The Therapeutic Efficacy and Mechanism of Action of Gnetin C, a Natural Compound from the Melinjo Plant, in a Preclinical Mouse Model of Advanced Prostate Cancer.

The metastasis-associated protein 1/protein kinase B (MTA1/AKT) signaling pathway has been shown to cooperate in promoting prostate tumor growth. Targeted interception strategies by plant-based polyphenols, specifically stilbenes, have shown great promise against MTA1-mediated prostate cancer progression. In this study, we employed a prostate-specific transgenic mouse model with MTA1 overexpression on the background of phosphatase and tensin homolog (Pten) null (R26MTA1; Ptenf/f) and PC3M prostate cancer cells which recapitulate altered molecular pathways in advanced prostate cancer. Mechanistically, the MTA1 knockdown or pharmacological inhibition of MTA1 by gnetin C (dimer resveratrol) in cultured PC3M cells resulted in the marked inactivation of mammalian target of rapamycin (mTOR) signaling. In vivo, mice tolerated a daily intraperitoneal treatment of gnetin C (7 mg/kg bw) for 12 weeks without any sign of toxicity. Treatment with gnetin C markedly reduced cell proliferation and angiogenesis and promoted apoptosis in mice with advanced prostate cancer. Further, in addition to decreasing MTA1 levels in prostate epithelial cells, gnetin C significantly reduced mTOR signaling activity in prostate tissues, including the activity of mTOR-target proteins: p70 ribosomal protein S6 kinase (S6K) and eukaryotic translational initiation factor 4E (elF4E)-binding protein 1 (4EBP1). Collectively, these findings established gnetin C as a new natural compound with anticancer properties against MTA1/AKT/mTOR-activated prostate cancer, with potential as monotherapy and as a possible adjunct to clinically approved mTOR pathway inhibitors in the future.

Adverse Cardiac Events of Hypercholesterolemia Are Enhanced by Sitagliptin Administration in Sprague Dawley Rats.

Background: Cardiovascular disease (CVD) affects millions worldwide and is the leading cause of death among non-communicable diseases. Western diets typically comprise of meat and dairy products, both of which are rich in cholesterol (Cho) and methionine (Met), two well-known compounds with atherogenic capabilities. Despite their individual effects, literature on a dietary combination of the two in the context of CVD are limited. An additional interest was to investigate the cardioprotective potential of sitagliptin, an anti-type 2 diabetic drug. Thus, we hypothesized that atherogenic feeding would result in adverse cardiac effects and would attenuate upon sitagliptin administration. Methods: Six-week-old adult male Sprague-Dawley rats were fed either a control (Con), high Met (1.5%), high Cho (2.0%), or high Met (1.5%) + high Cho (2.0%) diet for 35 days. They were orally gavaged with vehicle (water) or sitagliptin (100 mg/kg/d) from day 10 through 35. On day 36, rats were euthanized, and tissues were collected for analysis. Results: Histopathological evaluation revealed a reduction in myocardial striations and increased collagen deposition in hypercholesterolemia (HChol), responses that became exacerbated upon sitagliptin administration. Cardiac pro-inflammatory and pro-fibrotic responses were adversely impacted in similar fashion. The addition of Met to Cho (MC) attenuated all adverse structural and biochemical responses, with or without sitagliptin. Conclusion: Adverse cardiac outcomes in HChol were enhanced with sitagliptin administration and such effects were alleviated by Met. Our findings could be significant for understanding the risk-benefit of sitagliptin in type 2 diabetics who are known to consume atherogenic diets.

Ion-Solvent Interactions under Confinement Hold the Key to Tuning the DNA Translocation Speeds in Polyelectrolyte-Functionalized Nanopores.

