Short-Term Myopic Defocus and Choroidal Thickness in Children and Adults.

Purpose: Studies report conflicting findings regarding choroidal thickness changes in response to myopic defocus in humans. This study aimed to investigate the choroidal response to myopic defocus in children and adults using automated analysis. Methods: Participants (N = 46) were distance-corrected in both eyes and viewed a movie on a screen for 10 minutes. Two optical coherence tomography (OCT) radial scans were collected for each eye, then +3 diopters was added to one eye. Participants continued to watch the movie, OCT scans were repeated every 10 minutes for 50 minutes, and then recovery was assessed at 60 and 70 minutes. Defocus was interrupted for approximately two out of each 10 minutes for OCT imaging. OCT images were analyzed using an automated algorithm and trained neural network implemented in MATLAB to determine choroidal thickness at each time point. Repeated-measures ANOVA was used to assess changes with time in three age groups (6-17, 18-30, and 31-45 years) and by refractive error group (myopic and nonmyopic). Results: Choroidal thickness was significantly associated with spherical equivalent refraction, with the myopic group having a thinner choroid than the nonmyopic group (P < 0.001). With imposed myopic defocus, there were no significant changes in choroidal thickness at any time point for any age group and for either refractive error group (P > 0.05 for all). Conclusions: Findings demonstrate that, using the described protocol, the choroidal thickness of children and adults does not significantly change in response to short-term, full-field myopic defocus, in contrast to several previously published studies.

Gut microbiome as therapeutic target for diabesity management: opportunity for nanonutraceuticals and associated challenges.

Diabesity is showing rising prevalence. Current treatment modalities include pharmacological and non-pharmacological approaches, yet associated with various drawbacks. Recently, gut microbial dysbiosis is documented as a crucial factor in the pathogenesis of diabesity. Targeting gut microbiome using modulators shows promising therapeutic strategy for diabesity management. In this line, nanonutraceuticals represent new class of gut microbial modulators. The present article explores the potential of nanonutraceuticals including nanoprobiotics, nanoprebiotics, and plant-derived nanovesicles that are fabricated on the ecofriendly food based scaffold with gut microbial modulatory potential for diabesity management. A number of compelling evidences from different studies support Bifidobacterium, Enterococcus, and Bacteroides genera and Lactobacillus plantarum and Akkermansia muciniphila species significant in diabesity management. The probable mechanisms reported for gut microbial dysbiosis-induced diabesity are mentioned. The review findings suggest gut microbiome as significant therapeutic target for diabesity management. Moreover, ecofriendly nanonutraceuticals developed using natural products including food-grade materials are efficient modulators of gut microbiome and indicate next-generation diabesity therapeutics. Clinical studies are imperative as further exploration may provide new dimensions to the future research.

Awareness Regarding Antimicrobial Resistance and Antibiotic Prescribing Behavior among Physicians: Results from a Nationwide Cross-Sectional Survey in India.

(1) Background: Understanding the physicians' knowledge, attitudes, and antimicrobial prescribing behavior is a crucial step towards designing strategies for the optimal use of these agents. (2) Methods: A cross-sectional online survey was conducted among clinicians across India between May and July 2022 using a self-administered questionnaire in English comprising 35 questions pertaining to demographic characteristics, knowledge, attitude, and practices domains. (3) Results: A total of 544 responses were received from 710 physicians contacted. Sixty percent of participants were males, with mean age of 34.7 years. Mean ± Standard Deviation scores for knowledge, attitude, and practices domains were 8 ± 1.6, 20.2 ± 3.5, and 15.3 ± 2.1, respectively. Higher scores were associated with basic [odds ratio (95% Confidence Interval), p value: 2.95 (1.21, 7.2), 0.02], medical and allied sciences [2.71 (1.09, 6.67), 0.03], and central zone [3.75 (1.39, 10.12), 0.009]. A substantial proportion of dissatisfactory responses were found regarding hospital antibiograms, antibiotics effective against anaerobes, WHO AWaRe (access, watch, and reserve) classification of antibiotics, and the role of infection prevention and control (IPC) measures in the containment of antimicrobial resistance (AMR). (4) Conclusions: There is a need to sensitize and educate clinicians on various issues related to antimicrobial use, such as antibiograms, double anaerobic cover, IPC practices, and guideline-based recommendations, to curb the AMR pandemic.

