Quantification of feline herpesvirus 1 DNA in ocular fluid samples of clinically diseased cats by real-time TaqMan PCR

A fluorogenic PCR was established for the quantification of feline herpesvirus 1 (FeHV-1) DNA in ocular fluid samples of clinically diseased cats. The new assay was specific for FeHV-1 and sensitive. The 100% detection rate ranged from 0.6 to 6 50% tissue culture infective doses per sample. When spiked samples with known quantities of virus were used, infectious virus titers and quantification of viral DNA by PCR correlated to each other in a linear fashion (R(2) = 0.9858) over a range of 4 orders of magnitude. Within this range, it was possible to calculate the FeHV-1 DNA content from a given infectious dose, and vice versa. The new diagnostic procedure was applied to ocular fluid samples from cats experimentally infected with FeHV-1 and specific FeHV-1-free cats. A good correlation between virus titer and quantitative PCR was observed, although only early in infection. In a second stage, the titer of infectious virus collapsed, while the PCR signal remained high. A constantly decreasing PCR signal accompanied by negative virus isolation was characteristic for a final stage of the infection. Finally, clinical samples from 20 cats that were suspected to suffer from FeHV-1 infection were analyzed. By comparing virus titers and quantitative PCR signals, it was possible to determine the current stage of the ongoing infection. Based on these findings, comparison of the results of consecutive samples allows the tracking of the course of the infection. Therefore, the new method combines the advantages of the two previously established conventional methods, qualitative PCR and virus isolation and titration.     

Ontogeny of pulmonary cyclooxygenase-1 (COX-1) and -2 (COX-2)

Prostaglandins synthesized by enzymatic reactions such as cyclooxygenases have been implicated in lung pathophysiology. The goal of this study was to delineate the pulmonary ontogeny of cyclooxygenase enzymes (COX-1 and COX-2) immunohistochemical expression and cellular localization in various microanatomic locations of lungs from pre-term, term, and post-natal lambs. Lung tissues were obtained at 115 and 130 days of gestation from pre-term lambs, 145 days (term; complete gestation), and 15 days post-natally. No significant differences were seen in lung COX-1 expression at various microanatomic locations during pre-term, term, or postnatally. Moderate to strong COX-1 expression was present in macrophages, alveolar septa, bronchial smooth muscle cells, bronchiolar smooth muscle cells, vascular endothelial cells, and vascular smooth muscle cells. Minimal COX-1 expression was present in bronchial and bronchiolar epithelial cells. Most microanatomic locations lacked COX-2 expression with the exception of weak expression that was present in bronchial and bronchiolar epithelial cells at 145 days of full gestation and 15 days post-natally. This work suggests that: (a) COX-1 is constitutively expressed in lungs from pre-term, term, and post-natal lambs in various microanatomic pulmonary locations, (b) there is differential expression of COX-1 and COX-2 in the developing lung, and (c) COX-2 does not appear to play a role in lung fetal development, at least in neonatal lambs.     

H‐Phosphonic Acid Derivatives as Catalysts for Reversible Chain Transfer Catalyzed Polymerization (RTCP) at Ambient and High Pressure

Several H‐phosphonic acid derivatives (PADs) were analyzed regarding their ability to perform as effective catalysts in reversible chain transfer catalyzed polymerization (RTCP), a relatively new type of controlled radical polymerization. Therefore, bulk polymerizations of styrene at 100 °C using different PADs were studied in detail. The obtained number‐average molar masses and polydispersities of the polymeric products showed that a cyclic PAD derived from pinacol was found to be a very well suited catalyst for RTCP under the chosen conditions. In addition, RTCPs at high pressure up to 2000 bar were conducted indicating poor molar mass control compared to the systems at ambient pressure.

     

Cyclin D1 overexpression lacks prognostic significance in superficial urinary bladder cancer

The biological behaviour of urinary bladder neoplasms cannot be adequately predicted by histological criteria alone. Cyclin D1 is a cell-cycle regulating protein known to be overexpressed in a proportion of bladder carcinomas. To evaluate the prognostic significance of cyclin D1 expression and its relationship with tumour phenotype, 392 bladder carcinomas were analysed by immunohistochemistry. Clinical follow-up information was available in 337 patients with superficial bladder tumours (stages pTa/pT1). Cyclin D1 positivity was seen in 176 of 392 carcinomas. Cyclin D1 overexpression was strongly linked to papillary tumour growth, low stage, and low histological grade (p<0·005 each). Multivariate analysis showed that papillary tumour growth was the only parameter which was independently linked to cyclin D1 positivity. There was no significant difference in proliferative activity (Ki67 labelling index) between cyclin D1-negative and -positive tumours. Cyclin D1 positivity was not linked to the risk of recurrence or tumour progression, either in pTa or in pT1 carcinomas. It is concluded that cyclin D1 positivity distinguishes a large subgroup of papillary bladder tumours, but there is no evidence of prognostic significance for increased cyclin D1 expression. Copyright © 1999 John Wiley & Sons, Ltd.