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Edible oil industries generate wastewater consists of long-chain fatty acids (LCFA), such as linoleic acid, which causes inhibition during anaerobic digestion. In the current study, the performance of a laboratory-scale anaerobic fluidized bed reactor (AFBR) was investigated for the treatment of linoleic acid under anaerobic digestion. The AFBR was fabricated with a plexi glass column (60 mm diameter, 160 cm height, and volume of 2.95 L). The amount of biomass was increased within the AFBR using polyvinyl chloride (PVC) chips as a carrier medium. During the start-up, the AFBR was operated at hydraulic retention time (HRT) of 24 h, OLR = 0.50 g/L/d, flowrate 5 L/d, and an up-flow velocity of 9 × 10-4 m/min. The chemical oxygen demand (COD) values were decreased up to 76 mg/L, with a removal efficiency of 65.4%. However, the optimized conditions were achieved during the operational period when the influent flowrate was set at 15 L/d, and HRT was set at 6 h, corresponding up-flow velocity of 4.4 × 10-3 m/min and organic loading rate of 1.13 g/L/d. The values of COD were reduced up to 51.5 ± 1 mg/L. Consequently, the reactor efficiency was increased from 65.4% to 82.6% in terms of COD removal. Moreover, different linoleic acid concentrations were spiked in the AFBR (i.e., 100, 150, 200, and 250 mg/L) during the optimized condition and the linoleic acid removal was observed up to 91.2%. Therefore, the AFBR with utilized biomass media (PVC chips) seems to have the promising potential of the high strength wastewater treatment by the degradation of LCFA and the reduction of organic pollutants. [ABSTRACT FROM AUTHOR]

Rice straw (RS) residue consists of lignocellulosic biomass and is being mostly burned in the open air after harvesting in Pakistan. The atmospheric burning of RS is causing environmental degradation. While, the buffalo dung (BD) is suitable for production of methane because of containing various microbes and nutrients. In this study, the methane potential of anaerobic co-digestion of RS and BD was investigated. The RS was pretreated with hydrogen peroxide (H2O2) and calcium hydroxide (Ca(OH)2) prior to use in anaerobic digestion (AD) batch reactors. The ratio of RS to BD on the basis of the volatile solids (VS) was taken as 30:70, whereas the RS was shredded to particle size of 4 mm. The batch reactors were pretreated in serum cultural bottles with the separate H2O2 and Ca(OH)2 concentrations of 0.1, 0.2, 0.3, 0.4 0.5 and 0.6%. The co-digestion experiment was conducted in Semi-Automatic Methane Potential Test System (SAMPTS) under mesophilic conditions i.e., 37 ± 1 °C. The results show that both pretreatments remarkably degrade the RS and increases the production of methane as compared to the control. The highest methane was observed from the pretreatment concentration of 0.3% in case of H2O2 and 0.4% in the case of Ca(OH)2, whereas the methane production of 331.6 and 346.7 mL CH4 g−1 VS was observed, respectively. Moreover, in order the check the AD process dynamics, the kinetic coefficients were determined by using S-Gompertz and Cone models. On the basis of coefficient of determination, S-Gompertz model was better fitted as compared to the Cone model. [ABSTRACT FROM AUTHOR]

In more than a few regions of Pakistan, crop residues are being wasted and are leading to a solid waste problem. Anaerobic co-digestion of crop residues along with animal dung is one of the appropriate method to convert wasted crop residues into the renewable energy. In the present study, the anaerobic co-digestion of canola straw long with buffalo dung was carried out in continuously stirred tank reactor (CSTR) for optimization of organic loading rate (OLR) and hydraulic retention time (HRT). The various organic loads were added in the range of 2-4 gVS L-1day-1, whereas various HRTs were kept from 20-35 days. Results reveal that on the increase of the OLR, the volumetric methane production increases, without significantly compromising methane yield. On the contrary, by increasing the HRT, the volumetric methane production decreases, whereas the specific methane production increases. The optimum OLR and HRT were observed as 2.66 gVS L-1day-1 and 30 days respectively, where maximum specific methane production was observed as 143 NmL g-1VSadded. Moreover, the experimental data were simulated, which reveals that and the reaction kinetic follow the first order CSTR model efficiently. [ABSTRACT FROM AUTHOR]

The purpose of this study was to enhance the methane generation from the rice straw by anaerobically co-digestion with the buffalo dung. The study was carried out into three phases and it includes determination of ratio of rice straw and buffalo dung, specific mass of sodium bicarbonate (NaHCOs) as buffer and alkaline pretreatment agent and appropriate size of the rice straw particle size that yields maximum methane. The higher generation of methane from anaerobic co-digestion of rice straw and buffalo dung could be achieved by taking the ratio of rice straw and buffalo dung as 3:7 on the basis of volatile solids, the specific mass of NaHCO[sub 3] of 0.5 g/gVS and the rice straw particle size of 2 mm. Considering the above parameters, methane generation was obtained about 184 NmL/gVS at anaerobic biodegradability of about 48%. By using particle size of 4mm, methane generation was obtained about 172 NmL/gVS. However, because of about double energy consumption to produce rice straw of particle size of 2 mm than to the particle size of 4 mm, thus on the basis of energy consumption later one is suggested as appropriate particle size. [ABSTRACT FROM AUTHOR]

