Histological and Immunohistochemical Evaluation of Phosphorylated Mechanistic Target of Rapamycin in Canine Skin Tumours.

The mechanistic target of rapamycin (mTOR) plays an important role in cellular functions, including growth and metabolism. Recently, mTOR and the activated phosphorylated form of mTOR (p-mTOR) have been reported as potential prognostic markers in many human tumours. However, there are few studies on its activation in canine tumours. We investigated the expression of p-mTOR in 17 canine skin tumours (CSTs), of which 58.8% were epithelial and melanocytic and 41.2% were mesenchymal tumours. Seventy-six per cent of the CSTs had high or moderate expression of p-mTOR. Mean p-mTOR expression in the epithelial and melanocytic tumours (5.7 ± 0.56) was significantly higher (P <0.05) than that of the mesenchymal tumours (3.14 ± 0.55). The age of the animals had no influence on p-mTOR expression. These findings suggest that activation of m-TOR is important in the development of skin tumours in dogs and the study might form the basis for further research on utilizing m-TOR inhibitors as improved therapeutic modalities in canine skin tumours.

Quality laboratory issues in bleeding disorders.

Selected quality issues pertinent to the determination of accurate results in the haemostasis laboratory are discussed. Specifically, the implementation of a successful external quality-assessment scheme is described, including its impact on result accuracy as well as the programme's unique challenges and opportunities. Errors in the preanalytical phase of laboratory testing represent the greatest source for reporting incorrect test results. Some of the most common preanalytical errors are described including those that necessitate sample rejection. Analytical means to identify potential sources of error and analytical means to overcome particular interferences are described. Representing the most important clinical complication in the treatment of patients with haemophilia, quality issues related to determination of the presence of inhibitory antibodies against factor VIII (FVIII) are reviewed. Heat treatment of patient plasma prior to testing, particularly in patients receiving replacement FVIII concentrate or during induction of immune tolerance to achieve more accurate results is recommended, while screening activated partial thromboplastin time-based mixing tests to rule out inhibitor presence is discouraged. The initiatives presented in this review can be implemented in robust and resource restricted settings to improve the quality of laboratory testing in patients with bleeding disorders.

Utility of neutrophil volume conductivity scatter (VCS) parameter changes as sepsis screen in neonates.

OBJECTIVE: The objective of this study was to determine changes in neutrophil volume conductivity scatter (VCS) parameters and their distribution widths (DW) in neonatal sepsis and to estimate their optimal cutoff levels using receiver operating characteristic (ROC) curves. STUDY DESIGN: In a cohort of neonates evaluated for sepsis, blood counts and blood culture were performed initially, with repeat counts and C-reactive protein (CRP) done after 24 to 48 h. Neutrophil VCS parameters from both the initial and repeat blood counts were analyzed. Babies were classified as having blood culture-positive sepsis, probable sepsis (clinical course consistent with sepsis and CRP-positive, but culture-negative) and no sepsis (clinical course not compatible with sepsis, culture- and CRP-negative). RESULTS: A total of 600 babies were included: 240 (40%) babies in the sepsis group and 360 (60%) babies in the control group. All the neutrophil VCS parameters and their DWs (except for low angle light scatter in the repeat counts) were significantly different between the two groups, with an area under curve in the ROC curve of >0.6 for most parameters. The five most significant VCS parameters (mean neutrophil volume (MNV), median angle light scatter (MALS), lower median angle light scatter (LMALS), MNV-DW and ALL-DW) had around 65 to 75% sensitivity and specificity. A combination of leukopenia, thrombocytopenia, MNV and LMALS had a likelihood ratio (LR)+ of 15.3 and LR- of 0.17. With a pre-test probability of 40%, post-test probability increased to 91% for a positive test and decreased to 10% for a negative test. A prospective validation study was performed recruiting an additional 60 babies, which showed similar results, assuring that the cutoffs were robust. CONCLUSION: Neutrophil VCS parameters cannot be considered as stand-alone tests to diagnose or rule out neonatal sepsis, but can be used in combination with other hematological screening tests to improve the diagnostic accuracy of the neonatal sepsis screen.

Characteristics in response rates for surveys administered to surgery residents.

