Renal donor implication in the origin of BK infection: analysis of genomic viral subtypes.

UNLABELLED: BK virus (BKV) reactivation in immunocompromised kidney transplant patients can produce a tubulointerstitial nephropathy (BKVN). Molecular tools that test for DNA-BKV provide early detection and assist in management, but some aspects of the pathogenesis of this infection, such as donor causality, remain unclear. MATERIALS AND METHODS: Between November 2004 and January 2006, 55 Spanish kidney donors were studied for BK infection. A quantitative PCR assay was performed on urine and serum to detect BKV. To determine the origin of the viral infection, a transcription control region of the BK polymorphism sequence was designed to identify the viral subtype. RESULTS: Fifteen of 55 (27%) donors were BK-PCR positive: 13 in urine and 2 in serum and urine. Moreover, monitoring of recipient pairs detected BK-PCR positivity in 14 of 73 recipients. We studied eight BK-PCR positive recipients (corresponding to four pairs) and their respective donors. The same viral genome was observed in the four pairs, namely, the A250-1-a, WW-like, AS, and JL genotypes. Interestingly, one of the four pairs showed the donor and the two recipients to display exactly the same JL genotype. CONCLUSION: On the basis of our preliminary results analyzing the molecular fingerprints of donor and recipient pairs, we have presented new data implicating the donor, in at least some cases, as the source of BK infection.

[Small cell carcinoma of the urinary bladder in a young man. A case report and literature review]. / Carcinoma vesical de células pequeñas en un varón joven. Presentación de un caso y revisión de la literatura.

Small cell carcinoma of the bladder is a rare entity characterized by an aggressive clinical behaviour with a high incidence of systemic metastases. We report a case of small cell carcinoma of the bladder in a young man. The primary local tumour was treated by radical surgery, pelvic radiation therapy and polychemotherapy according CDDP protocol. The patient died six months after surgery because disease progression. We also review and update the literature concerning this infrequently tumour.

A risk model for non-small cell lung cancer using clinicopathological variables, angiogenesis and oncoprotein expression.

BACKGROUND: The aim of this study was to investigate new prognostic factors, by using a prognostic model, that could help to identify the patient group with the greatest probability of death. PATIENTS AND METHODS: First, the clinicopathological variables were analyzed. Second, microvessels were immunohistochemically (IHC) stained with factor VIII-related antibody and then counted in the most intense vascularization area or hotspot, using an automatic image analyzer. In addition, biological angiogenesis-related factors such as vascular endothelial growth factor (VEGF) and inducible nitric oxide synthase expression (iNOS) were also studied. Finally, we evaluated the IHC expression of p53 and p21WAF1 tumor supressor proteins. RESULTS: The significant independent predictors were: tumor size (p=0.0063), angiogenesis (p=0.0271) and p21WAF1 (p=0.0478). Thus, the most important factor was tumor size 2.7462 [95% CI=1.3307-5.6673]. Finally, these variables were included in a risk model, in order to identify the group with the highest associated probability of death. CONCLUSION: The analysis of several prognostic factors could establish a more accurate patient risk profile.

Polymerase chain reaction detection of BK virus and monitoring of BK nephropathy in renal transplant recipients at the University Hospital La Fe.