This article discusses the medicolegal implications of a recent judgment in relation to a patient who suffered significant morbidity as a result of patient positioning during an operative procedure. The patient developed an unexpected serious complication following surgery, in the context of a preoperative consent that did not cover every potential complication or contingency. The court held that the failure to warn of a particular risk that would have prevented the patient from undergoing a procedure but did not occur will not necessarily result in a finding of negligence in relation to another risk where the harm did occur. This finding is well aligned to current clinical practice and at the same time does not abrogate the practitioner's duty to provide a comprehensive list of possible complications during the consent process for any proceduralist. In the context of a procedure requiring anaesthesia, the importance of communication and understanding between the anaesthetist and proceduralist as to which aspects of the consent process are undertaken by whom, and to ensure the process is done comprehensively, is of great importance and is indirectly highlighted by this recent judgment. [ABSTRACT FROM AUTHOR]

Metabolic dysfunction-associated steatotic liver disease (MASLD) prevalence is increasing annually and affects over a third of US adults. MASLD can progress to metabolic dysfunction-associated steatohepatitis (MASH), characterized by severe hepatocyte injury, inflammation, and eventual advanced fibrosis or cirrhosis. MASH is predicted to become the primary cause of liver transplant by 2030. Although the etiology of MASLD/MASH is incompletely understood, dysregulated fatty acid oxidation is implicated in disease pathogenesis. Here, we develop a method for estimating hepatic β-oxidation from the metabolism of [D 15 ]octanoate to deuterated water and detection with deuterium magnetic resonance methods. Perfused livers from a mouse model of MASLD reveal dysregulated hepatic β-oxidation, findings that corroborate in vivo imaging. The high-fat-diet-induced MASLD mouse studies indicate that decreased β-oxidative efficiency in the fatty liver could serve as an indicator of MASLD progression. Furthermore, our method provides a clinically translatable imaging approach for determining hepatic β-oxidation efficiency., Competing Interests: Declaration of interests M.E.M. and R.M. are holders of patent ##T17984US002 related to metabolic imaging of HDO., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Cyclodextrins (CDs) are unique cyclic compounds that can form inclusion complexes via host-guest complexation with a wide range of molecules, thereby altering their physicochemical properties. These molecules offer the formation of inclusion complexes without the formation of covalent bonds, making them suitable for a variety of applications in pharmaceutical and biomedical fields. Due to their supramolecular host-guest properties, CDs are being utilized in the fabrication of biomaterials, metal-organic frameworks, and nano-drug carriers. Additionally, CDs in combination with biomolecules are biocompatible and can deliver nano to macromolecules at the site of drug actions. However, the availability of free hydroxyl groups and a simple crosslinking process for supramolecular fabrication show immense opportunities for researchers in the field of tissue engineering and biomedical applications. In this review article, we have covered the historical development, various types of chemical frameworks, unique chemical and physical properties, and important applications of CDs in drug delivery and biomedical sciences., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

β-lactams are a chemically diverse group of molecules with a wide range of biological activities. Having recently observed curious trends in 2 J HH coupling values in studies on this structural class, we sought to obtain a more comprehensive understanding of these diagnostic NMR parameters, specifically interrogating 1 J CH , 2 J CH , and 2 J HH , to differentiate 3- and 4-monosubstituted β-lactams. Further investigation using computational chemistry methods was employed to explore the geometric and electronic origins for the observed and calculated differences between the two substitution patterns., (© 2024 The Authors. Magnetic Resonance in Chemistry published by John Wiley & Sons Ltd.)