BACKGROUND: Surveys are important research tools that permit the accumulation of information from large samples that would otherwise be impractical to collect. Resident surveys have been used frequently to monitor the quality of postgraduate training. Low response rates threaten the utility of this research tool. The purpose of this study was to determine the standard response rate of surveys administered to surgery residents and identify characteristics associated with achieving greater response rates. METHODS: A search of peer-reviewed literature published between September 2003 and June 2011 was performed with the use of PubMed with Medical Subject Headings: "internship and residency," "surgery," "data collection," and "questionnaires." For inclusion, articles must have described a survey given to active surgery residents within the United States. Surveys were evaluated based on the following criteria: population size, response rate, incentive use, follow-up use, survey format (online vs paper), and institution versus national. RESULTS: Of 433 initial results, 47 met inclusion criteria with a mean response rate of 65.3%. Surveys administered in paper format had a greater response rate compared with those given electronically (mean 78.6% vs 36.4%, respectively, P < .001). Greatest mean response rates were seen for institutional surveys compared with those given nationally (83.1% vs 42% respectively, P < .001). CONCLUSION: Our review demonstrated that paper surveys administered at the institutional level and during assemblies integrated into residents' schedules demonstrated enhanced response rates. The validity and generalizability of data collected through such surveys will improve as the aspects which dictate response rate are better understood and implemented.

Underlying prothrombotic states in pregnancy associated cerebral venous thrombosis.

BACKGROUND: The exact pathogenesis of pregnancy associated cerebral venous thrombois is still unsettled. Aims : To identify possible inherited and acquired prothrombotic risk factors and also identify the factors associated with mortality in pregnancy associated CVT. SETTINGS AND DESIGN: Prospective cohort study to identify prothrombotic risk factors and case control study of influence of local traditional practice of puerperal water restriction on postpartum CVT. MATERIALS AND METHODS: Consecutive patients with pregnancy associated CVT seen over a period of three years. Thrombotic workup included genetic markers, protein assays, and other factors. STATISTICAL ANALYSIS: Univariate and chi-square analysis. RESULTS: Of the 41 patients studied during the study period, 71% of patient had a single and 34% had multiple prothrombotic risk factors. Methylene tetrahydro-folate reductase (MTHFR) heterozygosity (19.5%) and factor V Leiden heterozygous (7.3%) were the commonest genetic markers. Hyperhomocysteinemia (34%) and elevated factor VIII levels (14.6%) were the other important risk factors. In this cohort the mortality was 17%. Mortality increased by odds of 1.3 for every additional prothrombotic marker. The factors associated with increased mortality included: status epileptics (P = 0.05, OR 13.2, 95% CI 1.002 - 173), deep venous system involvement (P = 0.016, OR 9.64, 95% CI 1.53 - 60.6), presence of midline shift (P = 0.012, OR 24.7, 95% CI 2.05 - 29.8) and diffuse cerebral edema (P = 0.006, OR 14.5, 95% CI 2.18- 96.4). The traditional practice of decrease intake of water during puerperium was significant in woman with pregnancy associated CVT when compared to control subjects (P < 0.02). CONCLUSION: In patients with pregnancy associated CVT, prothrombotic markers can be multiple and are associated with increased odds of mortality. Deep venous system involvement, presence of midline shift and diffuse cerebral edema increased mortality. Peuperial water restriction may be a modifiable risk factor.

Successful surgical haemostasis in patients with von Willebrand disease with Koate DVI.

This report describes our experience with Koate DVI, a factor VIII (FVIII) concentrate containing von Willebrand factor (VWF) for surgery in patients with von Willebrand's disease (VWD). Twenty-one patients underwent 26 procedures, 10 of which were major and 16 were minor. The median age was 27 years (3-55) and the mean weight was 52 kg (16-88). Among the ten patients (type 2-5; type 3-5) who underwent major procedures, the pre-operative dose was 35 IU kg(-1) of FVIII followed by 10-20 IU kg(-1) once daily depending on FVIII:C levels. The mean total dose of FVIII used per procedures was 106 IU kg(-1) (30-190) over a mean duration of 7 days (3-11). In this group, pre-infusion FVIII:C, VWF:Ag and VWF: ristocetin cofactor (RCoF) level that were 19.5% (1-64), 20 U dL(-1) (0-96) and 12% (0-66) increased to 72% (54-198), 131 U dL(-1) (68-206) and 68% (27-108) postinfusion, respectively. Sixteen minor procedures were performed in 11 patients (type 1-3, type 2-6, type 3-2). The preparative dose of FVIII was 10-20 IU kg(-1). The average duration of factor support was 2 days (1-3) for a mean total dose of 23 IU kg(-1) (9-60). The pre-infusion levels of FVIII:C, VWF:Ag and VWF:ristocetin cofactor (RCo) which were 31% (22-64), 25.5 U dL(-1) (0-63) and 21% (0-76), respectively, increased to 76% (27-111), 73 U dL(-1) (30-137) and 45% (2-106) postinfusion. Whereas surgical haemostasis was achieved in all patients, minor postoperative bleeding occurred after one procedure in each group. Both were controlled with additional doses of factor replacement. We conclude that Koate DVI in modest doses provide adequate haemostasis for surgery in patients with VWD.

