Identification of microbial communities and multi-species biofilms contamination in seafood processing environments with different hygiene conditions.

Biofilms formed by spoilage and pathogenic bacteria increase microbial persistence, causing an adverse influence on the quality of seafood. The mono-species biofilms are widely reported, however, the contamination of multi-species biofilms and their matrix in food environments are still not fully understood. Here, we assessed the contamination of multi-species biofilms in three seafood processing environments with different hygiene levels by detecting bacterial number and three biofilm matrix components (carbohydrates, extracellular DNA (eDNA), and proteins). Samples comprising seven food matrix surfaces and eight food processing equipment surfaces were collected from two seafood processing plants (XY and XC) and one seafood market (CC). The results showed that the bacterial counts ranged from 1.89 to 4.91 CFU/cm2 and 5.68 to 9.15 BCE/cm2 in these surfaces by cultivation and real-time PCR, respectively. Six biofilm hotspots were identified, including four in CC and two in XY. Among the three processing environments, the amplicon sequence variants (ASVs) of Proteobacteria, Bacteroidetes, and Actinobacteria decreased with improved processing hygiene, while Firmicutes showed a decrease in the four most abundant phyla. The most prevalent bacteria belonged to genera Psychrobacter, Acinetobacter, and Pseudomonas, demonstrating the significant differences and alteration in bacterial community composition during different environments. From the biofilm hotspots, 15 isolates with strong biofilm forming ability were identified, including 7 Pseudomonas, 7 Acinetobacter, and 1 Psychrobacter. The Pseudomonas isolates exhibited the highest production of EPS components and three strong motilities, whose characteristics were positively correlated. Thus, this study verified the presence of multi-species biofilms in seafood processing environments, offering preliminary insights into the diversity of microbial communities during processing. It highlights potential contamination sources and emphasizes the importance of understanding biofilms composition to control biofilms formation in seafood processing environments.

Robust layout optimization for intelligent reflecting surfaces-based visible light communication systems.

Due to the inherent tendency to be blocked by obstacles, reliability is a major challenge for visible light communication (VLC). The intelligent reflective surface (IRS) is an effective way to reduce or eliminate the influence of blockage in the VLC system. However, the complexity increases correspondingly due to complex shadow analysis when access points (APs), IRSs, and obstacles coexist. We proposed a robust layout optimization scheme for the IRS-VLC system to resist blockages. First, we model the random obstructions based on spatial geometry methods. Second, we build the optimization problem model considering consistent illumination, achievable data rate (ADR), the positions of the APs, and the directions of the IRS array. In order to achieve this objective, we develop an anti-occlusion method based on the non-dominated sorting genetic algorithm-II (NSGA-II) to get the Pareto front and use enhanced measurement functions (ASFs) to extract the best solution. Simulation results show that the proposed scheme works well in the IRS-VLC system. It is noteworthy that the rectangle layout has always demonstrated superior performance in the IRS-VLC system compared to other traditional layouts.

