TargetPlex FFPE-Direct DNA Library Preparation Kit for SiRe NGS panel: an international performance evaluation study.

AIM: Next generation sequencing (NGS) represents a key diagnostic tool to identify clinically relevant gene alterations for treatment-decision making in cancer care. However, the complex manual workflow required for NGS has limited its implementation in routine clinical practice. In this worldwide study, we validated the clinical performance of the TargetPlex FFPE-Direct DNA Library Preparation Kit for NGS analysis. Impressively, this new assay obviates the need for separate, labour intensive and time-consuming pre-analytical steps of DNA extraction, purification and isolation from formalin-fixed paraffin embedded (FFPE) specimens in the NGS workflow. METHODS: The TargetPlex FFPE-Direct DNA Library Preparation Kit, which enables NGS analysis directly from FFPE, was specifically developed for this study by TargetPlex Genomics Pleasanton, California. Eleven institutions agreed to take part in the study coordinated by the Molecular Cytopathology Meeting Group (University of Naples Federico II, Naples, Italy). All participating institutions received a specific Library Preparation Kit to test eight FFPE samples previously assessed with standard protocols. The analytical parameters and mutations detected in each sample were then compared with those previously obtained with standard protocols. RESULTS: Overall, 92.8% of the samples were successfully analysed with the TargetPlex FFPE-Direct DNA Library Preparation Kit on Thermo Fisher Scientific and Illumina platforms. Altogether, in comparison with the standard workflow, the TargetPlex FFPE-Direct DNA Library Preparation Kit was able to detect 90.5% of the variants. CONCLUSION: The TargetPlex FFPE-Direct DNA Library Preparation Kit combined with the SiRe panel constitutes a convenient, practical and robust cost-saving solution for FFPE NGS analysis in routine practice.

The impact of managed care and practice size on primary care physicians' perceived ability to refer.

OBJECTIVES: To identify the determinants of primary care physicians' perceived ability to refer patients, to compare perceived ability to refer between solo/two-physician practices and group practices, and to determine the impact of managed care on perceived ability to refer. METHODS: Multivariate analysis using a dataset derived from the Community Tracking Study Physician Survey, 1996-1997. The variables used to explain physicians' perceived ability to refer included physician and practice characteristics as well as aspects of the financial arrangements of managed care. The sample was stratified by practice size. A likelihood ratio test was performed to determine whether there were differences in practice characteristics and managed care financial arrangements that could explain variations in perceived ability to refer between physicians in solo/two-physician and group practices. RESULTS: Perceived ability to refer did not vary much between physicians in solo/two-physician practices and those in group practices. However, the determinants of perceived ability to refer did vary by practice size. The effects of physicians' characteristics were more pronounced among physicians in group practice, whereas the effects of financial arrangements were significant for physicians in solo/two-physician practices. The most significant determinant of perceived ability to refer was primary care physicians' satisfaction in their communication with specialists. CONCLUSION: Group practices are more immune than solo/two-physician practices to external financial arrangements from managed care contracts, possibly through their ability to take advantage of economies of scale and to diversify their sources of funds.

Targeted serum miRNA (TSmiR) test for diagnosis and follow-up of (testicular) germ cell cancer patients: a proof of principle.

Germ cell cancers (GCC) are the most frequent malignancy in young Caucasian males. GCC can consist of seminomas (SE) and non-seminomas (malignant NS: embryonal carcinoma (EC), yolk sac tumor (YS), choriocarcinoma (CH) and teratoma (TE)). Current serum-markers used for diagnosis and follow-up (AFP, hCG) are predominantly related to YS and CH and marker positivity can vary during disease. Therefore, stable markers consistently identifying more GCC components, specifically the stem cell components SE and EC, are of interest. Expression of the embryonic stem cell miR-371-3 and miR-302/367 clusters in SE/EC/YS suggest possible application of these micro-RNAs as GCC tumor-markers. The TSmiR protocol constitutes a complete, quality-controlled pipeline for the detection of miRs in serum, based on magnetic bead-based purification and qPCR quantification. As a proof of principle, TSmiR was applied to five independent serum sample series including 80 GCCs, 47 controls, 11 matched pre/post orchidectomy samples and 12 no-GCC testicular masses. GCC serum samples showed a consistent, significant (p < 0.0064) increase of miR-371/372/373/367 levels. Analogous, serum levels returned to baseline after orchidectomy (stage-I disease). Moreover, there was a trend toward higher miR levels in patients with metastasis. These results imply suitability for diagnosis and follow-up. TSmiR showed an overall sensitivity of 98%, clearly outperforming the traditional serum markers AFP/hCG (36%/57%, sensitivity(AFP) = 3%/45%; sensitivity(hCG) = 62%/66%, SE/NS). TSmiR misclassified one tumor as a control. Serum AFP/hCG and TSmiR combined identified all T samples correctly. In conclusion, TSmiR constitutes a highly sensitive and reproducible serum test for GCC patients, suitable to be prospectively tested for diagnostic and follow-up purposes.

Correlation of body mass index and blood loss during total knee and total hip arthroplasty.

