Preparative biocatalytic synthesis of substituted pyrroles remains challenging despite recent advances in enzymatic α-aminoketone generation. In this study, we report a concurrent chemoenzymatic cascade to disubstituted pyrroles based on threonine dehydrogenase (ThrDH)-catalyzed generation of aminoacetone from l-threonine that is intercepted in situ by Knorr pyrrole condensation with β-dicarbonyl compounds. To overcome cofactor limitations at high substrate concentrations, Escherichia coli ThrDH was coupled to an alcohol dehydrogenase from Rhodococcus ruber DSM 44541 (ADH-A), enabling dual-function of the β-dicarbonyl cosubstrate as both a hydride acceptor and pyrrole building-block. The whole-cell catalyst tolerated up to 8% (v/v) β-dicarbonyl and afforded pyrrole products in up to 90% yield and space-time yields of 4.5 g L[-1] h[-1]. Preparative synthesis on a 100 mL scale furnished 4.3 g of pyrrole (279 mM, 93% isolated yield). Among the products obtained, the cascade provides direct access to a reported sunitinib intermediate, illustrating its potential utility for pharmaceutical synthesis. This work establishes ThrDHs as outstanding biocatalysts for heterocycle synthesis and demonstrates that amino acid feedstocks can be efficiently converted into substituted pyrroles through a scalable chemoenzymatic cascade.

Sheep- and goatpox are highly contagious, transboundary viral diseases caused by capripoxviruses (CaPV), severely impacting small ruminant production and resulting in significant economic losses. Our study aimed to analyze the spatiotemporal SGP distribution using the available genome sequence data and to evaluate the recombination occurrence. For this, the WAHIS, WOAH and FAO databases were utilized for the epidemiological analysis of SGP outbreaks. The cross-correlation was calculated to assess the impact of massive animal movements associated with the Islamic holiday of Eid al-Adha on the SGP epizootic situation, and phylodynamic and phylogeographic analysis, as well as a recombination analysis were performed. A total of 1629 SGP outbreaks were reported to WAHIS during 2010-2024, with the majority occurring in Mongolia, the Balkan countries, Russia and the Middle East. Over 60% of all SGP outbreaks were associated with croplands or grasslands, with the highest proportion corresponding to animal densities of 10-50 head/km[2]. Statistically significant positive cross-correlation (p < 0.05) was identified between the month of the Eid al-Adha celebration and the number of SGP outbreaks in Russia, Mongolia, Bulgaria and Tajikistan, while in Greece and China no significant correlation was found. The inferred goat pox virus (GTPV) transmission pathways from China to Vietnam and from India to Bangladesh; for the sheeppox virus SPPV, the routes between Kazakhstan and Russia, Kazakhstan and India, as well as between Russia and China, had the strongest Bayes factor support. Intra-specific recombination events were not detected for the SSPV and GTPV datasets. However, inter-specific CaPV recombination analysis identified a single recombination event in GTPV. Therefore, the use of molecular epidemiological tools, along with the time-calibrated phylodynamic and phylogeographic analyses, has significant applications in the local and international surveillance of the occurrence of SGP outbreaks and for identification of potential recombination events.

Human orf is a zoonotic parapoxvirus infection that is typically self-limiting with resolution in 3-6 weeks but may cause severe disease in immunocompromised patients. We report a kidney transplant recipient who developed a rapidly progressive fingertip lesion following contact with raw meat during Eid al-Adha. Initial misdiagnosis and surgical excision were followed by marked lesion progression, mimicking a vascular tumor. Histopathology confirmed orf virus infection. However, PCR testing was not done as the test was not available. Topical imiquimod was not initiated due to an active wound. Weekly liquid nitrogen cryotherapy as monotherapy resulted in complete resolution after nine sessions without recurrence. Further modifications/reduction in immunosuppressive medication was not needed. This case highlights cryotherapy as a potential treatment option for orf infection in transplant recipients.

Thermoanaerobacterium saccharolyticum is an anaerobic, thermophilic bacterium that has been proposed for use in consolidated bioprocessing in coculture with cellulolytic bacteria such as Clostridium thermocellum for ethanol production. Although the mixed acid fermentation of both species has been engineered to produce ethanol as the major fermentation product, the maximum titer produced thus far by C. thermocellum is half that produced by T. saccharolyticum. This has motivated us to understand the mechanistic basis of the robust T. saccharolyticum ethanol pathway, so that key features can be recapitulated in C. thermocellum. Previously, we characterized the individual role of the main genes responsible for electron transfer in the ethanol production of T. saccharolyticum. However, the consequences of the combined loss of function of these genes has not been investigated, nor has the way in which fermentative metabolism adapts to such constraints. Here, we combined knockouts of ferredoxin nicotinamide oxidoreductases (nfnA and nfnB) and hydrogenases (hydA and hfsD). We showed that these genetic modifications together impair growth and decrease electron transfer from reduced ferredoxin, thereby redirecting flux from the pyruvate ferredoxin oxidoreductase enzyme to the pyruvate formate lyase enzyme. We also performed adaptive evolution of these mutants to rescue their growth and observed a mutation in the alcohol dehydrogenase adhA gene. We determined that this point mutation causes a structural change that impairs the AdhA specificity for the NADPH cofactor and increases NADH-linked activity to restore redox balance. These findings consolidate our understanding of electron transfer pathways in this organism.IMPORTANCEThermoanaerobacterium saccharolyticum has the potential to be used for the conversion of lignocellulose-derived sugars into bioethanol via consolidated bioprocessing in cocultures with Clostridium thermocellum or via transfer of its ethanol production pathway to the same bacterium. However, attempts to transfer the pathway to increase ethanol titer have not yet been successful. A deeper understanding of how electron transfer pathways operate in this thermophile and how they adapt to metabolic disturbances (such as the absence of metabolic pathways for ferredoxin oxidation, for example) will improve our ability to engineer this organism for increased product formation and improve our ability to transfer its remarkable ethanol production phenotype to other microbes. These, in turn, have potential benefits for sustainable production of fuels and chemicals.

