Inhibition and eradication of Pseudomonas aeruginosa biofilms by secondary metabolites of Nocardiopsis lucentensis EMB25.

Millions of people worldwide have been impacted by biofilm-associated disorders, which are impregnable owing to frequent changes in surface antigens and gene expression. Globally, about 11% of nosocomial infections, including cystic fibrosis, chronic wound infections, and post-surgical infections, are caused by Pseudomonas aeruginosa, the most prevalent Gram-negative bacterial species. Moreover, biofilms are highly resistant to the host's immune system, and exhibit increased tolerance to stress factors such as starvation, dehydration, and antimicrobials. Here, we have isolated a rare halophilic actinobacteria, Nocardiopsis lucentensis EMB25, and utilized the secondary metabolites for inhibition and eradication of P. aeruginosa biofilm. For the first time, N. lucentensis EMB25 bacteria was explored to study the anti-effect of secondary metabolites on pre-established biofilm. The secondary metabolites targeted the quorum sensing pathway and were found to bind to LasR and RhlR, as confirmed via molecular docking. Also, the reduction in virulence factors, rhamnolipids and pyocyanin further supported the study as these two are regulated by LasR and RhlR. In addition, the downregulation of various QS system genes lasA, lasB, rhlA, rhlB, and pqsA confirmed that the secondary metabolites act on two main regulators of the quorum sensing pathway, LasR, and RhlR. The findings of this study support the bioprospecting of previously unknown and extreme-condition actinobacteria as a rich source of novel bioactives against infections caused by bacterial biofilms.

Profile of comorbidity and multimorbidity among women attending antenatal clinics: An exploratory cross-sectional study from Odisha, India.

Background: Multimorbidity is being recognized as a crucial maternal health challenge in India. However, pregnancy remains an exclusion criterion in most multimorbidity estimation studies resulting in a deficient understanding of the problem in this population. The present study aims to estimate the prevalence of multimorbidity, identify its correlates, and assess healthcare utilization and expenditure outcomes among women availing of antenatal care clinics. Methods: An exploratory cross-sectional study was conducted among 127 pregnant women attending three antenatal clinics of Bhubaneswar, Odisha, from February to July 2016. Data were collected using a "multimorbidity assessment questionnaire for primary care," contextualized for antenatal settings. The prevalence and correlates were determined using descriptive analysis, and the outcomes were measured by the number of healthcare visits, medications, and healthcare expenditure. About 15% of antenatal women were multimorbid. Result: Anemia (52.6%), hypertension (31.6%), acid-peptic diseases (26.3%), and thyroid (26.3%) were the leading chronic conditions. Statistically, significant association was found between multimorbidity and medical consultation, the medicines consumed, and functional limitation. The healthcare expenditure was significantly higher among the multimorbidity groups. Conclusion: Our findings suggest the inclusion of comprehensive multimorbidity assessment in routine antenatal screening. Preconception care should establish linkages between maternal and reproductive health with chronic disease prevention, and identify ways to reduce healthcare utilization and expenditure. Longitudinal studies to assess the trajectory and impact of multimorbidity on maternal and infant health are warranted.

Artificial intelligence and synthetic biology approaches for human gut microbiome.

The gut microbiome comprises a variety of microorganisms whose genes encode proteins to carry out crucial metabolic functions that are responsible for the majority of health-related issues in human beings. The advent of the technological revolution in artificial intelligence (AI) assisted synthetic biology (SB) approaches will play a vital role in the modulating the therapeutic and nutritive potential of probiotics. This can turn human gut as a reservoir of beneficial bacterial colonies having an immense role in immunity, digestion, brain function, and other health benefits. Hence, in the present review, we have discussed the role of several gene editing tools and approaches in synthetic biology that have equipped us with novel tools like Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR-Cas) systems to precisely engineer probiotics for diagnostic, therapeutic and nutritive value. A brief discussion over the AI techniques to understand the metagenomic data from the healthy and diseased gut microbiome is also presented. Further, the role of AI in potentially impacting the pace of developments in SB and its current challenges is also discussed. The review also describes the health benefits conferred by engineered microbes through the production of biochemicals, nutraceuticals, drugs or biotherapeutics molecules etc. Finally, the review concludes with the challenges and regulatory concerns in adopting synthetic biology engineered microbes for clinical applications. Thus, the review presents a synergistic approach of AI and SB toward human gut microbiome for better health which will provide interesting clues to researchers working in the area of rapidly evolving food and nutrition science.

