Designing Configurable Soft Microgelsomes as a Smart Biomimetic Protocell.

Self-assembly is an intriguing aspect of primitive cells. The construction of a semipermeable compartment with a robust framework of soft material capable of housing an array of functional components for chemical changes is essential for the fabrication of synthetic protocells. Microgels, loosely cross-linked polymer networks, are suitable building blocks for protocell capsule generation due to their porous structure, tunable properties, and assembly at the emulsion interface. Here, we present an interfacial assembly of microgel-based microcompartments (microgelsomes, MGC) that are defined by a semipermeable, temperature-responsive elastic membrane formed by densely packed microgels in a monolayer. The water-dispersible microgelsomes can thermally shuttle between 10 and 95 °C while retaining their structural integrity. Importantly, the microgelsomes exhibited distinct properties of protocells, such as cargo encapsulation, semipermeable membrane, DNA amplification, and membrane-gated compartmentalized enzymatic cascade reaction. This versatile approach for the construction of biomimetic microcompartments augments the protocell library and paves the way for programmable synthetic cells.

Bienzymatic Sequential Reaction on Microgel Particles and Their Cofactor Dependent Applications.

We report, the preparation and characterization of bioconjugates, wherein enzymes pyruvate kinase (Pk) and l-lactic dehydrogenase (Ldh) were covalently bound to poly(N-isopropylacrylamide)-poly(ethylenimine) (PNIPAm-PEI) microgel support using glutaraldehyde (GA) as the cross-linker. The effects of different arrangements of enzymes on the microgels were investigated for the enzymatic behavior and to obtain maximum Pk-Ldh sequential reaction. The dual enzyme bioconjugates prepared by simultaneous addition of both the enzymes immobilized on the same microgel particles (PL), and PiLi, that is, dual enzyme bioconjugate obtained by combining single-enzyme bioconjugates (immobilized pyruvate kinase (Pi) and immobilized lactate dehydrogenase (Li)), were used to study the effect of the assembly of dual enzymes systems on the microgels. The kinetic parameters (Km, kcat), reaction parameters (temperature, pH), stability (thermal and storage), and cofactor dependent applications were studied for the dual enzymes conjugates. The kinetic results indicated an improved turn over number (kcat) for PL, while the kcat and catalytic efficiency was significantly decreased in case of PiLi. For cofactor dependent application, in which the ability of ADP monitoring and ATP synthesis by the conjugates were studied, the activity of PL was found to be nearly 2-fold better than that of PiLi. These results indicated that the influence of spacing between the enzymes is an important factor in optimization of multienzyme immobilization on the support.

Maternal nutrient restriction during pregnancy impairs an endothelium-derived hyperpolarizing factor-like pathway in sheep fetal coronary arteries.

The mechanisms underlying developmental programming are poorly understood but may be associated with adaptations by the fetus in response to changes in the maternal environment during pregnancy. We hypothesized that maternal nutrient restriction during pregnancy alters vasodilator responses in fetal coronary arteries. Pregnant ewes were fed a control [100% U.S. National Research Council (NRC)] or nutrient-restricted (60% NRC) diet from days 50 to 130 of gestation (term = 145 days); fetal tissues were collected at day 130. In coronary arteries isolated from control fetal lambs, relaxation to bradykinin was unaffected by nitro-l-arginine (NLA). Iberiotoxin or contraction with KCl abolished the NLA-resistant response to bradykinin. In fetal coronary arteries from nutrient-restricted ewes, relaxation to bradykinin was fully suppressed by NLA. Large-conductance, calcium-activated potassium channel (BKCa) currents did not differ in coronary smooth muscle cells from control and nutrient-restricted animals. The BKCa openers, BMS 191011 and NS1619, and 14,15-epoxyeicosatrienoic acid [a putative endothelium-derived hyperpolarizing factor (EDHF)] each caused fetal coronary artery relaxation and BKCa current activation that was unaffected by maternal nutrient restriction. Expression of BKCa-channel subunits did not differ in fetal coronary arteries from control or undernourished ewes. The results indicate that maternal undernutrition during pregnancy results in loss of the EDHF-like pathway in fetal coronary arteries in response to bradykinin, an effect that cannot be explained by a decreased number or activity of BKCa channels or by decreased sensitivity to mediators that activate BKCa channels in vascular smooth muscle cells. Under these conditions, bradykinin-induced relaxation is completely dependent on nitric oxide, which may represent an adaptive response to compensate for the absence of the EDHF-like pathway.