A novel strain with strong antagonistic and anti-aflatoxigenic activity against Aspergillus flavus was isolated from a marine environment, highlighting its potential for biocontrol applications in food and agriculture.

Membership with professional organizations, including the American Dental Hygienists' Association (ADHA), plays an important role in supporting dental hygienists through continuing education, networking, advocacy, and professional identity development. Despite these established benefits, membership rates in professional associations remain low. The purpose of this pilot study was to examine the trends influencing membership decisions among licensed dental hygienists to inform the design of a larger, more representative national investigation. A cross-sectional electronic survey was pilot tested for content validity and distributed to dental hygienists in four states using convenience sampling. Participants provided demographic information, current membership status in any professional organizations (including the ADHA), and perceptions of benefits and barriers to engagement. Descriptive statistics and one-way ANOVA were used to analyze initial trends. Among the respondents (n=250), 25% reported current ADHA membership; most identified advocacy (82%), access to continuing education (76%), and networking (61%) as meaningful benefits of belonging to a professional organization. In contrast, non-members cited high costs of membership (52%), lack of employer reimbursement (38%), and time constraints (12%) as primary barriers. Non-members were significantly younger (p<0.001) and graduated more recently (p<0.001) than members. Individuals with higher educational attainment were more likely to maintain membership in a professional organization (p<0.001). Open-ended responses revealed varied perceptions of organizational relevance, accessibility, and visibility. These preliminary findings highlight key factors that may shape engagement with professional organizations, including the ADHA. The results underscore the need for expanded sampling, strengthened early-career exposure, and further qualitative exploration. Findings from this pilot will guide the development of a larger national study aimed at understanding and improving professional organization membership among dental hygienists.

Hemostatic fabric is widely employed as a clinical hemostatic agent, yet its clinical efficacy is significantly limited by blood permeation through the fabric, leading to persistent hemorrhage. To address this critical challenge, an effective hemostatic nanofabric composed of catechol-modified oxidized hyaluronic acid (COHA) and adipic dihydrazide modified hyaluronic acid (ADHA) has been reported. The reported COHA/ADHA nanofabric exhibits exceptional breathability and mechanical flexibility while demonstrating unique hemostatic functionality. Upon blood contact, Schiff base-mediated crosslinking between COHA and ADHA nanofibers induces rapid inter-fiber bonding, creating a dense network that establishes superior blood barrier functionality while simultaneously enhancing erythrocyte and platelet aggregation. Moreover, the COHA/ADHA nanofabric can adhere to the tissue to effectively seal the wound. Through integrated ex vivo and in vivo assessments it was found that the COHA/ADHA nanofabric exhibits enhanced hemostatic efficacy compared to conventional materials including cotton gauze, Combat Gauze™, and Surgicel® Original. Furthermore, the nanofabric demonstrates biodegradability, histocompatibility, and accelerated wound healing rates. This biomimetic design strategy creates a new pathway for developing high-performance hemostatic materials with integrated barrier function and physiological compatibility.

Chronic respiratory diseases, including chronic obstructive pulmonary disease (COPD), asthma, pneumoconiosis, interstitial lung disease (ILD) and pulmonary sarcoidosis, are major global causes of mortality and morbidity. Although the COVID-19 pandemic has influenced acute respiratory health, its impact on chronic respiratory conditions remains unclear. We estimated the global, regional and national burden of chronic respiratory diseases from 1990 to 2023, including risk factors, and evaluated how these burdens have shifted during the COVID-19 pandemic using the Global Burden of Disease Study 2023. In 2023, chronic respiratory diseases accounted for 569.2 million (95% uncertainty interval (UI), 508.8-639.8) cases and 4.2 million (3.6-5.1) deaths. The age-standardized death rate declined by 25.7% globally from 1990 to 2023 despite an increase in ILD and pulmonary sarcoidosis. Mortality declined in younger males, especially for asthma, whereas older adults experienced a rise in ILD and pulmonary sarcoidosis. Smoking was the primary risk factor for COPD, whereas high body mass index and silica exposure were key risk factors for asthma and pneumoconiosis. During the pandemic, the incidence of chronic respiratory diseases increased modestly, but the decline in mortality rates became more pronounced, highlighting the need for sustained global attention and action to address their long-term burden.