Profile of multimorbidity in outpatients attending public healthcare settings: A descriptive cross-sectional study from Odisha, India.

BACKGROUND: Multimorbidity, the co-occurrence of two or more long-term conditions (LTC) in individuals, is associated with greater healthcare utilization, expenditure, and premature mortality, thus positing a challenge for patients and healthcare providers. Given its sparsely available epidemiological evidence, we aimed to describe the profile of multimorbidity in a representative sample of public healthcare outpatients in India. METHODS: A facility-based cross-sectional study was conducted from 1st July to 31st December 2015 in Odisha, India. Fifteen public healthcare facilities were selected by stratified random sampling. Data was collected from 1,870 adult outpatients attending these settings using Multimorbidity Assessment Questionnaire for Primary Care (MAQ-PC) tool. RESULT: Nearly 3/4th of both women and men outpatients were either obese or overweight. >1/2 had multimorbidity (≥2 LTC) while 1/3rd had ≥3 LTC. Most prevalent condition was hypertension (63%), followed by chronic backache and arthritis. Cancer and psychiatric illness were least reported. Multimorbidity increased with age group, socioeconomic status, and education level. Females across all age groups had higher reported multimorbidity than males. Diabetes--hypertension was frequently occurring dyad. Both physical and mental component of quality of life was reduced in multimorbidity. CONCLUSION: Multimorbidity is becoming a norm in healthcare practice with high prevalence in females and older adults. Health services for non-communicable diseases need to include commonly occurring dyads along with health promotion. Higher prevalence in females reinforces the need to incorporate gender differences while studying multimorbidity. Analysis of multimorbidity epidemiology through an equity lens could illuminate the underpinning complexities and heterogeneities of this phenomenon.

Reach, accessibility and acceptance of different communication channels for health promotion: a community-based analysis in Odisha, India.

INTRODUCTION: To achieve universal health coverage, improving demand generation at community is necessary. Media plays an important role by acting as a linking pin between health service providers and the community. This study intended to assess the penetration and acceptability of various forms of media for health communication in Odisha, India. METHODS: A cross-sectional mixed method study was conducted in 2016 in four districts. Following a desk review, a situational analysis was done at state, district and sub-district level. Data was collected through direct observation of study sites using a predefined checklist on knowledge awareness and practice, focussed group discussion and in-depth interviews using semi-structured questionnaire. Qualitative data was analyzed using framework approach while for quantitative data, we used SPSS 20.0. RESULTS: Major identified media houses were television (TV), radio and newspaper. Many health programs were being broadcasted in regional TV channels of the state, whereas leading public radio channel broadcasted highest number of health programs almost daily. The major source for information on disease symptoms and prevention was television (63.6%), remove hyphen (36.6%), newspaper (21.6%), health facility/service providers (17.7%), radio (9.2%), and other media like posters, pamphlets and folk dance (5.5%). Information on disease treatment or management was received mostly from television (61.2%), poster/leaflets (39.2%), remove hyphen (35.2%) and newspaper (19.7%). Only 8% of people received any health related message in mobile in past one year. Boards and hoarding provided information to 16.5% of study population. Nearly 36% respondents got information from health-wall, which are used to promote health awareness through wall paintings, graffiti etc. For immunization related information, interpersonal communication through frontline health workers was the most preferred. CONCLUSION: Interpersonal communication is believed to be most acceptable source of information on maternal and child health, immunization and neonatal care. For people with low literacy, remove hyphen campaign, folk media and interpersonal communication were found to be effective.

Recent strategies for inhibiting multidrug-resistant and ß-lactamase producing bacteria: A review.

ß-lactam antibiotics are one of the most commonly used drugs for treating bacterial infections, but their clinical effectiveness has been severely affected with bacteria developing resistance against their action. Production of ß-lactamase enzymes by bacteria that can degrade ß-lactams is the most common mechanism of acquiring such resistance, leading to the emergence of multiple-drug resistance in them. Therefore, the development of efficient approaches to combat infections caused by ß-lactamase producing and multidrug-resistant bacteria is the need of the hour. The present review attempts to understand such recent strategies that are in line for development as potential alternatives to conventional antibiotics. We find that apart from efforts being made to develop new antibiotics, several other approaches are being explored, which can help tackle infections caused by resistant bacteria. This includes the development of plant-based drugs, antimicrobial peptides, nano-formulations, bacteriophage therapy, use of CRISPR-Cas9, RNA silencing and antibiotic conjugates with nanoparticles of antimicrobial peptides. The mechanism of action of these novel approaches and potential issues limiting their translation from laboratory to clinics is also discussed. The review is important from an interesting knowledge base which can be useful for researchers working in this domain.

