Two new 22-membered macrolides, dactylide D (1) and dactylide E (2), featuring substituted amino acid or a contiguous dipropionate unit, along with the known dactylosporolide B (3), were purified from Dactylosporangium aurantiacum ATCC 23491. Their structures were determined through comprehensive spectroscopic analysis and comparison with related congeners. Compound 1 features an N-acetylalanyl side chain, while compound 2 bears an unusual (E)-7-amino-oxohept-5-enoic acid moiety, along with C-7 oxidation and loss of the tetrahydropyran ring. NOE correlations indicated that the stereochemistry of the macrolactone cores in 1 and 2 is conserved relative to dactylide B. These findings expand the structural diversity of polyol macrolides from D. aurantiacum, underscoring the strain’s potential for natural product discovery.

This study explores the chemical ecology of the halophytic plant Salicornia brachiata and its associated Bacillus endophytes using LC-MS-based metabolomics. A total of 43 Bacillus strains were isolated and screened for the production of bioactive lipopeptides, with 26 strains showing both lipopeptide production and antimicrobial activity, particularly against Mycobacterium smegmatis MTCC6. Fermentation of a potent endophyte, Bacillus subtilis NPRoot-3, yielded purified surfactins and fengycins, which displayed significant inhibitory activity with low IC50 values against Mycobacterium smegmatis. LC-MS analysis of aqueous and methanolic extracts of S. brachiata revealed molecular features putatively corresponding to surfactins, fengycins, and bacillibactins, indicating the presence of similar bioactive compounds in both the host plant and its endophytic microbiome. These findings suggest that the antimycobacterial properties of S. brachiata may be partly attributed to metabolites produced by its endophytes. Overall, this study highlights cooperative defense mechanisms between the plant and its endophytes, confirms the role of endophyte-derived secondary metabolites in shaping plant bioactivity and offers promising leads for the discovery of novel antimycobacterial agents.

Outbreaks of Bordetella pertussis (BP), the causative agent of whooping cough, continue despite broad vaccination coverage and have been increasing since vaccination switched from whole-BP (wP) to acellular BP (aP) vaccines. wP vaccination has been associated with more durable protective immunity and an induced Th1 polarized memory T cell response. Here, we profile, by a multi-omics approach, the immune response of 30 wP and 31 aP-primed individuals and identify correlates of T cell polarization before and after Tdap booster vaccination. We find that early transcriptional changes indicating an interferon response, followed by an increase in plasma IFN-γ and interferon-induced chemokine levels (peaking at day 1-3 post-booster), correlate best with the Th1 polarization of the vaccine-induced memory T cell response on day 28. Our studies indicate that wP-primed individuals maintain their Th1 polarization through this early memory interferon response. This suggests that stimulating the interferon pathway during vaccination could be an effective strategy to elicit a predominant Th1 response in aP-primed individuals that protects better against infection.

Seaweed are abundant in primary and secondary metabolites, including phenolics, flavonoids, proteins, carbohydrates, and amino acids. Beyond their traditional use as food, their diverse chemical composition makes them promising candidates for pharmaceutical and nutraceutical applications. This study investigates antioxidant potential, phytochemical composition, and bioactive metabolites of six abundant seaweed species collected off the Indian coastline. Comprehensive phytochemical analyses were conducted to quantify phenolics, flavonoids, carbohydrates, proteins, amino acids, and ionomic composition to assess macronutrient and micronutrient content. Gas Chromatography-Mass Spectrometry (GC-MS) was employed to profile bioactive compounds in the seaweed extracts. Among the seaweeds analysed, Grateloupia indica (Rhodophyta) revealed the highest antioxidant activity (94 % at 1 mg mL-1), along with the highest phenolic content of 19.04 +/- 3.05 Tannic acid equivalent (TAE) g-1 of dry weight (DW) of extract and flavonoid content of 9.59 +/- 0.73 Rutin equivalent (RE) g-1 bohydrates, and beneficial elemental composition. GC-MS analysis revealed the presence of various metabolites with known antioxidant, anticancer, and antimicrobial properties. Multivariate analysis facilitated the understanding of seaweed chemosystematics and phytochemical profiling of the species. The study highlights the significant nutraceutical potential of seaweeds and provides a foundation for their further exploration into their pharmaceutical and nutraceutical applications. of DW of extract. All six seaweeds demonstrated substantial levels of proteins, essential amino acids, car-

