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.

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.

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.

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.

Expression of p53 protein and tumor angiogenesis as prognostic factors in nasopharyngeal carcinoma patients.

The objective of this study was to evaluate the possible prognostic significance of p53 protein overexpression and tumor angiogenesis (TA) in nasopharyngeal carcinoma (NPC) patients, together with other clinicopathological variables. Forty-two NPC patients were evaluated in relation to survival. Nuclear p53 overexpression in neoplastic and endothelial cells was detected by immunohistochemistry (IHC) with the monoclonal antibody DO-7 and the polyclonal antibody against factor VIII-related antigen, respectively. Thereafter, we evaluated p53 cases in order to determine their nuclear immunoreactivity from negative (-) to positive (+, ++, +++). In addition, microvessels were counted in the most active areas of tumor neovascularization or hotspots using an image computer analyzer (MicroImage). A Cox multiple regression survival analysis was used to determine the best prognostic indicators in NPC patients. As a result, tumor microvessel count, considered as a continuous variable, was the most important independent prognostic indicator in relation to survival (p = 0.0273), with a relative risk of death of 2,4399 [95% confidence interval = 1.1051 ; 5.3871] associated with the highest microvessel counts. Moreover, the only clinicopathological variable that demonstrated prognostic value in a Cox multiple regression survival analysis was histological type (p = 0.05). In addition, we did not observe any statistical association between intratumoral microvessel density (IMD), clinicopathological variables and p53 protein expression.

Primary intracranial malignant tumour mistaken for a postburn scalp Marjolin's ulcer.

This report presents an apparent squamous cell carcinoma over a burn scar in the scalp (Marjolin's ulcer) that was resected and soft tissue expanders inserted in the hair-bearing scalp. In the postoperative period the patient developed neurological impairment with a CT scan showing a parasagittal brain tumour infiltrating the skin. The tumour was resected and the resultant skin and cranial defect were covered with an expanded scalp flap. Histological examination showed malignant angioblastic meningioma. The tumour recurred aggressively, leading to the death of the patient. The case is considered very unusual, as a brain tumour does not seem to be a reasonable suspect in the differential diagnosis of a chronic ulcer over a burn-induced scalp scar.

Prefabrication of a free flap for tracheal reconstruction: an experimental study. Preliminary report.

The development of a prefabricated free flap that could have potential for tracheal reconstruction has been investigated in the goat model. Through a staged procedure, a composite cutaneous-chondromucosal premolded, prevascularized flap was obtained by prefabrication techniques. The procedure comprised three surgical stages. In the first stage, on day 0, the cartilaginous frame-work was constructed, along with the vascular pedicle (implantation of an arteriovenous fistula as a vascular carrier). In the second stage, on day 50, the inner surface of the neotrachea was lined with nasal mucosa. In the third stage, on day 60, the flap was elevated and free transferred to reconstruct a 15-cm circumferential defect in the cervical trachea. Ten animals were operated on, and the results were one infection, three early deaths, one free-flap failure with early tracheal stenosis, and five long-term survivors without significant stenosis. The structure of the neotracheal flap closely resembled that of the native trachea: internal respiratory epithelial lining, cartilage rings, and fibrovascular tissue. Fiberoptic bronchoscopy was done to all the animals at 10 and 60 days, revealing no significant stenosis in the long-term survivors.

Psammomatous melanotic schwannoma localized in the mediastinum: diagnosis by fine-needle aspiration cytology.

The cytological characteristics are presented in a case of psammomatous melanotic schwannoma localized in the mediastinum, diagnosed by fine-needle aspiration cytology (FNAC), with later histological confirmation. This lesion affected a 38-year-old male, with a tumor measuring 5 cm in diameter located in the posterior mediastinum in contact with the spinal cord, with erosion of the adjacent vertebra (T5), that caused constant pain localized in the right scapular region with an evolution of several weeks. The cytological characteristics of psammomatous melanotic schwannoma are discussed, along with the histological and immunohistochemical features of this infrequent neural tumor, which aid in establishing a differential diagnosis. The importance of a correct histological diagnosis of this tumor is of particular clinical relevance after having been identified as one of the possible components of Carney's complex. This possibility demands a meticulous patient exploration in search of any other components that make up this complex, given the aggressive character of some of them. These lesions could, if not detected early enough, produce fatal consequences for the patient, fundamentally in those patients with cardiac myxomas. In this case, following an exhaustive clinical examination there was no pathology associated with this complex. It is therefore considered to be an isolated psammomatous melanotic schwannoma, and not a part of Carney's complex.

[Pulmonary inflammatory pseudotumor. Its diagnosis by fine-needle aspiration puncture]. / Seudotumor inflamatorio pulmonar. Diagnóstico mediante PAAF.

Cytological characteristics are reported in the case of a 20-years-old male with pulmonary inflammatory pseudotumor (PIP) diagnosed by fine-needle aspiration (FNA) and later confirmed histologically. The patient presented a large tumor on the lower lobe of the right lung with hemoptysis, hemothorax and secondary acute anemia. The clinical-pathological profile of PIP are reviewed, along with diagnostic problems arising from the morpho-structural characteristics of the disease.

[Bronchial granular cell myoblastoma associated with epidermoid carcinoma]. / Mioblastoma bronquial de células granulares asociado a carcinoma epidermoide.

A 43-year-old male simultaneously presented a bronchial granular cell myoblastoma affecting the bronchi of the lower right lobe, and a bronchogenic epidermoid carcinoma with mediastinal spread. In both cases diagnosis was established through bronchial and mediastinal biopsies, with posterior immunohistochemical evaluation to confirm the nature of the neoplastic lesions. The clinicopathological characteristics of this rare observation not previously reported in our country are commented.