Comprehensive identification of pathogenic microbes and antimicrobial resistance genes in food products using nanopore sequencing-based metagenomics.

Foodborne pathogens, particularly antimicrobial-resistant (AMR) bacteria, remain a significant threat to global health. Given the limitations of conventional culture-based approaches, which are limited in scope and time-consuming, metagenomic sequencing of food products emerges as a promising solution. This method provides a fast and comprehensive way to detect the presence of pathogenic microbes and antimicrobial resistance genes (ARGs). Notably, nanopore long-read sequencing provides more accurate bacterial taxonomic classification in comparison to short-read sequencing. Here, we revealed the impact of food types and attributes (origin, retail place, and food processing methods) on microbial communities and the AMR profile using nanopore metagenomic sequencing. We analyzed a total of 260 food products, including raw meat, sashimi, and ready-to-eat (RTE) vegetables. Clostridium botulinum, Acinetobacter baumannii, and Vibrio parahaemolyticus were identified as the top three foodborne pathogens in raw meat and sashimi. Importantly, even with low pathogen abundance, higher percentages of samples containing carbapenem and cephalosporin resistance genes were identified in chicken and RTE vegetables, respectively. In parallel, our results demonstrated that fresh, peeled, and minced foods exhibited higher levels of pathogenic bacteria. In conclusion, this comprehensive study offers invaluable data that can contribute to food safety assessments and serve as a basis for quality indicators.

Ecology and risks of the global plastisphere as a newly expanding microbial habitat.

Plastic offers a new niche for microorganisms, the plastisphere. The ever-increasing emission of plastic waste makes it critical to understand the microbial ecology of the plastisphere and associated effects. Here, we present a global fingerprint of the plastisphere, analyzing samples collected from freshwater, seawater, and terrestrial ecosystems. The plastisphere assembles a distinct microbial community that has a clearly higher heterogeneity and a more deterministically dominated assembly compared to natural habitats. New coexistence patterns-loose and fragile networks with mostly specialist linkages among microorganisms that are rarely found in natural habitats-are seen in the plastisphere. Plastisphere microbiomes generally have a great potential to metabolize organic compounds, which could accelerate carbon turnover. Microorganisms involved in the nitrogen cycle are also altered in the plastisphere, especially in freshwater plastispheres, where a high abundance of denitrifiers may increase the release of nitrite (aquatic toxicant) and nitrous oxide (greenhouse gas). Enrichment of animal, plant, and human pathogens means that the plastisphere could become an increasingly mobile reservoir of harmful microorganisms. Our findings highlight that if the trajectory of plastic emissions is not reversed, the expanding plastisphere could pose critical planetary health challenges.

Role of Legionella pneumophila outer membrane vesicles in host-pathogen interaction.

Legionella pneumophila is an opportunistic intracellular pathogen that inhabits artificial water systems and can be transmitted to human hosts by contaminated aerosols. Upon inhalation, it colonizes and grows inside the alveolar macrophages and causes Legionnaires' disease. To effectively control and manage Legionnaires' disease, a deep understanding of the host-pathogen interaction is crucial. Bacterial extracellular vesicles, particularly outer membrane vesicles (OMVs) have emerged as mediators of intercellular communication between bacteria and host cells. These OMVs carry a diverse cargo, including proteins, toxins, virulence factors, and nucleic acids. OMVs play a pivotal role in disease pathogenesis by helping bacteria in colonization, delivering virulence factors into host cells, and modulating host immune responses. This review highlights the role of OMVs in the context of host-pathogen interaction shedding light on the pathogenesis of L. pneumophila. Understanding the functions of OMVs and their cargo provides valuable insights into potential therapeutic targets and interventions for combating Legionnaires' disease.

A Biomimetic Adhesive and Robust Janus Patch with Anti-Oxidative, Anti-Inflammatory, and Anti-Bacterial Activities for Tendon Repair.

