Evaluation of mobile digital light-emitting diode fluorescence microscopy in Hanoi, Viet Nam.

SETTING: Hanoi Lung Hospital, Hanoi, Viet Nam. OBJECTIVE: To compare the accuracy of CellScopeTB, a manually operated mobile digital fluorescence microscope, with conventional microscopy techniques. DESIGN: Patients referred for sputum smear microscopy to the Hanoi Lung Hospital from May to September 2013 were included. Ziehl-Neelsen (ZN) smear microscopy, conventional light-emitting diode (LED) fluorescence microscopy (FM), CellScopeTB-based LED FM and Xpert(®) MTB/RIF were performed on sputum samples. The sensitivity and specificity of microscopy techniques were determined in reference to Xpert results, and differences were compared using McNemar's paired test of proportions. RESULTS: Of 326 patients enrolled, 93 (28.5%) were Xpert-positive for TB. The sensitivity of ZN microscopy, conventional LED FM, and CellScopeTB-based LED FM was respectively 37.6% (95%CI 27.8-48.3), 41.9% (95%CI 31.8-52.6), and 35.5% (95%CI 25.8-46.1). The sensitivity of CellScopeTB was similar to that of conventional LED FM (difference -6.5%, 95%CI -18.2 to 5.3, P = 0.33) and ZN microscopy (difference -2.2%, 95%CI -9.2 to 4.9, P = 0.73). The specificity was >99% for all three techniques. DISCUSSION: CellScopeTB performed similarly to conventional microscopy techniques in the hands of experienced TB microscopists. However, the sensitivity of all sputum microscopy techniques was low. Options enabled by digital microscopy, such as automated imaging with real-time computerized analysis, should be explored to increase sensitivity.

Dynamic two-photon imaging of the immune response to Toxoplasma gondii infection.

Toxoplasma gondii is a highly successful parasite that can manipulate host immune responses to optimize its persistence and spread. As a result, a highly complex relationship exists between T. gondii and the immune system of the host. Advances in imaging techniques, and in particular, the application of two-photon microscopy to mouse infection models, have made it possible to directly visualize interactions between parasites and the host immune system as they occur in living tissues. Here, we will discuss how dynamic imaging techniques have provided unexpected new insight into (i) how immune responses are dynamically regulated by cells and structures in the local tissue environment, (ii) how protective responses to T. gondii are generated and (iii) how the parasite exploits the immune system for its own benefit.

The loss of Sirt1 in mouse pancreatic beta cells impairs insulin secretion by disrupting glucose sensing.

AIMS/HYPOTHESIS: Sirtuin 1 (SIRT1) has emerged as a key metabolic regulator of glucose homeostasis and insulin secretion. Enhanced SIRT1 activity has been shown to be protective against diabetes, although the mechanisms remain largely unknown. The aim of this study was to determine how SIRT1 regulates insulin secretion in the pancreatic beta cell. METHODS: Pancreatic beta cell-specific Sirt1 deletion was induced by tamoxifen injection in 9-week-old Pdx1CreER:floxSirt1 mice (Sirt1BKO). Controls were injected with vehicle. Mice were assessed metabolically via glucose challenge, insulin tolerance tests and physical variables. In parallel, Sirt1 short interfering RNA-treated MIN6 cells (SIRT1KD) and isolated Sirt1BKO islets were used to investigate the effect of SIRT1 inactivation on insulin secretion and gene expression. RESULTS: OGTTs showed impaired glucose disposal in Sirt1BKO mice due to insufficient insulin secretion. Isolated Sirt1BKO islets and SIRT1KD MIN6 cells also exhibited impaired glucose-stimulated insulin secretion. Subsequent analyses revealed impaired α-ketoisocaproic acid-induced insulin secretion and attenuated glucose-induced Ca(2+) influx, but normal insulin granule exocytosis in Sirt1BKO beta cells. Microarray studies revealed a large cluster of mitochondria-related genes, the expression of which was dysregulated in SIRT1KD MIN6 cells. Upon further analysis, we demonstrated an explicit defect in mitochondrial function: the inability to couple nutrient metabolism to mitochondrial membrane hyperpolarisation and reduced oxygen consumption rates. CONCLUSIONS/INTERPRETATION: Taken together, these findings indicate that in beta cells the deacetylase SIRT1 regulates the expression of specific mitochondria-related genes that control metabolic coupling, and that a decrease in beta cell Sirt1 expression impairs glucose sensing and insulin secretion.

[Increase of entomological indices during the pre-epidemic period of dengue in Ben Tre, South Vietnam]. / Augmentation des indices vectoriels en période pré-épidémique de dengue à Ben Tre, Sud Vietnam.