Antimicrobial resistance in biofilms: Exploring marine actinobacteria as a potential source of antibiotics and biofilm inhibitors.

Antimicrobial resistance (AMR) is one of the serious global public health threats that require immediate action. With the emergence of new resistance mechanisms in infection-causing microorganisms such as bacteria, fungi, and viruses, AMR threatens the effective prevention and treatment of diseases caused by them. This has resulted in prolonged illness, disability, and death. It has been predicted that AMR will lead to over ten million deaths by 2050. The rapid spread of multidrug-resistant bacteria is also causing old antibiotics to become ineffective. Among the diverse factors contributing to AMR, intrinsic biofilm development has been highlighted as an essential contributing facet. Moreover, biofilm-derived antibiotic tolerance leads to serious recurrent chronic infections. Therefore, the discovery of novel bioactive molecules is a potential solution that can help combat AMR. To achieve this, sustained mining of novel antimicrobial leads from actinobacteria, particularly marine actinobacteria, can be a promising strategy. Given their vast diversity and different habitats, the extraordinary capacity of actinobacteria can be tapped to synthesize new antibiotics or bioactive molecules for biofilm inhibition. Advanced screening strategies and novel approaches in the field of modern biochemical and molecular biology can be used to detect such new compounds. In view of this, the present review focuses on understanding some of the recent strategies to inhibit biofilm formation and explores the potential role of marine actinobacteria as sources of novel antibiotics and biofilm inhibitor molecules.

Recent metabolomics and gene editing approaches for synthesis of microbial secondary metabolites for drug discovery and development.

Microbial secondary metabolites (SMs) have been identified as an important natural source of drugs for several metabolic and neurological diseases. Along with biomedical applications, SMs are also used in the food and biochemical industries. SMs include natural products such as pigments, alkaloids, toxins, antimicrobials obtained from cultured microorganisms, while other non-cultivable microorganisms have also acted as a rich source of SMs. But, the isolation of SMs from these sources is a very tedious task. Metabolomics provides complete identification and structural information about the entire cellular metabolome under specific conditions using highly sophisticated instrumentation. Further, gene editing techniques such as cloning and gene refactoring, including advanced CRISPR-Cas, can be used for engineering microbes that have the potential to produce natural SMs that were not produced in native microbial strain. The present review describes integrated metabolomics and gene editing approaches for the synthesis of novel microbial SMs and their potential application towards drug discovery and development.

Protein Engineering for Improved Health: Technological Perspectives.

Protein engineering has enabled development of novel proteins aimed at disease diagnosis, alleviation and improved health attributes. The present article provides an overview of recent approaches and techniques used to modify proteins at diverse levels, which find therapeutically relevant applications. There is immense interest among researchers to discover new and increasingly valuable solutions for various health related issues and protein engineering could be a possible venue to sort out such problems. In this mini review we have tried to decipher some of the novel aspects of protein engineering in terms of protein-based therapeutics and diagnostics, in-silico tools and related approaches. A special emphasis has been given for some innovative aspects of protein-nanoparticle conjugates; use of artificial intelligence (AI)- based tools and post-translational modifications. Utilization of such approaches in protein engineering might be ground breaking in future research endeavor of researchers across the world.

Non-communicable Disease Risk Reduction Teaching in India: A Curricular Landscape.

Non-communicable diseases (NCDs) have today become a growing epidemic, surpassing infectious diseases and adversely impacting national health systems, policies, and socio-economic developments. Management of NCD and its risk factors such as tobacco use, poor diet, physical inactivity, and obesity has garnered paramount importance under the Sustainable Development Goals (SDGs). However, the educational settings for healthcare professionals to prepare them for delivery of preventive interventions w.r.t. to NCD risk factors require a stronger foundation. In the present work, a landscaping exercise of the teaching of NCD risk factors in the extant Bachelor of Medicine and Bachelor of Surgery (MBBS) curricula of healthcare professionals from select medical colleges in India has been undertaken. Our assessment indicates that in the present MBBS curriculum, effort is largely directed upon teaching only specific aspects of NCDs, such as hypertension, diabetes, mental illness, blindness, and cancer with greater inclination toward clinical aspects. With regard to NCD risk factors, there is inadequate focus on the healthcare promotion aspects. The overall syllabus also does not address long-term health implications of tobacco smoking, lack of proper diet or physical activity, excessive alcohol consumption etc. It also does not reflect any practical trainings or sessions related to health promotion or behavior change activities. As a way forward, we suggest that NCD risk reduction aspects be incorporated extensively into existing health professional education through the development of a curricular framework and an intervention plan contextualized to India. Findings from this study are expected to help provide critical inputs for the design and development of a curricular plan with thrust on tobacco cessation, physical activity, healthy diet, and restricted alcohol consumption, leading to creation of a health competent workforce engaged in NCD prevention and health promotion.