Early stage oxidative stress, inflammatory response, and infection after tendon surgery are highly associated with the subsequent peritendinous adhesion formation, which may diminish the quality and function of the repaired tendon. Although various anti-inflammatory and/or antibacterial grafts have been proposed to turn the scale, most of them suffer from the uncertainty of drug-induced adverse effects, low mechanical strength, and tissue adhesiveness. Here, inspired by the tendon anatomy and pathophysiology of adhesion development, an adhesive and robust dual-layer Janus patch is developed, whose inner layer facing the operated tendon is a multifunctional electrospun hydrogel patch (MEHP), encircled further by a poly-l-lactic acid (PLLA) fibrous outer layer facing the surrounding tissue. Specifically, MEHP is prepared by gelatin methacryloyl (GelMA) and zinc oxide (ZnO) nanoparticles, which are co-electrospun first and then treated by tannic acid (TA). The inner MEHP exhibits superior mechanical performance, adhesion strength, and outstanding antioxidation, anti-inflammation, and antibacterial properties, and it can adhere to the injury site offering a favorable microenvironment for tendon regeneration. Meanwhile, the outer PLLA acts as a physical barrier that prevents extrinsic cells and tissues from invading the defect site, reducing peritendinous adhesion formation. This work presents a proof-of-concept of a drug-free graft with anisotropic adhesive and biological functions to concert the healing phases of injured tendon by alleviating incipient inflammation and oxidative damage but supporting tissue regeneration and reducing tendon adhesion in the later phase of repair and remodeling. It is envisioned that this Janus patch could offer a promising strategy for safe and efficient tendon therapy.

Enzyme-Assisted Nucleic Acid Amplification in Molecular Diagnosis: A Review.

Infectious diseases and tumors have become the biggest medical challenges in the 21st century. They are driven by multiple factors such as population growth, aging, climate change, genetic predispositions and more. Nucleic acid amplification technologies (NAATs) are used for rapid and accurate diagnostic testing, providing critical information in order to facilitate better follow-up treatment and prognosis. NAATs are widely used due their high sensitivity, specificity, rapid amplification and detection. It should be noted that different NAATs can be selected according to different environments and research fields; for example, isothermal amplification with a simple operation can be preferred in developing countries or resource-poor areas. In the field of translational medicine, CRISPR has shown great prospects. The core component of NAAT lies in the activity of different enzymes. As the most critical material of nucleic acid amplification, the key role of the enzyme is self-evident, playing the upmost important role in molecular diagnosis. In this review, several common enzymes used in NAATs are compared and described in detail. Furthermore, we summarize both the advances and common issues of NAATs in clinical application.

Synthesis of Polylactic Acid Oligomers for Broad-Spectrum Antimicrobials.

Infectious microbial diseases are a major public health hazard, calling for more innovative antimicrobials. Herein, polylactic acid (PLA) oligomers have been explored and reported as a bio-safe and eco-friendly functional antimicrobial agent against pathogens, such as viruses (H1N1, H3N2, and SARS-CoV-2), bacteria (E. coli, S. aureus, K. pneumoniae, MRSA), and fungi (C. albicans). The PLA oligomers were prepared by direct catalyst-free condensation polymerization of l-lactic acid monomers and characterized by FT-IR and 1H-NMR. The antiviral results demonstrate that PLA oligomers possess robust (inhibiting rate > 99%) and rapid (<20 min) antiviral activity against two pandemic ssRNA viruses, including influenza A virus (IAV) and coronavirus (CoV). Furthermore, the PLA oligomers exhibit high antibacterial activities against both Gram negative (G−) and Gram positive (G+) bacteria. The PLA oligomers also perform efficiently in killing a large amount of C. albicans as high as 105 cfu/mL down to zero at the concentration of 10 mg/mL. Thus, the broad-spectrum antimicrobial activity endowed the PLA oligomers with a promising biocidal option, except antibiotics in a wide range of applications, such as medical textiles, food preservation, water disinfection, and personal hygiene, in light of their unique biodegradability and biocompatibility.

A Community-Based Participatory Research Approach to Developing and Testing Social and Behavioural Interventions to Reduce the Spread of SARS-CoV-2: A Protocol for the 'COPAR for COVID' Programme of Research with Five Interconnected Studies in the Hong Kong Context.