DNA sequencing and sensing using nanopore technology delves critically into the alterations in the measurable electrical signal as single-stranded DNA is drawn through a tiny passage. To make such precise measurements, however, slowing down the DNA in the tightly confined passage is a key requirement, which may be achieved by grafting the nanopore walls with a polyelectrolyte layer (PEL). This soft functional layer at the wall, under an off-design condition, however, may block the DNA passage completely, leading to the complete loss of output signal from the nanobio sensor. Whereas theoretical postulates have previously been put forward to explain the essential physics of DNA translocation in nanopores, these have turned out to be somewhat inadequate when confronted with the experimental findings on functionalized nanopores, including the prediction of the events of complete signal losses. Circumventing these constraints, herein we bring out a possible decisive role of the interplay between the inevitable variabilities in the ionic distribution along the nanopore axis due to its finite length as opposed to its idealized "infinite" limit as well as the differential permittivity of PEL and bulk solution that cannot be captured by the commonly used one-dimensional variant of the electrical double layer theory. Our analysis, for the first time, captures variations in the ionic concentration distribution across multidimensional physical space and delineates its impact on the DNA translocation characteristics that have hitherto remained unaddressed. Our results reveal possible complete blockages of DNA translocation as influenced by less-than-threshold permittivity values or greater-than-threshold grafting densities of the PEL. In addition, electrohydrodynamic blocking is witnessed due to the ion-selective nature of the nanopore at low ionic concentrations. Hence, our study establishes a functionally active regime over which the PEL layer in a finite-length nanopore facilitates controllable DNA translocation, enabling successful sequencing and sensing through the explicit modulation of translocation speed.

Tomosyns attenuate SNARE assembly and synaptic depression by binding to VAMP2-containing template complexes.

Tomosyns are widely thought to attenuate membrane fusion by competing with synaptobrevin-2/VAMP2 for SNARE-complex assembly. Here, we present evidence against this scenario. In a novel mouse model, tomosyn-1/2 deficiency lowered the fusion barrier and enhanced the probability that synaptic vesicles fuse, resulting in stronger synapses with faster depression and slower recovery. While wild-type tomosyn-1m rescued these phenotypes, substitution of its SNARE motif with that of synaptobrevin-2/VAMP2 did not. Single-molecule force measurements indeed revealed that tomosyn's SNARE motif cannot substitute synaptobrevin-2/VAMP2 to form template complexes with Munc18-1 and syntaxin-1, an essential intermediate for SNARE assembly. Instead, tomosyns extensively bind synaptobrevin-2/VAMP2-containing template complexes and prevent SNAP-25 association. Structure-function analyses indicate that the C-terminal polybasic region contributes to tomosyn's inhibitory function. These results reveal that tomosyns regulate synaptic transmission by cooperating with synaptobrevin-2/VAMP2 to prevent SNAP-25 binding during SNARE assembly, thereby limiting initial synaptic strength and equalizing it during repetitive stimulation.

Studies on α-amylase inhibition by acarbose and quercetin using fluorescence, circular dichroism, docking, and dynamics simulations.

This study looked at the effects of acarbose (ACA) and quercetin (QUE) on α-amylase activity, employing QUE and ACA to measure enzyme activity. The study observed that both drugs suppressed α-amylase activity, with greater inhibition reported at higher concentrations. The use of tryptophan residues as an intrinsic fluorescence probe permitted the observation of conformational changes in α-amylase, with CD measurements utilized to explore the secondary structure in the presence of QUE and ACA. Docking studies revealed an effective interaction between α-amylase, quercetin and acarbose, with a higher binding energy. Finally, a trajectory analysis was done to establish the stability and volatility of these complexes. These findings have potential significance for the development of new α-amylase-related therapeutics.

Utility of three-dimensional proton density-weighted sequence MRI in knee for the assessment of Anterolateral complex in Anterior cruciate ligament injury.

OBJECTIVES: To assess the proportion and pattern of injury of the anterolateral ligament (ALL) and the Kaplan fibre (KF) complex in knees with anterior cruciate ligament (ACL) injuries on MRI using three-dimensional (3D) proton density (PD) sequences. METHODS: A total of 88 patients having ACL injury were included in this cross-sectional study. 3D PD sequences were used to assess injury of ALL and the KF complex and were graded on a scale of 0 to 3. MR images were evaluated by two radiologists. Interobserver agreement was determined using Cohen Kappa. RESULTS: Femoral, meniscal, and tibial portions of ALL were visualized in 90.9%, 92%, and 94.3% of the study subjects, respectively. Proximal and distal KF were visualized in 92% and 93.2% of patients, respectively. Injury to ALL and KF was seen in 63.6% and 17% of the patients, respectively. Excellent interobserver agreement was noted for the identification and grading of ALL and KF complex injuries. CONCLUSIONS: Oblique reformatted 3DPD MRI reliably detect ALC; however, ALL injury is better characterized than KF injury using this sequence. ADVANCES IN KNOWLEDGE: Given the potential role of anterolateral complex (ALC) in maintaining the rotational stability of the knee, ALC assessment using the 3D PD sequences and their oblique reformatted images should be incorporated as routine review area of the knee MRI, particularly in the setting of ACL tear.

