Is there an association between coronary atherosclerosis and carcinoma of the prostate in men aged 50 years and older? An autopsy and coroner based post-mortem study.

BACKGROUND: Atherosclerotic disease is the most common cause of death in the United States and prostate cancer has the highest incidence among males in the United States. Reports have indicated that atherosclerosis and cancers my share common pathoetiologic and pathogenetic cascades. If atherosclerosis and cancers have common pathoetiologic and pathogenetic cascades, both diseases will co-occur and patients may represent a potential target group for cancer screening interventions. MATERIALS AND METHODS: Prostates and coronary vessels were examined from 37 deceased men, aged 50 years and older, who died unexpectedly and suddenly from traumatic causes. Tissue sections of the entire prostate were examined for benign and malignant lesions. Analysis of Variance was used to compare mean coronary artery atherosclerosis scores among groups of men with diagnosis of adenocarcinoma, intraepithelial neoplasm, benign hyperplasia and normal prostate glands. RESULTS: Twelve prostates (32.5%) showed adenocarcinoma of the prostate, four with Gleason score 7 and eight with Gleason score 6. After adjustment for age and race, there remained no statistical difference between prostate pathology groups and atherosclerosis score (F = 0.72; P = 0.55). CONCLUSIONS: To our knowledge, ours is the first study to use direct pathological examination of tissues for definitive identification of atherosclerosis and prostate cancer. In our case series, the occurrence and progression of coronary atherosclerotic disease and cancer of the prostate were not associated.

Associations between hepatocyte growth factor, c-Met, and basic fibroblast growth factor and survival in endometrial cancer patients.

BACKGROUND: Hepatocyte growth factor (HGF), c-Met, and basic fibroblast growth factor (bFGF) are molecular markers that contribute to angiogenesis and proliferation in numerous cancers. We assessed the prognostic significance of these factors in tumour and stroma of endometrial cancer (EC) patients (n=211). METHODS: Immunohistochemistry (IHC) was used to detect tumour and stromal protein expression of the biomarkers. Associations between expression and clinicopathological factors were assessed using Chi-square tests. Kaplan-Meier curves, log-rank tests, and Cox regression were used to summarise associations between biomarker expression and overall survival (OS) and recurrence-free survival (RFS). RESULTS: Tumour bFGF was significantly associated with high-grade endometrioid and clear cell histology (P<0.001), advanced stage (P=0.008), positive lymph-node involvement (P=0.002), poor OS (log-rank test, P=0.009), and poor RFS (P<0.001). In multivariable analyses, cases with HGF-positive, stromal bFGF-positive tumours had a lower risk of death compared with cases with HGF-positive, stromal bFGF-negative tumours (hazard ratio (HR): 0.14, 95% CI: 0.03, 0.60). Cases with HGF-positive, bFGF-positive tumours had a higher risk of recurrence compared with cases with negative expression of both markers (HR: 9.88, 95% CI: 2.63, 37.16). CONCLUSION: These IHC data show that tumour and stromal bFGF expression have opposite associations with survival outcomes in EC patients. If confirmed in larger studies, tumour-derived bFGF could be an attractive target in EC therapy.

Low-grade myxoid renal epithelial neoplasms with distal nephron differentiation.

We report 4 distinctive renal epithelial neoplasms that are essentially identical at the morphologic and immunohistochemical levels and do not fit an accepted category in the existing classification of these lesions. The patients were all females, with ages ranging from 32 to 79 years (mean, 50 years). The tumors were well circumscribed and were composed of uniform, predominantly low cuboidal cells with eosinophilic, focally vacuolated cytoplasm. Tumor cells generally formed interconnecting tubules, with smaller areas of cordlike growth and spindling in a bubbly, myxoid stroma. All tumors were confined to the kidney, and all were immunoreactive for high-molecular-weight cytokeratin 34betaE12, cytokeratin 7, epithelial membrane antigen, and cytokeratin cocktail AE1/3. Only 1 tumor was focally immunoreactive for Ulex europaeus agglutinin. Ultrastructural study showed tumor cells forming tubular structures reminiscent of the loop of Henle or distal convoluted tubule. Follow-up in all 4 cases was benign. These distinctive tumors may be confused with aggressive sarcomatoid renal cell carcinomas because of their spindled morphology. The morphologic, immunohistochemical, and ultrastructural features of these lesions indicate differentiation toward distal nephron segments. Similar tumors probably have been reported among low-grade collecting duct carcinomas or tumors "possibly related to the loop of Henle."