Proteomic profiling of Sporotrichum thermophile under the effect of ionic liquids: manifestation of an oxidative stress response.

Sporotrichum thermophile, a known producer of industrial enzymes exhibited stability in the presence of ionic liquids (ILs).The study reports, for the first time, the stress response of S. thermophile upon exposure to ILs. In vitro assay showed increased anti-oxidative enzyme levels indicating ROS-mediated oxidative stress by ILs. The proteomic profile and identification of differential proteins confirmed the fungal adaptations by (i) increased expression of glycolytic enzymes and ATP synthases (ii) downregulation of TCA cycle and protein synthesis machinery components (iii) expression of HSP70 and catalase/peroxidase. These changes are indicative of metabolic regulation of many important pathways and how ILs can be used to manipulate protein behavior.

Parental care-seeking pathway and challenges for autistic spectrum disorders children: A mixed method study from Bhubaneswar, Odisha.

BACKGROUND: Autism spectrum disorder (ASD) is globally a major cause of childhood disability. It is estimated that in India approximately 1.7-2 million children are affected with this disorder. Early diagnosis is an important criterion to start early interventions for ASD treatment. However, for every childhood problem that receives a medical diagnosis, there is a period between a parents' first recognition of unusual symptoms and the eventual diagnosis. Several factors influence this initial symptoms recognition and final diagnosis. AIMS: The present study attempts to decipher parents' perception regarding ASD and to understand the early signs of ASD recognition among parents, treatment-seeking pathways adopted their experiences and challenges in the overall process. SETTING AND DESIGN: A facility-based cross-sectional study was undertaken at Bhubaneswar, Odisha. METHODOLOGY: Parents of children with ASD were recruited for this study. Seventy-six interviews were conducted, and the data were analyzed. RESULTS: The major symptom identified by the majority of the parents was social difficulty among their children. Maximum parents had not heard about ASD before their child was diagnosed with the disorder. Parents consulted multiple professionals or traveled long distances to confirm the diagnosis. Child psychiatrists often confirmed the diagnosis and referred patients to rehabilitation centers. CONCLUSION: Lack of awareness, unavailability of services, and stress contributed to the delay in ASD diagnosis. There is a need for designing proper awareness and genuine center for the treatment. Similarly, referral and counter-referral mechanism is also required to be established to save delay in diagnosis and initiate prompt treatment in the field of ASD.

Antimicrobial Peptides: Recent Insights on Biotechnological Interventions and Future Perspectives.

With the unprecedented rise of drug-resistant pathogens, particularly antibiotic-resistant bacteria, and no new antibiotics in the pipeline over the last three decades, the issue of antimicrobial resistance has emerged as a critical public health threat. Antimicrobial Peptides (AMP) have garnered interest as a viable solution to this grave issue and are being explored for their potential antimicrobial applications. Given their low bioavailability in nature, tailoring new AMPs or strategizing approaches for increasing the yield of AMPs, therefore, becomes pertinent. The present review focuses on biotechnological interventions directed towards enhanced AMP synthesis and revisits existing genetic engineering and synthetic biology strategies for production of AMPs. This review further underscores the importance and potential applications of advanced gene editing technologies for the synthesis of novel AMPs in future.

Current Trends in Protein Engineering: Updates and Progress.

Proteins are one of the most important and resourceful biomolecules that find applications in health, industry, medicine, research, and biotechnology. Given its tremendous relevance, protein engineering has emerged as significant biotechnological intervention in this area. Strategic utilization of protein engineering methods and approaches has enabled better enzymatic properties, better stability, increased catalytic activity and most importantly, interesting and wide range applicability of proteins. In fact, the commercialization of engineered proteins have manifested in economically beneficial and viable solutions for industry and healthcare sector. Protein engineering has also evolved to become a powerful tool contributing significantly to the developments in both synthetic biology and metabolic engineering. The present review revisits the current trends in protein engineering approaches such as rational design, directed evolution, de novo design, computational approaches etc. and encompasses the recent progresses made in this field over the last few years. The review also throws light on advanced or futuristic protein engineering aspects, which are being explored for design and development of novel proteins with improved properties or advanced applications.

