Spatially-resolved transcriptomics reveal macrophage heterogeneity and prognostic significance in diffuse large B-cell lymphoma.

Macrophages are abundant immune cells in the microenvironment of diffuse large B-cell lymphoma (DLBCL). Macrophage estimation by immunohistochemistry shows varying prognostic significance across studies in DLBCL, and does not provide a comprehensive analysis of macrophage subtypes. Here, using digital spatial profiling with whole transcriptome analysis of CD68+ cells, we characterize macrophages in distinct spatial niches of reactive lymphoid tissues (RLTs) and DLBCL. We reveal transcriptomic differences between macrophages within RLTs (light zone /dark zone, germinal center/ interfollicular), and between disease states (RLTs/ DLBCL), which we then use to generate six spatially-derived macrophage signatures (MacroSigs). We proceed to interrogate these MacroSigs in macrophage and DLBCL single-cell RNA-sequencing datasets, and in gene-expression data from multiple DLBCL cohorts. We show that specific MacroSigs are associated with cell-of-origin subtypes and overall survival in DLBCL. This study provides a spatially-resolved whole-transcriptome atlas of macrophages in reactive and malignant lymphoid tissues, showing biological and clinical significance.

Patterns of Oncogene Coexpression at Single-Cell Resolution Influence Survival in Lymphoma.

Cancers often overexpress multiple clinically relevant oncogenes, but it is not known if combinations of oncogenes in cellular subpopulations within a cancer influence clinical outcomes. Using quantitative multispectral imaging of the prognostically relevant oncogenes MYC, BCL2, and BCL6 in diffuse large B-cell lymphoma (DLBCL), we show that the percentage of cells with a unique combination MYC+BCL2+BCL6- (M+2+6-) consistently predicts survival across four independent cohorts (n = 449), an effect not observed with other combinations including M+2+6+. We show that the M+2+6- percentage can be mathematically derived from quantitative measurements of the individual oncogenes and correlates with survival in IHC (n = 316) and gene expression (n = 2,521) datasets. Comparative bulk/single-cell transcriptomic analyses of DLBCL samples and MYC/BCL2/BCL6-transformed primary B cells identify molecular features, including cyclin D2 and PI3K/AKT as candidate regulators of M+2+6- unfavorable biology. Similar analyses evaluating oncogenic combinations at single-cell resolution in other cancers may facilitate an understanding of cancer evolution and therapy resistance. SIGNIFICANCE: Using single-cell-resolved multiplexed imaging, we show that selected subpopulations of cells expressing specific combinations of oncogenes influence clinical outcomes in lymphoma. We describe a probabilistic metric for the estimation of cellular oncogenic coexpression from IHC or bulk transcriptomes, with possible implications for prognostication and therapeutic target discovery in cancer. This article is highlighted in the In This Issue feature, p. 1027.

Molecular docking studies of Indian variants of pathophysiological proteins of SARS-CoV-2 with selected drug candidates.

SARS-CoV-2 pandemic has recently made the entire world come to a standstill. The number of cases in the world, especially India, have been increasing exponentially. The need of the hour is to assimilate as much data as possible to fast track the pipeline of bringing in new therapeutic tools against this fatal virus. In this brief communication, we aim to throw light on the various variants of the proteins involved heavily in the pathophysiology of COVID-19, namely Spike protein, ACE2, GRP78, TMPRSS2 and NSP-12. We also portray the molecular docking studies of these proteins with specific drugs that are currently being associated with the same. In our brief study, we come across a few key findings. First of all the combinations of the variants of spike protein and ACE2 binding show overall 25% unfavourable ΔΔG. Second, NSP12 is the most mutation prone among all the NSPs of the SARS-CoV-2 genome and the most common mutations are P323L and A97V. Third, we discovered the variants found in the Indian subpopulation that have greater binding with the currently investigated drugs.

Protein binding studies with human serum albumin, molecular docking and in vitro cytotoxicity studies using HeLa cervical carcinoma cells of Cu(ii)/Zn(ii) complexes containing a carbohydrazone ligand.

