Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Nasopharyngeal density of respiratory viruses in childhood pneumonia in a highly vaccinated setting: findings from a case-control study

BACKGROUND: Detection of pneumonia-causing respiratory viruses in the nasopharynx of asymptomatic children has made their actual contribution to pneumonia unclear. We compared nasopharyngeal viral density between children with and without pneumonia to understand if viral density could be used to diagnose pneumonia. METHODS: Nasopharyngeal swabs (NPS) were collected from hospitalised pneumonia cases at Princess Margaret Hospital (PMH) and contemporaneous age-matched controls at PMH outpatient clinics and a local immunisation clinic in Perth, Australia. The density (copies/mL) of respiratory syncytial virus (RSV), influenza A virus (InfA), human metapneumovirus (HMPV) and rhinovirus in NPS was determined using quantitative PCR. Linear regression analysis was done to assess the trend between viral density and age in months. The association between viral density and disease status was examined using logistic regression. Area under receiver operating characteristic (AUROC) curves were assessed to determine optimal discriminatory viral density cut-offs. RESULTS: Through May 2015 to October 2017, 230 pneumonia cases and 230 controls were enrolled. Median nasopharyngeal density for any respiratory virus was not substantially higher in cases than controls (p>0.05 for each). A decreasing density trend with increasing age was observed-the trend was statistically significant for RSV (regression coefficient -0.04, p=0.004) but not for other viruses. After adjusting for demographics and other viral densities, for every log10 copies/mL density increase, the odds of being a case increased by six times for RSV, three times for HMPV and two times for InfA. The AUROC curves were <0.70 for each virus, suggesting poor case-control discrimination based on viral density. CONCLUSION: The nasopharyngeal density of respiratory viruses was not significantly higher in children with pneumonia than those without; however, the odds of being a case increased with increased density for some viruses. The utility of viral density, alone, in defining pneumonia was limited

Factors driving customers to seek health care from pharmacies for acute respiratory illness and treatment recommendations from drug sellers in Dhaka city, Bangladesh

Fahmida Chowdhury,1 Katharine Sturm-Ramirez,1,2 Abdullah Al Mamun,1 A Danielle Iuliano,2 Mejbah Uddin Bhuiyan,1 Mohammod Jobayer Chisti,1 Makhdum Ahmed,1 Sabbir Haider,3 Mahmudur Rahman,3 Eduardo Azziz-Baumgartner2 1Infectious Diseases Division, International Centre for Diarrhoeal Disease Research, Bangladesh, Dhaka, Bangladesh; 2Influenza Division, Centers for Disease Control and Prevention, Atlanta, GA, USA; 3Institute of Epidemiology, Disease Control and Research, Dhaka, Bangladesh Background: Pharmacies in Bangladesh serve as an important source of health service. A survey in Dhaka reported that 48% of respondents with symptoms of acute respiratory illness (ARI) identified local pharmacies as their first point of care. This study explores the factors driving urban customers to seek health care from pharmacies for ARI, their treatment adherence, and outcome.Methods: A cross-sectional study was conducted among 100 selected pharmacies within Dhaka from June to December 2012. Study participants were patients or patients&rsquo; relatives aged &gt;18&nbsp;years seeking care for ARI from pharmacies without prescription. Structured interviews were conducted with customers after they sought health service from drug sellers and again over phone 5&nbsp;days postinterview to discuss treatment adherence and outcome.Results: We interviewed 302 customers patronizing 76 pharmacies; 186 (62%) sought care for themselves and 116 (38%) sought care for a sick relative. Most customers (215; 71%) were males. The majority (90%) of customers sought care from the study pharmacy as their first point of care, while 18 (6%) had previously sought care from another pharmacy and 11 (4%) from a physician for their illness episodes. The most frequently reported reasons for seeking care from pharmacies were ease of access to pharmacies (86%), lower cost (46%), availability of medicine (33%), knowing the drug seller (20%), and convenient hours of operation (19%). The most commonly recommended drugs were acetaminophen dispensed in 76% (228) of visits, antihistamine in 69% (208), and antibiotics in 42% (126). On follow-up, most (86%) of the customers had recovered and 12% had sought further treatment.Conclusion: People with ARI preferred to seek care at pharmacies rather than clinics because these pharmacies were more accessible and provided prompt treatment and medicine with no service charge. We recommend raising awareness among drug sellers on proper dispensing practices and enforcement of laws and regulations for drug sales. Keywords: drug sellers, pharmacy, acute respiratory illness, dispensing practice, health care seeking, customer

Sexually transmitted infections and sexual behaviour among youth clients of hotel-based female sex workers in Dhaka, Bangladesh.