Detection and differentiation of bovine group A rotavirus serotypes using polymerase chain reaction-generated probes to the VP7 gene.

Dot and Northern blot hybridization assays were developed to detect and differentiate group A bovine rotavirus serotypes using radiolabeled serotype 6 (Nebraska calf diarrhea virus [NCDV] and United Kingdom [UK] strains) or serotype 10 (Crocker [Cr] strain) VP7 gene probes. Partial length VP7-specific cDNA encompassing areas of major sequence diversity were generated by the polymerase chain reaction (PCR) using either cloned VP7 genes (NCDV and UK strains) or reverse transcribed mRNA (Cr strain) as templates. Radiolabeled probes prepared from the PCR-generated cDNA were tested at various stringency conditions to optimize the hybridization assays. At high stringency conditions (52 C, 50% formamide, 5 x standard saline citrate), the NCDV, UK, and Cr probes serotypically differentiated bovine rotavirus isolates in RNA samples prepared from cell culture propagated viruses or in fecal specimens from infected gnotobiotic calves. The sensitivity and specificity of NCDV and Cr VP7 probes were characterized in dot blot hybridization assays, and the probes were estimated to detect at least 1 ng of viral RNA. The serotyping results obtained using VP7 probes were similar to those obtained using serologic assays. Further development of these assays may provide a useful means for the rapid detection and differentiation of bovine rotavirus serotypes in fecal samples from calves in the field.

A survey of G6 and G10 serotypes of group A bovine rotaviruses from diarrheic beef and dairy calves using monoclonal antibodies in ELISA.

Group A bovine rotaviruses (BRV) have been identified worldwide as a major cause of diarrhea in the young of many species, including humans. Group A rotaviruses are classified into serotypes on the basis of the outer capsid proteins, VP7 (G types) and VP4 (P types). To date, there are 14 G types of group A rotaviruses, with G1, G6, G8, and G10 described for BRV isolates. In this study, G6- and G10-specific monoclonal antibodies (MAbs) were used in an enzyme-linked immunosorbent assay (ELISA) for the G typing of BRV-positive stool samples from diarrheic beef and dairy calves from South Dakota, Ohio, Michigan, Nebraska, and Washington, USA, and Ontario, Canada. ELISA plates were coated using a broadly reactive VP7 MAb (Common 60) or with G6- or G10-specific MAbs. BRV-positive fecal samples were diluted and added to duplicate wells, followed by the addition of polyclonal guinea pig anti-group A rotavirus serum as the secondary antibody. Several reference G6 and G10 BRV strains as well as other G types previously reported in cattle (G1, G2, G3, G8) and BRV-negative samples were included as G type specificity and negative controls. From a total of 308 field samples analyzed, 79% (244/308) tested positive by the broadly reactive VP7 MAb; of these, 54% (131/244) were G6 positive, 14% (35/244) were G10 positive, 4% (9/244) were both G6 and G10 positive, and 28% (69/244) were G6 and G10 negative.(ABSTRACT TRUNCATED AT 250 WORDS)

Single-stranded genomic RNA from turkey enterovirus-like virus.

Previously, we described a disease syndrome in young turkeys caused by an enterovirus-like virus. The virus was designated an enterovirus-like virus based on size, morphology, and intracytoplasmic crystalline arrays of virus. In the present study, further characterization of the virus was performed to ascertain its classification. The virus has a buoyant density of 1.33 g/ml in CsCl and single-stranded RNA genome of approximately 7.5 kilobases. These combined characteristics indicate that this agent is an enterovirus.

Primary renal epithelioid hemangioendothelioma.