Organic Bio-Based Aerogel from Food Waste: Preparation and Hydrophobization.

In this work, organic aerogels from spent ground coffee and apple pomace were prepared and characterized for the first time. Apple aerogel was found to be much lighter than that from coffee (0.19 vs. 0.016 g/cm3, whereas the specific surface areas are comparable (229 vs. 208 m2/g). Being intrinsically hydrophilic, these aerogels were silanized, both in liquid and gas phase, to increase stability in aqueous media. The latter modification method allowed chemical grafting of the silane to the aerogel surface (evidenced by FTIR and TGA) and resulted in certain hydrophobicity, as was evidenced via contact angle measurements: both aerogels possess a contact angle of ca. 100° after the gas hydrophobization, while for the pristine aerogels it was 50°. Furthermore, it was observed that the gas-phase silanization process is more applicable to apple aerogels.

Study of chloroquine susceptibility potential of plants using pseudomonas aeruginosa as in vitro model.

Chloroquine (CQ) is mainly known for antimalarial activity but due to lower sensitivity, it has not been well explored in the microbial disease treatment. In the present investigation, we attempted to enhance the CQ sensitivity in Pseudomonas aeruginosa. Presence of efflux pump is well demonstrated in bacterial system which plays an important role in drug sensitivity and resistance in bacteria and also serves other functions. Taking the advantage of presence of efflux pump in Pseudomonas aeruginosa, we made an attempt to sensitize the Pseudomonas aeruginosa with various plant extracts and phytochemicals for the development of CQ sensitivity. Ten rationally selected plant extracts were screened for the development of chloroquine sensitivity in P. aeruginosa. The chloroquine susceptibility assay was demonstrated by combining CQ and verapamil (a known efflux pump inhibitor) as a standard in an in vitro assay system. Results were quite encouraging as methanolic extracts of Syzygium aromaticum, Zingiber officinale and Curcuma longa were able to enhance chloroquine sensitivity in P. aeruginosa by increasing the zone of inhibition in well-defined assay system. These plant extracts were finally analysed for the presence of various phytochemicals. The Syzygium aromaticum extract showed the presence of phytochemicals, such as quinones, phenol, triterpenoid, saponins, tannins, alkaloids and flavonoids. On the other hand, the methanolic extract of Zingiber officinale and Curcuma longa showed the presence of saponins, tannins, alkaloids and flavonoids in the extract. Towards the identification of active principle of selected plant extract for CQ sensitivity enhancement, thin-layer chromatography was performed and various phytocomponent bands were isolated. Flavonoid (R f 0.44) in Syzygium aromaticum, alkaloid (R f 0.43) in Zingiber officinale and phenol (R f 0.62) in Curcuma longa were found responsible for the enhancement of CQ susceptibility in P. aeruginosa. This interesting finding confirmed the concept that a prior course or combination of plant extracts or phytochemicals with chloroquine can be effective against P. aeruginosa. Present investigation successfully presented the proof of concept for the enhancement of chloroquine sensitivity in bacterial system by modulating an efflux pump. Concept can be explored for repurposing chloroquine for new applications. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03382-1.

Conversion of Whey Protein Aerogel Particles into Oleogels: Effect of Oil Type on Structural Features.

Protein aerogel particles prepared by supercritical-CO2-drying (SCD) of ground whey protein (WP) hydrogels (20% w/w, pH 5.7) were converted into oleogels by dispersion in selected edible oils (castor, cod liver, corn, flaxseed, MCT, peanut and sunflower oil). The obtained oleogels were analysed for oil content, microstructure, rheological properties, and ATR-FTIR spectra. Except for castor oil, solid-like, plastic materials with comparable composition (80% oil, 20% WP) and rheological properties (G'~3.5 × 105 Pa, G″~0.20 × 105 Pa, critical stress~800 Pa, tanδ~0.060) were obtained. Optical and confocal microscopy showed that the generated structure was associated with the capillary-driven absorption of oil into the porous aerogel particles interconnected via particle-particle interactions. In this structure, the oil was stably entrapped. Results evidenced the reduced role of edible oil characteristics with the exception of castor oil, whose high polarity probably favoured particle-oil interactions hindering particle networking. This work demonstrates that WP aerogels could be regarded as versatile oleogel templates allowing the structuring of many edible oils into solid-like materials.