The present study highlights the detailed investigation of the physico-chemical characteristics of the anti-cancer enzyme L-aspraginase (LA) from the red seaweed Gracilaria dura-associated Bacillus licheniformis strain. We performed the enzyme purification, cytotoxicity assays, toxicity analysis on Caenorhabditis elegans, and kinetics characterisation. We standardized a two-step enzyme purification strategy, involving ammonium sulfate precipitation followed by Sephadex column chromatography in this study to achieve high enzyme purity. The purified LA exhibited high specificity for L-asparagine with a low Km (0.014 ± 0.0006 mM), absence of L-glutaminase activity; and a high level of enzyme activity (229 IU ml−1). This LA variant demonstrated cytotoxicity against HEK 293 cells and it did not affect the normal growth and life span of C. elegans. There fore this study underpins the possibility of utilising this LA variant for clinical applications. The enzyme is functional at a wide pH and temperature range, with optimal conditions aligning with the physiological levels of human blood (pH 7.37 °C). Moreover, the enzyme is secreted extracellularly leading to a simple downstream processing. We assume that the potential immunogenecity of this enzyme in humans to be minimal or absent, however it should be substantiated through assessments on higher organisms. Our findings not only provides useful insights into the potential of marine LA in human therapeutics but also shed light into the possibility of obtaining high value products from seaweed associated bacteria. This study enhances the scope of carrying out research on the seaweed ecology and interactions in the marine environment.

Systems vaccinology studies have been used to build computational models that predict individual vaccine responses and identify the factors contributing to differences in outcome. Comparing such models is challenging due to variability in study designs. To address this, we established a community resource to compare models predicting B. pertussis booster responses and generate experimental data for the explicit purpose of model evaluation. We here describe our second computational prediction challenge using this resource, where we benchmarked 49 algorithms from 53 scientists. We found that the most successful models stood out in their handling of nonlinearities, reducing large feature sets to representative subsets, and advanced data preprocessing. In contrast, we found that models adopted from literature that were developed to predict vaccine antibody responses in other settings performed poorly, reinforcing the need for purpose-built models. Overall, this demonstrates the value of purpose-generated datasets for rigorous and open model evaluations to identify features that improve the reliability and applicability of computational models in vaccine response prediction.

Diabetes mellitus and obesity are prevalent lifestyle-related diseases worldwide. Salicornia brachiata, a halophytic plant usually found in salt marshes with a succulent stem portion comprising seeds and roots beneath the ground. Tender succulent stem part has been consumed as a salad, and vegetable traditionally with reported antioxidant, anti-inflammatory, and cytotoxic biological activities. Previous reports described a presence of oil content and fatty acid composition in the seeds of Salicornia brachiata without highlighting its pharmacological effects. Since the fatty acids are ascribes to the anti-diabetic and lipid lowering potential, the current study aims to analyse the seed oil of edible halophyte Salicornia brachiata to explore its potential in managing diabetes and obesity through in silico, in vitro, and in vivo methodologies. Soxhlet extracted seed oil was analysed by gas chromatography-mass spectrometry which revealed significant presence of linoleic acid. The in silico analysis of linoleic acid with α-glucosidase enzyme, indicated a promising binding affinity to amino acid residues of enzyme, suggesting the formation of a stable protein-ligand complex. Subsequently, the in vitro studies demonstrated that the extracted oil inhibited α-glucosidase enzyme, corroborating the in silico findings. In vivo experiments conducted on two months old C57BL/6J male mice (n = 6) with 20–25 g body weight provided with a normal diet at a dosage of 10 mg/kg body weight described significant reductions in blood parameters, such as glucose, cholesterol, high-density lipoprotein, low-density lipoprotein, and triglycerides, highlighting the oil’s anti-hyperglycaemic and anti-hyperlipidaemic properties. To the best of our knowledge, this is the first comprehensive report paving the way for further research into its health benefits.