The expansion of the nuclear industry has led to various radioactive effluents, originating from routine operations or catastrophic incidents such as those at Three Mile Island (USA), Chernobyl (Ukraine), and Fukushima (Japan). Research conducted after these events emphasizes Cesium-137 (137Cs) and iodine 131 (131I) as major contributors to harmful airborne dispersion and fallout. These isotopes infiltrate the human body via inhalation, ingestion, or wounds, posing significant health risks. Understanding contamination mechanisms and devising effective countermeasures are crucial in mitigating nuclear incident consequences. We propose that concurrent administration of Pru-Decorp™/Pru-Decorp-MG and potassium iodide (KI) could synergistically reduce the levels of 137Cs and block uptake of 131I, respectively, in nuclear incident scenarios. Pru-Decorp™ capsules contain insoluble ferric hexacyanoferrate(II) and are equivalent to USFDA-approved Radiogardase®-Cs, offering radiation exposure mitigation for Cs and Tl contamination. Pru-Decorp-MG capsules consist of insoluble PB and magnesium hydroxide, serving as a prophylactic measure to reduce the risk of internal Cs and Tl contamination for rescue responders. Pru-Decorp™/Pru-Decorp-MG binds Cs/Tl ions in the gastrointestinal tract, hindering absorption and promoting excretion, while KI saturates the thyroid gland with stable iodine, decreasing the uptake of radioactive iodine isotopes. Our hypothesis is supported by studies demonstrating the effectiveness of combination therapies, such as calcium alginate, iron(III) ferrocyanide, and KI, in decreasing the retention of radioisotopes in vital organs. To test this hypothesis, we propose a comprehensive research plan, including in vitro studies simulating gastrointestinal conditions, animal studies to evaluate the efficacy of both drugs simultaneously, and safety clinical trials comparing Pru-Decorp™/Pru-Decorp-MG alone, KI alone, and their combination. Expected outcomes include insights into the synergistic effects of Pru-Decorp™/Pru-Decorp-MG and KI, guiding the development of optimized treatment protocols for simultaneous administration during radioactive contamination incidents. This research aims to address significant critical gaps in nuclear incident preparedness by providing evidence-based recommendations for concurrent antidote use in scenarios involving multiple isotope contamination. Ultimately, this will enhance public health and safety during nuclear emergencies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Obesity is a growing epidemic affecting millions of people worldwide and a major risk factor for a multitude of chronic diseases and premature mortality. Accumulating evidence suggests that mitochondria have a profound role in diet-induced obesity and the associated metabolic changes, but the molecular mechanisms linking mitochondria to obesity remain poorly understood. Our studies have identified a new function for mitochondrial MUL1 E3 ubiquitin ligase, a protein known to regulate mitochondrial dynamics and mitophagy, in the control of energy metabolism and lipogenesis. Genetic deletion of Mul1 in mice impedes mitophagy and presents a metabolic phenotype that is resistant to high-fat diet (HFD)-induced obesity and metabolic syndrome. Several metabolic and lipidomic pathways are perturbed in the liver and white adipose tissue (WAT) of Mul1(-/-) animals on HFD, including the one driven by Stearoyl-CoA Desaturase 1 (SCD1), a pivotal regulator of lipid metabolism and obesity. In addition, key enzymes crucial for lipogenesis and fatty acid oxidation such as ACC1, FASN, AMPK, and CPT1 are also modulated in the absence of MUL1. The concerted action of these enzymes, in the absence of MUL1, results in diminished fat storage and heightened fatty acid oxidation. Our findings underscore the significance of MUL1-mediated mitophagy in regulating lipogenesis and adiposity, particularly in the context of HFD. Consequently, our data advocate the potential of MUL1 as a therapeutic target for drug development in the treatment of obesity, insulin resistance, NAFLD, and cardiometabolic diseases., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Cilenti, Di Gregorio, Mahar, Liu, Ambivero, Periasamy, Merritt and Zervos.)

We report a new class of highly effective, benzooxaphosphole-based, water-soluble ligands in the application of Suzuki-Miyaura cross-coupling reactions for sterically hindered substrates in aqueous media. The catalytic activities of the coupling reactions were greatly enhanced by the addition of catalytic amounts of organic phase transfer reagents, such as tetraglyme and tetrabutylammonium bromide. The optimized general protocol can be conducted with a low catalyst load, thereby providing a practical solution for these reactions. The viability of this new Suzuki-Miyaura protocol was demonstrated with various substrates to generate important building blocks, including heterocycles, for the synthesis of biologically active compounds.

Introduction: The differential expression of emotional reactivity from early to late adulthood may involve maturation of prefrontal cortical responses to negative valence stimuli. In mice, age-related changes in affective behaviors have been reported, but the functional neural circuitry warrants further investigation., Methods: We assessed age variations in affective behaviors and functional connectivity in male and female C57BL6/J mice. Mice aged 10, 30 and 60 weeks (wo) were tested over 8 weeks for open field activity, sucrose preference, social interactions, fear conditioning, and functional neuroimaging. Prefrontal cortical and hippocampal tissues were excised for metabolomics., Results: Our results indicate that young and old mice differ significantly in affective behavioral, functional connectome and prefrontal cortical-hippocampal metabolome. Young mice show a greater responsivity to novel environmental and social stimuli compared to older mice. Conversely, late middle-aged mice (60wo group) display variable patterns of fear conditioning and during re-testing in a modified context. Functional connectivity between a temporal cortical/auditory cortex network and subregions of the anterior cingulate cortex and ventral hippocampus, and a greater network modularity and assortative mixing of nodes was stronger in young versus older adult mice. Metabolome analyses identified differences in several essential amino acids between 10wo mice and the other age groups., Discussion: The results support differential expression of 'emotionality' across distinct stages of the mouse lifespan involving greater prefrontal-hippocampal connectivity and neurochemistry., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Febo, Mahar, Rodriguez, Buraima, Pompilus, Pinto, Grudny, Bruijnzeel and Merritt.)