Mechanism of drug resistance in bacteria: efflux pump modulation for designing of new antibiotic enhancers.

Drug resistance has now become a serious concern in the domain of microbial infection. Bacteria are becoming smarter by displaying a variety of mechanisms during drug resistance. It is not only helping bacteria to adapt nicely in adverse environment but it also makes a smart system for better availability of nutritional status for microorganisms. In this domain, pathogenic bacteria are extensively studied and their mechanism for drug resistance is well explored. The common modes in bacterial resistance include degradation of antibiotics by enzymes, antibiotic target modification or inactivation by enzymatic actions, complete replacement of antibiotic targets, quorum sensing (QS) mechanism, and efflux pump-based extrusion of antibiotics. In this review, various mechanisms of drug resistance in bacteria have been highlighted with giving the importance of efflux pumps. This can be explored as a knowledge source for the management of a variety of bacterial infections, related disease and vibrant clue for next-generation drug development.

Prevalence of tuberculosis and diabetes comorbidity in patients attending secondary healthcare hospital in south India: A retrospective study.

BACKGROUND AND OBJECTIVES: Tuberculosis, a communicable disease and diabetes, a non-communicable disease together has a bidirectional relationship toward each other withsignificant morbidity and delayed treatment outcome. Therefore, there is a need to identify the prevalence of both these diseases in a community. A retrospective study was planned to identify the prevalence of both diseases among the patients attending secondary hospitals for 3 years. METHODS: The study was conducted in the chest diseases department in a secondary care hospital after obtaining approval from the institute ethics committee and RNTCP. The retrospective data in the hospital register was used to identify various parameters. The data for basic demographic characteristics, number of new cases, previously treated cases, pulmonary/extrapulmonary cases, drug resistance cases, and DM/TB cases were entered in Microsoft excel and were analyzed. RESULTS: The prevalence of TB among the patients attending the chest diseases department was 2.9%, 2.5%, and 3% for the years 2016, 2017, and 2018, respectively. The prevalence of DM/TB ranged between 8.5-11%, which is a lesser range when compared with many other studies. INTERPRETATIONS AND CONCLUSION: There was no significant difference in the prevalence between the years. The screening of one disease in the presence of the other can reduce the prevalence and improve the prognosis.

Role of Natural Products in the Treatment of Obesity: Nanotechnological Perspectives.

Obesity is a major disorder characterized by excessive fat in the body. Various factors responsible for obesity are dietary, lifestyle, genetic, and environmental factors. The last two decades witnessed an enormous increase in obesity and its comorbidities among people worldwide, thus making it the fifth major cause of human death. As per the recent report of WHO, a total of 38 million children (age < 5 years) were overweight or obese in 2019, and according to the Organization for Economic Cooperation and Development (OECD) report, almost 1 in 4 people are obese. For the treatment of obesity, various synthetic drugs are available but on the other side, these are associated with severe adverse effects. To overcome this problem and in the view of the current situation, researchers emphasize more on the development of natural products for the management of obesity. Primary and secondary metabolites like polyphenols, alkaloids, saponins, and flavonoids derived from various plants worldwide are used to develop a formulation for the management of obesity. The phytoconstituents exert their action by suppressing the proliferation of adipocytes, inducing apoptosis of adipocytes, inhibiting lipogenesis, activating lipases, stimulating fatty acid ß-oxidation, diminishing inflammatory responses, or conquering oxidative stress. This review also highlights the importance of the nanoencapsulation technique which enhances the efficacy of phytoconstituents by improving solubility, stability, and bioavailability to fight against obesity and comorbidity.

Evaluation of Early Knee Osteoarthritis Using Biomechanical and Biochemical Markers.