The present study explores the microbial community associated with the industrially important red seaweed Gracilaria dura to determine the diversity and biotechnological potential through culture and metagenomics approaches. In the first part of the investigation, we isolated and characterized 75 bacterial morphotypes, with varied colony characteristics and metabolic diversity from the wild seaweed. Phylogenetic analysis identified isolates in Proteobacteria, Firmicutes, and Actinobacteria, with Bacillus sp. being prevalent. B. licheniformis and Streptomyces sp. were notable in producing important enzymes like L-asparaginase, and polysaccharide lyases. Antimicrobial activity was significant in 21

Tuberculosis (TB) is one of the oldest leading health diseases of mankind, caused by the bacteria Mycobacterium tuberculosis. It is still the major cause of high mortality and morbidity worldwide from a single infectious agent. Plant species belonging to the genus Halosarcia (formerly known as Salicornia ) are known for their various medicinal properties like antioxidant, hepatoprotective, immunomodulatory, antimicrobial, etc., but not evaluated for their antimycobacterial property. The present study aimed to investigate the potential of Halosarcia indica (Willd.) (formerly known as Salicornia brachiata Roxb.), a halophytic plant, a source of antitubercular agents. The study involved the preparation of various extracts from different parts of the plant and the evaluation of their inhibitory activities against Mycobacterium tuberculosis H37Rv using in vitro and in vivo assays. It was observed that the aqueous extract of the roots had the highest anti -TB activity with minimum inhibitory concentration (MIC) of 50 m g/mL and a ten -fold reduction of M. tuberculosis bacilli in the lungs of treated mice. Further fractionation of aqueous extract of roots using butanol, MeOH:CHCl 3 , MeOH, MeOH:H 2 O, and H 2 O yielded fractions with varying anti -TB activity. The subfraction SF3K obtained from fraction 2 (F-2) showed the highest potency, with MIC <= 3.125 m g/mL and an increase of 5 days in the mean survival time of M. tuberculosis -infected and treated (with SF3K) mice. This subfraction was found to be non-cytotoxic ( in vitro ) and non-toxic (during in vivo ef ficacy evaluation). The present study, for the first time, provides scienti fic evidence for the use of H. indica in the treatment of tuberculosis. (c) 2024 SAAB. Published by Elsevier B.V. All rights reserved.

Zinc oxide nanoparticles (ZnO-NPs) are recognized as highly promising materials with applications in diverse fields, from therapeutics to agriculture. Hence, the present study aimed to biosynthesize ZnO-NPs employing the endophytic Streptomyces sp. 4VPT5-9 isolated from the halophytic plant Salicornia brachiata and evaluate the biological activity against plant pathogen Fusarium oxysporum NCIM 1008 and human breast (MDA-MB-231) and colorectal cancer (HCT-116) cell lines. The cell-free supernatant of the strain 4VPT5-9 was used as a greener attempt for the biosynthesis of ZnO-NPs. Biosynthesized ZnO-NPs exhibited an absorption peak at 287 nm. The hexagonal crystalline structure of the biosynthesized ZnO-NPs was validated through X-ray diffraction studies. These biosynthesized ZnO-NPs exhibited an irregular shape and varied in size, ranging between 120 and 150 nm with d-spacing value 0.26 nm. Moreover, FI-IR analysis showed different absorption peaks confirming the presence of different functional groups and formation of biosynthesized ZnO-NPs. XPS analysis confirmed the presence of Zn(II)O at different varied bending energies. The antifungal assay revealed that biosynthesized ZnO-NPs showed a pronounced inhibitory zone (12 mm) and the least MIC value of 60 µg/mL against plant pathogen F. oxysporum NCIM 1008. Moreover, the biosynthesized ZnO-NPs showed significant anticancer activity against MDA-MB-231 human breast cancer and HCT-116 human colorectal cell lines in a dose- and time-dependent manner. From this study, it is evident that an eco-conscious approach could pave a novel path for biosynthesis of ZnO-NPs through a halophytic plant-associated actinomycetes.