Secondary metabolites are essential components for the survival of plants. Secondary metabolites in complex mixtures from plants have been adopted and documented by different traditional medicinal systems worldwide for the treatment of various human diseases. The extraction strategies are the key components for therapeutic development from natural sources. Polarity-dependent solvent-selective extraction, acidic and basic solution-based extraction, and microwave- and ultrasound-assisted extraction are some of the most important strategies for the extraction of natural products from plants. The method needs to be optimized to isolate a specific class of compounds. Therefore, to establish the mechanism of action, the characterization of the secondary metabolites, in a mixture or in their pure forms, is equally important. LC-MS, GC-MS, and extensive NMR spectroscopic strategies are established techniques for the profiling of metabolites in crude extracts. Various protocols for the extraction and characterization of a wide range of classes of compounds have been developed by various research groups and are described in this review. Additionally, the possible means of characterizing the compounds in the mixture and their uniqueness are also discussed. Hyphenated techniques are crucial for profiling because of their ability to analyze a vast range of compounds. In contrast, inherent chemical shifts make NMR an indispensable tool for structure elucidation in complex mixtures.

Introduction: Informed consent forms (ICFs) for randomised clinical trials (RCTs) can be onerous and lengthy. The process has the potential to overwhelm patients with information, leading them to miss elements of the study that are critical for an informed decision. Specifically, overly long and complicated ICFs have the potential to increase barriers to trial participation for patients with mild cognitive impairment, those who do not speak English as a first language or among those with lower medical literacy. In turn, this can influence trial recruitment, completion and external validity., Methods and Analysis: SIMPLY-SNAP is a pragmatic, multicentre, open-label, two-arm parallel-group superiority RCT, nested within a larger trial, the Staphylococcus aureus Network Adaptive Platform (SNAP) trial. We will randomise potentially eligible participants of the SNAP trial 1:1 to a full-length ICF or a SIMPlified LaYered (SIMPLY) consent process where basic information is summarised with embedded hyperlinks to supplemental information and videos. The primary outcome is recruitment into the SNAP trial. Secondary outcomes include patient understanding of the clinical trial, patient and research staff satisfaction with the consent process, and time taken for consent. As an exploratory outcome, we will also compare measures of diversity (eg, gender, ethnicity), according to the consent process randomised to. The planned sample size will be 346 participants., Ethics and Dissemination: The study has been approved by the ethics review board (Sunnybrook Health Sciences Research Ethics Board) at sites in Ontario. We will disseminate study results via the SNAP trial group and other collaborating clinical trial networks., Trial Registration Number: ClinicalTrials.gov Registry (NCT06168474; www., Clinicaltrials: gov)., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Inhaled antibiotics delivered through dry powder inhalers (DPIs) effectively treat severe bacterial infections by directly targeting the lungs. Our study focused on developing inhalable dry powder microspheres of linezolid (LNZ) using biodegradable polycaprolactone (PCL) polymer. The LNZ-PCL microspheres were fabricated using a double emulsification solvent evaporation method. Optimization of formulation parameters was performed using a factorial design. Evaluation of the microspheres included size, shape, drug loading, entrapment efficiency, aerosolization, and drug release. The morphological analysis confirmed spherical-shaped rough particles within the inhalable size range. The encapsulation efficiency was determined to be 52.84%, indicating successful drug incorporation. Aerosolization efficiency was significantly enhanced when LNZ-PCL microspheres were combined with lactose as a carrier, achieving a fine particle fraction (FPF) value of 70.90%. In-vitro dissolution studies demonstrated sustained drug release for over 24 h under lung pH conditions. Overall, our study highlights the potential of inhalable LNZ-PCL microspheres as a targeted approach for treating pulmonary tuberculosis. Further research and in-vivo studies are needed to validate their effectiveness in life-threatening bacterial infections., Competing Interests: Declaration of competing interest There is no personal or financial conflict of interest among authors or with the institution with the publication of this manuscript., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Stable isotopes are powerful tools to assess metabolism. 13 C labeling is detected using nuclear magnetic resonance (NMR) spectroscopy or mass spectrometry (MS). MS has excellent sensitivity but generally cannot discriminate among different 13 C positions (isotopomers), whereas NMR is less sensitive but reports some isotopomers. Here, we develop an MS method that reports all 16 aspartate and 32 glutamate isotopomers while requiring less than 1% of the sample used for NMR. This method discriminates between pathways that result in the same number of 13 C labels in aspartate and glutamate, providing enhanced specificity over conventional MS. We demonstrate regional metabolic heterogeneity within human tumors, document the impact of fumarate hydratase (FH) deficiency in human renal cancers, and investigate the contributions of tricarboxylic acid (TCA) cycle turnover and CO 2 recycling to isotope labeling in vivo. This method can accompany NMR or standard MS to provide outstanding sensitivity in isotope-labeling experiments, particularly in vivo., Competing Interests: Declaration of interests R.J.D. is a founder and advisor at Atavistik Bio and serves on the Scientific Advisory Boards of Agios Pharmaceuticals, Vida Ventures, Droia Ventures, and Nirogy Therapeutics., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Resveratrol (RSV) is a bioactive phytoconstituent that has potential applications in respiratory diseases. However, poor oral bioavailability is the major hurdle to its clinical use. In the present work, resveratrol-loaded polycaprolactone (PCL) inhalable microspheres (MSs) were formulated to improve their therapeutic potential. The inhalable microspheres were formulated using the emulsion-solvent evaporation method. In this research, inhalable resveratrol microspheres were prepared using Tween 80 in place of polyvinyl alcohol which formed insoluble lumps. A 3 2 factorial design was applied taking polymer (PCL) and emulsifier (Tween 80) as independent variables and drug loading (DL) and encapsulation efficiency (EE) as dependent variables. The DL and EE of the optimized formulation were found to be 30.6% and 63.84% respectively. The in vitro aerosolization study performed using the Anderson cascade impactor showed that the fine particle fraction (FPF) of optimized resveratrol polycaprolactone microspheres (RSV-PCL-MSs) blended with lactose, and RSV-PCL-MSs were significantly higher than those of the pure drugs. The MMAD T (theoretical mass median aerodynamic diameter) of optimized RSV-PCL-MSs was found to be 3.25 ± 1.15. The particle size of microspheres was within the inhalable range, i.e., between 1 and 5 µm. The morphological analysis showed spherical-shaped particles with smooth surfaces. The in vitro release study showed sustained drug release from the microspheres for up to 12 h. The study concluded that resveratrol-loaded inhalable microspheres may be an efficient delivery system to treat COPD., (© 2023. The Author(s), under exclusive licence to American Association of Pharmaceutical Scientists.)