Smart core-shell microgel support for acetyl coenzyme A synthetase: a step toward efficient synthesis of polyketide-based drugs.

The flexibility in tuning the structure and charge properties of PNIPAm microgels during their synthesis makes them a suitable choice for various biological applications. Two-step free radical polymerization, a common method employed for synthesis of core-shell microgel has been well adopted to obtain cationic poly(N-isopropylacrylamide-aminoethyl methacrylate) (PNIPAm-AEMA) shell and PNIPAm core. Scanning electron microscopy (SEM), dynamic light scattering (DLS), zeta potential, and ninhydrin assay suggests nearly monodispersed particles of cationic nature. Amino groups on the microgel provides suitable attachment point for covalent immobilization of acetyl coenzyme A synthetase (Acs) via 1-ethyl-3-(3-N,N- dimethylaminopropyl) carbodiimide (EDC) chemistry. On immobilization, 61.55% of initial activity of Acs has been retained, while Michaelis-Menten kinetics of the immobilized Acs indicates identical K(m) (Michaelis constant) but decrease in the V(max) (maximum substrate conversion rate) compared to free enzyme. Immobilized Acs shows an improvement in activity at wide temperature and pH range and also demonstrates good thermal, storage, and operational stability. The Acs-microgel bioconjugate has been successfully reused for four consecutive operation cycles with more than 50% initial activity.

Targeting colorectal cancer using dietary flavonols.

Colorectal cancer is among the well-known forms of cancer and a prominent cause of cancer demises worldwide. In vitro experiments reinforced by animal studies, as well as epidemiological studies of human colorectal cancer propose that the growth of this disease can be moderated by eating aspects. Dietary intake including green vegetables and fruits may result in the reduction of colon cancer chances. The finding suggests that the combinations of dietary nutrients may deliver additive or synergistic effects and might be a powerful method to avoid or eradicate colon cancer beginning and/or development. Flavonols are one of the most widespread dietary nutrients of the polyphenols-flavonoids and major constituent of Allium and Brassicaceae vegetables. Flavonols present in vegetables of Allium and Brassicaceae family are kaempferol, myricetin, quercetin, and isorhamnetin. These flavonols are claimed to have antiproliferative activity in vivo and in vitro against colorectal cancer. The objective of this review is to summarize the role of flavonols obtained from dietary sources in the prevention and treatment of colorectal cancer.

Intraocular foreign body presenting as recalcitrant hypopyon anterior uveitis.

A 38-year-old male with a history of trauma presented to us with pain and decreased vision in the left eye (LE). Previously, the patient had been diagnosed with hypopyon uveitis and was prescribed topical steroids. We performed slit-lamp examination of the LE and found exudates in the anterior chamber (AC) extending from 6 to 9 o'clock, along with circumciliary congestion and pigmented keratic precipitates. We made a diagnosis of infectious anterior hypopyon uveitis; however, all of its causes were ruled out upon the necessary investigations, which returned normal results. The patient was started on topical and oral antibiotics, and subsequently, there was complete hypopyon resolution. Five months later, he presented with similar complaints. An examination revealed a black elevated lesion in the AC at 8 o'clock suspicious of an intraocular foreign body (IOFB), along with 1 mm hypopyon. An X-ray orbit examination confirmed the IOFB presence, and the IOFB was surgically removed from the AC. Three weeks after this surgery, the patient presented with a recurrence of similar symptoms. The nasal pterygium adjacent to the location of the previously removed IOFB was inflamed with an underlying black elevated limbal nodule, which was determined to likely be a remnant of the IOFB in the subconjunctival space. Here, we report a case of penetrating IOFB that was initially detected in the AC. Its remnant extraocular component persisted in the subconjunctival space, and its incomplete removal led to recurrent inflammation.

Advances in paper based isothermal nucleic acid amplification tests for water-related infectious diseases.