Immobilization of α-amylase onto functionalized molybdenum diselenide nanoflowers (MoSe2-NFs) as scaffolds: Characterization, kinetics, and potential applications in starch-based industries.

The current study presents the application of molybdenum diselenide nanoflowers (MoSe2-NFs) as an innovative substrate for immobilizing α-amylase by glutaraldehyde activation. This approach results in the development of a nanobiocatalyst that exhibits remarkable advantages compared to a standalone enzyme. Several physical methods, such as fluorescence microscopy, FT-IR, SEM, TEM, XRD, AFM, and Raman spectroscopy, were used to confirm that α-amylase was successfully attached to MoSe2-NFs. By employing the Box-Behnken design of the RSM, the parameters were optimized, resulting in an immobilization efficiency of roughly 87.33%. The immobilized variant of α-amylase demonstrated superior thermostability, pH stability, reusability, and storage stability in comparison to the soluble enzyme. The catalytic activity of α-amylase was highest when immobilized on MoSe2-NFs at the same pH and temperature as the soluble enzyme. However, there was an expansion in the range of parameters in which this activity was observed. Furthermore, the immobilized enzyme exhibited a retention of nearly 80% residual activity following 12 successive reuses. The immobilized enzyme exhibited around 82% residual activity after being stored for 120 days. It is possible that the immobilization process changed the Michaelis-Menten constant, which means that the substrate could no longer reach certain active sites on the enzyme because it had become longer. The study's findings suggest that the α-amylase-MoSe2-NFs system could be useful in industry because it can work in a wider range of temperature and pH conditions. Furthermore, the intrinsic non-toxic characteristics of the matrix, along with its ability to be kept for prolonged periods and recycled, render nano biocatalysts very well-suited for the effective synthesis of maltose in the food and pharmaceutical industries.

The Role of Gut Microbiota and Associated Compounds in Cardiovascular Health and its Therapeutic Implications.

It is possible that gut bacteria may have a beneficial effect on cardiovascular health in humans. It may play a major role in the progression of a variety of cardiovascular diseases, including Heart Failure (HF), Atherosclerosis, Coronary Arterial Disease (CAD), Ischemic Heart Disease (IHD), and Others. Dysbiosis of the gut microbiota, along with its direct and indirect impact on gut health, may induce cardiovascular disorders. Although advanced studies have demonstrated the relationship of various metabolites to cardiovascular diseases (CVD) in animals, translating their functional capacity to humans remains a significant area of research. This paper simplifies the demonstration of some compounds, pathways, and components like Trimethylamine N-oxide (TMAO), short-chain fatty acids (SCFAs), and butyrate production. It demonstrates how a change in eating habits causes TMAO and how the impact of different drugs on gut microbiota species and high consumption of Westernized food causes several heartrelated problems, such as atherosclerosis and inflammation that can even become the cause of heart failure. Modulation of the gut microbiome, on the other hand, is a novel therapeutic measure because it can be easily altered through diet and other lifestyle changes. It could then be used to lower the risk of several CVDs.

Investigation, scaffold hopping of novel donepezil-based compounds as anti-Alzhiemer's agents: synthesis, in-silico and pharmacological evaluations.

