Co-detection and discrimination of six human herpesviruses by multiplex PCR-ELAHA.

BACKGROUND: Herpesviruses are a significant cause of human morbidity. Traditional approaches to the identification of these viruses require infectious or at least antigenic virus. Multiplex PCR (mPCR) is capable of simultaneously amplifying a range of targets from a single preparation of nucleic acids and when combined with a suitable detection assay, it is capable of discriminating each of the amplicons. OBJECTIVES: Several methods have been described in the literature, however, they lack one or more significant design features required to suitably control a routinely applied nucleic acid amplification assay. We aimed to design a multiplex herpesvirus PCR that could co-amplify eight human herpesvirus targets plus an internal control (IC) molecule in a single tube. STUDY DESIGN: Primers were designed to target the DNA polymerase genes of each of the human herpesviruses. Synthetic controls were developed to act as templates for the evaluation of assay sensitivity and specificity and for development of an in-house competitive quantitative PCR. Amplicon was discriminated using a simplified enzyme linked amplicon hybridisation assay (ELAHA). RESULTS AND CONCLUSIONS: For routine diagnostic use we reduced the number of herpesviral targets from 8 to 6 in order to maintain adequate clinical sensitivity. The ELAHA proved more sensitive than agarose gel electrophoresis. Additionally, 36 cytomegalovirus positive patients were examined with an in-house quantitative PCR-ELAHA which was developed to confirm that that the mPCR's co-detection limit of 10(2) copy of synthetic template per millilitre was relevant for use in detecting virus from clinical samples. The mPCR-ELAHA was then applied to the screening of 174 patient specimens resulting in a specificity of 98% and a sensitivity of 93%. This preliminary study demonstrated that the mPCR-ELAHA was a complete approach to the detection of herpesviruses from a range of clinical samples and disease states.     

Predictors of fatigue in obstructive sleep apnea

Objective  The objective of this study was to determine potential inflammatory predictors of fatigue in obstructive sleep apnea (OSA). Materials and methods  Fifty-six women and men untreated OSA patients had their sleep monitored with polysomnography. Fatigue was assessed by the Multidimensional Fatigue Symptom Inventory-Short Form. Depressed mood was assessed by the Center for Epidemiologic Studies-Depression Scale. Blood was drawn to assess circulating levels of Interleukin-6 (IL-6) and soluble tumor necrosis factor receptor I (sTNF-RI). Age, gender, body mass index (BMI), blood pressure, OSA severity, depressed mood, and inflammatory biomarkers were entered into a hierarchical multiple linear regression analysis predicting self-reported fatigue. Results  Approximately 42% of the patients reported significant amounts of fatigue. Higher BMI (p = 0.014), greater depressed mood (p = 0.004), and higher sTNF-RI levels (p = 0.033) were independent predictors of fatigue in the final model (full model R ² = .571; p = .003). Age, gender, blood pressure and apnea severity were unrelated to fatigue. Conclusion  The findings suggest that in addition to depressed mood, fatigue in OSA may be associated with increased body weight and elevated levels of the proinflammatory cytokine receptor sTNF-RI. The findings support a linkage between the widely reported fatigue in OSA and a sleep-related component of inflammation.     

The Brazilian Founder Mutation TP53 p.R337H is Uncommon in Portuguese Women Diagnosed with Breast Cancer

Since the first studies reporting the TP53 p.R337H mutation as founder mutation in Southern and Southeastern Brazil, there has been controversy on its origin. Preliminary analysis of a small subset of Brazilian mutation carriers revealed that the haplotype incided on a Caucasian background. The vast majority of carriers identified today reside in Brazil or, if identified in other countries, are Brazilian immigrants. To our knowledge, the only two exceptions of carriers without a recognizable link with Brazil are two European families, from Portugal and Germany. Haplotype analysis in the Portuguese family revealed the same haplotype identified in Brazilian individuals, but in the German family, a distinct haplotype was found. Knowing that a significant proportion of women with breast cancer (BC) in Southern Brazil are p.R337H carriers, we analyzed p.R337H in a Portuguese cohort of women diagnosed with this disease. Median age at diagnosis among the first 573 patients tested was 60 years and 100 (17.4%) patients had been diagnosed at or under the age of 45 years. Mutation screening failed to identify the mutation in the 573 patients tested. These results are in contrast with the mutation frequency observed in a study including 815 BC‐affected women from Brazil, in which carrier frequencies of 12.1 and 5.1% in pre‐ and postmenopausal women were observed, respectively. These findings suggest that the Brazilian founder mutation p.R337H, the most frequent germline TP53 mutation reported to date, is not a common germline alteration in Portuguese women diagnosed with BC.     