Altered cellular mechano-transduction and biochemistry lead to degeneration of articular cartilage in people with knee osteoarthritis. However, the influence of low-moderate exposure to weight-bearing activity such as squatting on cartilage metabolism has not been adequately studied. The current study explored associations between knee adduction moment (KAM) during walking, biochemical markers and daily squat exposure. 3D gait analysis was used to determine external loads acting on the knee as indicators of joint compressive forces whereas biomarkers-Urine type-II-collagen-telopeptide (uCTxII), antioxidant and phospholipase A2 (PLA2) activity reflected on articular cartilage status. Following ethical approval, 66 participants with varying daily squat exposure (non-squatters [n = 21, exposure = 0 min]; activity of daily living [ADL] squatters [n = 16, exposure = 34 min]; occupational squatters [n = 13, exposure = 102 min]) and people with grade 2-3 knee osteoarthritis (n = 16, exposure = 28 min) were evaluated using 3D gait and biomarker analysis. The PLA2 activity was lowest in ADL squatters while occupational squatters demonstrated highest activity (p < 0.05). KAM and urine biomarker were similar among the groups. Moderate-strong positive association was observed between sweat PLA2 activity and age (r = 0.819, p = 0.004), daily squat exposure and biomarker uCTxII (r = 0.604, p = 0.013), antioxidant activity and Right-KAM (r = -0.917, p = 0.001), and Left-KAM (r = -0.767, p = 0.016), in people with knee OA. Healthy people demonstrated weak positive associations between KAM, uCTxII, and BMI. Associations between non-invasive biomechanical and biochemical markers indicate their potential use to identify early knee osteoarthritis. Studies with larger sample size are necessary to support prescription of body weight joint loading activities such as squatting in moderation, to delay functional decline caused by knee OA.

Epigenomic programming in early fetal brain development.

Aim: To provide a comprehensive understanding of gene regulatory networks in the developing human brain and a foundation for interpreting pathogenic deregulation. Materials & methods: We generated reference epigenomes and transcriptomes of dissected brain regions and primary neural progenitor cells (NPCs) derived from cortical and ganglionic eminence tissues of four normal human fetuses. Results: Integration of these data across developmental stages revealed a directional increase in active regulatory states, transcription factor activities and gene transcription with developmental stage. Consistent with differences in their biology, NPCs derived from cortical and ganglionic eminence regions contained common, region specific, and gestational week specific regulatory states. Conclusion: We provide a high-resolution regulatory network for NPCs from different brain regions as a comprehensive reference for future studies.

Rapid and accurate species identification for ecological studies and monitoring using CRISPR-based SHERLOCK.

One of the most fundamental aspects of ecological research and monitoring is accurate species identification, but cryptic speciation and observer error can confound phenotype-based identification. The CRISPR-Cas toolkit has facilitated remarkable advances in many scientific disciplines, but the fields of ecology and conservation biology have yet to fully embrace this powerful technology. The recently developed CRISPR-Cas13a platform SHERLOCK (Specific High-sensitivity Enzymatic Reporter unLOCKing) enables highly accurate taxonomic identification and has all the characteristics needed to transition to ecological and environmental disciplines. Here we conducted a series of "proof of principle" experiments to characterize SHERLOCK's ability to accurately, sensitively and rapidly distinguish three fish species of management interest co-occurring in the San Francisco Estuary that are easily misidentified in the field. We improved SHERLOCK's ease of field deployment by combining the previously demonstrated rapid isothermal amplification and CRISPR genetic identification with a minimally invasive and extraction-free DNA collection protocol, as well as the option of instrument-free lateral flow detection. This approach opens the door for redefining how, where and by whom genetic identifications occur in the future.

Non-invasive genetic monitoring for the threatened valley elderberry longhorn beetle.