Dengue has emerged in Vietnam 50 years ago and since has become endemo-epidemic throughout the whole country. Each year, major epidemics of dengue fever (DF) and dengue hemorrhagic fever (DHF) hit South Vietnam during the rainy season, causing significant morbidity and mortality, especially among young children. The only preventive measure is vector control, but it is often implemented too late or indiscriminately. The aim of this study was to investigate, in the pre-epidemic stage, the existence of significant changes in vector indices, which will predict DF/DHF outbreaks. We conducted a descriptive transversal study, repeated once a month for four months (March to June) in the village of Locthuan (province Ben Tre) in the Mekong's delta. Adult mosquitoes were caught in 30 houses, and larvae were collected in water holding containers of 50 houses. The houses were randomly selected. Vector densities were calculated according to the indices recommended by WHO. Virological analysis was carried out on lots of female Aedes and larvae in order to determine viral infection rates. Catches of adult mosquitoes collected 496 specimens including 329 Aedes, 139 Culex and 28 Anopheles. Aedes aegypti was present in 63% of visited homes that is an average density of 1.8 mosquitoes per house. The increase in imaginal indices during the 4 months was not significant. The survey of breeding sites of Ae. aegypti identified 1292 water containers in which 71,569 larval specimens were collected. The values of house index, container index [CI] and Breteau index [BI] increased each month, the latter from 166 to 442. This increase was significant for CI and BI. Breeding sites were mostly intra-home, mainly consisting of large and small ceramic jars. Larval density of Ae. aegypti in the containers also increased significantly over the 4 months. It was correlated with the lack of cover and predators such as Mesocyclops spp., Micronecta spp. and larvivorous fishes. Cultivation of 15 pools of 10 adult females and 29 pools of larvae (ie 1088 specimens) of Ae. aegypti failed to isolate dengue virus. The high Stegomyia indices measured in this South Vietnamese village and their increase before the rainy season reflect a situation at high risk of epidemics but cannot predict the occurrence of an outbreak in the absence of virus isolation from mosquitoes. They justify conducting an integrated vector control throughout the year.

Should moderate acute rejection of a cardiac transplant graft be treated?

UNLABELLED: Histologically proven, moderate acute rejection after orthotopic heart transplantation (OHT) is commonly treated with intravenous steroids. This regimen may result in severe metabolic and infectious side-effects. The purpose of this study was to assess and compare outcomes in treated (T) versus not treated (N-T) biopsy proven 3A rejection episodes in cardiac transplant recipients. METHODS: A retrospective analysis was conducted to identify all biopsy proven 3A rejection episodes that occurred over the time period 1995-2000 in patients (patients) >or= 6 months after OHT (n=48 episodes in 35 patients). Of the 48 episodes, 19 were N-T and 29 were T. Decision to treat 3A rejection was based on time after transplant, haemodynamic and/or clinical compromise and left ventricular (LV) dysfunction measured by 2D echo. Most N-T episodes received an increase in background immunotherapy. RESULTS: Time from transplant to index 3A episode in N-T patients was 4.2 versus 2.7 yr for the T patients (p=0.06). There were no differences seen between T and N-T groups for the first and second post-3A biopsy results or LV function post-3A. Presence of coronary disease or death were not different between groups. Of the 29 patients with T episodes, no differences in outcomes (death, first and second post-3A biopsy score, coronary disease, myocardial infarction, or LV function) were seen based on use of treatment with intravenous versus oral steroid. CONCLUSION: In patients more than 6 month after OHT, there were no differences in outcomes (ongoing rejection or LV function) between N-T episodes of 3A rejection and T episodes. In T patients the use of oral steroids was equally as effective for treatment of 3A episodes as intravenous steroids.

Multilocus genotyping indicates that the ability to invade the bloodstream is widespread among Candida albicans isolates.

Multilocus genotyping was used to compare populations of Candida albicans from oral mucosa and blood. No significant differences in allele frequencies between the two samples were detected, and in a dendrogram of genotypic similarities, genotypes from both types of samples were finely interspersed. This is evidence for widespread distribution of invasive potential.

Impact of injections during diagnostic coronary arteriography on coronary patency in the setting of acute myocardial infarction from the TIMI trials. Thrombolysis In Myocardial Infarction.

The mechanical force of injection at 90 minutes opens 13.4% of occluded arteries, but overall, only 2.4% of all culprit arteries (already open and occluded combined) are opened. Thus, although some arteries are opened by the force of hand injection, the frequency of mechanical opening among all arteries is low, and hand injections appear to alter current 80% patency rates by approximately 2.5%.