Water-associated or water-related infectious disease outbreaks are caused by pathogens such as bacteria, viruses, and protozoa, which can be transmitted through contaminated water sources, poor sanitation practices, or insect vectors. Low- and middle-income countries bear the major burden of these infections due to inadequate hygiene and subpar laboratory facilities, making it challenging to monitor and detect infections in a timely manner. However, even developed countries are not immune to these diseases, as inadequate wastewater management and contaminated drinking water supplies can also contribute to disease outbreaks. Nucleic acid amplification tests have proven to be effective for early disease intervention and surveillance of both new and existing diseases. In recent years, paper-based diagnostic devices have made significant progress and become an essential tool in detecting and managing water-associated infectious diseases. In this review, we have highlighted the importance of paper and its variants as a diagnostic tool and discussed the properties, designs, modifications, and various paper-based device formats developed and used for detecting water-associated pathogens.

Bactericidal and biocatalytic temperature responsive microgel based self-cleaning membranes for water purification.

The present study focuses on creating an antimicrobial and biocatalytic smart gating membrane by synthesizing unique core-shell microgels. The core-shell microgels are synthesized by grafting short chains of poly(ethylenimine) (PEI) onto a poly((N-isopropyl acrylamide)-co-glycidyl methacrylate)) (P(NIPAm-co-GMA)) core. Subsequently, the produced microgels are utilized as a substrate for synthesizing and stabilizing silver nanoparticles (Ag NPs) through an in-situ approach. These Ag NPs immobilized microgels are then suction filtered over a polyethylene terephthalate (PET) track-etched support to create cross-linked composite microgel membranes (CMMs). After structural and permeation characterization of the prepared CMMs, the laccase enzyme is then covalently grafted to the surface of the membrane and tested for its effectiveness in degrading Reactive red-120 dye. The laccase immobilized biocatalytic CMMs show effective degradation of the Reactive red-120 by 71%, 48%, and 34% at pH 3, 4, and 5, respectively. Furthermore, the immobilized laccase enzyme showed better activity and stability in terms of thermal, pH, and storage compared to the free laccase, leading to increased reusability. The unique combination of Ag NPs and laccase on a thermoresponsive microgel support resulted in a responsive self-cleaning membrane with excellent antimicrobial and dye degradation capabilities for environmentally friendly separation technology.

Exploring the Effects of Elevated Serum Uric Acid Levels on Hypertension: A Scoping Review of Hyperuricemia.

Hypertension (HTN) is a global health concern due to its increasing prevalence and association with life-threatening complications. An intriguing area of investigation in HTN research is the relationship between HTN and hyperuricemia. In light of this, we conducted a review to summarize the relevant studies exploring the link between elevated serum uric acid (sUA) concentration and new-onset HTN. Through a comprehensive search of PubMed Central, MEDLINE, and PubMed databases, we identified 20 studies that met our inclusion criteria. The research encompassed various study designs, including cohort studies, cross-sectional studies, reviews, and clinical trials. Pathologically, the elevated sUA levels activate the renin-angiotensin system and also cause the formation of urate crystals, triggering inflammation in the kidneys. Additionally, direct effects on the endothelium contribute to inflammation, oxidative stress, nitric oxide depletion, and smooth muscle cell proliferation, ultimately leading to atherosclerosis. These diverse mechanisms collectively play a role in the pathogenesis of HTN. Interestingly, lowering sUA has been shown to reverse early-stage HTN dependent on uric acid. However, this effect is not observed in the uric acid-independent second stage of HTN. Various studies have demonstrated an independent and dose-dependent association between sUA levels and the prevalence of HTN across different populations and genders. The review highlights the potential role of uric acid-lowering drugs, like allopurinol, in the prevention and early-stage management of HTN. However, there is scarce research on the efficacy of other uric acid-lowering agents and combination therapies. We believe our review provides compelling evidence of the association between elevated sUA concentration and new-onset HTN. Identifying and managing hyperuricemia can provide a preventive approach to reducing the burden of HTN and its associated complications.

Unlocking the Potential of Left Cardiac Sympathetic Denervation: A Scoping Review of a Promising Approach for Long QT Syndrome.