Systems vaccinology studies have identified factors affecting individual vaccine responses, but comparing these findings is challenging due to varying study designs. To address this lack of reproducibility, we established a community resource for comparing Bordetella pertussis booster responses and to host annual contests for predicting patients' vaccination outcomes. We report here on our experiences with the “dry-run” prediction contest. We found that, among 20+ models adopted from the literature, the most successful model predicting vaccination outcome was based on age alone. This confirms our concerns about the reproducibility of conclusions between different vaccinology studies. Further, we found that, for newly trained models, handling of baseline information on the target variables was crucial. Overall, multiple co-inertia analysis gave the best results of the tested modeling approaches. Our goal is to engage community in these prediction challenges by making data and models available and opening a public contest in August 2024.

BACKGROUNDPatients hospitalized for COVID-19 exhibit diverse clinical outcomes, with outcomes for some individuals diverging over time even though their initial disease severity appears similar to that of other patients. A systematic evaluation of molecular and cellular profiles over the full disease course can link immune programs and their coordination with progression heterogeneity.METHODSWe performed deep immunophenotyping and conducted longitudinal multiomics modeling, integrating 10 assays for 1,152 Immunophenotyping Assessment in a COVID-19 Cohort (IMPACC) study participants and identifying several immune cascades that were significant drivers of differential clinical outcomes.RESULTSIncreasing disease severity was driven by a temporal pattern that began with the early upregulation of immunosuppressive metabolites and then elevated levels of inflammatory cytokines, signatures of coagulation, formation of neutrophil extracellular traps, and T cell functional dysregulation. A second immune cascade, predictive of 28-day mortality among critically ill patients, was characterized by reduced total plasma Igs and B cells and dysregulated IFN responsiveness. We demonstrated that the balance disruption between IFN-stimulated genes and IFN inhibitors is a crucial biomarker of COVID-19 mortality, potentially contributing to failure of viral clearance in patients with fatal illness.CONCLUSIONOur longitudinal multiomics profiling study revealed temporal coordination across diverse omics that potentially explain the disease progression, providing insights that can inform the targeted development of therapies for patients hospitalized with COVID-19, especially those who are critically ill.TRIAL REGISTRATIONClinicalTrials.gov NCT04378777.FUNDINGNIH (5R01AI135803-03, 5U19AI118608-04, 5U19AI128910-04, 4U19AI090023-11, 4U19AI118610-06, R01AI145835-01A1S1, 5U19AI062629-17, 5U19AI057229-17, 5U19AI125357-05, 5U19AI128913-03, 3U19AI077439-13, 5U54AI142766-03, 5R01AI104870-07, 3U19AI089992-09, 3U19AI128913-03, and 5T32DA018926-18); NIAID, NIH (3U19AI1289130, U19AI128913-04S1, and R01AI122220); and National Science Foundation (DMS2310836).

The discovery of new natural products has become more challenging because of the re-isolation of compounds and the lack of new sources. Microbes dwelling in extreme conditions of high salinity and temperature are huge prospects for interesting natural metabolites. In this study, the endophytic bacteria Bacillus velezensis 7NPB-3B isolated from the halophyte Salicornia brachiata was screened for its biofilm inhibition against methicillin-resistant Staphylococcus aureus (MRSA). The fractionation of the crude extract was guided by bioassay and LC-HRMS-based metabolomics using multivariate analysis. The 37 fractions obtained by high-throughput chromatography were dereplicated using an in-house MS-Excel macro coupled with the Dictionary of Natural Products database. Successive bioactivity-guided separation yielded one novel compound (1), a diketopiperazine (m/z 469.258 [M − H]−) with an attached saturated decanoic acid chain, and four known compounds (2–5). The compounds were identified based on 1D- and 2D-NMR and mass spectrometry. Compounds 1 and 5 exhibited excellent biofilm inhibition properties of >90% against the MRSA pathogen at minimum inhibition concentrations of 25 and 35 µg/mL, respectively. The investigation resulted in the isolation of a novel diketopiperazine from a bacterial endophyte of an untapped plant using an omics approach.