BACKGROUND: While a number of population preventive measures for COVID-19 exist that help to decrease the spread of the virus in the community, there are still many areas in preventative efforts that need improvement or refinement, particularly as new strains of the virus develop. Some of the key issues currently include incorrect and/or inconsistent use of face masks, low acceptance of early screening or vaccination for COVID-19, vaccine hesitance, and misinformation. This is particularly the case in some vulnerable populations, such as older people with chronic illnesses, ethnic minorities who may not speak the mainstream language well and children. The current protocol introduces a large programme of research through five interrelated studies that all focus on social and behavioural interventions to improve different aspects of community-related preventative indicators. Hence, the specific objectives of the overall programme are to (1) increase early testing for COVID-19 and promote the uptake of COVID-19 vaccines in the community (Study 1); (2) increase COVID-19-related health literacy and vaccine literacy and promote improved preventative measures in minority ethnic groups, chronically ill populations and caregivers (Study 2); (3) strengthen the public's motivation to stay at home and avoid nonessential high-risk activities (Study 3); (4) decrease COVID-19 vaccine hesitancy (Study 4); and (5) enhance the adherence to COVID-19-related hygiene practices and the uptake of early testing in school children (Study 5). METHODS: We will utilise a community-based participatory research (CBPR) approach in the proposed studies. All studies will incorporate an intervention development phase in conjunction with key community stakeholders, a feasibility study and an execution stage. A variety of self-reported and objective-based measures will be used to assess various outcomes, based on the focus of each study, in both the short- and long-term, including, for example, the 8-item self-reported eHealth Literacy Scale (eHEAL) and objective measures such as vaccine uptake. DISCUSSION: Theory-driven interventions will address each study's focus (e.g., social distancing, promotion of vaccine uptake, eHealth education, preventive measures and early detection). Improvements are expected to be seen in the outcomes of vulnerable and high-risk groups. Decreased infection rates are expected due to improved preventative behaviours and increased vaccine uptake. Long-term sustainability of the approach will be achieved through the CBPR model. The publication of this protocol can assist not only in sharing a large-scale and complex community-based design, but will also allow all to learn from this, so that we will have better insight in the future whether sharing of study designs can elicit timely research initiatives.

A universal biocompatible coating for enhanced lubrication and bacterial inhibition.

Antibacterial coatings that inhibit bacterial adhesion are essential for many implanted medical devices. A variety of antibacterial strategies, such as repelling or killing bacteria, have been developed, but not yet been completely successful. Here, we develop a universal biocompatible coating for enhanced lubrication and bacterial inhibition. The coating is designed based on mussel-inspired surface-attachable dopamine bases and consists of lubricating zwitterionic polymers poly(2-methacryloxyethyl phosphorylcholine) (MPC) and a bacterial membrane destroying anti-bacteria molecule poly(3-hydroxybutyric acid) (PHB). The coating boasts strong adhesion to surfaces of various materials (such as polydimethylsiloxane (PDMS)/ceramic/316L stainless steel (316L SS); it is biocompatible, and cell/platelet/bacteria repelling, significantly inhibiting bacterial growth. We envision that our strategy represents a universal strategy for surface functionalization of a variety of biomedical devices and implants.

Time-Variant Positive Air Pressure in Drainage Stacks as a Pathogen Transmission Pathway of COVID-19.

Time-variant positive air pressure in a drainage stack poses a risk of pathogenic virus transmission into a habitable space, however, the excessive risk and its significance have not yet been sufficiently addressed for drainage system designs. This study proposes a novel measure for the probable pathogenic virus transmission risk of a high-rise drainage stack with the occurrence of positive air pressure. The proposed approach is based on time-variant positive air pressures measured in a 38 m high drainage stack of a full-scale experimental tower under steady flow conditions of flow rate 1-4 Ls-1 discharging at a height between 15 m to 33 m above the stack base. The maximum pressure and probabilistic positive air pressures in the discharging stack ventilation section with no water (Zone A of the discharging drainage stack) were determined. It was demonstrated that the positive air pressures were lower in frequency as compared with those in other stack zones and could propagate along the upper 1/3 portion of the ventilation pipe (H' ≥ 0.63) towards the ventilation opening at the rooftop. As the probabilistic positive pressures at a stack height were found to be related to the water discharging height and flow rate, a risk model of positive air pressure is proposed. Taking the 119th, 124th, 140th and 11,547th COVID-19 cases of an epidemiological investigation in Hong Kong as a baseline of concern, excessive risk of system overuse was evaluated. The results showed that for a 20-80% increase in the frequency of discharge flow rate, the number of floors identified at risk increased from 1 to 9 and 1 to 6 in the 34- and 25-storey residential buildings, respectively. The outcome can apply to facilities planning for self-quarantine arrangements in high-rise buildings where pathogenic virus transmission associated with drainage system overuse is a concern.

