Determining methylation status of methylguanine DNA methyl transferase (MGMT) from formalin-fixed, paraffin embedded tumor tissue.

O-6-methylguanine-DNA methyltransferase (MGMT) has been associated with resistance to alkylating agent cancer therapy in Glioblastoma (GBM), the most common and aggressive primary brain tumor in adults. Lower expression or silencing of the MGMT protein by promoter methylation has been reported to improve survival in patients with GBM [1]. This protocol describes bisulfite conversion, methylation sensitive PCR amplification and data analysis/interpretation.

Quantification of magnetic nanoparticles with low frequency magnetic fields: compensating for relaxation effects.

Quantifying the number of nanoparticles present in tissue is central to many in vivo and in vitro applications. Magnetic nanoparticles can be detected with high sensitivity both in vivo and in vitro using the harmonics of their magnetization produced in a sinusoidal magnetic field. However, relaxation effects damp the magnetic harmonics rendering them of limited use in quantification.

Immunotherapy for the treatment of glioblastoma.

Glioblastoma, the most aggressive primary brain tumor, thrives in a microenvironment of relative immunosuppression within the relatively immune-privileged central nervous system. Despite treatments with surgery, radiation therapy, and chemotherapy, prognosis remains poor. The recent success of immunotherapy in the treatment of other cancers has renewed interest in vaccine therapy for the treatment of gliomas.

A review of current and emerging approaches to address failure-to-rescue.

Failure-to-Rescue, defined as hospital deaths after adverse events, is an established measure of patient safety and hospital quality. Until recently, approaches used to address failure-to-rescue have been focused primarily on improvement of response to a recognized patient crisis, with limited success in terms of patient outcomes. Less attention has been paid to improving the detection of the crisis.

Meal-induced increases in C-reactive protein, interleukin-6 and tumour necrosis factor α are attenuated by prandial + basal insulin in patients with Type 2 diabetes.

Aim: To determine if a regimen with prandial + basal insulin compared with basal insulin attenuates post-meal inflammatory and glycative biomarkers in patients with Type 2 diabetes.

Methods: This test-meal sub-study in the USA is from a previously reported clinical trial comparing the effect on glycaemic control of 24 weeks of thrice-daily pre-meal insulin lispro mix 50 (50% insulin lispro, 50% insulin lispro protamine suspension) or bedtime insulin glargine, both plus metformin. In the sub-study, glucose, insulin, triglycerides, high-sensitivity C-reactive protein, tumour necrosis factor α, interleukin-6, methylglyoxal and 3-deoxyglucosone were measured during the post-meal period of a mixed-meal breakfast at the final visit.

Evaluation of a BK virus real-time PCR assay designed using novel bioinformatics technology.

Monitoring of active polyomavirus BK (BKV) infections by quantitative real-time PCR is becoming a progressively more routine practice in the care of renal transplant patients due to the potential for these infections to injure transplanted kidneys. Quantitative BKV results from a previously validated, laboratory-developed real-time PCR assay based on commercially available MGB Alert reagents were compared to results obtained from the same urine and plasma specimens using a commercially designed real-time PCR assay by IntelligentMDx. When compared qualitatively, the two assays performed identically with the exception of one urine specimen in which BKV DNA was detected near the lower limit of quantification by the MGB Alert assay.

Reproductive-endocrine point-of-care testing: current status and limitations.

Point-of-care (POC) testing for the detection of pregnancy and the prediction of ovulation has grown and evolved since the introduction of the first qualitative pregnancy test marketed directly to the consumer over three decades ago. Numerous publications have analyzed both pregnancy and ovulation prediction devices for their sensitivity, accuracy and general utility. Despite vast improvements in ease-of-use and sensitivity from their earlier forms, the primary literature regarding the utility of these devices is at times incomplete.

A randomized clinical trial to evaluate optimal treatment for unexplained infertility: the fast track and standard treatment (FASTT) trial.

Objective: To determine the value of gonadotropin/intrauterine insemination (FSH/IUI) therapy for infertile women aged 21-39 years.

Design: Randomized controlled trial.

Setting: Academic medical center associated with a private infertility center.

Sublethal concentrations of gemcitabine (2',2'-difluorodeoxycytidine) alter mitochondrial ultrastructure and function without reducing mitochondrial DNA content in BxPC-3 human pancreatic carcinoma cells.

2',2'-Difluorodeoxycytidine (gemcitabine), a pyrimidine nucleoside analog, is used therapeutically in the treatment of pancreatic, non-small cell lung, and breast cancer. The cytotoxic effect of gemcitabine is thought to be due to masked chain termination after the triphosphorylated anabolite of the drug is incorporated into nascent DNA strands. We tested the hypothesis that sublethal concentrations of gemcitabine inhibit DNA polymerase gamma and reduce mitochondrial DNA content in BxPC-3 and MOLT-4 cell lines, and we used 2',3'-dideoxycytidine, a known inhibitor of DNA polymerase gamma as a positive control.

