Rapid Detection of Human Cytomegalovirus UL97 and UL54 Mutations Directly From Patient Samples.

Rapid Detection of Human Cytomegalovirus UL97 and UL54 Mutations Directly from Patient Samples.

J Clin Microbiol. 2013 May 15;
Sedlak RH, Castor J, Butler-Wu SM, Chan E, Cook L, Limaye AP, Jerome KR

Human cytomegalovirus (CMV) is a significant contributor to morbidity and mortality in immunocompromised patients, particularly in the transplant setting. The availability of anti-CMV drugs has improved treatment, but drug resistance is an emerging problem. Here we describe an improved, rapid sequencing-based assay for the two genes in CMV where drug resistance occurs, UL97 and UL54. This assay is performed in 96-well format with a single master mix and provides clinical results within two days. It sequences codons 440-645 in UL97 and codons 255-1028 in UL54 with a limit of detection of 240 IU/ml. With this assay, we tested 43 specimens that had previously been tested for UL97 drug resistance and identified 3 with UL54 mutations. One of these patients had no concurrent UL97 mutation, pointing towards the need for an assay that facilitates dual UL97/UL54 testing for complete resistance profiling. HubMed – drug


Anti-inflammatory salicylate treatment alters the metabolic adaptations to lactation in dairy cattle.

Am J Physiol Regul Integr Comp Physiol. 2013 May 15;
Farney JK, Mamedova LK, Coetzee JF, Kukanich B, Sordillo LM, Stoakes SK, Minton JE, Hollis LC, Bradford BJ

Adapting to the lactating state requires metabolic adjustments in multiple tissues, especially in the dairy cow, which must meet glucose demands that can exceed 5 kg/day in the face of negligible gastrointestinal glucose absorption. These challenges are met through the process of homeorhesis, the alteration of metabolic setpoints to adapt to a shift in physiological state. To investigate the role of inflammation-associated pathways in these homeorhetic adaptations, we treated cows with the non-steroidal anti-inflammatory drug sodium salicylate (SS) for the first 7 days of lactation. Administration of SS decreased liver TNF? mRNA and marginally decreased plasma TNF? concentration, but plasma eicosanoids and liver NF-?B activity were unaltered during treatment. Despite the mild impact on these inflammatory markers, SS clearly altered metabolic function. Plasma glucose concentration was decreased by SS, but this was not explained by a shift in hepatic gluconeogenic gene expression or by altered milk lactose secretion. Insulin concentrations decreased in SS-treated cows on day 7 compared with controls, which was consistent with the decline in plasma glucose concentration. The revised quantitative insulin sensitivity check index (RQUICKI) was then used to assess whether altered insulin sensitivity may have influenced glucose utilization rate with SS. The RQUICKI estimate of insulin sensitivity was significantly elevated by SS on day 7, coincident with the decline in plasma glucose concentration. Salicylate prevented postpartum insulin resistance, likely causing excessive glucose utilization in peripheral tissues and hypoglycemia. These results represent the first evidence that inflammation-associated pathways are involved in homeorhetic adaptations to lactation. HubMed – drug


Differential regulation of c-Jun plays an instrumental role in chemoresistance of cancer cells.

J Biol Chem. 2013 May 15;
Xia Y, Yang W, Bu W, Ji H, Zhao X, Zheng Y, Lin X, Li Y, Lu Z

The chemotherapeutic drug cisplatin (CDDP) is widely used in treatment of human cancers. However, the mechanism underlying intrinsic tumor resistance to CDDP remains elusive. Here we demonstrate that treatment with CDDP results in downregulation of cJun expression via caspase9 dependent cleavage of cJun at D65 and MEKK1 mediated ubiquitylation and degradation of cJun in CDDP sensitive cancer cells. In contrast, activation of JNK2 but not JNK1 phosphorylates and upregulates the expression of cJun in CDDP resistant cells. Activated cJun binds to the promoter regions of the MDR1 gene and promotes the expression of MDR1. Expression of a D65A cleavage-resistant cJun mutant suppresses CDDP induced apoptosis of CDDP sensitive cells, whereas depletion of JNK2, cJun, or MDR1 in CDDP resistant cancer cells promotes apoptosis upon CDDP treatment. In addition, mammary gland tumors induced by polyoma virus middle T antigen in jnk2-/- mice are more sensitive to CDDP than are those in jnk2+/+ mice. These findings highlight the instrumental role of c-Jun in the resistance of tumors to treatment with CDDP and indicate that cJun is a molecular target for improving cancer therapy. HubMed – drug


Cross-reactivity of designer drugs, including cathinone derivatives, in commercial enzyme-linked immunosorbent assays.

Drug Test Anal. 2013 May 15;
Swortwood MJ, Hearn WL, Decaprio AP

Since the introduction of synthetic heroin, designer drugs have been increasing in prevalence in the United States drug market over the past few decades. Recently, ‘legal highs’ sold as ‘bath salts’ have become a household term for one such class of designer drugs. While a number of federal and state bans have been enacted, the abuse of these designer drugs still continues. Few assays have been developed for the comprehensive detection of such compounds, so it is important to investigate how they may or may not react in presumptive screens, i.e. pre-existing commercial immunoassays. In this experiment, 16 different ELISA reagents were evaluated to determine the cross-reactivity of 30 designer drugs, including 24 phenylethylamines (including 8 cathinone derivatives), 3 piperazines, and 3 tryptamines. Cross-reactivity towards most drugs was <4% in assays targeting amphetamine or methamphetamine. Compounds such as MDA, MDMA, ethylamphetamine, and ?-methyltryptamine demonstrated cross-reactivities in the range of 30-250%, but data were consistent with both manufacturer's inserts and published literature. When tested against the Randox Mephedrone/Methcathinone kit, cathinone derivatives demonstrated cross-reactivity at concentrations as low as 150 ng/ml. Since this same reagent did not cross-react with other amphetamine-like compounds, it opens the possibility to screen post-mortem specimens without the interference of putrefactive amines. All other assays demonstrated essentially no cross-reactivity towards any of the analytes evaluated. Given these results, a great need exists for more broad-range screening techniques to be applied when analyzing biological specimens by immunoassays for drugs of abuse, specifically the more recent designer drugs. Copyright © 2013 John Wiley & Sons, Ltd. HubMed – drug