Left cardiac sympathetic denervation (LCSD) has emerged as an alternative therapy for individuals diagnosed with long QT syndrome (LQTS), a genetic disorder characterized by abnormal electrical activity in the heart and sudden cardiac death (SCD). This review examines the history and rationale behind LCSD in LQTS treatment, as well as the procedure, its efficacy, and indications along with the adverse effects that may be associated with it. LQTS presents with prolonged QT intervals on an electrocardiogram and can manifest as seizures, fainting, and SCD. Beta-blockers are the primary treatment for LQTS but some patients do not respond well to these medications or experience side effects. Additionally, implantable cardioverter-defibrillators (ICDs) are not always effective in preventing arrhythmias and can lead to complications. LCSD might offer an alternative approach by disrupting sympathetic activity in the heart. In humans, LCSD reduces the release of norepinephrine, normalizes the QT interval, and decreases the likelihood of life-threatening heart rhythms. The procedure does not impair heart rate or cardiac function due to the compensatory effects of the right cardiac sympathetic nerves. The surgical procedure for LCSD involves the removal of the lower half of the stellate ganglion and thoracic ganglia. Complete denervation is essential for optimal outcomes, while incomplete procedures are considered unacceptable. Traditional and minimally invasive approaches, such as video-assisted thoracic surgery (VATS), are available, with VATS offering shorter hospital stays and fewer complications. In conclusion, LCSD provides a viable treatment option for individuals with LQTS who do not respond well to beta-blockers or require additional protection beyond medication or ICDs. Further research and clinical experience are needed to enhance its acceptance and implementation in routine practice.

Understanding the Link Between Adult Asthma and Coronary Artery Disease: A Narrative Review.

Asthma is a common pathology worldwide that occurs due to chronic inflammation of the respiratory airways. Persistent pulmonary inflammation leads to low-grade systemic inflammation, influencing blood vessels and triggering coronary artery disease (CAD) events. This review's objectives include discussing the susceptible population for CAD, the mechanism underlying CAD creation in asthma patients, the characteristics of asthma, and the influence of anti-asthmatic medications on CAD development. Adult-onset asthma is strongly linked to CAD and stroke. Future research may shed light on these disparities. Atherosclerosis and asthma are linked through both intrinsic and extrinsic pathways, with inflammation being the intrinsic pathway and hypoxia and tachyarrhythmia being the extrinsic pathways. The most probable mechanisms for increased coronary vasospastic angina (CVsA) incidence in asthmatic patients are vascular smooth muscle cell hypercontraction and endothelial dysfunction. Studies have shown a dose-response relationship between asthma control and myocardial infarction (MI) risk, with uncontrolled asthma at the highest risk. Impairment of ventilatory function is a distinct risk factor for lethal MI and cardiovascular death (CVD). The use of beta-2-agonists and chronic oral glucocorticoid therapy in severe asthmatics has been linked to increasing the risk for CAD. However, some studies have shown that the risk of MI among patients with active asthma is not related to the use of asthma medications. Further research is needed to determine the involvement of adult asthma features and their treatments in the development of CAD.

Validation of HPTLC and HPLC methods for the quantitative determination of allyl disulfide in some polyherbal oils.

Allium sativum L (garlic) is an essential component of many polyherbal oils used in traditional systems of medicine. Allyl disulfide has been a major component found in vegetable oil macerate of garlic, and can be used as reliable marker for determination of garlic in oil macerates of garlic. The HPLC separation of allyl disulfide was achieved on a Phenomenex Luna C18 (25 cm x 4.6 mm id x 5 pm particle size) column using acetonitrile-water-tetrahydrofuran (70 + 27 + 3, v/v/v) mobile phase at a flow rate of 1.0 mL/min. Quantitation was achieved with UV detection at 298 nm over the concentration range 8-48 microg/mL. HPTLC separation of allyl disulfide was achieved on an aluminum-backed layer of silica gel 60 F254 using n-hexane mobile phase. Quantitation was achieved by densitometric analysis at 298 nm over the 200-1200 ng/band concentration range. The methods were validated according to International Conference on Harmonization guidelines.

Biocatalytic self-assembled synthetic vesicles and coacervates: From single compartment to artificial cells.