Bioactives and metabolites of Tetrastigma hemsleyanum root extract alleviate DSS-induced ulcerative colitis by targeting the SYK protein in the B cell receptor signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Tetrastigma hemsleyanum is an endemic Chinese herb with a wide range of pharmacological activities, including anti-inflammatory, antiviral, antioxidant, antitumor, and immunomodulatory activities. However, the effect and mechanisms of the anti-inflammatory activity of T. hemsleyanum root extract against dextran sodium sulfate (DSS)-induced ulcerative colitis (UC) have not yet been fully investigated. AIM OF THE STUDY: This study aimed to explore the therapeutic effect and molecular mechanisms of T. hemsleyanum root extract in DSS-induced UC mice and knockdown cells. MATERIALS AND METHODS: T. hemsleyanum root extract was obtained and analyzed by high-performance liquid chromatography (HPLC). The therapeutic effects of T. hemsleyanum root extract on DSS-induced UC mice were evaluated by the disease activity index (DAI) score, colon length, serum inflammatory cytokines and oxidant/antioxidant levels, and histopathological features of the ileum and colon. Genome-wide gene expression profiles of ileal and colonic tissues were collected by transcriptomics, and signaling pathways were analyzed by the KEGG database. UC-related pathways were uploaded to the STRING database, then the protein-protein interactions (PPIs) were determined by Cytoscape, and the enriched genes were evaluated by real-time quantitative PCR (qPCR). The protein-ligand complexes were docked by AutoDock, and the genes were knocked down in Caco-2 cells by shRNA. The non-targeted metabolomic profiling of ileal contents was analyzed by ultra-high-performance liquid chromatography (UHPLC), and gut microflora were sequenced by an Illumina MiSeq System. RESULTS: Ten components that alleviated UC symptoms in mice by decreasing the DAI and serum inflammatory cytokines and oxidant levels, promoting intestinal development, and increasing serum antioxidant levels were identified in T. hemsleyanum root extract. T. hemsleyanum root extract activated the B cell receptor signaling pathway in the colon tissue of UC mice, in which two components, rutin and astragaline, bound to the spleen tyrosine kinase (SYK) protein but also restored gut microflora diversity and increased the proportion of probiotics. Furthermore, metabolites of T. hemsleyanum root extract were involved in vitamin metabolism, fatty acid metabolism, and ferroptosis. CONCLUSIONS: The rutin and astragaline components of T. hemsleyanum root extract, by binding to SYK protein, activated the B cell receptor signaling pathway and restored gut microflora diversity to alleviate UC symptoms in mice.

Extracellular matrix affects mature biofilm and stress resistance of psychrotrophic spoilage Pseudomonas at cold temperature.

Psychrotrophic Pseudomonas as the dominant spoilage bacteria, have biofilm forming ability, increasing persistence and contamination in the chilled food. Biofilm formation of spoilage Pseudomonas at cold temperature was documented, however, role of extracellular matrix in mature biofilm and stress resistance of psychrotrophic Pseudomonas are much less abundant. The aim of this study was to investigate the biofilm forming characteristics of three spoilers P. fluorescens PF07, P. lundensis PL28, and P. psychrophile PP26 at 25 °C, 15 °C and 4 °C, and to explore their stress resistance to chemical and thermal treatments of mature biofilms. The results showed that biofilm biomass of three Pseudomonas at 4 °C was significantly higher than that at 15 °C and 25 °C. The secretion of extracellular polymeric substances (EPS) greatly increased in those Pseudomonas under low temperature, of which extracellular protein constituted about 71.03%-77.44%. Compared to 25 °C, the mature biofilms were observed to more aggregation and thicker spatial structure at 4 °C ranging from 42.7 to 54.6 µm, in contrast to 25.0-29.8 µm at 25 °C, especially strain PF07. These Pseudomonas biofilms switched into moderate hydrophobicity, and their swarming and swimming were significantly inhibited at low temperature. Furthermore, the resistance to NaClO and heating at 65 °C apparently enhanced for mature biofilm formed at 4 °C, indicating the difference in EPS matrix production influenced the stress resistance of biofilm. In addition, three strains contained alg and psl operons for exopolysaccharide biosynthesis, and biofilm related genes of algK, pslA, rpoS, and luxR were significantly up-regulated, while flgA gene was down-regulated at 4 °C compared to 25 °C, consistent with the above phenotype changes. Thus, the dramatic increase of mature biofilm and their stress resistance in psychrotrophic Pseudomonas were associated with large secretion and protection of extracellular matrix under low temperature, which provide a theoretical basis for subsequent biofilm control during cold chain.

Osteogenic and long-term antibacterial properties of Sr/Ag-containing TiO2 microporous coating in vitro and in vivo.