The valley elderberry longhorn beetle (VELB), Desmocerus californicus dimorphus (Coleoptera: Cerambycidae), is a federally threatened subspecies endemic to the Central Valley of California. The VELB range partially overlaps with that of its morphologically similar sister taxon, the California elderberry longhorn beetle (CELB), Desmocerus californicus californicus (Coleoptera: Cerambycidae). Current surveying methods are limited to visual identification of larval exit holes in the VELB/CELB host plant, elderberry (Sambucus spp.), into which larvae bore and excavate feeding galleries. Unbiased genetic approaches could provide a much-needed complementary approach that has more precision than relying on visual inspection of exit holes. In this study we developed a DNA sequencing-based method for indirect detection of VELB/CELB from frass (insect fecal matter), which can be easily and non-invasively collected from exit holes. Frass samples were collected from 37 locations and the 12S and 16S mitochondrial genes were partially sequenced using nested PCR amplification. Three frass-derived sequences showed 100% sequence identity to VELB/CELB barcode references from museum specimens sequenced for this study. Database queries of frass-derived sequences also revealed high similarity to common occupants of old VELB feeding galleries, including earwigs, flies, and other beetles. Overall, this non-invasive approach is a first step towards a genetic assay that could augment existing VELB monitoring and accurately discriminate between VELB, CELB, and other insects. Furthermore, a phylogenetic analysis of 12S and 16S data from museum specimens revealed evidence for the existence of a previously unrecognized, genetically distinct CELB subpopulation in southern California.

Cardiopulmonary resuscitation-induced consciousness.

Cardiopulmonary resuscitation-induced consciousness (CPRIC) is a rare yet confusing event that can cause management issues during resuscitation. The reported incidence of CPRIC is increasing. Being aware of this phenomenon and developing a standardized approach for its management is important to prevent both delay and interruptions during resuscitation efforts. We present a patient who demonstrated purposeful movements suggesting consciousness during cardiopulmonary resuscitation that would repeatedly abate with cessation of chest compressions.

The Kuala Pilah cluster cataract study: Accessible eye care reduces cataract blindness.

OBJECTIVE: The Ministry of Health introduced the cluster hospital project in Kuala Pilah district in 2016 to allow sharing of resources between the hospitals in the same vicinity. The aim of this study is to compare the demographic profile, prevalence of cataract blindness and low vision among patients who presented for cataract surgery before and after the programme. METHODOLOGY: This is a retrospective cohort study of patients who underwent cataract surgery in Kuala Pilah Cluster Hospitals between 2010 and 2017. A total of 2539 records of patients were reviewed. Patients were assigned into two groups: Group 1 (2010-2012)- before the programme (2010-2012) and Group 2 (2015-2017) after the introduction of the programme. RESULTS: There was a significant increase in number of cataract cases in the district hospital after the cluster initiative. The mean age of patients undergoing cataract surgery was similar in both groups. The common comorbidities were hypertension (Group 1=57.3%; Group 2=70.8%) and diabetes mellitus (Group 1=40.6%; Group 2=51.1%). In 2010-2012, most of the patients were one eye blind (34.4%), whereas in 2015-2017 majority of patients presented with vision better than 6/18 (43.5%). The proportion of patients with cataract blindness reduced from 6% in 2010-2012 to 4.3% in 2015-2017 (p<0.01). CONCLUSION: There is a significant decrease in percentage of patients with cataract blindness and low vision after the introduction of Kuala Pilah Cluster Hospital Program. We believe that that cluster hospital system is effective in improving accessibility to eye care and therefore increases the cataract detection rate.

Proteinaceous Pancreatic Lipase Inhibitor from the Seed of Litchi chinensis.

A study of the pancreatic lipase inhibitory activity of a protein from the seed of Litchi chinensis was carried out. Protein was isolated by 70% ammonium sulphate precipitation followed by dialysis. Lipase inhibitory activity of the protein was evaluated using both synthetic (p-nitrophenyl palmitate) and natural (olive oil) substrates. Protein at the final concentration of 100 µg/mL was able to inhibit 68.2% pancreatic lipase on synthetic substrate and 60.0% on natural substrate. Proteinaceous nature of the inhibitor was determined using trypsinization assay. Pancreatic lipase inhibitory protein was sensitive to 0.05% trypsin treatment with the loss of 61.9% activity. IC50 of this proteinaceous pancreatic lipase inhibitor was 73.1 µg/mL using synthetic substrate. This inhibitory protein was sensitive to pH, with the highest inhibitory activity at pH=8.0 and the lowest at pH=3.0. Protein was further analyzed using 10% non-reducing sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and, interestingly, it showed the presence of a single band of (61±2) kDa when stained with Coomassie brilliant blue. The isolated protein was finally crystallized to see its homogeneity by batch crystallization method. Crystals were well formed with distinct edges. The isolated protein showed good pancreatic lipase inhibitory activity.

