Sorting Nexin 24 is required for α-granule biogenesis and cargo delivery in megakaryocytes

Germline defects affecting the DNA-binding domain of the transcription factor FLI1 are associated with a bleeding disorder that is characterised by the presence of large, fused α-granules in platelets. We investigated whether the genes showing abnormal expression in FLI1-deficient platelets could be involved in platelet α-granule biogenesis by undertaking transcriptome analysis of control platelets and platelets harbouring a DNA-binding variant of FLI1. Our analysis identified 2276 transcripts that were differentially expressed in FLI1- deficient platelets. Functional annotation clustering of the coding transcripts revealed significant enrichment for gene annotations relating to protein transport, and identified Sorting nexin 24 (SNX24) as a candidate for further investigation. Using an iPSC-derived megakaryocyte model, SNX24 expression was found to be increased during the early stages of megakaryocyte differentiation and downregulated during proplatelet formation, indicating tight regulatory control during megakaryopoiesis. CRISPR-Cas9 mediated knockout (KO) of SNX24 led to decreased expression of immature megakaryocyte markers, CD41 and CD61, and increased expression of the mature megakaryocyte marker CD42b (p=0.0001), without affecting megakaryocyte polyploidisation, or proplatelet formation. Electron microscopic analysis revealed an increase in empty membrane-bound organelles in SNX24 KO megakaryocytes, a reduction in α-granules and an absence of immature and mature multivesicular bodies, consistent with a defect in the intermediate stage of α-granule maturation. Co-localisation studies showed that SNX24 associates with each compartment of α-granule maturation. Reduced expression of CD62P and VWF was observed in SNX24 KO megakaryocytes. We conclude that SNX24 is required for α-granule biogenesis and intracellular trafficking of α-granule cargo within megakaryocytes.

Evolution of a cis-acting SNP that controls Type VI Secretion in Vibrio cholerae

Mutations in regulatory mechanisms that control gene expression contribute to phenotypic diversity and thus facilitate the adaptation of microbes to new niches. Regulatory architecture is often inferred from transcription factor identification and genome analysis using purely computational approaches. However, there are few examples of phenotypic divergence that arise from the rewiring of bacterial regulatory circuity by mutations in intergenic regions, because locating regulatory elements within regions of DNA that do not code for protein requires genomic and experimental data. We identify a single cis-acting single nucleotide polymorphism (SNP) dramatically alters control of the type VI secretion system (T6), a common weapon for inter-bacterial competition. Tight T6 regulatory control is necessary for adaptation of the waterborne pathogen Vibrio cholerae to in vivo conditions within the human gut, which we show can be altered by this single non-coding SNP that results in constitutive expression in vitro. Our results support a model of pathogen evolution through cis-regulatory mutation and preexisting, active transcription factors, thus conferring different fitness advantages to tightly regulated strains inside a human host and unfettered strains adapted to environmental niches.

Status of probiotic regulations in Southeast Asia countries

This review summarises the key components of the available probiotic regulations in six Southeast Asia countries (Indonesia, Malaysia, Philippines, Singapore, Thailand and Vietnam). Diverse approaches have been undertaken by the countries in regulating the marketing and sale of probiotics in foods and health supplements. Only Indonesia, Malaysia, Philippines and Thailand have enacted specific regulations which include their respective legal definition of probiotics. Only Malaysia, Philippines and Thailand publish a list of microorganisms permitted to be used as probiotics in foods or health supplements; the approved microorganisms are not harmonised among these countries. All six countries allow the application for new microorganisms to be used, but have adopted differing requirements and approaches. A common requirement is that all applications must be accompanied by scientific data to demonstrate clinically that the microorganisms are safe and provide health benefits. All the countries, except Indonesia and Vietnam permit the use of a small number of pre-approved generic function claims. It is noted that the countries have different specific labelling requirement for products containing probiotics. The divergent probiotic regulations in the region, either for foods or health supplements, creates inconsistencies and difficulties for all stakeholders including regulators, academia, industries and consumers, as well as impacting trade among countries. This review highlights the importance of having regulatory control to ensure consumers have access to safe, genuine and efficacious probiotic products. We propose working towards a harmonised probiotics regulation in the region to enable further development and progress of probiotics in the region.