Deuterated water ( 2 H 2 O) is a widely used tracer of carbohydrate biosynthesis in both preclinical and clinical settings, but the significant kinetic isotope effects (KIE) of 2 H can distort metabolic information and mediate toxicity. 18 O-water (H 2 18 O) has no significant KIE and is incorporated into specific carbohydrate oxygens via well-defined mechanisms, but to date it has not been evaluated in any animal model. Mice were given H 2 18 O during overnight feeding and 18 O-enrichments of liver glycogen, triglyceride glycerol (TG), and blood glucose were quantified by 13 C NMR and mass spectrometry (MS). Enrichment of oxygens 5 and 6 relative to body water informed indirect pathway contributions from the Krebs cycle and triose phosphate sources. Compared with mice fed normal chow (NC), mice whose NC was supplemented with a fructose/glucose mix (i.e., a high sugar [HS] diet) had significantly higher indirect pathway contributions from triose phosphate sources, consistent with fructose glycogenesis. Blood glucose and liver TG 18 O-enrichments were quantified by MS. Blood glucose 18 O-enrichment was significantly higher for HS versus NC mice and was consistent with gluconeogenic fructose metabolism. TG 18 O-enrichment was extensive for both NC and HS mice, indicating a high turnover of liver triglyceride, independent of diet. Thus H 2 18 O informs hepatic carbohydrate biosynthesis in similar detail to 2 H 2 O but without KIE-associated risks., (© 2022 The Authors. NMR in Biomedicine published by John Wiley & Sons Ltd.)

Pulmonary administration of biodegradable polymeric formulation is beneficial in the treatment of various respiratory diseases. For respiratory delivery, the polymer must be non-toxic, biodegradable, biocompatible, and stable. Poly D, L-lactic-co-glycolic acid (PLGA) is a widely used polymer for inhalable formulations because of its attractive mechanical and processing characteristics which give great opportunities to pharmaceutical industries to formulate novel inhalable products. PLGA has many pharmaceutical applications and its biocompatible nature produces non-toxic degradation products. The degradation of PLGA takes place through the non-enzymatic hydrolytic breakdown of ester bonds to produce free lactic acid and glycolic acid. The biodegradation products of PLGA are eliminated in the form of carbon dioxide (CO 2 ) and water (H 2 O) by the Krebs cycle. The biocompatible properties of PLGA are investigated in various in vivo and in vitro studies. The high structural integrity of PLGA particles provides better stability, excellent drug loading, and sustained drug release. This review provides detailed information about PLGA as an inhalable grade polymer, its synthesis, advantages, physicochemical properties, biodegradability, and biocompatible characteristics. The important formulation aspects that must be considered during the manufacturing of inhalable PLGA formulations and the toxicity of PLGA in the lungs are also discussed in this paper. Additionally, a thorough overview is given on the application of PLGA as a particulate carrier in the treatment of major respiratory diseases, such as cystic fibrosis, lung cancer, tuberculosis, asthma, and pulmonary hypertension., (© 2023. The Author(s), under exclusive licence to American Association of Pharmaceutical Scientists.)