Biofilm-Induced Antibiotic Resistance in Clinical Acinetobacter baumannii Isolates.

In order to understand the role of biofilm in the emergence of antibiotic resistance, a total of 104 clinical Acinetobacter baumannii strains were investigated for their biofilm-forming capacities and genes associated with biofilm formation. Selected biofilm-formers were tested for antibiotic susceptibilities when grown in biofilm phase. Reversibility of antibiotic susceptibility in planktonic cells regrown from biofilm were investigated. We found 59.6% of the strains were biofilm-formers, among which, 66.1% were non-multidrug resistant (MDR) strains. Presence of virulence genes bap, csuE, and abaI was significantly associated with biofilm-forming capacities. When strains were grown in biofilm state, the minimum biofilm eradication concentrations were 44, 407, and 364 times higher than the minimum bactericidal concentrations (MBC) for colistin, ciprofloxacin, and imipenem, respectively. Persisters were detected after treating the biofilm at 32-256 times the MBC of planktonic cells. Reversibility test for antibiotic susceptibility showed that biofilm formation induced reversible antibiotic tolerance in the non-MDR strains but a higher level of irreversible resistance in the extensively drug-resistant (XDR) strain. In summary, we showed that the non-MDR strains were strong biofilm-formers. Presence of persisters in biofilm contributed to the reduced antibiotic susceptibilities. Biofilm-grown Acinetobacter baumannii has induced antibiotic tolerance in non-MDR strains and increased resistance levels in XDR strains. To address the regulatory mechanisms of biofilm-specific resistance, thorough investigations at genome and transcription levels are warranted.

Durable Antimicrobial Behaviour from Silver-Graphene Coated Medical Textile Composites.

Silver nanoparticle (AgNP) and AgNP/reduced graphene oxide (rGO) nanocomposite impregnated medical grade polyviscose textile pads were formed using a facile, surface-mediated wet chemical solution-dipping process, without further annealing. Surfaces were sequentially treated in situ with a sodium borohydride (NaBH4) reducing agent, prior to formation, deposition, and fixation of Ag nanostructures and/or rGO nanosheets throughout porous non-woven (i.e., randomly interwoven) fibrous scaffolds. There was no need for stabilising agent use. The surface morphology of the treated fabrics and the reaction mechanism were characterised by Fourier transform infrared (FTIR) spectra, ultraviolet-visible (UV-Vis) absorption spectra, X-ray diffraction (XRD), Raman spectroscopy, dynamic light scattering (DLS) energy-dispersive X-ray analysis (EDS), and scanning electron microscopic (SEM). XRD and EDS confirmed the presence of pure-phase metallic silver. Variation of reducing agent concentration allowed control over characteristic plasmon absorption of AgNP while SEM imaging, EDS, and DLS confirmed the presence of and dispersion of Ag particles, with smaller agglomerates existing with concurrent rGO use, which also coincided with enhanced AgNP loading. The composites demonstrated potent antimicrobial activity against the clinically relevant gram-negative Escherichia coli (a key causative bacterial agent of healthcare-associated infections; HAIs). The best antibacterial rate achieved for treated substrates was 100% with only a slight decrease (to 90.1%) after 12 equivalent laundering cycles of standard washing. Investigation of silver ion release behaviours through inductively coupled plasmon optical emission spectroscopy (ICP-OES) and laundering durability tests showed that AgNP adhesion was aided by the presence of the rGO host matrix allowing for robust immobilisation of silver nanostructures with relatively high stability, which offered a rapid, convenient, scalable route to conformal NP-decorated and nanocomposite soft matter coatings.