INTRODUCTION: Reactivation of BK infection occurs in immunocompromised hosts causing tubulointerstitial nephropathy (BKVN). Approximately 5% of kidney transplant recipients (KTR) develop BKVN, special half of whom lose their grafts. However, BKVN morphologic diagnosis on a renal biopsy is complicated, because the cytopathic changes can sometimes mimic rejection. Thus, BKV DNA-polymerase chain reaction (PCR) assay on serum, urine, and renal tissue is useful for early detection and monitoring of BKV. MATERIALS AND METHODS: We performed routine monthly urine cytologies looking for decoy cells as a marker of virus replication. Then, we performed a qualitative PCR on urine and serum in all recipients (independently of positive or negative cytology). We amplified 3 BK viral genome regions, LT (early transcription region) and VP1 (late transcription region) seeking a more accurate virus detection, and the TCR (control transcription region) region to perform a polymorphism sequence analysis to identify the BK genomic variant. Finally, the BKVN diagnosis was confirmed using renal biopsy. RESULTS: At present, 132 patients have been monitored. Thirteen of 40 (33%) were PCR-urine-positive cases (5 LT+/VP1- and 8 LT+/VP1+), and 10 of 132 (7.5%) were PCR-serum-positive cases (7 LT+/VP1- and 3 LT+/VP1+). When we compared PCR-urine and cytology results, 11 of 40 (27.5%) patients showed a positive cytology, 6 of whom were PCR- urine-positive (1 LT+/VP1- and 5 LT+/VP1+); whereas, 29 patients showed a negative cytology, 7 of whom were PCR-urine-positive(3 LT+/VP1- and 4 LT+/VP1+). Thus, comparison of PCR- urine and cytology results revealed false-positive and false-negative cases. Finally, TCR sequence analysis was performed in 9 patients to identify the BK genomic variants. CONCLUSION: Testing for BKV DNA in urine and serum is a noninvasive early detection assay and monitoring tool.

Ameloblastoma. Diagnosis by means of FNAB. Report of two cases.

INTRODUCTION: Ameloblastomas are the most frequent odontogenic tumors of the maxilla. In spite of their benign cytohistological appearance, they behave as invasive recurring tumors, with the possibility of metastasis. FNAB is a rapid, bloodless test that provides a pre-surgical diagnosis, thus, on occasions avoiding the need for diagnostic biopsies. We present the cytological characteristics of two cases of jugal recurrences of mandibular ameloblastomas diagnosed by FNAB, as well as their cytohistological correlation. CLINICAL CASES: Two patients, a 36-year-old woman, and a 62-year-old male who both attended with mandibular swelling of a few months evolution. In both cases the first diagnostic approximation was the histological study of the tumoral mass, together with the radiological studies. Following therapeutic extirpation both cases recurred. The diagnosis of the recurrences was established cytologically by means of FNAB. The cytologic smears revealed a granular background with isolated macrophages and giant multinucleate cells and an abundant epithelial cellularity of basaloid appearance arranged in cohesive groups forming images of peripheral palidasing, as well as small groups of squamous metaplastic cells. DISCUSSION: FNAB is considered to be a rapid, bloodless and reliable method for the diagnosis of ameloblastoma. The cytology of these tumors reveals components of the lesion that, in general, are sufficient for the diagnosis of ameloblastoma, especially in cases of recurrence.

[Prognostic value for progression of the regulating proteins of the cellular cycle in PT1G3 bladder tumours]. / Valor pronóstico de progresión de las moléculas reguladoras del ciclo celular en tumores vesicales T1G3.

INTRODUCTION AND OBJECTIVE: Bladder tumor T1G3 constitutes the group of superficial tumors more aggressive. New prognostic factors in the field of the cytogenetics and molecular biology have been analyzed, with often contradictory results, being little the specific works in tumors T1G3. Our objective is to determine if in this group of tumors the immunohistochemical markers present predictive value of clinically useful progression, and therefore with validity to indicate more suitable a precocious therapeutic attitude. MATERIAL AND METHODS: Retrospective study of a series of 83 patients affected of bladder tumor T1G3, on which we analyzed a total of 14 variables; between the new predictive factors: the immunohistochemical determination of regulating proteins of the cellular cycle: p53, p21 and bcl-2, as well as the Ki-67 protein like marker of cellular proliferation. By means of logistic regression analysis we establish the independent prognostic variables for tumorlike progression. RESULTS: The cut point established for Ki67 and p53 was 40% of inmmunomarked cells, 20% for p21 and 10% for Bcl-2. The univariant analysis showed different rates from progression and free times of progression based on the immunohistochemistry of Ki67 and p53: nevertheless, the logistic regression demonstrated that single the immunohistochemistry of p53 presented independent predictive value. CONCLUSIONS: The determination of p53 presents predictive value of clinically useful progression in bledder tumors T1G3, so that its determination can constitute a essential factor in the strategies of treatment of these tumors.

[Predictive value of progression of muscularis mucosae and percentage G3 cells in vesical TIG3 tumours]. / Valor predictivo de progresin de la muscularis mucosae y del porcentaje de células G3 en tumores vesicales T1G3.