The time–space tactics of Chinese Buddhist and Taoist believers under state–religion tension

This study explores the experience of elderly rural Buddhist and Taoist believers in communist China where the ruling party has maintained decades-long regulatory control over religion. Based on ethnographic observation and oral histories, the analysis begins with how the actors made sense of and coped in their relationship with the state during the fieldwork period (May–June 2020) when state regulations restricted public religious practice because of COVID-19. The analysis then looks back on how practitioners experienced tightening state ideological control from the early 2010s to before COVID-19; further back at the religious revival during the opening and reform (1980s–2010s); and finally, the Cultural Revolution period (1960s–70s) when strict atheistic measures were imposed. Their narratives reveal the practical logic (habitus) which practitioners used to mediate their resistance against and compromise with the authoritarian state. Specifically, four logical modes that involve actors’ different time–space tactics were identified, namely state–religion disengagement, state–religion enhancement, religious (dis)enlightenment, and karma. The implications of these ostensibly conflicting modes of thinking in mediating the actors’ resistance–compliance interface in contemporary China are discussed.

Identification of hepatozon spp. In the population of target rods of the Chernobyl radiation and ecological biosphere reserve

The ecosystems that have developed in the exclusion zone after the accident at the Chernobyl nuclear power plant in 1986 have a number of features. Parasites, as part of an ecosystem, can act as indicators of the processes taking place in it. Mouse rodents play an important role in the preservation and transmission of pathogens of some, including zoonotic, diseases in the wild. Therefore, the study of parasites in murine rodents, especially those transmitted transmissively, makes it possible to understand the characteristics of the distribution and the level of threat of their transmission to other susceptible animals, including agricultural, small domestic animals and humans. We carried out a study of blood smears taken from murine rodents for the presence of pathogens of transmissible parasitic diseases. According to the results of the studies, the pathogen Hepatozoon spp. Was detected for the first time in Ukraine. in the population of murine rodents. Creating an exclusion zone is one of the most obvious and long-lasting consequences of the Chernobyl Nuclear Power Plant (CNPS) accident. Because of 90-95% of this territory lacks systematic human activity and regime equal to the reserve. Ecosystems formed in the exclusion zone are affected a number of key factors as radioactive contamination, succession, wills and climate change. Spatial heterogeneity of density, radionuclide composition and physical chemical forms of precipitation are a characteristic feature of radioactive pollution in the Chernobyl Zone. Much of the aquatic and terrestrial ecosystems in the Chernobyl Zone as of 1986 was an artificial or semi-artificial systems that were under the regulatory control of man. The removal of regulatory control has led to the inclusion of natural influences mechanisms. That is, a significant part of ecosystems is in a state far from equilibrium, where the processes of succession are active. The testamentary effect is the result of a radical economic contraction activities and the creation of a strict security regime. Given this dynamics ecosystems in the Exclusion Zone has a unique character. Parasites are part of the ecosystem, so they can act as indicators of the processes occurring in it. Mouse-like rodents play an important role in the preservation of agents some infectious diseases in the wild. It is known that rodents are reservoirs hosts at least 60 zoonotic diseases, playing an important role in their transmission and distribution. Not the last role in it is played by features biology and way of life of these animals. Study of parasites in murine rodents, especially those that transmitted transmissively, will make it possible to understand the peculiarities of distribution and the level of threat of their transmission to other susceptible animals, including agricultural, small pets and humans. The object of our study was blood smears taken from murine rodents: Apodemus agrarius, Apodemus flavicollis, Myodes glareolus and Apodemus spp. Catching mice for further sampling conducted at 3 three landfills. Mouse-like rodents are significantly affected by pathogens of infectious diseases, including those transmitted through blood-sucking arthropods. Therefore, the study of the presence and prevalence of transmissible agents in murine rodents is important in order to prevent the disease in other species of animals or humans. The purpose of the study was to study the role of rodents Chornobyl Excusion Zone as a reservoir of blood-borne diseases. The study was conducted during 2020-2021. Blood smears from rodents Apodemus agrarius, Apodemus flavicollis, Myodes glareolus and Apodemus spp, which were selected during the expedition from three landfills and transferred to the Department of Pharmacology, Parasitology and Tropical Veterinary Medicine of NULES of Ukraine, where they conducted further research. Blood smears were stained by the method of Romanowski-Gimza, Leuko-Diff and examined under an immersion microscope system. A total of 117 blood smears were collected from 117 murine rodents from three landfills. According to the results of microscopic examination in blood smears revealed parasites, which morphologically we attributed to Hepatozoon spp. This agent was detected in Ukraine for the first time. Agents had an oval body shape with a large nucleus and were located in the cytoplasm of leukocytes (mostly neutrophils, rarely lymphocytes). In total, hepatozoan agents were detected in blood smears from 13 studied animals, which was 11.11%. And the invasion of Hepatozoon spp. was found in murine rodents of the first landfill, in 7 rodents. The smallest of the murine rodents of the second landfill was found only in blood smears from 2 animals. In addition to hepatozoons in murine rodents, we also found agents of other diseases, such as Babesia spp., Rickettsia spp., Borrelia spp. and Trypanosoma spp.