A cross-sectional study was conducted among youth clients of hotel-based female sex workers (YCHBFSWs) in nine randomly selected hotels in Bangladesh to examine sexual-risk behaviour, condom use and determinants of condom use in last sex, knowledge of HIV, sexually transmitted infection (STI) prevalence and STI care-seeking behaviour. A prestructured questionnaire was used to collect sociodemographic, behavioural, clinical information; urine specimens (before sex) and blood were collected for diagnosis of Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, syphilis and herpes simplex virus 2 (HSV2) infection. One thousand and thirteen participants were enroled in the study. Approximately half of them reported visiting female sex workers (FSWs) at least once a month and 25% visited FSWs at least once a week. Only 12% of participants reported regular condom use. The prevalence of N. gonorrhoeae, C. trachomatis, T. vaginalis, syphilis and HSV2 was 2.2%, 3.9%, 7.2%, 2.6% and 12.9%, respectively. Only 15.3% of the YCHBFSW sought STI care in the past year. Negotiation of condom use with FSWs was the main determinant (odds ratio = 17.95) for condom use at last sex. Male clients of FSWs, including YCHBFSW, are an important bridge population for HIV transmission in Bangladesh and HIV interventions should be designed and implemented for them

Computer aided diagnostic tool for osteoporosis estimation

Quantitative estimation of osteoporosis is the need of the hour. A majority of the older population worldwide is suffering from this disease. X-ray examinations are the most widely used method for osteoporosis estimation. It has severe limitation of examination being qualitative. The paper presents a novel approach by digitising the X-ray films using an X-ray digitiser and analysing the digitised films using LabVIEW Software. The method calculates the Osteoporosis Estimation Index on the basis of the ratio of histogram calculated for selected reference ROI and measurand ROI on the digitised patient images. The results indicate the accuracy and reliability of the technique over the present methods use

Costs of hospitalization with respiratory syncytial virus illness among children aged &lt; 5 years and the financial impact on households in Bangladesh, 2010

Background Respiratory syncytial virus (RSV) is the leading cause of acute respiratory illness in young children and results in significant economic burden. There is no vaccine to prevent RSV illness but a number of vaccines are in development. We conducted this study to estimate the costs of severe RSV illness requiring hospitalization among children 50% families borrowed money to meet treatment cost. We estimated that the median direct cost of RSV-associated hospitalization in children aged < 5 years in Bangladesh was US$10 million (IQR: US$ 7-16 million), the median indirect cost was US$ 3.0 million (IQR: 2-5 million) in 2010. Conclusion: RSV-associated hospitalization among children aged < 5 years represents a substantial economic burden in Bangladesh. Affected families frequently incurred considerable out of pocket and indirect costs for treatment that resulted in financial hardship

Synthesis and characterization of highly organized crystalline rutile nanoparticles by low-temperature dissolution-reprecipitation process

Rutile nanoparticles have been synthesized by acid hydrolysis of titanium isopropoxide by low-temperature dissolution-reprecipitation process. High-resolution transmission electron micrographs of the rutile colloidal solution show needle-shaped rutile nanoparticles with the dimensions of 10-30 nm in diameter and 100-150 nm in length. X-ray diffraction (XRD) data show the existence of only the rutile polymorph in TiO<inf>2</inf> powder with a crystallite size of 11.3 nm. The dielectric constant of rutile nanoparticles has been found to be 57 at 10 MHz AC frequency and DC conductance as 2.3 7 10-6 S/cm. Transmission electron micrographs and XRD data analysis imply that the rutile crystallites are self-organized in a regular fashion to produce multilayer three-dimensional linear clusters. The clusters have been found to be microporous (average porosity 1.4 nm) with high specific surface area (132.2 m2/g). At higher concentration, the clusters aggregate to produce interconnected network of star- or flower-like structures. This organized crystalline microporous metal-oxide semiconductor might find various practical applications.Peer reviewed: YesNRC publication: Ye