Investigating a clinically actionable BRAF mutation for monitoring low-grade serous ovarian cancer: A case report.

Low-grade serous ovarian cancer (LGSOC) poses a specific clinical challenge due to advanced presentation at diagnosis and the lack of effective systemic treatments. The aim of this study was to use a precision medicine approach to identify clinically actionable mutations in a patient with recurrent LGSOC. Primary, metastatic and recurrence tissue, and blood samples were collected from a stage IV LGSOC patient. Single-gene testing for clinically actionable mutations (BRAF V600, KRAS and NRAS) and subsequent whole-exome sequencing (WES) were performed. Droplet digital PCR was used to evaluate the presence of an identified BRAF D594G mutation in the matched plasma cell-free DNA (cfDNA). No clinically actionable mutations were identified using single-gene testing. WES identified a BRAF D594G mutation in six of seven tumor samples. The patient was commenced on a MEK inhibitor, trametinib, but with minimal clinical response. A newly designed ddPCR assay detected the BRAF alteration in the matched tissues and liquid biopsy cfDNA. The identification and sensitive plasma detection of a common "druggable" target emphasises the impact of precision medicine on the management of rare tumors and its potential contribution to novel monitoring regimens in this field.

The emergence of DNA methylation as a key modulator of aberrant cell death in prostate cancer.

It is now well established that cancer cells exhibit a number of genetic defects in the machinery that governs programmed cell death and that sabotage of apoptosis is one of the principal factors aiding in the evolution of the carcinogenic phenotype. A number of studies have implicated aberrant DNA methylation as a key survival mechanism in cancer, whereby promoter hypermethylation silences genes essential for many processes including apoptosis. To date, studies on the methylation profile of apoptotic genes have largely focused on cancers of the breast, colon and stomach, with only limited data available on prostate cancer. Here we discuss the major developments in the field of DNA methylation and its role in the regulation of aberrant apoptosis in prostate cancer. The most significant advances have involved the discovery of apoptotic gene targets of methylation, including XAF1, (fragile histidine triad (FHIT ), cellular retinol binding protein 1 (CRBP1), decoy receptor 1(DCR1), decoy receptor 2 (DCR2 ), target of methylation-induced silenceing 1 (TMS1), TNF receptor superfamily, member 6 (FAS), Reprimo (RPRM) and GLI pathogenesis-related 1 (GLIPR1). These genes are reported to be hypermethylated in prostate cancer and some offer potential as diagnostic and prognostic markers. We also introduce the concept of an 'apoptotic methylation signature' for prostate cancer and evaluate its potential in a diagnostic, prognostic and therapeutic setting.

In silico mining identifies IGFBP3 as a novel target of methylation in prostate cancer.

Promoter hypermethylation is central in deregulating gene expression in cancer. Identification of novel methylation targets in specific cancers provides a basis for their use as biomarkers of disease occurrence and progression. We developed an in silico strategy to globally identify potential targets of promoter hypermethylation in prostate cancer by screening for 5' CpG islands in 631 genes that were reported as downregulated in prostate cancer. A virtual archive of 338 potential targets of methylation was produced. One candidate, IGFBP3, was selected for investigation, along with glutathione-S-transferase pi (GSTP1), a well-known methylation target in prostate cancer. Methylation of IGFBP3 was detected by quantitative methylation-specific PCR in 49/79 primary prostate adenocarcinoma and 7/14 adjacent preinvasive high-grade prostatic intraepithelial neoplasia, but in only 5/37 benign prostatic hyperplasia (P < 0.0001) and in 0/39 histologically normal adjacent prostate tissue, which implies that methylation of IGFBP3 may be involved in the early stages of prostate cancer development. Hypermethylation of IGFBP3 was only detected in samples that also demonstrated methylation of GSTP1 and was also correlated with Gleason score > or =7 (P=0.01), indicating that it has potential as a prognostic marker. In addition, pharmacological demethylation induced strong expression of IGFBP3 in LNCaP prostate cancer cells. Our concept of a methylation candidate gene bank was successful in identifying a novel target of frequent hypermethylation in early-stage prostate cancer. Evaluation of further relevant genes could contribute towards a methylation signature of this disease.

Cost-effectiveness analysis of inhaled zanamivir in the treatment of influenza A and B in high-risk patients.

OBJECTIVE: To evaluate the cost effectiveness of zanamivir 10 mg twice daily for 5 days in the treatment of influenza in high-risk patients. DESIGN: Bootstrap cost-effectiveness analysis incorporating within-trial analysis of pooled patient-level cost and effect data. SETTING: UK unit costs and utilities applied to high-risk patients drawn from 6 multinational clinical trials. PATIENTS: A total of 154 zanamivir and 167 placebo high-risk patients were included in the analysis. MAIN OUTCOME MEASURES: Cost per day of normal activities; cost per symptom-free day; cost per complication averted; cost per quality-adjusted life-year (QALY). RESULTS: The mean benefit was estimated to be 2.5 days [95% confidence interval (CI): 0.68 to 4.27] of normal activities gained; 2.0 (95% CI: 0.56 to 3.51) symptom-free days; and a 9% reduction in complications (95% CI: 0 to 18%). Excluding the effect of rare hospitalisation costs, the cost (1999 values) of gaining a day of normal activities was 9.50 Pounds (95% CI: 5 Pounds to 39 Pounds); cost per symptom-free day was 11.56 Pounds (95% CI: 6 Pounds to 43 Pounds); cost per complication averted was 262 Pounds (95% CI: 90 Pounds to 1574 Pounds). Influenza was estimated to reduce utility by 0.883 per day, demonstrating the debilitating effect of the disease. Extrapolating a day of normal activities to a standard utility measure resulted in a cost per QALY of 3900 Pounds excluding inpatient costs (7490 Pounds including inpatient costs). Cost-effectiveness acceptability curves demonstrated 90% certainty that zanamivir would be cost effective at 8000 Pounds per QALY. CONCLUSIONS: Significant health benefits can be obtained with zanamivir treatment in high-risk patients. The cost per QALY for zanamivir in these patients compares well with that of other commonly used pharmacological interventions.