Epithelioid hemangioendothelioma is a rare vascular tumor with intermediate biologic behavior and metastatic potential. Primary renal epithelioid hemangioendothelioma is extremely rare and we present the second report of this rare tumor in an interesting clinical scenario. A 59-year-old male with established history of widely metastatic high grade esophageal adenocarcinoma was found to have an isolated renal nodule on a followup computed tomography (CT) scan. Surgical excision, with the suspicion of metastatic carcinoma, and subsequent pathologic examination revealed an epithelioid hemangioendothelioma. The various differential diagnoses and use of morphological clues and immunohistochemistry are discussed.

Ewing sarcoma/primitive neuroectodermal tumor of the kidney: two unusual presentations of a rare tumor.

Only few cases of primary renal Ewing's sarcoma have been reported in the literature to date. We present here two cases of renal ES/PNET with an uncanny presentation. The first case was discovered after the patient presented clinically with irradiating flank pain, mimicking the pain related with kidney stones. The second case had clinical presentation of pulmonary thromboembolism after the patient was involved in an automobilist accident. The tumors were mainly composed of small blue cells which by immunohistochemical were positive for neural markers, and FISH revealed the translocation 22q12 for the EWSR1 gene. The diagnosis of renal primitive neuroectodermal tumor/EWING tumor is very rare and usually involves several different diagnostic techniques. The differential diagnosis is usually broad with frequent overlapping features between the entities. The cases presented in this paper illustrated the difficulties with which routine anatomical pathologist is faced when dealing with rare renal poorly differentiated neoplasm in adults.

Comparative nucleotide and deduced amino acid sequence analysis of VP7 gene of the NCDV Cody (I-801) strain of group A bovine rotavirus.

The prevalent G serotypes of group A bovine rotavirus (BRV) reported are G6, G10, and less commonly, G8. Neonatal Calf Diarrhea Virus (NCDV), Lincoln and Cody strains were first isolated from diarrheic calves in Nebraska. The NCDV Lincoln strain is the currently used U.S. vaccine strain and has a G6 serotype. In this study, the complete nucleotide sequence of the VP7 gene of NCDV Cody (I-801 strain) was determined using the primer extension method. The VP7 gene nucleotide sequence homologies between Cody I-801 and established G8 rotaviruses, A5 (Thailand BRV), 678 (UK BRV), B37 (human RV) and 69M (human RV) were 84.7%, 86.4%, 84.7% and 85.9%, respectively. The deduced VP7 amino acid sequence of Cody I-801 was similar to that of A5, 678, B37 and 69M (93.6%, 95.7%, 92.6% and 95.1%, respectively). The VP7 gene nucleic acid sequence homologies between NCDV Cody (I-801) and NCDV Lincoln or B223 (G10) was 76.2% and the deduced VP7 amino acid sequence homologies between Cody I-801 and NCDV Lincoln or B223 were 82.5% and 81.3%, respectively. Thus, our sequence data suggests that the VP7 gene of Cody I-801 strain of BRV is genetically most similar to G8 rotaviruses and unrelated to the NCDV Lincoln G6 rotavirus strain.

The characterization of VP7 (G type) and VP4 (P type) genes of bovine group A rotaviruses from field samples using RT-PCR and RFLP analysis.