Disruption of c-di-GMP signaling networks unlocks cryptic expression of secondary metabolites during biofilm growth in Burkholderia pseudomallei

The regulation and production of secondary metabolites during biofilm growth of Burkholderia spp. is not well understood. To learn more about the crucial role and regulatory control of cryptic molecules produced during biofilm growth, we disrupted c-di-GMP signaling in Burkholderia pseudomallei, a soil-borne bacterial saprophyte and the etiologic agent of melioidosis. Our approach to these studies combined transcriptional profiling with genetic deletions that targeted key c-di-GMP regulatory components to characterize responses to changes in temperature. Mutational analyses and conditional expression studies of c-di-GMP genes demonstrates their contribution to phenotypes such as biofilm formation, colony morphology, motility, and expression of secondary metabolite biosynthesis when grown as a biofilm at different temperatures. RNA-seq analysis was performed at varying temperatures in a ΔII2523 mutant background that is responsive to temperature alterations resulting in hypo- and hyper- biofilm forming phenotypes. Differential regulation of genes was observed for polysaccharide biosynthesis, secretion systems, and nonribosomal peptide and polyketide synthase (NRPS/PKS) clusters in response to temperature changes. Deletion mutations of biosynthetic gene clusters (BGCs) clusters 2, 11, 14 (syrbactin), and 15 (malleipeptin) in wild-type and ΔII2523 backgrounds also reveals the contribution of these BGCs to biofilm formation and colony morphology in addition to inhibition of Bacillus subtilis and Rhizoctonia solani. Our findings suggest that II2523 impacts the regulation of genes that contribute to biofilm formation and competition. Characterization of cryptic BGCs under differing environmental conditions will allow for a better understanding of the role of secondary metabolites in the context of biofilm formation and microbe-microbe interactions.

Early life energy expenditure deficits drive obesity in a mouse model of Alström syndrome

Alström syndrome is an extremely rare multi-system disorder for which early-onset childhood obesity is one of the cardinal features. Similar to humans with Alström syndrome, animal models with Alms1 loss of function mutations develop obesity, strongly supporting the notion that ALMS1/Alms1 is required for the regulatory control of energy balance across species. To determine which component(s) of energy balance are reliant on functional Alms1, we performed comprehensive energy balance phenotyping of the tvrm102 mouse model of Alström syndrome. We found that that adiposity gains occurred early and rapidly in male mice but much later in females. Rapid increases in body fat in males were, at least in part, due to a marked reduction in energy expenditure during early life and not due to any genotype-specific influence over energy intake. Energy intake did increase in a genotype-specific manner when mice were provided a palatable, high-energy diet, although this was not necessary for the initial establishment of obesity. Interestingly, the energy expenditure deficit observed in male Alms1-/-mice did not persist as mice age, suggesting that loss of Alms1 either causes a developmental delay in the mechanisms controlling early life energy expenditure, or that there is activation of compensatory mechanisms after obesity is established. Future studies will tease out how ALMS1/Alms1 modulates energy expenditure in early life and how sex moderates this process.

Development of Silica Nanoparticle Supported Imprinted Polymers for Selective Lysozyme Recognition

Protein imprinted MIPs show notable potential for applications in many analytical areas such as clinical analysis, medical diagnostics and environmental monitoring, but also in drug delivery scenarios. In this study, we present various modifications of two different synthesis routes to create imprinted core-shell particles serving as a synthetic recognition material for the protein hen egg white (HEW) lysozyme. HEW lysozyme is used as food additive E 1105 for preservation due to its antibacterial effects. For facilitating quality and regulatory control analysis in food matrices, it is necessary to apply suitable isolation methods as potentially provided by molecularly imprinted materials. The highest binding capacity achieved herein was 58.82 mg/g with imprinting factors ranging up to 2.74, rendering these materials exceptionally suitable for selectively isolating HEW lysozyme.