Alzheimer's disease (AD) is a multifaceted neurodegenerative condition. The pathogenesis of AD is highly intricate and the disease is apparent in the aged population ~ 50-70 years old. Even after > 100 years of research, the root origin of AD and its pathogenesis is unclear, complex and multifaceted. Herein, we have designed and synthesized 9 novel molecules with three different heterocyclic scaffolds namely pyrrolidone-2-one, quinoline & indoline-2-one to imitate and explore the novel chemical space around donepezil. The synthesized molecules were evaluated for their potential as anti-Alzheimer's agents through in-vitro and in-vivo studies in appropriate animal models. To further understand their interaction with acetylcholinesterase enzyme (AChE), extra-precision docking, and molecular dynamics simulation studies were carried out. As the number of compounds was limited to thoroughly explore the structure-activity relationship, atom-based 3D-quantitative structure-activity relationships (QSAR) studies were carried out to get more insights. All the designed compounds were found to inhibit AChE with IC50 in the micromolar range. From pyrrolidone-2-one series, 6-chloro-N-(1-(1-(3,4-dimethoxybenzyl)-2-oxopyrrolidin-3-yl)piperidin-4-yl)pyridine-3-sulfonamide (9), 2-(1-benzylpiperidin-4-yl)-6,7-dimethoxy-4-(4-methoxyphenyl)quinoline (18) from quinoline series and N-(1-benzylpiperidin-4-yl)-2-(2-oxoindolin-3-yl)acetamide (23) from indolin-2-one series inhibited AChE with an IC50 value of 0.01 µM. Based on other biochemical studies like lipid peroxidation, reduced glutathione, superoxide dismutase, catalase, nitrite, and behavioural studies (Morris water maze), compound 9 was found to be a potent AChE inhibitor which can be further explored as a lead molecule to design more potent and effective anti-Alzheimer's agents.

The Marketing and Perceptions of Non-Tobacco Blunt Wraps on Twitter.

OBJECTIVE: Non-tobacco blunt wraps (N-TBWs), which entered the marketplace in 2017, are being promoted as an alternative to traditional TBWs (e.g., cigarillos) for blunt smoking. The lack of studies on these novel products warrants an investigation. This study was the first to explore blunt smokers' perceptions about N-TBWs and the extent of product marketing on Twitter. METHODS: A corpus of tweets from Twitter, posted between January 2017 and November 2021, were identified by a Boolean search string (N = 149,343), where 48,695 tweets were classified as relevant by a machine learning algorithm. These relevant tweets were further screened and labeled as promotional or organic based on product URLs, usernames, keywords, or hashtags. Topic modeling using Dirichlet Allocation was then employed for identifying latent patterns of words among relevant tweets. The Social Networking Potential (SNP) score was employed for identifying influential accounts. RESULTS: Most relevant tweets (89%) were organic, non-promotional expressions about N-TBWs. Account users who only posted non-promotional tweets had a significantly higher SNP than those who only posted promotional tweets. Yet, neither of the two groups of account users consisted of known celebrities. Topic modeling revealed three broad groups of topics (7 in total) denoting the attributes of hemp N-TBWs, interest in non-hemp N-TBWs, and product marketing. CONCLUSIONS: The large proportion of organic tweets is indicative of the nascency of N-TBWs, which will need to be marketed more extensively if they are to replace cigar products used by blunt smokers.

L-Isoleucine reverses hyperammonemia-induced myotube mitochondrial dysfunction and post-mitotic senescence.

Perturbations in the metabolism of ammonia, a cytotoxic endogenous metabolite, occur in a number of chronic diseases, with consequent hyperammonemia. Increased skeletal muscle ammonia uptake causes metabolic, molecular, and phenotype alterations including cataplerosis of (loss of tricarboxylic acid cycle (TCA) cycle intermediate) α-ketoglutarate (αKG), mitochondrial oxidative dysfunction, and senescence-associated molecular phenotype (SAMP). L-Isoleucine (Ile) is an essential, branched-chain amino acid (BCAA) that simultaneously provides acetyl-CoA as an oxidative substrate and succinyl-CoA for anaplerosis (providing TCA cycle intermediates). Our multiomics analyses in myotubes and skeletal muscle from hyperammonemic mice and human patients with cirrhosis showed perturbations in BCAA transporters and catabolism. We, therefore, determined if Ile reverses hyperammonemia-induced impaired mitochondrial oxidative function and SAMP. Studies were performed in differentiated murine C2C12 myotubes that were early passage, late passage (senescent), or those depleted of LAT1/SLC7A5 and human induced pluripotent stem cell-derived myotubes (hiPSCM). Ile reverses hyperammonemia-induced reduction in the maximum respiratory capacity, complex I, II, and III functions in early passage murine myotubes and hiPSCM. Consistently, low ATP content and impaired global protein synthesis (high energy requiring cellular process) during hyperammonemia are reversed by Ile in murine myotubes and hiPSCM. Lower abundance of critical regulators of protein synthesis in mTORC1 signaling, and increased phosphorylation of eukaryotic initiation factor 2α are also reversed by Ile. Genetic depletion studies showed that Ile responses are independent of the amino acid transporter LAT1/SLC7A5. Our studies show that Ile reverses the hyperammonemia-induced impaired mitochondrial oxidative function, cataplerosis, and SAMP in a LAT1/SLC7A5 transporter-independent manner.