Almost one-third of Americans older than 20 years are considered obese. Excessive weight has been linked to faster destruction of weight-bearing joints, which may then need to be replaced. Joint replacement surgeons disagree about an association between obesity and increased blood loss during hip or knee joint replacement. In this retrospective study, we examined the effect of body mass index (BMI), operative time (length of procedure), and anesthesia time on total blood loss during primary total knee arthroplasty (TKA) and primary total hip arthroplasty (THA). Intraoperative data from 94 primary TKAs and 78 primary THAs were reviewed, and divided into obese and nonobese groups on the basis of calculated BMI. Regression analysis was used to compare intraoperative blood loss amounts to patient characteristics. TKA and THA groups were analyzed separately. Obesity did not correlate with increased intraoperative blood loss in the TKA or THA group. However, operative time correlated with increased intraoperative blood loss. A 1-minute increase in anesthesia time resulted in total blood loss increases of 3.167 mL during TKA and 1.552 mL during THA.

A study to examine the attitudes, knowledge, and utilization of CAM by primary care professional in West Texas.

OBJECTIVE: This study examined the attitudes, knowledge, and utilization of CAM among primary care providers at two campuses of the Texas Tech University Health Sciences Center (TTUHSC). DESIGN AND SETTING: A cross-sectional study design and a convenient sampling method were used. This study employed the questionnaire adapted from the Wahner-Roedler's study to survey participants in TTUHSC. Primary survey collection was conducted at the two campuses where all Family Medicine healthcare professionals were recruited. MAIN OUTCOME MEASURES: We measured participants' knowledge of, their familiarity and experience with, their attitudes towards and utilization of CAM. RESULTS: Of the 69 respondents, more than half (56.5%) were female and younger than 36 years. Overall, our study revealed a positive attitude towards CAM. More than 60% of the providers would like to refer a patient to a CAM practitioner and about 75% of them believed that incorporation of CAM therapies into the practice would have a positive impact. Providers were most familiar with and felt most comfortable counseling their patients about massage therapy and St. John's Wort among all CAM modalities. About 70% of the participants believed that the institution should offer proven CAM therapies to patients. CONCLUSIONS: This study provides some preliminary findings that may lead to further exploration of healthcare professional's attitudes towards CAM.

Combined hepatocellular cholangiocarcinomas; analysis of a large database.

AIM: Combined hepatocellular cholangiocarcinoma (combined tumor) has been described as either a variant of hepatoma or a variant of cholangiocarcinoma. Prior studies evaluated fewer than 50 patients with combined tumors, precluding multivariate analyses. Posited was the notion that analysis of a large database would yield more definite answers. METHODS: This study used SEER (Surveillance, Epidemiology, and End Results Program of the National Cancer Institute) to analyze 282 combined tumors, 2,035 intrahepatic cholangiocarcinomas, and 19,336 hepatomas between the years 1973-2003. Multinomial logit regression calculated point estimates and 95% confidence intervals (c.i.) for relative risk (rr). Cox regression calculated point estimates and 95% confidence intervals (c.i.) for hazard ratios (h). RESULTS: Men less often had cholangiocarcinomas than they had combined tumors (rr = 0.63, c.i. = 0.49-0.81). Hepatomas less often than combined tumors presented with distant spread (rr = 0.56, c.i. = 0.43-0.72). Men (rr = 1.50, c.i. = 1.17-1.93) and patients with a known Asian or Pacific birthplace (rr = 2.36, c.i. = 1.56-3.56) more often had hepatomas than they had combined tumors. Among patients not known to have an Asian/Pacific birthplace, a diagnosis of cholangiocarcinoma (h = 0.72, c.i. = 0.63-0.82) or hepatoma (h = 0.75, c.i. = 0.66-0.86) provided a better prognosis than did a diagnosis of combined tumor. CONCLUSION: Combined tumors differ from hepatomas and cholangiocarcinomas in terms of distribution and survival patterns in the population; they should be considered neither cholangiocarcinomas nor hepatomas.

Cost-effectiveness of intermediate or long-acting insulin versus Exenatide in type 2 diabetes mellitus patients not optimally controlled on dual oral diabetes medications.

OBJECTIVE: To better understand exenatide's role in the treatment of type 2 diabetes, this analysis assessed its cost-effectiveness in comparison to an intermediate (NPH) and long-acting insulin (glargine). Exenatide is a recently approved medication for the treatment of type 2 diabetes for use in addition to frequently used oral diabetes medications. METHODS: Two studies were identified by a Medline search (1996-Oct 2005) that were similar in study duration, baseline glycemic control, population size, and primary outcomes to appropriately assess the cost-effectiveness of either insulin in comparison to exenatide on both glycemic and weight control. RESULTS: Both NPH and glargine appear to be more cost effective than exenatide with respect to glycemic control (incremental CE ratios -1,968 and - 65,520 respectively). Exenatide appears to be more cost effective for reductions in body weight than either NPH (CE ratio 235) or glargine (CE ratio 128). CONCLUSIONS: Compared to intermediate and long-acting insulin therapies, exenatide does not appear to be as cost effective for the treatment of type 2 diabetes.