Building an accurate lipid inventory relies on coordinated information from orthogonal analytical capabilities. Integrating the familiar workflow of liquid chromatography (LC), high-resolution mass spectrometry (HRMS), and tandem mass spectrometry (MS/MS) with proton nuclear magnetic resonance spectroscopy ( 1 H NMR) would be ideal for building that inventory. For absolute lipid structural elucidation, LC-HRMS/MS can provide lower-level structural information with superior sensitivity, while 1 H NMR can provide invaluable higher-order structural information for the disambiguation of isomers with absolute chemical specificity. Digitization of the LC eluent followed by splitting the microfractions into two flow paths in a defined ratio for HRMS/MS and NMR would be the ideal strategy to permit correlation of the MS and NMR data as a function of chromatographic retention time. Here, we report an active segmentation platform to transform analytical flow rate LC eluent into parallel microliter segmented flow queues for high confidence correlation of the MS, MS/MS, and NMR data. The practical details in implementing this strategy to achieve an integrated LC-MS-NMR platform are presented, including the development of an active segmentation technology using a four-port two-way valve to transform the LC eluent into parallel segmented flows for online MS analysis followed by offline segment-specific 1 H NMR and optimization of the detector response toward segmented flow. To demonstrate the practicality of this novel platform, it was tested using lipid mixture samples.

Oncolytic viral therapy is a recent advance in cancer treatment, demonstrating promise as a primary treatment option. To date, the secondary metabolic effects of viral infection in cancer cells has not been extensively studied. In this work, we have analyzed early-stage metabolic changes in cancer cells associated with oncolytic myxoma virus infection. Using GC-MS based metabolomics, we characterized the myxoma virus infection induced metabolic changes in three cancer cell lines-small cell (H446) and non-small cell (A549) lung cancers, and glioblastoma (SFxL). We show that even at an early stage (6 and 12 h) myxoma infection causes profound changes in cancer cell metabolism spanning several important pathways such as the citric acid cycle, fatty acid metabolism, and amino acid metabolism. In general, the metabolic effects of viral infection across cell lines are not conserved. However, we have identified several candidate metabolites that can potentially serve as biomarkers for monitoring oncolytic viral action in general., (© 2022. The Author(s).)

The compound β-lapachone, a naturally derived naphthoquinone, has been utilized as a potent medicinal nutrient to improve health. Over the last twelve years, numerous reports have demonstrated distinct associations of β-lapachone and NAD(P)H: quinone oxidoreductase 1 (NQO1) protein in the amelioration of various diseases. Comprehensive research of NQO1 bioactivity has clearly confirmed the tumoricidal effects of β-lapachone action through NAD + -keresis, in which severe DNA damage from reactive oxygen species (ROS) production triggers a poly-ADP-ribose polymerase-I (PARP1) hyperactivation cascade, culminating in NAD + /ATP depletion. Here, we report a novel combination strategy with aminooxyacetic acid (AOA), an aspartate aminotransferase inhibitor that blocks the malate-aspartate shuttle (MAS) and synergistically enhances the efficacy of β-lapachone metabolic perturbation in NQO1 + breast cancer. We evaluated metabolic turnover in MDA-MB-231 NQO1 + , MDA-MB-231 NQO1 - , MDA-MB-468, and T47D cancer cells by measuring the isotopic labeling of metabolites from a [U- 13 C]glucose tracer. We show that β-lapachone treatment significantly hampers lactate secretion by ~85% in NQO1 + cells. Our data demonstrate that combinatorial treatment decreases citrate, glutamate, and succinate enrichment by ~14%, ~50%, and ~65%, respectively. Differences in citrate, glutamate, and succinate fractional enrichments indicate synergistic effects on central metabolism based on the coefficient of drug interaction. Metabolic modeling suggests that increased glutamine anaplerosis is protective in the case of MAS inhibition.

Alstonia scholaris is a well-known source of alkaloids and widely recognized for therapeutic purposes to treat the ailments in human and livestock. However, the composition and production of alkaloids vary due to tissue specific metabolism and seasonal variation. This study investigated alkaloids in leaves, stems, trunk barks, fruits, and flowers of A. scholaris . The impact of seasonal changes on the production of alkaloids in the leaves of A. scholaris was also investigated. One and two-dimensional Nuclear Magnetic Resonance (NMR) experiments were utilized for the characterization of alkaloids and total eight alkaloids (picrinine, picralinal, akuammidine, 19 S scholaricine, 19,20 E vallesamine, Nb-demethylalstogustine N-Oxide, Nb-demethylalstogustine, and echitamine) were characterized and quantified. Quantitative and multivariate analysis suggested that the alkaloids content is tissue specific, illustrating the effect of plant tissue organization on alkaloidal production in A. scholaris . The results suggest that the best part to obtain alkaloids is trunk barks, since it contains 7 alkaloids. However, the best part for isolating picrinine, picralinal, akuammidine, 19 S scholaricine, and 19,20 E vallesamine is fruit, since it shows highest amount of these alkaloids. Undoubtedly, NMR and statistical methods are very helpful to differentiate the profile of alkaloids in A. scholaris .