Characterization of the VP7 (G type) and VP4 (P type) genes of bovine group A rotaviruses (BRV) from field samples was performed using RT-PCR and RFLP analysis. After RT-PCR amplification of the full length VP7 genes and partial length VP4 genes (nucleotides 1 to 1096), four enzymes, EcoRV, NlaIV, BamHI and HpaII were used for digestion analysis. For VP7, four RFLP profiles were observed after analysis of the digests: they were designated as G6, G6s (subtype, showed about 86% nucleotide and 90% amino acid identity to reference G6 strains), G8 and G10. For VP4, three RFLP profiles were observed: designated as P[1], P[5] and P[11]. The G typing analysis of 86 BRV fecal samples from 5 states, representing at least 11 different herds revealed that 60.5% (52/86) were G6, which included G6s (9/52); 19.8% (17/86) were G10; 7% (6/86) were G8; 10.4% (9/86) were G6 and G10 mixtures including two G6s samples; and 2.3% (2/86) were G6 and G6s mixtures. The P typing analysis of the same 86 fecal samples revealed that 64% (55/86) were P[5]; 28% (24/86) were P[11]; 1.2% (1/86) were P[1] and 6 samples (7%) were mixtures of either P[11] or P[5]. When the same samples were analyzed according to G and P type specificity, all possible combinations of G and P types existed in the field. The G6P[5] type was most prevalent and accounted for 46.7% (41/86) of the samples; 12.8% (11/86) were G10P[11]; 7% (6/86) were G10P[5] and an equal number were G6sP[11]. The G6P[11] (n = 2), G8P[1] (n = 1), G8P[5] (n = 1) and G8P[11] (n = 3) combinations were also observed. The following mixed BRV infections were observed in the field samples; G6sP[5 + 11] (n = 1), G8P[5 + 11] (n = 1), G6 + G10P[5] (n = 1) G6 + G10P[5 + 11] (n = 2), G6 + G6sP[11] (n = 1), G6 + G6sP[1 + 11] (n = 1), G6s + G10P[11] (n = 1) and G6s + G10P[5 + 11] (n = 1). Information on the G and P types and G/P combinations in the field samples should be useful for understanding the epidemiology of BRV and designing vaccination strategies to control BRV in the field.

Biochemical characterization of porcine enteric calicivirus: analysis of structural and nonstructural viral proteins.

In this report, the molecular weight and antigenicity of the proteins of a porcine enteric calicivirus (PEC) were characterized. The PEC virions were purified from intestinal contents of infected pigs and from infected cell culture lysates. The average buoyant density of the purified virus was 1.37 gm/cm3 in cesium chloride. One major structural protein with a molecular weight of approximately 58 k was found in the gut and cell culture-passaged PEC using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Using immunoblotting techniques only one immunoreactive protein (58 k) ws identified. The PEC and a prototype calicivirus, feline calicivirus (FCV) were propagated in pig kidney and feline kidney (Crandell) cell lines, respectively and intrinsically labeled using [35S]methionine at various times post-inoculation (PI). SDS-PAGE of the radiolabeled proteins indicated the presence of the major structural protein (58 k) and one probable nonstructural protein (28 k) synthesized in the PEC-infected cell lysates by 12 h PI. Other minor protein bands were also evident by 24 h PI (32 k and 82 k). Only the 58 k major protein was detected by radioimmunoprecipitation (RIP) analysis using hyperimmune anti-PEC serum. SDS-PAGE and RIP analysis of FCV-infected cell lysates using hyperimmune anti-FCV serum identified a single major protein of approximately 64 k. No antigenic relationship between PEC and FCV proteins was detected by RIP analysis. The single major structural protein of PEC, the morphological appearance and size of the virus, and its average density of 1.37 gm/cm3 in cesium chloride are consistent with properties of other members of the family Caliciviridae.

Identification of group B rotaviruses with short genome electropherotypes from adult cows with diarrhea.

Two field strains (BB-RVLV and KD) of group B rotaviruses from adult dairy cows with diarrhea displayed short genome electropherotypes. Gnotobiotic calves inoculated with fecal filtrates of each group B rotavirus developed diarrhea, and only group B rotaviruses or antigens were detected in the feces by immunoelectron microscopy and in intestinal epithelial cells by immunofluorescent staining, respectively. The feces or intestinal contents of the cows and inoculated calves were negative for group A and C rotaviruses by enzyme-linked immunosorbent assay, immunoelectron microscopy, or cell culture immunofluorescence assays. Comparison of the genome electropherotypes of the calf-passaged BB-RVLV and KD strains with the original samples and reference bovine group A, B, and C rotaviruses revealed conservative of their short-genome electropherotypes and double-stranded RNA migration patterns characteristics of group B rotaviruses. To our knowledge, our previous study (L.J. Saif, K.V. Brock, D.R. Redman, and E.M. Kohler, Vet. Rec. 128:447-449, 1991) and this report are the first description of bovine group B rotaviruses (in a mixed infection with bovine coronavirus or singly in fecal contents) in adult cows with diarrhea and this is the first report of short-genome electropherotypes among group B rotaviruses.