The study explored Simarouba glauca DC. for mosquito larvicidal potential by performing bioactivity-guided chemical investigation of its root extract resulting in isolation of the known bioactive metabolite glaucarubinone (1). Mosquito larvicidal activity of glaucarubinone (1) against the three vector species viz. Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus was determined using a modified WHO 2005 protocol. It was observed that Culex quinquefasciatus larvae were the most susceptible species with LC50 13.88 ppm and LC90 70.01 ppm followed by Aedes aegypti and Anopheles stephensi at 24 h of exposure. The mode of action as observed microscopically is the lysis of midgut and thorax cells of the third instar larvae. The crystal structure of the glaucarubinone (1) is reported for the first time using X-ray crystallography. This phytochemical product has the potential to act as a green alternative to existing chemical-based insecticides for integrated vector management.

The brain’s ability to adapt through structural rewiring during developmental transitions is a fundamental aspect of neuroscience. Our study conducts a detailed comparison of Drosophila melanogaster ‘s brain networks during larval and adult stages, revealing significant changes in neuronal wiring during developmental phases. The degree distribution of the larval brain deviates significantly from power-law behavior and fits well with the Weibull distribution. In contrast, the adult brain exhibits power-law behavior in its degree distribution, with the exponent for the out-degree distribution lying in the scale-free regime and the exponent for the in-degree distribution being close to this regime. This difference reflects a change in the robustness of brain development from larval to adult phases. The core of these networks also changes during development in terms of their cell composition and topological influence. The larval core comprises Mushroom Body neurons, while the adult core mainly has Antennal Lobe neurons. Moreover, all the core neurons in the larval brain are also part of the rich-club neurons, a group of neurons with high in/out degrees that are well connected, whereas the same is not true for the adult brain network. Additionally, the core of the larval brain displays a more heterogeneous connectivity profile in its second-order neighbors compared to adult brain neurons, indicating greater diversity in larval brain connectivity. Our work stands as a step forward in understanding the rewiring of brain networks across the life stages of Drosophila melanogaster . ### Competing Interest Statement The authors have declared no competing interest.

Three new 22-membered polyol macrolides, dactylides A−C ( 1−3 ), were isolated from Dactylosporangium aurantiacum ATCC 23491 employing repeated chromatographic separations, and their structures were established based on detailed analysis of NMR and MS data. The relative configurations at the stereocenters were established via vicinal 1 H– 1 H coupling constants, NOE correlations, and by application of Kishi’s universal NMR database. In order to get insights into the biosynthetic pathway of 1−3 , the genome sequence of the producer strain D. aurantiacum was obtained and the putative biosynthetic gene cluster encoding their biosynthesis was identified through bioinformatic analysis using antiSMASH. Compounds 1−3 showed significant in-vitro antimycobacterial and cytotoxic activity.

Antibiotics are required for everyday life functions, and most of them are naturally decorated with various sugars. Glycosylation of natural products leads to enhanced physicochemical and pharmacological properties such as solubility, bioavailability, stability, and bioactivity. Glycosyltransferase (GT) enzymes catalyze glycosidic bond formation between a donor sugar molecule and hydroxyl group of acceptor molecules. The structural basis of enzymes provides an excellent opportunity for the genetic engineering of these enzymes. Diversifying natural products through glycosyltransferases catalyzed by glycosylation is exceptional, which is impossible to achieve using chemical synthesis. This chapter thus tries to give an insight into the classification of GTs, their role in glycosylated antibiotics, and different strategies for glycosylation.

Gracilaria dura associated bacterial diversity revealed 75 isolates, which were screened for L-asparaginase (LA) production. Out of the three positive isolates, upon secondary screening, the strain 4T1C26E identified as Bacillus licheniformis registered the highest (25.2 IU ml- 1) LA activity. This isolate was further subjected to yield improvement by optimizing process parameters through Response Surface Methodology (RSM) and Artificial Neural Networking-Particle Swarm Optimization (ANN-PSO). A 2.5-fold increase in production by using RSM while 4-fold increase by using ANN with respect to un-optimized conditions could be achieved. In the preliminary toxicity analysis on the standard animal model Caenorhabditis elegans, the worms treated with crude LA showed similar growth pattern to the control worms marking the enzyme safe for human applications. The important benefit of LA obtained here is that it possesses no L-glutaminase activity and is free of endotoxins which is suitable for downstream processing thereby reducing chances of the immunogenic responses.