Compartmentalization is an intrinsic feature of living cells that allows spatiotemporal control over the biochemical pathways expressed in them. Over the years, a library of compartmentalized systems has been generated, which includes nano to micrometer sized biomimetic vesicles derived from lipids, amphiphilic block copolymers, peptides, and nanoparticles. Biocatalytic vesicles have been developed using a simple bag containing enzyme design of liposomes to multienzymes immobilized multi-vesicular compartments for artificial cell generation. Additionally, enzymes were also entrapped in membrane-less coacervate droplets to mimic the cytoplasmic macromolecular crowding mechanisms. Here, we have discussed different types of single and multicompartment systems, emphasizing their recent developments as biocatalytic self-assembled structures using recent examples. Importantly, we have summarized the strategies in the development of the self-assembled structure to improvise their adaptivity and flexibility for enzyme immobilization. Finally, we have presented the use of biocatalytic assemblies in mimicking different aspects of living cells, which further carves the path for the engineering of a minimal cell.

Nature Inspired Multienzyme Immobilization: Strategies and Concepts.

In a biological system, the spatiotemporal arrangement of enzymes in a dense cellular milieu, subcellular compartments, membrane-associated enzyme complexes on cell surfaces, scaffold-organized proteins, protein clusters, and modular enzymes have presented many paradigms for possible multienzyme immobilization designs that were adapted artificially. In metabolic channeling, the catalytic sites of participating enzymes are close enough to channelize the transient compound, creating a high local concentration of the metabolite and minimizing the interference of a competing pathway for the same precursor. Over the years, these phenomena had motivated researchers to make their immobilization approach naturally realistic by generating multienzyme fusion, cluster formation via affinity domain-ligand binding, cross-linking, conjugation on/in the biomolecular scaffold of the protein and nucleic acids, and self-assembly of amphiphilic molecules. This review begins with the discussion of substrate channeling strategies and recent empirical efforts to build it synthetically. After that, an elaborate discussion covering prevalent concepts related to the enhancement of immobilized enzymes' catalytic performance is presented. Further, the central part of the review summarizes the progress in nature motivated multienzyme assembly over the past decade. In this section, special attention has been rendered by classifying the nature-inspired strategies into three main categories: (i) multienzyme/domain complex mimic (scaffold-free), (ii) immobilization on the biomolecular scaffold, and (iii) compartmentalization. In particular, a detailed overview is correlated to the natural counterpart with advances made in the field. We have then discussed the beneficial account of coassembly of multienzymes and provided a synopsis of the essential parameters in the rational coimmobilization design.

Pediatric uveitis: a retrospective analysis at a tertiary eye care hospital in South India.

PURPOSE: To analyze the demographics, etiology, complications, treatment modalities, and visual outcomes in pediatric uveitis patients at a tertiary eye care hospital. METHODS: A retrospective review of medical records of pediatric uveitis patients who presented with us from January 2014 to January 2020 was evaluated. RESULTS: Out of the 178 pediatric uveitis patients, 65 children were included in the study. The most common age group was 6-10 years (46%). Of the included patients, 36 (55.4%) were male and 29 (44.6%) were female. Presentation was bilateral in 39 (60%) and unilateral in 26 (40%). Anterior uveitis was seen in 19 (29.23%), intermediate in 18 (27.69%), posterior in 16 (24.62%), and panuveitis in 12 (18.46%) patients. There were 2 cases of masquerades. Non-infectious uveitis was the most commonly seen, in 48 (73.84%) of total cases, among which 21 (43.75%) were idiopathic and 7 (14.58%) were associated with juvenile idiopathic (JIA) arthritis. Infectious uveitis was present in 17 (26.15%); the most common etiology was toxoplasmosis. Baseline visual acuity was low in 22 (33.84%) children. After initiating treatment, 37 (56.92%) showed improvement in vision and 10 (15.38%) had worsening of vision. Intraocular pressure (IOP) rise was seen in 5 (7.69%) children; 51 (78.46%) children required medical management and 16 (24.61%) children required surgical intervention; 46 (70.76%) children had uveitis related complications out of which most of them 30 (65.21%) were present at baseline. CONCLUSIONS: Anterior and intermediate uveitis were the most common types observed in our study. Toxoplasmosis was the most common type of infectious uveitis and JIA the most common cause in non-infectious type apart from idiopathic uveitis. Posterior uveitis had low visual acuity at baseline and follow-up. Children presented to us with poor visual acuity and complications at baseline, hence an early referral to a tertiary eye hospital and management accordingly can improve the quality of vision and visual rehabilitation.

Selective determination of aconitine in polyherbal oils containing Aconitum chasmanthum using high-performance thin-layer chromatography.