INTRODUCTION AND OBJECTIVE: Vesical tumor T1G3 constitutes the border between the superficial tumor and the infiltrante tumor. Some of these tumors do not respond to BCG and progress, with cystectomy that present poor results, patients who would benefit from a precocious and aggressive treatment if we could identify them in an preinvasive stage. New predictive factors try to select to these tumors, being little the works that consider anatomo-pathological meticulous study (substanding of the T1 in T1a and T1b and percentage of present G3 cells in the tumor). Our objective is to analyze the value of these anatomo-pathological considerations like predictive factors of progression. MATERIAL AND METHODS: Retrospective study of a series of 91 patient affection of vesical tumor T1G3 with initial treatment by means of RTU and BCG. We analyzed 12 variables. The new predictive factors: the level of invasion respect to muscularis mucosae and the percentage of G3 cells. By means of logistic regression analisys we establish the independent pronostic factors for tumoral progression. RESULTS: A total of 31 patients presented infiltration of detrusor, passing away 17 of tumoral cause, after an average time of pursuit of 57.8 +/- 28.2 months. In 8 cases (9%) the substanding could not be determined. The rate of progression for T1a tumors was of 20% (8/40) and for T1b 53% (23/43). Presented independent predictive value of progression the multiplicity (odds: 7.26), the size (odds: 2.14), the presence of Cis (odds: 1.42) and the subestanding (odds: 6.81). CONCLUSION: The substanding is a predictive factor of progression clinically useful in vesical tumors T1G3, reason why we considered habitual clinical introduction.

Liver disease as primary manifestation of multiple myeloma in a young man.

We report a 27-year-old man who presented with fatigue, moderate weight loss and progressive abdominal distension as primary manifestations of a light-chain multiple myeloma (MM). Liver scan showed an enlarged liver with multiple low attenuation areas. Liver biopsy revealed sinusoidal infiltration by small size cells identified as Kappa light chain-producing primitive plasma cells by immunohistochemistry. The patient responded to three courses of EDAP. Subsequently he received intensive therapy with busulfan/melfalan and a peripheral blood stem cell transplantation enriched for CD34+ cells from his HLA-identical brother. No acute graft-versus-host disease was detected. Now, 12 months after transplant, the patient is asymptomatic.

Counts and areas of S-100-positive epidermal dendritic cells in atypical molluscum contagiosum affecting HIV+ patients.

Molluscum contagiosum is a common and self-limiting viral infection, that in HIV+ patients courses as an opportunist affection with atypical clinical features. Impaired cell-mediated immune response could be involved in such atypical growth. We evaluated the density and area of Langerhans cells (LC) using S-100 immunohistochemistry in seven atypical molluscum contagiosum. LC density was quantified by three different methods using computer-assisted morphometry as well as estimating the relative area of LC with respect to epidermal area. Results were compared with two control groups (normal skin specimens and molluscum contagiosum affecting non-AIDS healthy patients). We found a virtual absence of LC in areas of molluscum lesions affecting both HIV+ and non-AIDS patients. Likewise we observed an evident decrease in LC density in perilesional epidermis of atypical molluscum with respect to both control groups. Upon comparing the counts and areas, we observed that this reduction in LC count was statistically significant only when considering LC related to length of basement membrane in atypical molluscum with respect to normal skin specimens. Our finding of a reduced number of LC in the perilesional epidermis of HIV+ patients with atypical molluscum could explain the high frequency and clinical challenge of molluscum contagiosum in immunocompromised people. In spite of these results, further studies of LC kinetics and functions are required to precisely elucidate their role in the course of molluscum contagiosum in HIV+ patients.

Immunohistochemical expression of Bcl-2 oncoprotein in EBV-associated nasopharyngeal carcinoma correlated to histological type and survival.