MUL1 is a multifunctional E3 ubiquitin ligase that is involved in various pathophysiological processes including apoptosis, mitophagy, mitochondrial dynamics, and innate immune response. We uncovered a new function for MUL1 in the regulation of mitochondrial metabolism. We characterized the metabolic phenotype of MUL1(-/-) cells using metabolomic, lipidomic, gene expression profiling, metabolic flux, and mitochondrial respiration analyses. In addition, the mechanism by which MUL1 regulates metabolism was investigated, and the transcription factor HIF-1α, as well as the serine/threonine kinase Akt2, were identified as the mediators of the MUL1 function. MUL1 ligase, through K48-specific polyubiquitination, regulates both Akt2 and HIF-1α protein level, and the absence of MUL1 leads to the accumulation and activation of both substrates. We used specific chemical inhibitors and activators of HIF-1α and Akt2 proteins, as well as Akt2(-/-) cells, to investigate the individual contribution of HIF-1α and Akt2 proteins to the MUL1-specific phenotype. This study describes a new function of MUL1 in the regulation of mitochondrial metabolism and reveals how its downregulation/inactivation can affect mitochondrial respiration and cause a shift to a new metabolic and lipidomic state., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Cilenti, Mahar, Di Gregorio, Ambivero, Merritt and Zervos.)

Malignant tumors exhibit altered metabolism resulting in a highly acidic extracellular microenvironment. Here, we show that cytoplasmic lipid droplet (LD) accumulation, indicative of a lipogenic phenotype, is a cellular adaption to extracellular acidity. LD marker PLIN2 is strongly associated with poor overall survival in breast cancer patients. Acid-induced LD accumulation is triggered by activation of the acid-sensing G-protein-coupled receptor (GPCR) OGR1, which is expressed highly in breast tumors. OGR1 depletion inhibits acid-induced lipid accumulation, while activation by a synthetic agonist triggers LD formation. Inhibition of OGR1 downstream signaling abrogates the lipogenic phenotype, which can be rescued with OGR1 ectopic expression. OGR1-depleted cells show growth inhibition under acidic growth conditions in vitro and tumor formation in vivo. Isotope tracing shows that the source of lipid precursors is primarily autophagy-derived ketogenic amino acids. OGR1-depleted cells are defective in endoplasmic reticulum stress response and autophagy and hence fail to accumulate LDs affecting survival under acidic stress., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Murraya koenigii (L.) Spreng (Curry leaf) is a commercially important medicinal plant in South Asia, containing therapeutically valuable carbazole alkaloids (CAs). Thus, the quantitative evaluation of these compounds from different climatic zones of India are an important aspect for quality assessment and economic isolation of targeted compounds from the plant. In this study, quantitative estimation of CAs among 34 Indian natural populations of M. koenigii was assessed using UPLC/MS/MS. The collected populations represent the humid subtropical, tropical wet & dry, tropical wet, semi-arid, arid, and montane climatic zones of India. A total of 11 CAs viz. koenine-I, murrayamine A, koenigine, koenimbidine, koenimbine, O-methylmurrayamine A, girinimbine, mahanine, 8,8''-biskoenigine, isomahanimbine, and mahanimbine were quantified using multiple reaction monitoring (MRM) experiments within 5.0 min. The respective range for natural abundance of CAs were observed as 0.097-1.222, 0.092-5.014, 0.034-0.661, 0.010-1.673, 0.013-7.336, 0.010-0.310, 0.010-0.114, 0.049-5.288, 0.031-1.731, 0.491-3.791, and 0.492-5.399 mg/g in leaves of M. koenigii. The developed method shown linearity regression coefficient (r 2 >0.9995), LOD (0.003-0.248 ng/mL), LOQ (0.009-0.754 ng/mL), and the recovery was between 88.803-103.729 %. The bulk of these CAs were recorded in their highest concentrations in the humid subtropical zone, followed by the tropical wet & dry zones of India. Further, principal component analysis (PCA) was performed which differentiated the climatic zones according to the dominant and significant CAs contents within the populations. The study concludes that the method established is simple, rapid, with high sample throughput, and can be used as a tool for commercial purposes and quality control of M. koenigii., (© 2021 Wiley-VHCA AG, Zurich, Switzerland.)

