Development of the first PCR for detection of Psoroptes ovis var. cuniculi infection and its comparison to microscopic examination and serological assay in rabbits.

Psoroptes mites are the common ecto-parasites of wild and domestic animals worldwide, which causes considerable economic losses in livestock industry. Microscopy is deemed to be the 'gold standard' for the diagnosis of Psoroptes mite infection but it has poor sensitivity for low mite infections and/or sub-clinical infections. To overcome these shortcomings, we screened four genes to develop a sensitive and specific PCR for the detection of Psoroptes mite infection in rabbits, and confirmed its practicability in detecting early infection and monitoring treatment outcome with traditional microscopy and serological tests. Results showed that PCR assay targeting ITS2 (ITS2-PCR) had a high specificity and sensitivity (detection limit: 40.3 pg/µL DNA) for detecting P. ovis DNA. In rabbits artificially infected with P. ovis, all three diagnostic tests showed the same detection rate from 14 days post infection (dpi) to 42 days dpi. However, these diagnostic tests behave differently at 7 dpi and after treatment: at 7 dpi, the detection rate of ITS2-PCR was higher than rPsoSP3-based iELISA and traditional microscopy (ITS2-PCR: 88.9%, rPsoSP3-iELISA: 77.7%, microscopy: 33.3%); at 7 days post treatment (dpt), positivity rates of ITS2-PCR and microscopy rapidly decreased to 0.00% and 11.1%, whereas rPsoSP3-iELISA remained 100% positive rate. Furthermore, the comprehensive comparisons of diagnostic performance and features of three diagnostic tests at 7 dpi were performed. Compared to ITS2-PCR or rPsoSP3-iELISA, microscopy had the lowest sensitivity, and the agreement between these assays was low (κ < 0.3). Field study showed that ITS2-PCR showed a higher detection rate than microscopy (19.4% and 11.1%, respectively). Our results suggested that the ITS2-PCR developed in this study provided a new laboratory tool for diagnosis of P. ovis var. cuniculi infection, and it had advantages over microscopic examination in detection low-level mite infections and serological assay in monitoring treatment outcome.

Association between urothelial carcinoma after kidney transplantation and aristolochic acid exposure: the potential role of aristolochic acid in HRas and TP53 gene mutations.

OBJECTIVE: We sought to investigate the association between urothelial carcinoma (UC) after kidney transplantation and aristolochic acid (AA) exposure, and to explore the potential role of AA in HRas and TP53 gene mutations. MATERIALS AND METHODS: UC was confirmed in 100 of 3790 patients who underwent kidney transplantation between January 1974 and May 2011. We retrospectively analyzed clinical data for these patients. Malignancies were identified in 136 patients (15 males and 85 females, of age range 27-71 years mean, 53.2±6.3). UC was the most common diagnosis (100/136; 73.5%). The median time from transplantation to the first confirmed diagnosis of a tumor was 34.5 months (range, 2-273). Polymerase chain reaction-based resequencing methods were used to detect mutations in the target regions of exons 2 and 3 of the HRas gene and exons 5, 6, 7, and 8 of the TP53 gene in 90 of 100 samples. RESULTS: UC was identified in 53 of 380 patients exposed to AA and 47 of 3410 patients not exposed to AA (P<0.05). Pathologic examination of the UC specimens from AA-exposed patients identified heterozygous HRas changes in 3 cases, and deletion or replacement mutations in the TP53 gene in 4. No genetic mutations were observed among the control group. CONCLUSION: UC after kidney transplantation significantly correlated with AA exposure; however, there seemed to be no significant difference in mutations in exons 2 and 3 of the HRas gene and those in exons 5, 6, 7, and 8 of the TP53 gene in terms of tumor development, a result that is inconsistent with previous studies.

Association between urothelial carcinoma after renal transplantation and infection by human papillomavirus types 16 and 18.

OBJECTIVE: To explore the association between urothelial carcinoma following renal transplantation and infection by human papillomavirus (HPV) types 16 and 18. MATERIALS AND METHODS: Of 3780 patients who underwent renal transplantation, we identified 90 cases of urothelial carcinoma. Tumor tissues collected from the 90 renal transplant recipients were compared with those from 30 nontransplanted patients with bladder cancer (control group) for HPV types 16 and 18 using polymerase chain reactions. RESULTS: Seven transplanted patients were HPV positive: HPV-16 was detected in 3 patients with bladder cancer (3/90; 3.3%), and HPV-18 in 2 patients with bilateral pelvic ureteral carcinoma (2/90; 2.2%), and 2 patients with bladder cancer (2/90; 2.2%). Only 2 cases from the control group were HPV positive (both HPV-18; 2/30; 5%). The difference between the RTR and control groups was not significant (P > .05). CONCLUSION: Malignant tumors in the urinary system following renal transplantation did not seems to be associated with infection by HPV-16 or -18.

Androgen receptor isoforms in human and rat prostate.

AIM: To investigate the androgen receptor (AR) isoforms and its variability of expression in human and rat prostatic tissues. METHODS: Human benign prostatic hyperplasia (BPH) and prostatic cancer tissues were obtained from patients undergoing prostatectomy, and rat ventral prostate was incised 3 days after castration. Forty-one AR-positive BPH specimens, 3 prostatic cancer specimens, and 6 rat prostates were used. After processing at 4 degrees C, the tissues were examined by means of high resolution isoelectric focusing (IEF) technique to determine their AR isoforms. RESULTS: From the prostatic specimens, 3 types of AR isoforms were detected with pI values at 6.5, 6.0, and 5.3. In human BPH tissues, 15/41 (36.6%) specimens showed all the three types of isoforms, while 19/41 (46.3%) showed 2 isoforms at various combinations and 7/41 (17.1%), 1 isoform. For the 3 prostatic cancer specimens, one showed 3 isoforms, one, 2 isoforms, and the other failed to show any isoform. All rat prostatic tissues showed 2 isoforms at different combinations. Binding of 3H-dihydrotestosterone (DHT) to the isoforms was inhibited by the addition of 100-fold excess of DHT or testosterone, but not progesterone, oestradiol or diethylstilboestrol. CONCLUSION: AR isoforms are different in different patients. Although their genesis is not clear, the therapeutic implication of the present observation appears to be interesting, that may help clarifying the individual differences in the response to hormonal therapy.