Health at the time of demonetization.

Health care is one of the key essential services to be traditionally impacted by financial turbulences. The Government of India announced the demonetization of INR 500 and INR 1000 currency notes in November 2016 to curb corruption and introduce economic transparency. The present commentary analyzes the impact of this economic reform on the availability/delivery of health-care services and also its transient effect on the general population availing such services during the first 2 weeks post demonetization. While print and electronic media indicated initial setback and displeasure with reference to delivery and access of health-care services, personal interaction with caregivers or patients suggested that there was no lasting adverse effect on health-care delivery. In fact, the enthusiasm for a cleaner economy meant for the greater good of the country prevailed among the general public and allowed them to bear these hardships. Our assessment suggests that demonetization and its consequential transition were viewed favorably. Financial and economic reforms initiated in the national interest can therefore be managed well with public support.

Community Management of Acute Malnutrition (CMAM) in Odisha, India: A Multi-Stakeholder Perspective.

India remains home to nearly one-third of the world's children with severe and acute malnutrition (SAM). The present study looks at the function and implementation of a Community Management of Severe Acute Malnutrition (CMAM) programme for treatment of children with SAM in Odisha, an Indian state. A cross-sectional study design using qualitative techniques with direct observation of process and infrastructure was adopted to explore the views of stakeholders on the programme implementation. The study focuses on Kandhamal, a district in Odisha, and was conducted during June-August, 2015. Of the district and community level stakeholders involved in CMAM programme, 49 were selected as study participants using purposive sampling. In-depth interviews were conducted to obtain relevant information. Data was analyzed using data analysis software, atlas.ti version 7. The analysis demonstrated the overall acceptability, feasibility and economic viability of the programme. Additionally, the study identified several enablers (such as good response from child, village leadership involvement, multisectoral participation etc.) and barriers (such as limited awareness, increased work load, irregular staff payment etc.) linked to programme implementation. Interactions with beneficiaries and stakeholders also provided the real picture on the ground. The study emphasizes the need for stakeholders to work responsibly and in unison, and need for beneficiaries to accept, participate and contribute to the programme. In view of maximum impact, the study recommends that CMAM programmes be implemented with existing primary healthcare facilities. The study also outlines future scope for policy-level interventions and support to ensure sustainability of this healthcare delivery model.

Cell Line Techniques and Gene Editing Tools for Antibody Production: A Review.

The present day modern formulation practices for drugs are based on newer tools and techniques toward effective utilization. The methods of antibody formulations are to be revolutionized based on techniques of cell engineering and gene editing. In the present review, we have discussed innovations in cell engineering toward production of novel antibodies for therapeutic applications. Moreover, this review deciphers the use of RNAi, ribozyme engineering, CRISPR-Cas-based techniques for better strategies for antibody production. Overall, this review describes the multidisciplinary aspects of the production of therapeutic proteins that has gained more attention due to its increasing demand.

Structure and Functional Characterisation of a Distinctive ß-Lactamase from an Environmental Strain EMB20 of Bacillus cereus.

The rampant use and misuse of antibiotics in human medicine, agriculture and veterinary have become the key contributors to global antimicrobial resistance. One of the significant resistance mechanisms that inactivates antibiotics and impedes treatment of bacterial infections is the expression of ß-lactamases. Rising evidence of newer variants of ß-lactamases in the environment is therefore a serious threat to the presently available antibiotic armoury. The present work describes the purification of a variant ß-lactamase isolated from a soil strain EMB20 of Bacillus cereus. The lactamase was purified using three-phase partitioning and gel filtration chromatography to a 30-fold purification and 15% recovery yield. Contrary to the general trend, the lactamase was not a metalloenzyme, but its activity was enhanced in the presence of Mg2+ and Mn2+. The EMB20 lactamase exhibited improved stability against inhibitors and denaturing agents such as urea and GdmCl as compared to its commercial analogue. The improved stability of EMB20 lactamase was further validated by circular dichroism and fluorescence spectroscopy. This study reemphasizes the rising prevalence of environmental lactamase variants. Decoding the structure-function correlation of such lactamases in the presence of inhibitors will provide insights into the response of this enzyme towards inhibitors as well as its substrates.