Bacterial infection and poor osseointegration are two critical issues that need to be solved for long-term use of titanium implants. As such, Sr/Ag-containing TiO2 microporous coatings were prepared on a Ti alloy surface in the current study via a single-step microarc oxidation technique. The coatings showed both good cytocompatibility in vitro and biosafety in vivo. Sr/Ag incorporation brought no significant change in the surface micromorphology and physicochemical properties, but endowed the coating with strong osteogenic activity and long-term antibacterial capability in vitro. Furthermore, the osteogenic and antibacterial capability of the coating was also confirmed in vivo. In a rat osseointegration model, new bone formation, implant-bone contact, removal torque and bone mineralization were all significantly increased in the M-Sr/Ag group when compared with those in group M, although they were slightly lower than those in group M-Sr. In a periimplantitis model, no rats suffered infection in the M-Sr/Ag group after 3 months of osseointegration and 5 weeks of bacterial inoculation period, when compared to 100% and 75% infection rates in M and M-Sr groups, respectively. In addition, active bone remodeling and many mesenchymal cells were observed in the M-Sr group, suggesting good bone regeneration potential in Sr-containing coatings in the case of controlled periimplantitis. Overall, the Sr/Ag-containing TiO2 microporous coating is valuable for preventing periimplantitis and improving implant reosseointegration, and is therefore promising for long-term and high quality use of titanium implants.

Effect of oligosaccharides as lyoprotectants on the stability of curcumin-loaded nanoliposomes during lyophilization.

Nanoliposome is a promising delivery system, whereas its commercial application is limited by the structural instability, cargo leakage and particles aggregation during the processing such as freeze-drying. In this study, the effect of four oligosaccharides, fructo-oligosaccharides, lactose, inulin and sucrose (control), on the physicochemical properties, structural stability, and in vitro semi-dynamic digestion behavior of curcumin-loaded nanoliposomes were investigated before and after lyophilization. The results showed that the addition of the oligosaccharides inhibited the changes in particle size and reduced curcumin leakage from lyophilized nanoliposomes. Oligosaccharides significantly improved the physical stability of lyophilized nanoliposomes and delayed curcumin release during in vitro digestion. In addition, oligosaccharides could decrease the hydrophobicity of liposomal membrane and the tightness of phospholipid molecule arrangement, with the increase in micropolarity and fluidity of the bilayer membranes. These results suggested that fructo-oligosaccharides, lactose and inulin could be effective lyoprotectants for lyophilized nanoliposomes.

Angiogenic property of cobalt and calcium-phosphorus doped coating on titanium surfaces in vitro / 口腔疾病防治

Objective@# To investigate the effect of cobalt (Co) and calcium-phosphate (Ca/P) doped coating on titanium surfaces and their angiogenic effect.@*Methods @# Microarc oxidation (MAO) was used to prepare Co-Ca/P-doped and Co-doped coatings. Titanium (Ti) sheet without MAO treatment was used as control. Scanning electron microscopy (SEM) was used to observe the surface micromorphology of the coatings. Energy dispersive spectrometry (EDS) was also applied to detect the doped chemicals and their contents. Standard soaking solutions of these coatings were prepared using an endothelial cell medium (ECM) solution for subsequent angiogenesis experiments. Human umbilical vein endothelial cells (HUVECs) were cultured on Matrigel with ECM soaking solutions for 4 h and 8 h. The microvessels were observed under a microscope, and the number of microtubules and their interconnecting nodes were analyzed with Image J software. @*Results@# Co doped and Co-Ca/P-doped coatings were successfully prepared by MAO, which was demonstrated by both SEM observation and EDS analysis. SEM observation showed that irregular crystals of the above chemicals were present on both Co and Co-Ca/P-doped coatings, commonly with a diameter <2 μm. However, more crystals were observed on the Co-Ca/P coatings than on the Co coating, and the distribution of the crystals was more homogenous on the Co-Ca/P coatings. However, only polishing scratches were observed on the Ti sample surface. EDS analysis indicated that in contrast to only Co in the Co coating, Co, Ca and P were doped within the Co-Ca/P coating, and none of the three elements were observed on the Ti plate surface. The number of vascular rings and nodes formed by HUVECs in the extract of the Co-Ca/P group was significantly higher than that of the Co group (P<0.05), and the angiogenic effect of these two components was significantly better than that of the Ti group (P<0.05). @*Conclusion@#The Co-Ca/P coating exhibits good angiogenic properties in vitro and is valuable for the development of new titanium implants with high surface bioactivity.