Achieving and maintaining quality in the laboratory.

In order to ensure the delivery of a service of the highest possible quality, it is an essential requirement that laboratories undertake strict internal quality control (QC) measures as well as participate in external quality assessment (EQA) schemes. For any given test, a critical part of the internal QC process involves the establishment of reference intervals using samples taken from normal individuals, and then calculating limits representing the 95% range. This forms the basis for assessment of abnormal test results, which will in turn impact on laboratory performance in proficiency testing exercises in EQA programmes. Whereas for plasma-based assay systems, variability in performance in EQA exercises is usually determined by measurement of a coefficient of variation (CV), results of genetic testing is usually measured in absolute terms. Despite this, results of genetic EQA programmes confirm that errors in testing do occur, as much because of inadvertent sample switching and transcription errors as to analytical mistakes. EQA programmes involving identification of mutations by DNA sequencing, such as haemophilia, is made difficult by the high information content of sequence data. Nevertheless, results show that errors are usually made in the naming of the mutations, indicating that this is an evolving and poorly standardized area. Developing countries face particular challenges in the encouragement of laboratories to participate in local EQA programmes, as well as in relation to the logistical issues of sample provision, distribution and result collation in an effective and affordable manner.

Pre-transplant reduction of isohaemagglutinin titres by donor group plasma infusion does not reduce the incidence of pure red cell aplasia in major ABO-mismatched transplants.

Major ABO incompatibility in stem cell transplant recipients has been associated with pure red cell aplasia (PRCA). Reduction of incompatible isohaemagglutinin titres pre-transplant by various methods has been thought to reduce the incidence of PRCA. Our data suggest that pre-transplant reduction of incompatible isohaemagglutinin titres by donor group plasma infusion does not reduce the incidence of PRCA. We also failed to find any relationship between pre-transplant ABO isohaemagglutinin titre and the risk of developing PRCA.

Total synthesis of the calphostins: application of fischer carbene complexes and thermodynamic control of atropisomers.

The total syntheses of the potent protein kinase C inhibitors calphostins A, B, C, and D as well as a variety of structural analogues are reported. An aminobenzannulation reaction of an enantiopure chromium Fischer carbene complex is utilized to prepare a pentasubstituted naphthylamine. After optimization of side-chain substituents, conversion of the naphthylamine to an o-naphthoquinone was followed by biomimetic oxidative dimerization using trifluoroacetic acid and air yielding a 1:2 P/M mixture of atropisomeric perylenequinones. Thermal equilibration to a 3:1 P:M atropisomeric ratio and separation of the perylenequinones followed by side chain desymmetrization and functionalization led to the total synthesis of enantio- and diastereomerically pure calphostin C in only twelve steps from commercially available starting materials. In addition, calphostins A, B, D, and several structural analogues were prepared to evaluate biological activities.

Distribution of ABO and Rh-D blood groups among blood donors in a tertiary care centre in South India.

The distribution of ABO and Rh-D blood groups was studied among 150,536 blood donors screened at the Dr John Scudder Memorial Blood Bank, Christian Medical College Hospital, Vellore, over a period of 11 years (April 1988 to March 1999). The most common blood group was found to be group O [58,330 (38.75%)], followed by group B [49,202 (32.69%)], and group A [28,372 (18.85%)]. The least common blood group was AB group [7,930 (5.27%)]. A2 or A2B groups were found in 3.01% and 1.43% of donors, respectively. The prevalence of Rh-D negative group was found in 8,225 (5.47%) donors. Bombay group (H negative non-secretor, genotype hh phenotype Oh) was found in six donors (0.004%). Although the incidence of Rh-D negative group was identical to previously published data from North India, the most common blood group was O group in our study as opposed to B group.