Emergence of Carbapenem-Resistant Acinetobacter baumannii ST195 Harboring blaOXA-23 Isolated from Bacteremia in Hong Kong.

Aims: The aim of the study was to analyze the epidemiology of Acinetobacter baumannii and investigate the genetic characteristics of carbapenem-resistant A. baumannii (CRAB) isolates isolated from blood cultures in a regional hospital in Hong Kong. Results: Twenty blood culture isolates were collected from a regional hospital in Hong Kong from 2014 to 2017. Twenty isolates were grouped into five existing sequence types (STs) and five new STs within the following prevalence: ST195 was predominant with a prevalence of 45% (n = 9), followed by ST373 and ST447 (10%; n = 2 each), and ST176 and ST345 (5%; n = 1 each). Resistance to carbapenem antibiotics was 55% (n = 11). Six carbapenem-resistant isolates harbored blaOXA-23 genes and ISAba1 mobile elements. Polymerase chain reaction confirmed that ISAba1 is located upstream to the blaOXA-23 genes, suggesting an association between ISAba1 and blaOXA-23 genes with carbapenem resistance. Conclusion: This study is the first to report the emergence of CRAB ST195 harboring blaOXA-23 in Hong Kong.

Involvement of caspase-4 in IL-1 beta production and pyroptosis in human macrophages during dengue virus infection.

Caspase-4 physically interacts with caspase-1 and is believed to be a proinflammatory caspase that can induce the inflammatory form of programmed cell death (pyroptosis) and the release of mature interleukin (IL)-1ß. However, the function of caspase-4 in dengue virus infection is not yet fully understood. We examined the function of caspase-4 in IL-1ß production and pyroptosis during dengue virus serotype-2 (DENV-2) infection in human macrophages. In this study, DENV-2 infection increased IL-1ß protein level with activated caspase-4 activity. Using primary macrophages, we observed that caspase-4 induces activation of caspase-1 and secretion of IL-1ß in response to DENV-2 infection, without the need for secondary signals to stimulate the assembly of the inflammasome. These findings indicate that the regulation of caspase-1 activity by capsase-4 could represent a unique mechanism. Our data suggest that caspase-4 is upstream of caspase-1 in the pathway that regulates pyroptosis and IL-1ß synthesis in macrophages during DENV-2 infection.

Composite Membranes of Recombinant Silkworm Antimicrobial Peptide and Poly (L-lactic Acid) (PLLA) for biomedical application.

Antimicrobial peptides, produced by innate immune system of hosts in response to invading pathogens, are capable of fighting against a spectrum of bacteria, viruses, fungi, parasites and cancer cells. Here, a recombinant silkworm AMP Bmattacin2 from heterologous expression is studied, indicating a broad spectrum of antibacterial activity and showing selective killing ability towards skin and colon cancer cells over their normal cell counterparts. For the purpose of biomedical application, the electrospinning fabrication technique is employed to load Bmattacin2 into PLLA nanofibrous membrane. In addition to a good compatibility with the normal cells, Bmattacin2 loaded nanofibrous membranes demonstrate instant antibacterial effects and sustained anticancer effects. The cancer cell and bacteria targeting dynamics of recombinant Bmattacin2 are investigated. With these characteristics, PLLA/Bmattacin2 composite membranes have a great potential for developing novel biomedical applications such as cancer therapies and wound healing treatments.

Distinct expression of interferon-induced protein with tetratricopeptide repeats (IFIT) 1/2/3 and other antiviral genes between subsets of dendritic cells induced by dengue virus 2 infection.