Integrated visible light communication and positioning CDMA system employing modified ZCZ and Walsh code.

In this paper, an integrated visible light communication and positioning (VLCP) code division multiple access (CDMA) system is proposed for recovering original user data and obtaining positioning information simultaneously. A generalized modification method for balanced bipolar code set is presented, which can be applied to VLCP-CDMA system with intensity modulation. Both modified bipolar code sets, zero correlation zone (ZCZ) code set and Walsh code set, are employed and evaluated in VLCP-CDMA systems. When considering synchronous and quasi-synchronous (QS) systems, it is demonstrated that the modified ZCZ code set with ideal zero correlation zone properties performs better communication and positioning performance than the modified Walsh code set by system simulations and experiments. As a result, a bit error rate (BER) of 1.8×10-3 and an average positioning error of 1.50 cm are successfully obtained by adopting modified ZCZ code set in a 4-user real-time VLCP-CDMA system. It will offer a promising solution to meet both communication and positioning requirements of future intelligent systems.

Case report: Novel compound-heterozygous mutations in the TCN2 gene identified in a chinese girl with transcobalamin deficiency.

Transcobalamin (TC) deficiency is a rare autosomal recessive disease characterized by megaloblastic anemia. It is caused by cellular vitamin B12 depletion, which subsequently results in elevated levels of homocysteine and methylmalonic acid. This disease is usually diagnosed by genetic analysis of the TCN2 gene. Here, we described a 2.2-month-old Chinese girl with TC deficiency presenting with diarrhea, fever and poor feeding. Whole-exome sequencing detected a pair of compound-heterozygous mutations in TCN2 gene, c.754-12C>G and c.1031_1032delGA (p.R344Tfs*20). To our knowledge, it is the first time that they were identified and reported in TC deficiency. This study contributes to a better understanding of the TC deficiency, expanding the spectrum of TCN2 mutations in this disorder and also supporting the early diagnosis and proper treatment of similar cases in the future.

In vitro Digestion Characteristics of Hydrolyzed Infant Formula and Its Effects on the Growth and Development in Mice.

Infant formula, an important food for babies, is convenient and nutritious, and hydrolyzed formulas have attracted much attention due to their non-allergicity. However, it is uncertain whether hydrolyzed formulars cause obesity and other side effects in infants. Herein, three infant formulas, standard (sIF), partially hydrolyzed (pHIF), and extensively hydrolyzed (eHIF), were analyzed in an in vitro gastrointestinal digestion model. With increasing degree of hydrolysis, the protein moleculars, and allergenicity of the proteins decreased and the long-chain polyunsaturated fatty acid content increased. Moreover, the digestion model solutions quickly digested the small fat globules and proteins in the hydrolyzed formula, allowing it to become electrostatically stable sooner. The eHIF-fed mice presented larger body sizes, and exhibited excellent exploratory and spatial memory abilities in the maze test. Based on villus height and crypt depth histological characterizations and amplicon sequencing, eHIF promoted mouse small intestine development and changed the gut microbiota composition, eventually favoring weight gain. The mouse spleen index showed that long-term infant formula consumption might be detrimental to immune system development, and the weight-bearing swimming test showed that eHIF could cause severe physical strength decline. Therefore, long-term consumption of infant formula, especially eHIF, may have both positive and negative effects on mouse growth and development, and our results might shed light on feeding formula to infants.