Comparative sequence analysis of the VP7 genes of G6, G8 and G10 bovine group A rotarviruses and further characterization of G6 subtypes.

We previously reported the relatively high prevalence (15%) of bovine G6 subtypes (G6s) in the field using RT-PCR and restriction fragment length polymorphism (RFLP) analysis (Chang et al., Arch. Virol. 141: 1727-39). In the present study, we report the nucleotide and antigenic characterization of a G6s strain (C-8336). We also sequenced the VP7 genes of four additional bovine rotavirus (BRV) strains: another G6s (MC27), G6 (IND), G8 (C-8008) and G10 (2292B) and compared these with other bovine and human rotavirus strains. The C-8336 and MC27 strains were confirmed as P[11]G6s by RT-PCR and RFLP analysis. The VP7 genes of the C-8336 and MC27 strains showed high homology to each other (approximately 98%) and with other bovine G6s strains (greater than 95% homology in nucleotide and amino acid sequence with KN-4[P[11]G6s]) and also showed lower, but substantial sequence homology with human G6s strains and prototype G6 BRV (79-87% in nucleotide and 88-91% in amino acid). Serologic analysis of the cell culture adapted C-8336 strain showed that it was neutralized by a G6 monoclonal antibody (MAb IC3) to similar titers as the reference NCDV and IND G6 strains. In two-way cross-neutralization tests, strain C-8336 showed 4- to 16-fold differences in antibody titers with NCDV and IND G6 BRV. Moreover polyclonal antiserum against strain C-8336 neutralized the NCDV and IND strains weakly. Genetic variability was also observed among G8 and G10 bovine and human group A rotaviruses: the VP7 genes of the bovine C-8008 (P[5]G8) and 2292 B (P[11]G10) strains showed from 10 to 17% nucleotide divergence with those of Cody 1801 (P[1]G8, bovine), A5 (P[1]G8, bovine), 69M (P[10]G8, human) and Hal 1166 (P[14]G8, human), and I321(P[11]G10, human) and MC35 (P[14]G10, human) rotaviruses, respectively. The divergence of VP7 genes among bovine and human G6, G8 and G10 strains appears related to host species origin and their combination with VP4 (P type). The data presented in this report confirms the genetic variability among homotypic bovine and human strains and highlights the importance of continued monitoring of BRV G and P types circulating in the field for the future development and monitoring of effective vaccines.

Detection of rotavirus serotypes G1, G2, G3, and G11 in feces of diarrheic calves by using polymerase chain reaction-derived cDNA probes.

On the basis of antigenic variability in the VP7 outer capsid glycoprotein, at least 14 G serotypes exist for group A rotaviruses. Serotypic diversity exists among bovine rotaviruses (BRV), with serotypes G1, G6, G8, and G10 reported for cattle. Although G1 and G8 rotaviruses were originally described for humans, the recent isolation of G6 and G10 rotaviruses from humans further emphasizes the serotypic similarity between human and bovine rotaviruses and the possible zoonotic potential of rotaviruses. Results of our previous studies have indicated that more than 24% of BRV-positive field samples from diarrheic calves were nonreactive with cDNA probes or monoclonal antibodies to serotypes G6, G8, and G10. In this study, cDNA probes were prepared by polymerase chain reaction amplification of the hyperdivergent regions of the VP7 genes (nucleotides 51 to 392) from human (G1, G2, and G3) and porcine (G4, G5, and G11) rotaviruses. These probes were used in a dot blot hybridization assay to further characterize the G types of 59 BRV strains (fecal samples from diarrheic calves in Ohio, Nebraska, Washington, and South Dakota) that were nonreactive with cDNA probes to G6, G8, and G10. Rotaviruses belonging to serotypes G1 (n = 7), G2 (n = 1), G3 (n = 2), and G11 (n = 3) were identified among the BRV field samples. The BRV associated with these G types accounted for 22% of the samples tested; the other 78% of these samples remained untypeable with these probes. To our knowledge, this is the first report in the United States of the identification among BRV isolates of rotavirus serotypes G1, G2, G3, and G11.