Cashew is a key economic crop in coastal Kenya, yet its production faces significant challenges due to post-harvest fungal contamination, particularly by Aspergillus flavus and Aspergillus aculeatus. These fungi are known producers of mycotoxins, including aflatoxins, potent carcinogenic and mutagenic compounds that pose serious food safety and public health risks. Despite growing concerns, the genomic architecture, metabolic potential, and ecological adaptations of these fungi in cashew-growing regions remain poorly understood. This study investigated the genetic diversity, aflatoxin biosynthetic gene clusters (BGCs), secondary metabolite profiles, and carbohydrate-active enzyme (CAZyme) repertoires of aflatoxigenic A. flavus and compared them with non-aflatoxigenic A. aculeatus isolates obtained from cashew nuts collected in Kilifi, Kwale, and Lamu counties. A total of 18 fungal isolates (16 A. flavus, 2 A. aculeatus) were cultured and subjected to whole-genome sequencing using the Illumina platform. Genome mining and comparative analyses were performed using OrthoFinder, antiSMASH, and dbCAN. Phylogenomic analysis revealed five distinct clades among A. flavus isolates, suggesting substantial intraspecific diversity and potential regional adaptation, while A. aculeatus formed a single monophyletic clade. Comparative analysis of aflatoxin BGCs in A. flavus revealed notable structural variation, including gene deletions (aflT, omtA), insertions, and rearrangements, with conservation observed for core genes such as ordB, moxY, avfA, and adhA; however, the regulatory gene aflR was not conserved across all isolates. All the 2 A. aculeatus isolates lacked aflatoxin biosynthetic genes, indicating they are not aflatoxigenic. Both species harbored diverse SMBGCs, including PKS, NRPS, and strain-specific clusters such as YWA1, fusarin, and aspergillic acid, suggesting potential for co-production of multiple bioactive compounds. CAZyme profiling revealed abundant glycoside hydrolases and auxiliary activity enzymes, underscoring adaptation to pectin-rich cashew substrates. Despite similar genome sizes and GC content, species-specific differences in carbohydrate metabolism and secondary metabolite pathways indicate ecological plasticity. Our findings demonstrate that A. flavus and A. aculeatus populations in coastal Kenya are genetically diverse, metabolically versatile, and shaped by local environmental and anthropogenic pressures. The study provides essential genomic insights for region-specific aflatoxin risk assessment and highlights the need for careful screening of atoxigenic strains in biocontrol applications, as well as further exploration of cryptic BGCs and CAZyme functions in post-harvest fungal ecology.

1,4-Butanediol (1,4-BDO) is a key commodity chemical used in the production of various products, including plastics, fibers, and polyesters. Here, a potent microbial platform for 1,4-BDO production from glucose via the L-glutamate-derived CoA-independent pathway was developed using Corynebacterium glutamicum as the host strain. Introducing a CoA-independent 1,4-BDO biosynthesis pathway and removing a metabolic flux bypass, the resulting initial strain produced 2.20 g/L of 1,4-BDO from glucose. Upon prolonged cultivation, C. glutamicum showed an endogenous capacity to degrade 1,4-BDO. Transcriptomic analysis was conducted to identify genes potentially involved in this pathway. Among them, fadH, aldH, and adhA were verified as the key enzymes for 1,4-BDO degradation. The deletion of three dehydrogenase genes from the initial strain removed the product degradation and achieved an improved 1,4-BDO titer of 3.14 g/L. Finally, fed-batch fermentation of the final strain was conducted, resulting in 13.4 g/L of 1,4-BDO production. This work demonstrates the first successful development of C. glutamicum as a potent platform strain for the biosynthesis of 1,4-BDO.

Zymomonas mobilis has an extremely high specific glucose uptake rate and produces ethanol at high yields and productivities. For synthesizing products beyond ethanol in Z. mobilis, reducing ethanol yields is a major challenge. Previous efforts have sought to decrease PDC activity to minimize ethanol formation. Here, we sought to modulate ADH (encoded by adhA and adhB genes) activity to redirect flux away from ethanol. We found that deletion of adhB combined with co-expression of a heterologous NAD(P) regenerating enzyme (L-LDH) diverted more than 50 % of carbon flux away from ethanol formation. The sequential deletion of adhA in the adhB-deleted strain led to ∼90 % reduction in ADH activity. During batch growth, the strain showed ∼90 % reduction in ethanol titres (27 mM) compared to WT (209 mM) and significant flux redirection toward L-LA formation (169 mM). The results demonstrate that modulation of ADH activity by deletion of adhA and adhB genes is an effective strategy for rediverting flux toward products beyond ethanol in Z. mobilis.