Metabolomics reveals spoilage characteristics and interaction of Pseudomonas lundensis and Brochothrix thermosphacta in refrigerated beef.

Pseudomonas lundensis and Brochothrix thermosphacta are key spoilage microorganisms in aerobically stored chilled meat. The present study aimed to investigate the physicochemical and metabolomic profiles of refrigerated ground beef inoculated P. lundensis (PL) and B. thermosphacta (BT) as mono- or co-culture (BP). P. lundensis was the dominant spoilage strain in the co-culture of ground beef. A large amount of TCA-soluble peptide, TVB-N and TBA were formed in the PL and BP, while acetion was mainly produced in the BT, as accompanied by the different sensory and color changes. Meat metabolome indicated that 95, 396, and 409 metabolites with significant differences, were identified in ground beef inoculated BT, PL, and BP, respectively. These differential metabolites covered 58 metabolic pathways, in which histidine metabolism was identified as an important pathway related to spoilage in the three groups. Specifically, creatine, inosine, anserine, uracil, alanine, glutamine, 3-methylhistidine and 3-hydroxycapric acid were enriched as potential spoilage biomarkers. Taken together, those findings reveal the complex and competitive interactions of their co-culture of B. thermosphacta and P. lundensis, which provided a comprehensive insight into microbial spoilage mechanism in chilled beef.

AI Empowered Virtual Reality Integrated Systems for Sleep Stage Classification and Quality Enhancement.

Insomnia is a common public health problem and an open biomedical research topic. Insomnia results in various health problems, including memory decline, decreases concentration and weakens problem-solving ability. The insufficient sleep also leads to skin ageing, heart disease, high blood pressure, arrhythmia and stroke. While it remains as a global health concern, sleep quality improvement using modern technologies, such as machine learning, classification technologies, virtual reality (VR), becomes an open and hot research problem. These modern technologies offer new curing solutions under certain conditions. In this paper, we present a sleeping-aid system with a single-channel electroencephalogram (EEG) sleep stage classification algorithm to improve the sleep quality. The sleeping-aid system promotes machine learning integrated VR and multimedia technology for sleep improvement. Ninety participants were invited to test on three different systems with 3D VR, 2D video, and music only. An adequate stimulus of audio-vision can be a complement of the drug treatment. The experimental results showed that the proposed method demonstrated superior performance over existing methods.

Genetic Basis of High-Pressure Tolerance of a Vibrio parahaemolyticus Mutant and Its Pathogenicity.

Foodborne pathogens with high-pressure processing (HPP) tolerance and their pathogenicity have gained considerable attention in the field of food safety. However, tolerance to pressure treatment varies among microorganisms and growth phases, and the mechanism by which Vibrio parahaemolyticus can become tolerant of HPP is currently not known. In this study, 183 strains of V. parahaemolyticus were isolated from seafood products, and one strain, C4, carried a thermostable direct hemolysin (tdh) gene. A strain, N11, which was acquired from the C4 strain through adaptive laboratory evolution under HPP stress, could tolerate up to 200 MPa for 10 min. Compared with the C4 strain, the catalase and Na+/K+-ATPase activities in N11 strain were increased by about 2-3 times, and the cells maintained an intact cell membrane structure under HPP treatment. As shown by murine infection trials, the C4 and N11 strains impacted the physiological activities of mice and damaged liver and spleen cells. Comparative genomic analysis showed that 19 nucleotides were mutated in the N11 strain, which led to sustained high expression of mlaC and mlaD genes in this strain. Knockout of these genes confirmed that they were involved in the high-pressure stress response, and also related to pathogenicity of V. parahaemolyticus. Thereby, our findings revealed a HPP tolerance mechanism of V. parahaemolyticus, and the high-pressure-tolerant strain still retained pathogenicity in mice with skin and fur pleating and lethargy, indicating the pressure-tolerant foodborne pathogens present health risks.

