Comparative evaluation of three JAK2V617F mutation detection methods

The correlation of JAK2V617F with a proportion of chronic myeloproliferative disorders has generated numerous studies focused on the development of molecular-based assays for JAK2V617F detection. The current parallel study comparatively evaluated 3 JAK2V617F molecular detection methods. Genomic DNA from blood or bone marrow was assayed by 3 laboratories using allele-specific polymerase chain reaction (AS-PCR) or kit-based restriction fragment length polymorphism methods, which used polyacrylamide gel or capillary electrophoresis analysis. In addition, samples were sequenced in 2 of the laboratories. Results found 100% concordance among the 3 methods, with analytic sensitivities of 5% for both kit methods and 0.01% for AS-PCR. The kitbased assays detect JAK2V617F with equal sensitivity regardless of analysis method, and, despite greater sensitivity of AS-PCR, all 3 methods yielded 100% concordant results for this 36-sample set. Consistent with other reports, direct sequencing was insufficiently sensitive to serve as an initial diagnostic tool for JAK2V617F detection.     

Contribution of DOG1 expression to the diagnosis of gastrointestinal stromal tumors

Gastrointestinal stromal tumors (GIST) are the most common mesenchymal tumors of the gastrointestinal tract, and the majority contain KIT or PDGFRA-activating mutations. However, up to 10% of GISTs are c-kit-negative. Antibodies with increased sensitivity and specificity for the detection of c-kit-negative GIST cases may be of value, especially because some of these cases may also benefit from tyrosine kinase inhibitor therapy.     

The MHC influences NK and NKT cell functions associated with immune abnormalities and lifespan

The lifespans of H-2 congenic mice differ significantly. The B10.AKM (H-2m) strain has a median survival time (MST) of 15 months, whereas the B10.BR (H-2k) strain has an MST of 24 months. It was previously shown that B10.AKM mice at 13-15 months of age have immunological function comparable to those of B10.BR mice at 22-26 months of age. These functions include: a low proliferative response, reduced levels of intracellular calcium release [Ca2+]i, and an increase in the frequency of memory helper T-cells (CD4+ CD44hiCD45RBlo). In this report similar deficiencies were demonstrated in B10.AKM mice at 2-4 months of age and show that activated spleen NK1.1+CD4+ T (NKT) cells from young B10.AKM mice produce a significantly higher level of IL-4 but a lower level of IFN-gamma as compared to NKT cells from B10.BR mice of the same age. Also, the cytotoxic activity of natural killer (NK) cells from spleens of young (2-4 months) as well as adult (12-16 months) B10.AKM mice is significantly lower (P < 0.01) than that of NK cells from B10.BR mice. These findings suggest that the NKT activity in young B10.AKM mice is a factor for the early onset of immune dysfunction leading to a shorter lifespan.     

Disappearance of Female Genital Mutilation from the Bedouin Population of Southern Israel

IntroductionRecently, clinicians in Southern Israel perceived that the practice of female genital mutilation had disappeared entirely in the Bedouin population. We previously studied the prevalence of this practice in 1995.AimWe decided to survey again the Bedouin population focusing on those tribes previously reported to perform this practice.MethodsEighty percent of the interviews were done by an Arabic-speaking psychiatrist and 20% were done by an Arabic speaking nurse in the gynecologic clinic of a large Bedouin township or the gynecologic clinic of a smaller Bedouin township. Women were asked if they would be willing to answer a few questions about their past and if they were willing to have the gynecologist, with no additional procedure, note whether any operation had been performed on their genitalia.Main Outcome MeasuresPhysical examination by gynecologist and an oral questionnaire.ResultsOne hundred and thirty two women were examined. No cases of any scarring of the kind reported in the previous study were found on physical examination.ConclusionsFGM has apparently disappeared over 15 years in a population in which it was once prevalent. Halila S, Belmaker RH, Rabia YA, Froimovici M, and Applebaum J. Disappearance of female genital mutilation from the bedouin population of Southern Israel. J Sex Med 2009;6:70–73.     

Intravitreal trovafloxacin against experimental Staphylococcus epidermidis endophthalmitis

This study was designed to test the effects of intravitreal trovafloxacin on an experimental rabbit model of Staphylococcus epidermidis endophthalmitis. Out of 26 rabbits, 22 were given intravitreal S. epidermidis (100,000 CFU). At 24 h, group 1 (8 rabbits) and, at 48 h, group 2 (8 rabbits) received 100 microg intravitreal trovafloxacin. Group 3 (6 rabbits) was used as inoculated but untreated controls. Four rabbits (group 4) were used as uninfected controls. Clinical scores were calculated at 24, 48 and 72 h. Microbiological and histopathological scorings were made. Microbiological analysis showed that trovafloxacin administered at 24 or 48 h significantly reduced the number of bacteria compared to the untreated group. We conclude that trovafloxacin applied at 24 or 48 h is effective against S. epidermidis endophthalmitis in this experimental rabbit model.     

Automatic electronic laboratory-based reporting of notifiable infectious diseases at a large health system

Electronic laboratory-based reporting, developed by the UPMC Health System, Pittsburgh, Pennsylvania, was evaluated to determine if it could be integrated into the conventional paper-based reporting system. We reviewed reports of 10 infectious diseases from 8 UPMC hospitals that reported to the Allegheny County Health Department in southwestern Pennsylvania during January 1-November 26, 2000. Electronic reports were received a median of 4 days earlier than conventional reports. The completeness of reporting was 74% (95% confidence interval [CI] 66% to 81%) for the electronic laboratory-based reporting and 65% (95% CI 57% to 73%) for the conventional paper-based reporting system (p>0.05). Most reports (88%) missed by electronic laboratory-based reporting were caused by using free text. Automatic reporting was more rapid and as complete as conventional reporting. Using standardized coding and minimizing free text usage will increase the completeness of electronic laboratory-based reporting.     

Diagnosis of tuberculosis lymphadenitis using a polymerase chain reaction on peripheral blood mononuclear cells

Providing prompt and precise laboratory confirmation of a clinical diagnosis of tuberculous lymphadenitis is difficult given the paucibacillary nature of lymph node specimens. In this study carried out in Karachi, Pakistan, a polymerase chain reaction (PCR) assay aimed at detecting Mycobacterium tuberculosis DNA in peripheral blood mononuclear cells (PBMC-PCR) was standardized and compared with standard M. tuberculosis diagnostic techniques or a lymph node PCR (LN-PCR) for the diagnosis of tuberculosis lymphadenitis. Thirty-seven (77%) specimens from 48 patients with clinical or diagnosis of tuberculosis lymphadenitis were positive by cytology [17/48 (35%) with no acid fast bacilli (AFB) (suggestive), and 20/48 (42%) with AFB (positive) in direct smears], 30 (63%) by PBMC-PCR, 16 (33%) by LN-PCR, and 13 (27%) by culture. All controls were negative, with the exception of one false-positive LN-PCR. These data suggest the PBMC-PCR may be helpful in the diagnosis of tuberculous lymphadenitis.