Jump-landing mechanics in patellar tendinopathy in elite youth basketballers

Patellar tendinopathy (PT) is a leading cause of morbidity in jump-landing athletes. Landing mechanics are identified as a factor associated with PT and/or patellar tendon abnormality. This study aimed to identify key jump-landing variables associated with PT. Thirty-six junior elite basketball players (men n = 18, women n = 18) were recruited from a Basketball Australia development camp. Three-dimensional (3D) kinematic and ground reaction force (GRF) data during a stop-jump task were collected as well as ultrasound scans of the patellar tendons and recall history of training load data. Mixed-model factorial analyses of variance were used to determine any significant between-group differences. Of the 23 participants included for statistical analyses, 11 had normal bilateral patellar tendons (controls) and eight reported PT (currently symptomatic); however, the four participants categorized as asymptomatic with patellar tendon abnormality on diagnostic imaging were excluded from statistical analyses due to their small sample size. Athletes with PT displayed a similar knee flexion angle at initial foot-ground contact (IC) and hip extension strategy during a stop-jump horizontal landing. Despite a similar kinematic technique, athletes with PT utilized a strategy of a longer stance duration phase from IC to peak force. This strategy did not lead to those athletes with PT decreasing their peak vertical GRF nor patellar tendon force during landing but enabled these athletes to land with a lower rate of loading (control 59.2 ± 39.3 vs. PT 29.4 ± 33.7 BW.s-1). Athletes with PT still reported significantly reduced training volume (control 4.9 ± 1.8 vs PT 1.8 ± 1.1 sessions/wk; total training time/wk control 2.4 ± 1.0 vs PT 1.4 ± 1.1 h/wk).     

Visceral leishmaniasis and HIV in Tigray,Ethiopia

OBJECTIVES: To identify characteristics that increased the risk of mortality in Ethiopian visceral leishmaniasis patients in a treatment programme managed by Médecins sans Frontières, in Tigray, Northern Ethiopia. METHODS: Retrospective review of a cohort of 791 patients treated for visceral leishmaniasis. RESULTS: The cohort displayed all the classical signs and symptoms of the disease. The case fatality rate was 18.5% (146) (95% CI: 15.8-21.3%). Logistic regression showed that individuals who experienced at least one episode of vomiting or haemorrhage were more likely to die than those who did not. A subcohort of individuals who tested human immunodeficiency virus (HIV)-positive were more than four times more likely to die than those who tested HIV-negative (OR 4.5, 95% CI: 1.8-11.4). CONCLUSION: This study identifies characteristics associated with death in this population and highlights the devastating effect of co-infection with visceral leishmaniasis and HIV in the African context.     

Bronchial tear in a child after a liquid petroleum gas tank explosion

Humans have been exposed to blast effects since the invention of gunpowder and explosives. Bronchial injury because of an explosion is a rare but lethal injury that requires prompt recognition and treatment. In this article, we present a case of a bronchial tear after an explosion.     

Reduction of CO2-pneumoperitoneum-induced metabolic hypoxaemia by the addition of small amounts of O2 to the CO2 in a rabbit ventilated model. A preliminary study

Dear Sir, We read with great interest the recent paper (Mynbaev et al.,2002) on the reduction of CO₂-pneumoperitoneum-induced metabolichypoxaemia by the addition of small amounts of O₂ to the CO₂in a rabbit ventilated model. In the