Adaptive feedback threshold based demodulation for mobile visible light communication and positioning integrated system.

In this paper, a novel adaptive feedback threshold (AFT) based demodulation for mobile visible light communication and positioning (VLCP) integrated system is proposed. The AFT can vary with the received signal without excessive fluctuation and support communication and positioning in mobile environment. For the VLCP integrated system with single light emitting diodes (LED) and multiple photodetectors (PDs), maximal ratio combining (MRC) and received optical power ratio (ROPR) based on the AFT is further achieved for communication and positioning with high performance, respectively. It is demonstrated that high credible communication and high accuracy positioning for the mobile integrated VLCP system can be realized by using the proposed AFT based demodulation. As a result, the implemented VLCP system with a moving speed of 1 m/s is evaluated experimentally. Average positioning error of 3.43 cm with 800 mA current and the bit error ratio (BER) with different currents are also obtained.

Case Report: Next-Generation Sequencing Identified a Novel Pair of Compound-Heterozygous Mutations of LPL Gene Causing Lipoprotein Lipase Deficiency.

Lipoprotein lipase deficiency (LPLD) is a rare disease characterized by the accumulation of chylomicronemia with early-onset. Common symptoms are abdominal pain, hepatosplenomegaly, eruptive xanthomas and lipemia retinalis. Serious complications include acute pancreatitis. Gene LPL is one of causative factors of LPLD. Here, we report our experience on an asymptomatic 3.5-month-old Chinese girl with only milky blood. Whole-exome sequencing was performed and identified a pair of compound-heterozygous mutations in LPL gene, c.862G>A (p.A288T) and c.461A>G (p.H154R). Both variants are predicted "deleterious" and classified as "likely pathogenic". This study expanded the LPL mutation spectrum of disease LPLD, thereby offering exhaustive and valuable experience on early diagnosis and proper medication of LPLD.

Development of the digestive system in early infancy and nutritional management of digestive problems in breastfed and formula-fed infants.

Food digestion and absorption in infants are closely related to early growth and long-term health. Human milk and infant formula are the main food sources for 0-6 month-old infants. Due to the immature gastrointestinal tract of newborns, mild digestive problems, such as inefficient digestion and impaired absorption of proteins, lipids and lactose, and gut dysbiosis, are often seen in infancy. The differences in composition between infant formula and human milk make mild digestive problems more likely to occur in formula-fed infants. In recent years, several types of infant formulas have been developed to treat or reduce gastrointestinal digestive problems in infants. This review summarizes the gastrointestinal environment of infants and the digestion of human milk and different infant formulas. We particularly focus on the common digestive problems and appropriate nutritional solutions that may occur in healthy term infants during the first six months of life.

Correlation analysis of temporomandibular joint changes after maxillary protraction in adolescent patients / 口腔疾病防治

Objective @#To study the effect of anterior traction on the temporomandibular joint in adolescent patients with skeletal Class Ⅲ malocclusion.@*Methods@#Twenty-nine patients with early permanent dentition with skeletal class Ⅲ malocclusions were measured by cephalometry and a coordinate system before and after maxillary protraction. The correlation between dentofacial structures and the temporomandibular joint was analyzed. @*Results @# After maxillary protraction, cephalometric measurements showed that the dentofacial structure changed significantly; ANB increased by 3.97° ± 2.32° (P<0.001); U1-SN increased by 4.97° ± 5.51° (P<0.001); L1-MP decreased by 1.26° ± 1.41° (P = 0.008); and MP-SN increased by 1.02° ± 3.90° (P = 0.003). The coordinate system measurement showed that the S-Fpx was decreased by 0.16 ± 1.52 mm (P = 0.041), the S-Ciy distance was significantly decreased by 0.09 ± 2.03 mm (P = 0.028), and there was no significant change in the temporomandibular joint spaces (A, P, and C) (P>0.05). Correlation analysis showed a moderate negative correlation between the posterior margin of the temporomandibular joint fossa and U1-SN (r = -0.427, P = 0.042). There was a moderate positive correlation between the leading edge of the condyle and ANB (r = 0.425, P = 0.043); there was no correlation between the joint space and dentofacial changes. @* Conclusion@#After treatment with maxillary protraction for adolescent skeletal class Ⅲ malocclusion, maxillary protraction had some effect on changes in the temporomandibular joint fossa and condyle and had no effect on the joint space.