A versatile and narrow linewidth infrared radiation source for ro-vibration state-selected preparation of molecules in molecular beams.

We describe the design and characterization of a versatile pulsed (5 ns, 10 Hz repetition rate) optical parametric oscillator and amplifier system capable of generating single longitudinal mode, narrow linewidth (0.01 cm-1) radiations in the wavelength range of 680-870 nm and 1380-4650 nm. Using a combination of power-normalized photoacoustic signal and a Fizeau interferometer-based wavemeter, we are able to actively stabilize the output wavenumber to within 0.005 cm-1 (3σ) over a timescale longer than 1000 s. We demonstrate an application of this system by performing ro-vibration state-selected preparation of CO in the v = 2 state, via direct overtone excitation (v = 0 → 2 at 2346 nm) and subsequent state-selected detection in an internally cold molecular beam.

The Role of Indirect Immunofluorescence (IIF) and Line Immunoassay (LIA) in the Diagnosis of Autoimmune Diseases and Their Clinical Correlation: An Observational Study From a Tertiary Care Center in Bihar.

Background and aim The presence of distinct sets of autoantigens and autoantibodies bestow these autoimmune diseases (ADs) with specific immune profiles or fingerprints, which has cleared the diagnostic dilemma associated with these ADs. This study was planned to collate and compare the reporting of indirect immunofluorescence (IIF) with line immunoassay (LIA) and their clinical correlations. This study was conducted to investigate the association between the reporting of anti-nuclear antibody (ANA) screening by IIF and ANA profile reporting by LIA. Additionally, it aimed to explore the association of ANA pattern detection by IIF with the detection of autoantibodies against nuclear antigens by LIA and the association of autoantibody detection by LIA with clinical diagnosis. Methodology A total of 98 samples from patients suspected of having ADs were subjected to both IIF and LIA, and results were correlated with clinical diagnosis. Results In the homogenous pattern noted by IIF, the clustered antigens identified by LIA included dsDNA, Nucleosome, Histone, and Mi-2. In the speckled pattern, the identified antigens were SS-A/Ro52, P0, SS-A/Ro60, SS-B/La, and U1-snRNP. On the other hand, the nucleolar pattern revealed antigens AMA M2, PCNA, and CENP-B. The centromere pattern was mostly associated with CENP-B. The speckled pattern was found to be most commonly associated with systemic lupus erythematosus (SLE). The most common autoantibody found in total ANA profile-positive samples was anti-U1-snRNP followed by anti-SS-A/Ro60 and anti-SS-B/La, and all three were found to be associated with SLE. Conclusions SLE was the most common AD identified in our study samples, with the speckled pattern being the most common pattern on IIF and anti-U1-snRNP being the most common ANA identified by LIA. The fluorescence pattern of IIF predicts the presence of specific antibodies. LIA should be reserved for IIF-positive but dubious cases and whose signs and symptoms are nebulous and do not match the disease dictated by IIF.

Molecular Efficacy of Gnetin C as Dual-Targeted Therapy for Castrate-Resistant Prostate Cancer.