Treatment of cancers with β-lapachone causes NAD(P)H: quinone oxidoreductase 1 (NQO1) to generate an unstable hydroquinone that regenerates itself in a futile cycle while producing reactive oxygen species (ROS) in the form of superoxide and subsequently hydrogen peroxide. Rapid accumulation of ROS damages DNA, hyperactivates poly-ADP-ribose polymerase-I, causes massive depletion of NAD + /ATP, and hampers glycolysis. Cells overexpressing NQO1 subsequently die rapidly through an NAD + -keresis mechanism. Assessing changes in glycolytic rates caused by NQO1 bioactivation would provide a means of assessing treatment efficacy, potentially lowering the chemotherapeutic dosage, and reducing off-target toxicities. NQO1-mediated changes in glycolytic flux were readily detected in A549 (lung), MiaPaCa2 (pancreatic), and HCT-116 (colon) cancer cell lines by 2 H-NMR after administration of [ 2 H 7 ]glucose. The deuterated metabolic products 2 H-lactate and HDO were quantified, and linear relationships with glucose consumption for both products were observed. The higher concentration of HDO compared to 2 H-lactate allows for more sensitive measurement of the glycolytic flux in cancer. Gas chromatography-mass spectrometry analysis agreed with the NMR results and confirmed downregulated energy metabolism in NQO1 + cells after β-lapachone treatment. The demonstrated method is ideal for measuring glycolytic rates, the effects of chemotherapeutics that target glycolysis, and has the potential for in vivo translation.

Introduction: Despite carrying a disproportionately high burden of depression, patients in low-income countries lack access to effective care. The collaborative care model (CoCM) has robust evidence for clinical effectiveness in improving mental health outcomes. However, evidence from real-world implementation of CoCM is necessary to inform its expansion in low-resource settings., Methods: We conducted a 2-year mixed-methods study to assess the implementation and clinical impact of CoCM using the WHO Mental Health Gap Action Programme protocols in a primary care clinic in rural Nepal. We used the Capability Opportunity Motivation-Behaviour (COM-B) implementation research framework to adapt and study the intervention. To assess implementation factors, we qualitatively studied the impact on providers' behaviour to screen, diagnose and treat mental illness. To assess clinical impact, we followed a cohort of 201 patients with moderate to severe depression and determined the proportion of patients who had a substantial clinical response (defined as ≥50% decrease from baseline scores of Patient Health Questionnaire (PHQ) to measure depression) by the end of the study period., Results: Providers experienced improved capability (enhanced self-efficacy and knowledge), greater opportunity (via access to counsellors, psychiatrist, medications and diagnostic tests) and increased motivation (developing positive attitudes towards people with mental illness and seeing patients improve) to provide mental healthcare. We observed substantial clinical response in 99 (49%; 95% CI: 42% to 56%) of the 201 cohort patients, with a median seven point (Q1:-9, Q3:-2) decrease in PHQ-9 scores (p<0.0001)., Conclusion: Using the COM-B framework, we successfully adapted and implemented CoCM in rural Nepal, and found that it enhanced providers' positive perceptions of and engagement in delivering mental healthcare. We observed clinical improvement of depression comparable to controlled trials in high-resource settings. We recommend using implementation research to adapt and evaluate CoCM in other resource-constrained settings to help expand access to high-quality mental healthcare., Competing Interests: Competing interests: PR, PA, MB, BB, BG, PG, TKG, BK, RM, SP, JR, SSw and PT were employed by, and NC, DC, SH, DM, AR, DS, RS, SSa, SSh, AT and BA work in partnership with a non-profit healthcare company (Nyaya Health Nepal, with support from the US-based non-profit, Possible) that delivers free healthcare in rural Nepal using funds from the Government of Nepal and other public, philanthropic and private foundation sources. MB is a faculty at BP Koirala Institute of Health Sciences, Dharan, Nepal. NC, SH and DM are employed by, and DC, DM and SSa are faculty members at a private medical school (Icahn School of Medicine at Mount Sinai). DC is a faculty member at, DC and SH are employed part-time by and SH is a graduate student at a public university (University of Washington). SKD is a resident at an academic medical centre (Hurley Medical Center) that receives revenue through private sector fee-for-service medical transactions and a charitable private foundation. TKG is a fellow with a bidirectional fellowship program (HEAL Initiative) that is affiliated with a public university (University of California, San Francisco) that receives funding from public, philanthropic and private foundation sources. DM and BA are members on Possible’s Board of Directors, for which they receive no compensation. VN is employed at a public university (University of California, Los Angeles). DS and RS are employed at an academic medical centre (Brigham and Women’s Hospital) that receives public sector research funding, as well as revenue through private sector fee-for-service medical transactions and private foundation grants. DS and RS are faculty members at a private medical school (Harvard Medical School). DS is employed at an academic research centre (Ariadne Labs) that is jointly supported by an academic medical center (Brigham and Women’s Hospital) and a private university (Harvard TH Chan School of Public Health) via public sector research funding and private philanthropy. RS is employed at an academic medical centre (Massachusetts General Hospital) that receives public sector research funding, as well as revenue through private sector fee-for-service medical transactions and private foundation grants. SSh is a faculty member at a private college (Wheaton College). PT is a graduate student at a public university (University of New South Wales). BA is a faculty member at a public university (University of California, San Francisco). All authors have read and understood BMJ Open’s policy on declaration of interests, and declare that we have no competing financial interests. The authors do, however, believe strongly that healthcare is a public good, not a private commodity., (© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)