Dengue virus (DENV) infection is an emerging public health hazard threatening inhabitants of the tropics and sub-tropics. Dendritic cells (DCs) are one of the major targets of DENV and the initiators of the innate immune response against the virus. However, current in vitro research on the DENV-DC interaction is hampered by the low availability of ex vivo DCs and donor variation. In the current study, we attempted to develop a novel in vitro DC model using immature DCs derived from the myeloid leukaemia cell line MUTZ-3 (IMDCs) to investigate the DENV-DC interaction. The IMDCs morphologically and phenotypically resembled human immature monocyte-derived dendritic cells (IMMoDCs). However, the permissiveness of IMDCs to DENV2 was lower than that of IMMoDCs. RT-PCR arrays showed that a group of type I interferon (IFN) -inducible genes, especially IFIT1, IFITM1, and IFI27, were significantly up-regulated in IMMoDCs but not in IMDCs after DENV2 infection. Further investigation revealed that IFIT genes were spontaneously expressed at both transcriptional and protein levels in the naive IMDCs but not in the naive IMMoDCs. It is possible that the poor permissiveness of IMDCs to DENV2 was a result of the high basal levels of IFIT proteins. We conclude that the IMDC model, although less permissive to DENV2, is a useful platform for studying the suppression mechanism of DENV2 and we expand the knowledge of cellular factors that modulate DENV2 infection in the human body.

Temperature induced modulation of lipid oxidation and lipid accumulation in palmitate-mediated 3T3-L1 adipocytes and 3T3-L1 adipocytes.

Human skin temperature can vary widely depending on anatomical location and ambient temperature. It is also known that local changes in skin and subcutaneous temperature can affect fat metabolism. This study aimed to explore the potential effects of surrounding thermal environment on fat by investigating cell viability, lipid oxidation, and lipid accumulation in 3T3-L1 adipocytes and palmitate-treated adipocytes after 4h incubation. No significant differences of viability in 3T3-L1 adipocytes were detected under different temperature conditions. Despite no significant increase being observed under warm temperature (39°C) conditions, a similarly significant suppression of intracellular reactive oxygen species (ROS) and lipid peroxidation were found in 3T3-L1 adipocytes and palmitate-treated adipocytes under 4h exposure to cooler temperatures of 31-33°C (P<0.01). ROS, chemically reactive molecules containing oxygen, are currently understood to be a major contributor to oxidantive stress in obesity. Additionally, cooler temperatures (31-33°C) could improve the size of lipid droplets in 3T3-L1 adipocytes (P<0.01), but no significant effect was generated by temperature change on lipid droplets in palmitate-treated adipocytes. In the palmitate-induced adiposity model, although excessive ROS and lipid peroxidation has been attenuated by temperature decrease (P<0.01), it still does not positively modulate lipid droplet size (P>0.05) and remedy the palmitate damage induced cell death (P<0.01). These findings provide preliminary support for potential interventions based on temperature manipulation for cell metabolism of adipocytes.

Evaluation on masks with exhaust valves and with exhaust holes from physiological and subjective responses.

The purpose of this study was to compare the effects of wearing different kinds of masks on the ear canal temperature, heart rate, clothing microclimate, and subjective perception of discomfort. Ten subjects performed intermittent exercise on a treadmill while wearing the protective masks in a climatic chamber controlled at an air temperature of 25 degrees C and a relative humidity of 70%. Two types of mask-mask A, with exhaust valves and mask B, with exhaust holes-were used in the study. The results of this study indicated: (1) The subjects had a tendency toward lower maximum heart rate when wearing mask A than when wearing mask B. (2) Temperatures and absolute humidities (the outer surface of mask, the microclimate inside the mask, the chest wall skin and microclimate) of mask A were significantly lower than those of mask B. (3) The ear canal temperature increased significantly in mask B as compared to that in mask A. (4) The ear canal temperature showed significant augmentation along with increased temperature and humidity inside the mask microclimate. The mask microclimate temperature also affected significantly the chest microclimate temperature. (5) Mask A was rated significantly lower for perception of humidity, heat, breath resistance, tightness, unfitness, odor, fatigue, and offered less overall discomfort than mask B. (6) Subjective preference for mask A was higher. (7) The ratings of subjective overall discomfort showed significant augmentation along with increased wetness and fatigue. We discuss how the ventilation properties of masks A and B induce significantly different temperature and humidity in the microclimates of the masks and the heat loss of the body, which have profound influences on heart rate, thermal stress, and subjective perception of discomfort.