SCOPE: Resistance of castrate-resistant prostate cancer (CRPC) to enzalutamide (Enz) involves the expression of constitutively active androgen receptor splice variant (AR-V7). In addition to altered AR pathways, CRPC is characterized by "non-AR-driven" signaling, which includes an overexpression of metastasis-associated protein 1 (MTA1). Combining natural compounds with anticancer drugs may enhance drug effectiveness while reducing adverse effects. In this study, the in vitro and in vivo anticancer effects of Gnetin C (GnC) alone and in combination with Enz against CRPC are examined. METHODS AND RESULTS: The effects of GnC alone and in combination with Enz are assessed by cell viability, clonogenic survival, cell migration, and AR and MTA1 expression using 22Rv1 cells. The tumor growth in vivo is assessed by bioluminescent imaging, western blots, RT-PCR, and IHC. GnC alone and in combined treatment inhibit cell viability, clonogenic survival and migration, and AR and MTA1 expression in 22Rv1 cells. The underlying AR- and MTA1-targeted anticancer mechanisms of treatments in vivo involve inhibition of proliferation and angiogenesis, and induction of apoptosis. CONCLUSION: The findings demonstrate that GnC alone and GnC combined with Enz effectively inhibits AR- and MTA1-promoted tumor-progression in advanced CRPC, which indicates its potential as a novel therapeutic approach for CRPC.

Does the peroral chromium administration in young Hariana calves reduce the risk of calf diarrhea by ameliorating insulin response, lactose intolerance, antioxidant status, and immune response?

BACKGROUND: The reduction in insulin sensitivity during rumen development may predispose dairy calves towards lactose intolerance, which could be the reason behind neonatal calf diarrhea (NCD). Chromium (Cr) results in a range of effects when fed to ruminants, but most studies have shown improved insulin sensitivity. The aim of this study was to determine the effect of Cr supplementation on insulin sensitivity, lactose intolerance, diarrhoea and antioxidant, and immune response in young Hariana calves. METHODS: A total of 20 milk-fed Hariana calves were randomly assigned to 1 of 2 treatments, each consisted of 10 calves: (1) a control group without supplemental Cr and (2) a 0.15 mg Cr as Cr-picolinate (CrPic)/kg BW0.75 supplemented group (Cr0.15). RESULTS: A more rapid glucose disappearance with unaltered insulin kinetics during intravenous glucose tolerance test (IVGTT) and oral lactose tolerance test (OLTT) indicates greater insulin sensitivity in Cr supplemented calves. Better insulin sensitivity in Cr supplemented calves was further confirmed by higher values of the quantitative insulin sensitivity check index (QUICKI), revised quantitative insulin sensitivity check index (RQUICKI) and insulin receptor substrate-1 (IRS-1) and lower (P < 0.05) values of homeostasis model assessment-insulin resistance (HOMA-IR) and glucose-to-insulin ratio in Cr supplemented calves during IVGTT. Cr supplementation resulted in a lower (P < 0.05) serum cortisol concentration, whereas serum non-esterified fatty acid (NEFA) concentrations during IVGTT did not differ among the groups. The rise in serum glucose concentrations within 2 h post lactose infusion during OLTT peaked at more than twice the basal glucose concentration, therefore calves were not considered as lactose intolerant. Within monthly blood samples, concentrations of serum insulin were similar among treatments, whereas the Cr supplemented group had lower (P < 0.05) serum glucose concentration and glucose-to-insulin ratio compared with the control group. No treatment differences were detected in the biomarkers of antioxidant status and immunity. Serum Cr concentrations were higher (P < 0.05) in Cr supplemented calves while concentrations of other studied minerals were remained unaltered. The incidence, duration of diarrhea, and faecal score were better (P < 0.05) in calves fed on Cr supplemented diet whereas, no treatment effect was observed on average daily gain (ADG). During the study period, no calves died, and no calves were found to have pneumonia, navel or joint disease. CONCLUSION: Feeding a Cr-supplemented diet improved insulin sensitivity and reduced the risk of diarrhoea in milk-fed young calves, but had no or minimal effects on lactose intolerance, antioxidant status, immune response, and growth performance.

A systematic study on the kinetics of H-shift reactions in pristine acyl peroxy radicals.

A series of acyl peroxy radical H-shifts were systematically studied using computational approaches. Acyl peroxy radicals were categorized into small- (ethanal-pentanal), medium- (hexanal and heptanal) and large-sized (octanal and nonanal) molecules. The H-shifts spanning from 1,4 to 1,9 were inspected for each studied system. For all acyl peroxy radicals, it is the combination of barrier heights and quantum mechanical tunneling that explains the yield of the peracid alkyl radical product. We used the ROHF-ROCCSD(T)-F12a/VDZ-F12//ωB97X-D/aug-cc-pVTZ level of theory to estimate the barrier heights and the subsequent rate coefficients with the exception of the smallest acyl peroxy radical ethanal, for which MN15 density functional was applied. The estimated multiconformer H-shift rate coefficients were found to be in the range of 10-2 s-1 to 10-1 s-1 for the fastest H-migrations. The determined rates imply that these H-shift reactions are often competitive with other RO2 loss processes and should be considered as a path to functionalization in modelling not only rural but also urban air quality.