Expression of Bcl-2 is associated with inhibition of apoptosis and extension of cell survival. In vitro Bcl-2 protein expression is up-regulated by the EBV-latency associated antigen latent membrane protein (LMP-1). We have investigated the relationship between the presence of EBV-DNA screened by means of sensitive nested-PCR, nasopharyngeal carcinoma (NPC) histological types according to two different schemata (WHO and Micheau classifications) and Bcl-2-124 immunohistochemical expression in 55 biopsy samples of NPC. EBV genome was detected in 100% of samples with sufficient DNA quality to support the previous view that all types of NPC are variants of EBV-infected neoplasia. Bcl-2 was observed in the basal layer of normal nasopharyngeal mucosa and also at cytoplasmic level in 42 of 55 (76.4%) NPC cases. Mitotic neoplastic cells usually showed strong cytoplasmic and chromosomal staining, a finding not well referred to previously. Bcl-2 expression was significantly associated (p<0.05) to undifferentiated NPC (UNPC) when a histological classification with only two major microscopical types was applied. No close correlations were found between the presence of EBV-DNA, NPC location, clinical stage and age or sex of the patients in relation to Bcl-2 positive expression. However, when comparing Bcl-2 expression and known survival mean of the patients, significant differences were observed (p<0.001) so that mean survivals were 31.1, 24.4, 52.2 and 54.1 months respectively for NPC patients with -, +, ++ and Bcl-2 immunoreactivity. Nevertheless this better clinical outcome in Bcl-2 NPC positive cases may depend on the histological type due to close relationship with UNPC. Only studies of larger series with long-term follow-up and multivariate analyses may document whether Bcl-2 expression is an independent prognostic marker in the evolution of NPC patients.

Comparative analysis of Epstein-Barr virus (EBV) detection by nested-PCR and non-isotopic in situ hybridization in nasopharyngeal carcinoma (NPC).

The aims of this study were two-fold: first, to assess the relative diagnostic performance of non-isotopic in situ hybridization (ISH) and the nested polymerase chain reaction (nested-PCR) applied to Epstein-Barr virus (EBV) detection in a series of 55 unselected nasopharyngeal carcinoma (NPC) cases and, secondly, to correlate these data with histopathological classification. Our study shows that in 76.36% of NPC cases positive nuclear signals were observed using EBV-ISH. Overall, EBV-ISH positivity varied according to histological type, in that undifferentiated carcinomas showed a higher proportion of positive cases than differentiated cell carcinomas, although ISH results do not show significant differences in relation to histological types when employing two different schemes (WHO and Micheau). However, in adequate quality DNA samples (54 NPC cases), EBV-DNA was detected in 100% of cases using a nested-PCR, supporting the previous view that all histological types of NPC are in reality variants of EBV-infected neoplasia. ISH-negative cases probably reflect a lower sensitivity than PCR, particularly when a small number of viral copies are present, as well as a variable technical effectiveness for detected EBV, independent of the NPC histological type.

Relationship of p53 molecular abnormalities with flow cytometry and growth factor receptor content in lung cancer.

This study attempts of clarify the oncological significance of the p53 molecular abnormalities and p53 expression in lung cancer (LC) and their relationship with flow cytometry (FC) parameters and epidermal growth factor receptor (EGFR). The study includes 65 samples taken from both LC and normal lung (NL). The p53 molecular abnormalities of exons 4-8 were studied by single strand conformation polymorphisms (SSCP) and the loss of heterozygosity (LOH) of exon 4 by the Metzler method. P53 protein was detected by Western blot. EGFR was determined by a radioligand assay using [125I]EGF. The FC parameters S phase fraction (SPF), DNA index (D.I.), G1G0 and growth rate (G2M + SPF) were evaluated from cellular monosuspensions. The LC with SSCP p53 molecular abnormalities have a significantly higher EGFR content (P < 0.001), SPF (P < 0.007), D.I. (P < 0.017) and a lower proportion of G1G0 cells (P < 0.04) than LC with no molecular abnormalities. No relationship between p53 molecular abnormalities and tumor TN or evolutive events was found. Neither the relationship between the molecular results and p53 expression detected by Western blot nor that of the p53 expression detected by Western with FC parameters or EGFR could be shown. In NL the growth fraction cells decrease significantly (P < 0.05) with the intensity of p53 expression. The lack of biological functionality of p53 with molecular abnormalities seemed to relate to fast growing LC whereas p53 expression detected by Western seemed more related to the wild type of p53.

