Examining relationships between total silver concentration and Sil silver resistance genes in domestic wastewater treatment plants.

AIMS: The overarching aims of this study were to determine if microbes found in wastewater treatment plants (WWTPs) carry silver resistance genes and ascertain whether a relationship exists between silver loading and sil silver resistance gene copy numbers. METHODS AND RESULTS: Activated sludge samples were collected from 17 WWTPs across the United States and characterized for chemical oxygen demand (COD) and total silver concentration. Gene copy numbers were quantified using qPCR for four common silver resistance genes (silC, silP, silR and silS). Spearman correlation analyses were performed to examine associations between three WWTP characteristics (i.e. size, COD and total silver concentration) and sil gene copy numbers. sil genes were found in all activated sludge samples, however the quantity of gene copy numbers was inconsistent. Of the sil genes screened, silS was the most abundant, followed by silR. Gene copy numbers for silC and silP were generally similar and overall lower than silS and silR. CONCLUSIONS: Neither individual or combined sil gene copy numbers correlated significantly with total silver concentration, suggesting either the presence of alternate mechanisms of silver resistance or that a critical silver concentration threshold within WWTPs was not met that would require specific resistance against silver. SIGNIFICANCE AND IMPACT OF THIS STUDY: To the authors' knowledge, this is one of the first studies to measure the prevalence of sil genes in engineered systems. Results from this study suggest that at least four silver resistance genes are commonly found within WWTP-activated sludge, however it is unclear if these genes are being expressed under current total silver loadings.

In vivo reactivation of DNases in implanted human prostate tumors after administration of a vitamin C/K(3) combination.

Human prostate cancer cells (DU145) implanted into nude mice are deficient in DNase activity. After administration of a vitamin C/vitamin K(3) combination, both alkaline DNase (DNase I) and acid DNase (DNase II) activities were detected in cryosections with a histochemical lead nitrate technique. Alkaline DNase activity appeared 1 hr after vitamin administration, decreased slightly until 2 hr, and disappeared by 8 hr after treatment. Acid DNase activity appeared 2 hr after vitamin administration, reached its highest levels between 4 and 8 hr, and maintained its activity 24 hr after treatment. Methyl green staining indicated that DNase expression was accompanied by a decrease in DNA content of the tumor cells. Microscopic examination of 1-microm sections of the tumors indicated that DNase reactivation and the subsequent degradation of DNA induced multiple forms of tumor cell death, including apoptosis and necrosis. The primary form of vitamin-induced tumor cell death was autoschizis, which is characterized by membrane damage and the progressive loss of cytoplasm through a series of self-excisions. These self-excisions typically continue until the perikaryon consists of an apparently intact nucleus surrounded by a thin rim of cytoplasm that contains damaged organelles.