that 70% were papillary transitional carcinomas infiltrated in lamina propria (T1), and almost 22.85% represent non-invasive papillary carcinomas (Ta); we found only five cases in Tis stage (7.15%). The immunohistochemistry investigated three antibodies: p53 oncoprotein, bcl-2 oncoprotein and retinoblastoma protein (pRb). We noticed the antibodies distribution related to stage: carcinoma in situ (Cis or Tis) high percent of p53 (69) and bcl-2 (37.5%). Concerning the superficial tumors we found low values of p53 in T1 (45%) versus invasive tumors (51%); oncoprotein bcl-2 is higher in T1 (35%) versus non-invasive one (6%).