The interaction of two binuclear mixed ligand Cu(ii) complexes [Cu(o-phen)LCu(OAc)] (1) and [Cu(o-phen)LCu(o-phen)](OAc) (2) (H3L = o-HOC6H4C(H)[double bond, length as m-dash]N-NH-C(OH)[double bond, length as m-dash]N-N[double bond, length as m-dash]C(H)-C6H4OH-o) and a new mononuclear Zn(ii) complex [Zn(HL)(o-phen)(H2O)](OAc)·H2O (3) (H2L = o-HOC6H4-C(H)[double bond, length as m-dash]N-NH-C([double bond, length as m-dash]O)-NH-N[double bond, length as m-dash]C(H)-C6H4OH-o, o-phen = 1,10-phenanthroline, and OAc = CH3COO-) with human serum albumin (HSA) was studied using fluorescence quenching, synchronous and 3D fluorescence measurements and UV-vis spectroscopy. 3D fluorescence studies showed that the HSA structure was altered at the secondary and tertiary levels upon binding with the complexes. This was further supported by the electronic absorption spectral studies of HSA in the absence and presence of the compounds. The average binding distance (r) between HSA and the complexes was obtained by Förster's resonance energy transfer theory. Complex 3 was structurally characterized by X-ray crystallography. Molecular docking studies indicated that all three complexes primarily bind to HSA in subdomain IIA with amino acid residues such as Arg218 and Lys199 which are located at the entrance of Sudlow's site I. The in vitro cytotoxicities of complexes 1-3 against HeLa cells showed promising anticancer activity (IC50 = 3.5, 3.9 and 16.9 µM for 1, 2 and 3, respectively). Live cell time lapse imaging for 1 was done to capture the dynamic behavior of the cells upon treatment with the complex. Cell cycle analysis by flow cytometry with HeLa cells indicated that 1 and 2 induced cell cycle arrest in the G2/M phase while 3 induced arrest in the G0/G1 phase leading to cell death. Compounds 1 and 2 but not 3 induced apoptosis through the mitochondrial pathway as suggested from the relative p53, caspase3 and bcl2 mRNA levels measured by real-time quantitative PCR analysis.

Birth of Lasya: Emphasizing the Need for Support in Home Births.

This article explores the support structures required for home births. Written by a first-time mother who is a physician, it details her journey toward having a home birth and how these support structures shaped her confidence in having a natural birth. She also elaborates on her preparations during pregnancy, her labor, and the insights she gained during this process.

Methodologies for measuring travelers' risk perception of infectious diseases: A systematic review.

Numerous studies in the past have stressed the importance of travelers' psychology and perception in the implementation of preventive measures. The aim of this systematic review was to identify the methodologies used in studies reporting on travelers' risk perception of infectious diseases. A systematic search for relevant literature was conducted according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. There were 39 studies identified. In 35 of 39 studies, the methodology used was that of a knowledge, attitude and practice (KAP) survey based on questionnaires. One study used a combination of questionnaires and a visual psychometric measuring instrument called the 'pictorial representation of illness and self-measurement" or PRISM. One study used a self-representation model (SRM) method. Two studies measured psychosocial factors. Valuable information was obtained from KAP surveys showing an overall lack of knowledge among travelers about the most frequent travel-associated infections and associated preventive measures. This methodological approach however, is mainly descriptive, addressing knowledge, attitudes, and practices separately and lacking an examination of the interrelationships between these three components. Another limitation of the KAP method is underestimating psychosocial variables that have proved influential in health related behaviors, including perceived benefits and costs of preventive measures, perceived social pressure, perceived personal control, unrealistic optimism and risk propensity. Future risk perception studies in travel medicine should consider psychosocial variables with inferential and multivariate statistical analyses. The use of implicit measurements of attitudes could also provide new insights in the field of travelers' risk perception of travel-associated infectious diseases.

Global incidence of serogroup B invasive meningococcal disease: a systematic review.

Use of recently licensed vaccines against Neisseria meningitidis serogroup B (NmB) will depend partly on disease burden estimates. We systematically reviewed NmB incidence and mortality worldwide between January, 2000, and March, 2015, incorporating data from 37 articles and 12 websites. Most countries had a yearly invasive NmB incidence of less than 2 per 100,000 people. Within these relatively low incidence rates (compared with common causes of invasive bacterial diseases), substantial variation was detected between countries, with a notably higher incidence in Australia, Europe, North America, and South America. China and India had reports only of sporadic cases, and except for South Africa, sub-Saharan Africa showed a near absence of disease. In countries with consistently collected data, NmB incidence has tended to decrease, even as the proportion of invasive meningococcal disease cases caused by serogroup B has increased. With few exceptions, case-fatality ratios were fairly consistent, ranging between 3% and 10%. In high-income countries, incidence rates of NmB were relatively low compared with other vaccine-preventable diseases and might be decreasing. High case-fatality ratios, substantial disease-related morbidity, and the threat of outbreaks could nevertheless make NmB an attractive target for preventive and reactive immunisation programmes. The low availability of data from low-income and middle-income countries suggests the need for improved surveillance before vaccination strategies are designed.

The inevitable Hajj cough: Surveillance data in French pilgrims, 2012-2014.