Screening and diagnosis of gestational diabetes mellitus.

Screening, diagnosis, and treatment of gestational diabetes mellitus (GDM) are common practice, despite controversy regarding benefits. A review of the literature from 1950 to 2006 revealed 3 randomized controlled trials evaluated the treatment of GDM but 2 of these studies lacked power to detect a difference in outcomes. The single trial with sufficient power showed a 67% lower rate of serious perinatal complication (a composite of shoulder dystocia, nerve injury, fracture, and death) and a 53% lower rate of macrosomia with treatment of GDM.

Percutaneous coronary intervention in two occluded arms of a saphenous 'Y'-graft: the importance of protecting both of your arms.

Percutaneous intervention in saphenous vein grafts (SVG) carries a higher risk of distal embolization than intervention in a native vessel, and use of a distal protection device has been shown to improve the outcomes in SVG interventions. We describe an intervention done in an unexpected 'Y' SVG which required dual distal protection with Filterwires placed in both limbs of the diseased graft and which was performed via a 6 Fr guide catheter.

Molecular diagnostics: a historical perspective.

The rapid growth in molecular diagnostic testing, which has averaged between 10% and 20% per year for the past 5 years, is largely attributable to both breakthroughs in our basic understanding (i.e., the Human Genome Project) and in applied technology.

Negative urine opioid screening caused by rifampin-mediated induction of oxycodone hepatic metabolism.

Introduction: Oxycodone has become widely used in the clinic for the treatment of chronic pain. This reflects its favorable pharmacokinetics and side effect profiles.

Case Report: We report a 60-y-old man who had a clinically significant drug interaction between rifampin and oxycodone, resulting in 3 consecutive negative urine oxycodone screens in a 2-month period, suggesting non-adherence.

Susceptibility to diabetic nephropathy is related to dicarbonyl and oxidative stress.

Dicarbonyl and oxidative stress may play important roles in the development of diabetes complications, and their response to hyperglycemia could determine individual susceptibility to diabetic nephropathy. This study examines the relationship of methylglyoxal, 3-deoxyglucosone (3DG), and oxidative stress levels to diabetic nephropathy risk in three populations with diabetes. All subjects in the Overt Nephropathy Progressor/Nonprogressor (ONPN) cohort (n = 14), the Natural History of Diabetic Nephropathy study (NHS) cohort (n = 110), and the Pima Indian cohort (n = 45) were evaluated for clinical nephropathy, while renal structural measures of fractional mesangial volume [Vv(Mes/glom)] and glomerular basement membrane (GBM) width were determined by electron microscopy morphometry in the NHS and Pima Indian cohorts.

Clinical genotyping: the need for interrogation of single nucleotide polymorphisms and mutations in the clinical laboratory.

Background: Detection of single nucleotide polymorphisms (SNPs) and gene mutations is becoming more routine to the clinical laboratory.

Methods: Completion of the Human Genome Project has led to new scientific knowledge of human disease processes that has revealed the most fundamental of abnormalities in nucleic acids while at the same time bringing some of the most sophisticated diagnostic tools to the clinical laboratory. In addition, public awareness (both lay persons and healthcare providers) and sensitivity to human genetics has increased tremendously.

What diagnostic tests are useful for low back pain?

Low back pain (LBP) is a common problem that poses some interesting and difficult diagnostic problems. It is typically benign and self-limited, but it is occasionally the presenting symptom of serious systemic disease. The general diagnostic approach to low back pain is to check for 'red flags' in the history and physical that suggest the presence of malignancy, infection or spondyloarthridites, and for neurological compromise that could indicate that surgery is required (cauda equina syndrome) or may be beneficial (such as herniated discs or spinal stenosis that have not improved with conservative care).

Clerkship enhancement of interpersonal skills.

Background: The purpose of this research was to determine if students improve interpersonal skills as the third year progresses despite the lack of any specific curriculum or teaching methods.

Methods: Third-year students completed 1 of 3 16-week sequential clerkship blocks. Each student completed a clinical performance examination before and after clerkship consisting of a videotaped standardized patient interview and physical examination.

Poison ivy: an underreported cause of erythema multiforme.

The relationship between herpes simplex virus (HSV) and erythema multiforme (EM) has been well described. Many authors contend that EM (excluding Stevens-Johnson syndrome and toxic epidermal necrolysis) occurs almost exclusively as a response to HSV infection. During the past year, however, we have observed several cases of EM complicating severe Rhus allergic contact dermatitis.