C-erbB-2 expression and its relationship with ploidy, p53 abnormalities and epidermal growth factor receptor content in human non-small cell lung cancer.

This study attempts to clarify the role of c-erbB-2 overexpression in human non-small cell lung cancer (NSCLC) and relate it with the p53 alterations, DNA index (D.I.) and epidermal growth factor receptor (EGFR) content in sixty four patients with NSCLC. c-erbB-2 and EGFR quantification were carried out from tissue homogenates using quantitative ELISA procedures. p53 alterations were determined by immunohistochemical (IHC) detection with the monoclonal antibody DO-7 and analysis for p53 mutations on exons 4 to 8 by single strand conformation polymorphism (SSCP). The D.I. was performed by flow cytometry. c-erbB-2 hyperexpression was found in 13 of 58 LC (22%), and it was closely associated with hyperdiploid tumors (D.I. >1.3; P = 0.00). The p53 abnormalities detected by SSCP were statistically more frequent in hyperdiploid tumors (16/25; P = 0.015) than in diploid ones (8/30). No relationship between the results of IHC p53 and SSCP was found. The patients with c-erbB-2 hyperexpressing tumors were prone to have frequent relapses (P = 0.03), although the patients with hyperdiploid NSCLC are the ones with the highest relapse rate (P = 0.008). From the results obtained in this study the following conclusions can be drawn: (a) c-erbB-2 hyperexpressing NSCLC are associated with abnormalities in other biological markers and with a greater rate of relapses; (b) SSCP seemed to be more specific that IHC to detect p53 molecular abnormalities; and (c) the D.I. is the parameter more tightly related with relapse.

Analysis of EBV latency by EBER in situ hybridization in nasopharyngeal carcinoma Spanish patients.

BACKGROUND: The aim of this study was to analyse the Epstein-Barr virus (EBV) latency by detecting the EBV-associated latent small nuclear RNAs (EBER), in a group of biopsies from Spanish patients with diagnosed nasopharyngeal carcinoma (NPC). MATERIALS AND METHODS: NPC paraffin samples with the presence of EBV demonstrated by non-isotopic in situ hybridization (NISH) and nested-PCR, were analysed for EBV latency using EBER in situ hybridization (EBER-ISH). RESULTS: We detected EBER in 83.3% of samples (10 out of 12 cases), demonstrating the relationships between EBV genome presence with the latent viral infection. We correlated these results of EBV-DNA and -RNA presence with the immunoexpression of latent membrane protein-1 (LMP-1), a viral oncogenic protein (8 out of 12 cases or 66.6%). CONCLUSION: These results indicate that all the types of NPC are variants of an EBV-associated malignancy and that viral latency is a critical phenomenon in the development of this neoplasia.

Morphometric study of tumor angiogenesis as a new prognostic factor in nasopharyngeal carcinoma patients.

The aim of this study was to determine the possible prognostic significance of tumor angiogenesis (TA) in nasopharyngeal carcinoma (NPC) patients. Fifty-five NPC patients were evaluated in relation to survival. Endothelial cells were immunohistochemically stained with anti-von Willebrand factor (F-VIII), CD-31 and CD-34 antibodies, and microvessels counted in the most active areas of tumor neovascularization or hotspotsusing both a manual and an automatic method. Overall survival analysis calculated by the Kaplan Meiertest revealed that both methods were correlated with a statistical significance between intratumoral microvessel density (IMD) and overall survival, using either manual (p=0.0141) or automatic counting (p=0.0117). Other angiogenic parameters studied were perimeter, roundness and accumulative area of the microvessels using a morphometric analyzer. Moreover, our results show that cases with high IMD demonstrated a prognostic significance in relation to the accumulative area (p=0.0072).

Immunohistochemical absence of CD21 membrane receptor in nasopharyngeal carcinoma cells infected by Epstein-Barr virus in Spanish patients.