AI & experimental-based discovery and preclinical IND-enabling studies of selective BMX inhibitors for development of cancer therapeutics.

The current work aims to design and provide a preliminary IND-enabling study of selective BMX inhibitors for cancer therapeutics development. BMX is an emerging target, more notably in oncological and immunological diseases. In this work, we have employed a predictive AI-based platform to design the selective inhibitors considering the novelty, IP prior protection, and drug-likeness properties. Furthermore, selected top candidates from the initial iteration of the design were synthesized and chemically characterized utilizing 1H NMR and LC-MS. Employing a panel of biochemical (enzymatic) and cancer cell lines, the selected molecules were tested against these assays. In addition, we used artificial intelligence to predict and evaluate several critical IND-focused physicochemical and pharmacokinetics values of the selected molecules. A secondary objective of the current work was also to validate the sole role of BMX in animal models known to be mediated by BMX. More than 50 molecules were designed in the present study employing five novel discovered scaffolds. Two molecules were nominated for further IND-focused studies. Compound II showed promising in-vitro activity against BMX in both enzymatic assays compared to other kinases and in cancer cell lines with known BMX overexpression. Interestingly, compound II showed very favorable physicochemical and pharmacokinetics properties as predicted by the used platforms. The animal study further confirmed the sole role of BMX in the disease model. The current work provides promising data on a selective BMX inhibitor as a potential lead for therapeutics development, and the asset is currently in the optimization stage. Notably, the current study shows a framework for a combined approach employing both AI and experimentation that can be used by academic labs in their research programs to more streamline programs into IND-focused to be bridged easily for further clinical development with industrial partners.

α-Amylase purified and characterized from fenugreek (Trigonella foenum-graecum) showed substantial anti-biofilm activity against Staphylococcus aureus MTCC740.

Starch hydrolyzing α-amylase from germinated fenugreek (Trigonella foenum-graecum) has been purified 104-fold to apparent electrophoretic homogeneity with a final specific activity of 297.5 units/mg. SDS-PAGE of the final preparation revealed a single protein band of 47.5 kDa, supported by LC/MS analysis and size-exclusion chromatography on the Superdex 200 (ÄKTA-FPLC). α-Amylase exhibited maximum activity at pH 5.5. An activation energy (Ea) of 9.12 kcal/mol was found to exist in the temperature range of 20 to 90 °C. When substrate concentrations were evaluated between 0.5 and 10 mg/mL, the Km and Vmax values for starch were observed to be 1.12 mg/mL and 384.14 µmol/min/mg, respectively. The major substrate starch exhibited high specificity for fenugreek α-amylase. In the presence of EDTA (5 mM), the activity was lost, however, it could be largely reversed with the addition of calcium. Furthermore, an effort was made to assess the ability of fenugreek seed-derived partially purified (DEAE-cellulose enzyme) and purified α-amylase to disperse inside 48 h-old biofilms of Staphylococcus aureus MTCC740. The outcomes clearly demonstrated that the purified and partially purified α-amylase both exhibited strong biofilm dispersion activity.

Molecular rearrangement of bicyclic peroxy radicals is a key route to aerosol from aromatics.

The oxidation of aromatics contributes significantly to the formation of atmospheric aerosol. Using toluene as an example, we demonstrate the existence of a molecular rearrangement channel in the oxidation mechanism. Based on both flow reactor experiments and quantum chemical calculations, we show that the bicyclic peroxy radicals (BPRs) formed in OH-initiated aromatic oxidation are much less stable than previously thought, and in the case of the toluene derived ipso-BPRs, lead to aerosol-forming low-volatility products with up to 9 oxygen atoms on sub-second timescales. Similar results are predicted for ipso-BPRs formed from many other aromatic compounds. This reaction class is likely a key route for atmospheric aerosol formation, and including the molecular rearrangement of BPRs may be vital for accurate chemical modeling of the atmosphere.