BACKGROUND: Respiratory tract infections are the most common infection affecting Hajj pilgrims, and the ''Hajj cough'' is considered by pilgrims almost de rigueur. METHODS: French pilgrims were recruited between January 2012-December 2014 and information on demographics, medical history, compliance with preventive measures and health problems during travel were collected. RESULTS: A total of 382 pilgrims were included with 39.3% aged ≥65 years and 55.1% suffering from a chronic disease, most frequently hypertension and diabetes. The prevalence of cough was 80.9% and a high proportion presented with associated sore throat (91.0%), rhinitis (78.7%) and hoarseness (63.0%). Myalgia was reported in 48.3% of cases and subjective fever in 47.3%. The incubation time of respiratory symptoms was 7.7 days (range 0-25 days) and 51.9% of pilgrims presenting with a cough during their stay were still symptomatic on return. Among pilgrims with a cough, 69.4% took antibiotics. The prevalence of cough was significantly higher among females than men, but age, chronic conditions and preventive measures had no significant effect. CONCLUSIONS: The Hajj cough is highly common, likely a result of crowded conditions at religious places. Pilgrims should be advised to carry symptomatic relief for the Hajj cough such as cough suppressant, soothing throat lozenges and paracetamol. Use of antibiotics should be discouraged.

French Hajj pilgrims' experience with pneumococcal infection and vaccination: A knowledge, attitudes and practice (KAP) evaluation.

BACKGROUND: Transmission of respiratory infections poses a major public health challenge during the Hajj and Umrah in the Kingdom of Saudi Arabia. Acquisition of Streptococcus pneumoniae during Hajj has been studied in the past and recommendations for vaccination against S. pneumoniae have been made for high risk groups. METHODS: The purpose of this study was to assess the knowledge and attitudes of French Hajj pilgrims towards pneumococcal vaccination. Adult pilgrims departing from Marseille, France to Mecca for the 2014 Hajj season were administered a face-to-face questionnaire to ascertain their knowledge and attitudes towards pneumococcal vaccination before departing for Hajj. RESULTS: A total of 300 participants took part. Their overall knowledge about the severity of pneumonia and the existence of the vaccine was very low. Out of 101 participants who had an indication for pneumococcal vaccination, irrespective of their travel status, only 7% were advised to have the vaccine by their general practitioner. CONCLUSIONS: These results reinforce the need for better dissemination of information either before or during the pre-travel counselling. The visit to the travel clinic for receiving the mandatory meningococcal vaccination for Hajj is a good opportunity to update routine immunizations, including pneumococcal vaccination.

Diarrhea at the Hajj and Umrah.

This systematic review aims to summarize the incidence and etiology of diarrheal illness among pilgrims attending the Hajj and Umrah. Gastroenteritis and diarrhea have been potential threats during previous Hajj pilgrimages. The last cases of Hajj related cholera were reported in 1989. Currently, respiratory tract infections account for the majority of health problems during the Hajj. This shift in epidemiology reflects the improvement of sanitary conditions in Saudi Arabia in general, and at religious sites in particular. Nevertheless, gastrointestinal diseases, food-poisoning outbreaks, and diarrhea continue to occur among pilgrims. Available studies about diarrhea among Hajj pilgrims indicate a mean prevalence of 2% with the highest prevalence of 23% among a group of French pilgrims in 2013. There is an obvious lack of information about the etiology of diarrheal disease at the Hajj. Further studies addressing this issue in hospitalized patients as well as prospective cohort studies would be of interest. During the Hajj, hand washing is regularly carried out by pilgrims under a ritual purification, often called ablution. We recommend implementation of effective hand hygiene practices focusing on the regular use of alcohol-based hand rubs, as they require less time than traditional hand washing, act more rapidly, and contribute to sustained improvement in compliance associated with decreased infection rates.

Foot ailments during Hajj: A short report.

A study of ailments of the feet in pilgrims of Hajj revealed that 31% of them suffered from blisters, and the prevalence was five times higher in females. The presence of comorbidity (diabetes, obesity and advanced age) warrants immediate attention to them to avoid serious complications.

A systematic literature review of missed opportunities for immunization in low- and middle-income countries.

BACKGROUND: Missed opportunities for immunization (MOIs) may contribute to low coverage in diverse settings, including developing countries. METHODS: We conducted a systematic literature review on MOIs among children and women of childbearing age from 1991 to the present in low- and middle-income countries. We searched multiple databases and the references of retrieved articles. Meta-analysis provided a pooled prevalence estimate and both univariate and multivariate meta-regression analysis was done to explore heterogeneity of results across studies. RESULTS: We found 61 data points from 45 studies involving 41,310 participants. Of the 45 studies, 41 involved children and 10 involved women. The pooled MOI prevalence was 32.2% (95% CI: 26.8-37.7) among children - with no change during the study period - and 46.9% (95% CI: 29.7-64.0%) among women of child-bearing age. The prevalence varied by region and study methodology but these two variables together accounted for only 12% of study heterogeneity. Among 352 identified reasons for MOIs, the most common categories were health care practices, false contraindications, logistic issues related to vaccines, and organizational limitations, which did not vary by time or geographic region. CONCLUSIONS: MOI prevalence was high in low- and middle-income settings but the large number of identified reasons precludes standardized solutions.