OBJECTIVES/HYPOTHESIS: The aim of this study was to analyze the relevance of the CD21 membrane receptor in nasopharyngeal carcinoma (NPC). CD21 is implicated in the introduction of the Epstein-Barr virus (EBV) genome into epithelial cells and B lymphocytes. STUDY DESIGN: Immunohistochemical analysis of CD21 in NPC. METHODS: Paraffin-embedded samples of NPC of different histological types with demonstrated presence of EBV were analyzed for CD21 expression using immunohistochemistry. RESULTS: We detected EBV by non-isotopic in situ hybridization (NISH) and nested polymerase chain reaction (PCR) in 100% of samples, regardless of histological type, supporting the previous view that all the types of NPC are variants of an EBV-associated malignancy. CD21 was not detected in NPC, and this absence was a typical feature in our data group. CONCLUSIONS: The loss of the CD21 membrane receptor could be one of the immunophenotypical changes of the neoplastic cells that occur in the evolution of the NPC malignancy.

Diphenylhydantoin-induced gingival overgrowth in man: a clinico-pathological study.

In a cross-sectional, epidemiological study of diphenylhydantoin-induced gingival overgrowth (DGO) in 60 epileptic patients, gingival lesion severity was statistically compared with other clinical, laboratory, and histopathological findings. Evident DGO was observed in 50% of patients, and positive correlations were detected between overgrowth severity and oral debris, calculus accumulation, plaque score, gingival inflammation, and probing depth. However, no valid correlation was observed between lesion and patient age, mouth breathing, daily drug dose, plasma diphenylhydantoin level, or duration of drug intake. Also, there were no significant correlations between connective fiber richness and the intensity of epithelial hyperplasia and severity of gingival overgrowth.

Malignant fibrohistiocytoma (MFH) of the breast. Primary and postirradiation variants - an ultrastructural study.

Two cases of malignant fibrohistiocytoma (MFH) of the mammary gland are presented. Patient No. 1 had been previously locally irradiated for an infiltrating duct carcinoma (5600 rads) 5 years earlier. Prognosis of postirradiation MFH was poor and the patient died within the first year after diagnosis of the second malignancy. The primary MFH (Case No. 2) is free of disease after a 5-year follow-up period, having been submitted to a radical mastectomy. Histological appearance is in accordance with previously published cases. No marked differences existed between both cases at optical level. Lysozyme (PAP technique) was observed within isolated histiocytes. Electron microscopy distinguished up to 4 different cell types within the neoplasm: fibroblasts, histiocytes, mixed cells of fibro-histiocytic appearance, and more immature blastemal cells. Transitional forms existed among them.

Lipid-rich versus lipid-secreting carcinoma of the mammary gland.

An unusual case of a malignant breast tumor in a 70 year-old woman is reported. The histological picture displayed a mixed pattern of vacuolated and apocrine-like cells. At electron microscopical level the tumoral cells presented, in a very regular manner, lipidic intracytoplasmic vacuoles. These lipid droplets were secreted by the tumoral cells, and appeared surrounded by SER profiles. A differential diagnosis is discussed with other mammary gland neoplasms, such as histiocytoid carcinoma, apocrine cell carcinoma and other clear cell carcinomas. Based upon our findings we propose that this case should be considered as a lipid secreting carcinoma of the breast.

Warthin-like tumor of the thyroid: a papillary carcinoma with mitochondrion-rich cells and abundant lymphoid stroma. A case report.

Papillary carcinoma is the most common form of thyroid carcinoma and, generally, it has a more favorable prognosis than other carcinoma types, although diverse variants or subtypes have been described, some of which appear to have a less favorable prognosis. Recently, a new variant, the so-called "Warthin-like tumor" or "tall-cell variant with extensive lymphocytic infiltration of papillary thyroid carcinoma" with behavior similar to the usual papillary carcinoma, has been described. We present a case with the cytohistological pattern of "Warthin-like tumor" of the thyroid. Immunohistochemistry revealed reactivity at the epithelium lining papillae for antimitochondrial 113-I antibody and also membranous positivity for CD15 (Leu M1). Lymphoid stroma showed an immunoprofile similar to chronic lymphocytic thyroiditis. These findings point to the consideration of "Warthin-like tumor" as being a hybrid neoplasm between the tall-cell variant and oxyphilic papillary carcinoma associated to a lymphoid-rich stroma.