A High-Throughput Dose-Response Cellular Thermal Shift Assay for Rapid Screening of Drug Target Engagement in Living Cells, Exemplified Using SMYD3 and IDO1.

A persistent problem in early small-molecule drug discovery is the frequent lack of rank-order correlation between biochemical potencies derived from initial screens using purified proteins and the diminished potency and efficacy observed in subsequent disease-relevant cellular phenotypic assays. The introduction of the cellular thermal shift assay (CETSA) has bridged this gap by enabling assessment of drug target engagement directly in live cells based on ligand-induced changes in protein thermal stability. Initial success in applying CETSA across multiple drug target classes motivated our investigation into replacing the low-throughput, manually intensive Western blot readout with a quantitative, automated higher-throughput assay that would provide sufficient capacity to use CETSA as a primary hit qualification strategy. We introduce a high-throughput dose-response cellular thermal shift assay (HTDR-CETSA), a single-pot homogenous assay adapted for high-density microtiter plate format. The assay features titratable BacMam expression of full-length target proteins fused to the DiscoverX 42 amino acid ePL tag in HeLa suspension cells, facilitating enzyme fragment complementation-based chemiluminescent quantification of ligand-stabilized soluble protein. This simplified format can accommodate determination of full-dose CETSA curves for hundreds of individual compounds/analyst/day in replicates. HTDR-CETSA data generated for substrate site and alternate binding mode inhibitors of the histone-lysine N-methyltransferase SMYD3 in HeLa suspension cells demonstrate excellent correlation with rank-order potencies observed in cellular mechanistic assays and direct translation to target engagement of endogenous Smyd3 in cancer-relevant cell lines. We envision this workflow to be generically applicable to HTDR-CETSA screening spanning a wide variety of soluble intracellular protein target classes.

Thrombospondin-4 mediates TGF-ß-induced angiogenesis.

TGF-ß is a multifunctional cytokine affecting many cell types and implicated in tissue remodeling processes. Due to its many functions and cell-specific effects, the consequences of TGF-ß signaling are process-and stage-dependent, and it is not uncommon that TGF-ß exerts distinct and sometimes opposing effects on a disease progression depending on the stage and on the pathological changes associated with the stage. The mechanisms underlying cell- and process-specific effects of TGF-ß are poorly understood. We are describing a novel pathway that mediates induction of angiogenesis in response to TGF-ß1. We found that in endothelial cells (EC) thrombospondin-4 (TSP-4), a secreted extracellular matrix (ECM) protein, is upregulated in response to TGF-ß1 and mediates the effects of TGF-ß1 on angiogenesis. Upregulation of TSP-4 does not require the synthesis of new protein, is not caused by decreased secretion of TSP-4, and is mediated by activation of SMAD3. Using Thbs4-/- mice and TSP-4 shRNA, we found that TSP-4 mediated pro-angiogenic functions in cultured EC and angiogenesis in vivo in response to TGF-ß1. We observed~3-fold increases in tumor mass and levels of angiogenesis markers in animals injected with TGF-ß1, and these effects did not occur in Thbs4-/- animals. Injections of an inhibitor of TGF-ß1 signaling SB-431542 also decreased the weights of tumors and cancer angiogenesis. Our results from in vivo angiogenesis models and cultured EC document that TSP-4 mediates upregulation of angiogenesis by TGF-ß1. Upregulation of pro-angiogenic TSP-4 and selective effects of TSP-4 on EC may contribute to stimulation of tumor growth by TGF-ß despite the inhibition of cancer cell proliferation.

Brightness measurement of an electron impact gas ion source for proton beam writing applications.

We are developing a high brightness nano-aperture electron impact gas ion source, which can create ion beams from a miniature ionization chamber with relatively small virtual source sizes, typically around 100 nm. A prototype source of this kind was designed and successively micro-fabricated using integrated circuit technology. Experiments to measure source brightness were performed inside a field emission scanning electron microscope. The total output current was measured to be between 200 and 300 pA. The highest estimated reduced brightness was found to be comparable to the injecting focused electron beam reduced brightness. This translates into an ion reduced brightness that is significantly better than that of conventional radio frequency ion sources, currently used in single-ended MeV accelerators.