Many polyherbal oil formulations in traditional systems of medicine contain aconite root. This paper reports the development and validation of a simple, rapid, and sensitive HPTLC method for identification and quantification of aconitine from polyherbal oil formulations. Chromatography of methanolic extract of these formulations was performed on silica gel 60F254 aluminum-backed HPTLC plates with a 0.2 mm layer thickness. The plates were developed up to 85 mm with the binary mobile phase ethyl acetate-ethanol (7.5 + 2.5, v/v) at 22 +/- 2 degrees C with 20 min of chamber saturation. The system produced a compact band of the marker aconitine at an R(f) value of 0.33 that was quantified at its maximum absorbance of 238 nm. The LOD and LOQ values were found to be 20 and 70 ng/band, respectively. The linearity with respect to peak area was in the range of 300 to 1800 ng/band with an r of 0.9991. Polyherbal oil formulations were analyzed with reasonable accuracy, and no matrix interference was observed. The developed HPTLC method is accurate, precise, and cost-effective, and can be used for marker-based QA of polyherbal oil formulations containing Aconitum chasmanthum as one of the active ingredients.

Estimation of catechin in Ayurvedic oil formulations containing Acacia catechu.

A sensitive, simple, rapid, and efficient HPTLC method was developed and validated for the analysis of catechin in marketed Ayurvedic oil formulations containing Acacia catechu. Chromatography of methanolic-0.1% formic acid (7:3, v/v) extracts of these formulations was performed on silica gel 60 F254 aluminum-backed TLC plates of 0.2 mm layer thickness. The plate was developed up to 85 mm with the ternary-mobile phase chloroform-acetone-0.1% formic acid (7.7 + 1.5 + 0.8%, v/v/v) at 22 +/- 2 degrees C with 20 min of chamber saturation. The system produced compact spots of catechin at an Rf value of 0.36. The marker, catechin, was quantified at its maximum absorbance of 296 nm. The limit of detection and quantitation values were 6 and 20 ng/spot, respectively. The linear regression analysis data for the calibration plot showed a good linear relationship with a correlation coefficient of 0.9993 in the concentration range of 200-1200 ng/spot for catechin with respect to peak area. Repeatability of the method was 0.88% RSD. Recovery values from 97 to 102% indicate excellent accuracy of the method. The developed HPTLC method is accurate, precise, and cost-effective, and it can be successfully applied for the determination of catechin in marketed Ayurvedic oil formulations containing Acacia catechu.

Determination of psoralen and plumbagin from its polyherbal oil formulations by an HPTLC densitometric method.

Many polyherbal oil formulations in Indian and Chinese traditional systems of medicine used for control of skin diseases contain seeds of Psoralea corylifolia and roots of Plumbago zeylanica L. Psoralen and plumbagin are the reliable markers for Ps. corylifolia and Pl. zeylanica, respectively. However, no attempt is made to standardize the polyherbal oil formulations containing Ps. corylifolia and Pl. zeylanica in terms of their active ingredients or marker compounds. In this paper, a simple, rapid, and sensitive HPTLC method is described for the first time to identify and quantify psoralen and plumbagin from such polyherbal oil formulations. The methanolic extract of oil formulations was used for analysis of markers. Psoralen gives a sharp UV absorbance peak at 302 nm and plumbagin at 275 nm. Good resolution of psoralen (Rf = 0.37) and plumbagin (Rf = 0.77) was attained using toluene-ethyl acetate (7.5 + 2.5, v/v) mobile phase. The method was validated in terms of calibration curve, limits of detection and quantification, precision, accuracy, and robustness following a standard protocol. Polyherbal oil formulations were analyzed with reasonable accuracy, and no matrix interference was observed. The method developed can be used for marker-based quality assurance of oil formulations containing Ps. corylifolia and Pl. zeylanica as one active ingredient.

Characterization of aeromonads isolated from the river Narmada, India.

Thirty isolates of Aeromonas species were isolated from river Narmada at Jabalpur during the period of January 2002-December 2002. Diversity of these fresh water isolates were determined by randomly amplified polymorphic DNA (RAPD) analysis. These environmental isolates were found to be positive for virulence factors, i.e. protease, amylase, lipase and DNase. Isolates were also positive for beta-hemolytic activity. All Aeromonas species were tested for antibiotic resistance patterns and were found to be resistant to ampicillin and sensitive to gentamycin.