The Environmental Health Profile (EHP) for the Acre Valley (Israel): xenobiotics in animals and physiological evidence of stress.

The scope of this study was to analyse the levels of various xenobiotics in animals, wild and domestic, of the Acre Valley in the north of Israel, the most polluted and dense urbanised part of the country. The focus was placed upon the bioaccumulation of heavy metals, such as Pb, Hg, Cu, Zn, Al, Fe, as well as other elements and PCBs. Analysis was also performed on levels of cholinesterase and acetylcholinesterase in birds and man influenced by insecticides in the same region. For this study we used X-ray scanning electron microscopic (S.E.M.) electron probe, atomic absorption spectrophotometry, gas chromatography for PCBs and biochemical methods specific for enzymes. We studied four species of littoral molluscs, four species of birds and five species of mammals including dogs and cats. The results showed very high levels of metals and PCBs and enzyme deviations in birds and man around the sprayed cotton fields, as compared to man from the close to towns. All the collected data will serve as an Environmental Health Profile for immediate and future consideration.

Monooxygenase activity of rat liver microsomes immobilized by entrapment in a crosslinked prepolymerized polyacrylamide hydrazide.

Rat liver microsomes were immobilized by entrapment in a chemically crosslinked synthetic gel obtained by crosslinking prepolymerized polyacrylamide-hydrazide with glyoxal. Approximately 88% of the microsomal fraction was entrapped in the gel. The specific rate of O-demethylation of p-nitroanisole was used to assay the microsomal cytochrome P-450 activity of the immobilized microsomal preparations. The gel entrapped microsomes showed monooxygenase activity at 37 degrees C of Vmax = 2.3 nmol p-nitrophenol/min per nmol cytochrome P-450, similar to that of microsomes in suspension. The Km value for the p-nitroanisole-immobilized microsomal cytochrome P-450 system (1.2 X 10(-5) M) was rather close to that of microsomes in suspension (0.8 X 10(-5) M). Under the experimental conditions used the pH activity curve of the immobilized preparation was shifted towards more alkaline values by approx. 0.5 pH unit in comparison with microsomes in suspension. The rate of cytochrome c reduction by the immobilized microsomal system (11.7 nmol/min per mg protein) at 25 degrees C was considerably lower than that of the control (microsomes in suspension, 78 nmol/min per mg protein). Enzyme activity in both preparations showed the same temperature dependence at the temperature range of 10 to 37 degrees C. The immobilized microsomal monooxygenase system could be operated continuously for several hours at 37 degrees C provided that adequate amounts of an NADPH-generating system were added periodically. Under similar conditions a control microsomal suspension lost its enzymic activity within 90 min.

The effect of Aroclor 1254 (PCB) on the physiology of reproduction in the female rat--I.

Daily oral exposure of rats to 30 mg/kg of Aroclor 1254 for 1 month caused deleterious effects on the reproductive process, which were reflected in a decrease in the reproductive potential. The following disturbances were observed: prolongation of the estrous cycle; decrease in sexual receptivity; delay in timing of copulation; vaginal bleeding during gestation; decrease in litter size and delay in the time of parturition. The offspring, whether exposed to PCBs either prenatally and/or postnatally, showed a slower rate of body weight gain than controls. This was accompanied by high mortality until weaning of treated pups. Vaginal opening in the PCB-treated (as young) females occurred precociously, while other reproductive parameters were not affected at adulthood. Discontinuance of PCB treatment reversed the above symptoms.

Pesticide residues in fish and aquatic invertebrates.

Over the past 6 years a great number of fish and aquatic invertebrates from experimental and commercial fishponds have been analyzed for the presence of organochlorine insecticides and polychlorinated biphenyls (PCBs). Invariably, all specimens contained DDE and PCBs and in many instances also DDT, DDD and BHC isomers. A study of the kinetics of the organophosphorus compound parathion in an experimental fishpond ecosystem revealed a rapid bioconcentration of the chemical in algae, zooplankton, aquatic invertebrates and fish. The time-course of parathion disappearance showed an exponential type decline of the chemical over a period of three weeks.

Postnatal development of the mixed function oxidase system in nestling barn owls and baby chicks and induction of this system in the mature forms by Aroclor 1254.

Postnatal changes in content and activity of the mixed-function oxidase system in nestling barn owls and baby chicks showed the following: 1. Increase in liver weight in both. 2. Significantly higher cytochrome P-450 level in 1-day old chicks but lower in 1-day old owls than at any other age. 3. Ratio of cytochrome b5 to P-450 was lower than one in nestling owls but higher than one in chicks. 4. Aroclor 1254 (PCBs) increased the level and catalytic activity of cytochrome P-450 more in the owl than in the chick. 5. The ratio 455:430 nm characteristic of ethylisocyanide binding was not altered in the barn owl due to PCB treatment but changed significantly in the treated chicken.