Regulatory function of sigma factors RpoS/RpoN in adaptation and spoilage potential of Shewanella baltica.

Shewanella baltica is a typical specific spoilage organism causing the deterioration of seafood, but the exact regulation of its adaptive and competitive dominance in diverse environments remains undefined. In this study, the regulatory function of two sigma factors, RpoS and RpoN, in environmental adaptation and spoilage potential were evaluated in S. baltica SB02. Two in-frame deletion mutants, ΔrpoS and ΔrpoN, were constructed to explore the roles in their motility, biofilm formation, stress response and spoilage potential, as well as antibiotics by comparing the phenotypes and transcription with those of wild type (WT) strain. Compared with WT strain, the ΔrpoN showed the slower growth and weaker motility due to loss of flagella, while swimming of the ΔrpoS was increased. Deletion of rpoN significantly decreased biofilm biomass, and production of exopolysaccharide and pellicle, resulting in a thinner biofilm structure, while ΔrpoS formed the looser aggregation in biofilm. Resistance of S. baltica to NaCl, heat, ethanol and three oxidizing disinfectants apparently declined in the two mutants compared to WT strain. The ΔrpoN mutant decreased sensory score, accumulation of trimethylamine, putrescine and TVB-N and protease activity, while a weaker effect was observed in ΔrpoS. The two mutants had significantly higher susceptibility to antibiotics than WT strain, especially ΔrpoN. Deficiency of rpoN and rpoS significantly repressed the activities of two diketopiperazines related to quorum sensing (QS). Furthermore, transcriptome analyses revealed that RpoN was involved in the regulation of the expression of 143 genes, mostly including flagellar assembly, nitrogen and amino acid metabolism, ABC transporters. Transcript changes of seven differentially expressed coding sequences were in agreement with the phenotypes observed in the two mutants. Our findings reveal that RpoN, as a central regulator, controls the fitness and bacterial spoilage in S. baltica, while RpoS is a key regulatory factor of stress response. Characterization of these two sigma regulons in Shewanella has expanded current understanding of a possible co-regulatory mechanism with QS for adaptation and spoilage potential.

Recombinant unpurified rETXH106P/ CTB-rETXY196E protects rabbits against Clostridium perfringens epsilon toxin.

Epsilon toxin (ETX), produced by Clostridium perfringens types B and D, has been touted as a potential biological weapon and is known to induce fatal enterotoxemia in a variety of livestock animals. For the efficient production of recombinant proteins with the objective of investigating the effects of different recombinant vaccines against ETX, a bicistronic design (BCD) expression system including the ETX coding sequence with mutation of amino acid 106 from Histidine to Proline (ETXH106P) in the first cistron, followed by Cholera Toxin B (CTB) linked with the ETX coding sequence with mutation of amino acid 196 from Tyrosine to Glutamic acid (ETXY196E) in the second cistron, was generated under the control of a single promoter. Rabbits were immunized twice with five inactivated recombinant Escherichia coli (E. coli) vaccines containing 100 µg/ml of the recombinant mutant rETXH106P/CTB-rETXY196E proteins mixed with different adjuvants. Apart from rETXH106P/CTB-rETXY196E-IMS1313-vaccinated rabbits, the neutralizing antibody titers of rETXH106P/CTB-rETXY196E-vaccinated rabbits were higher after the initial immunization than those administered the ETX toxoid or current commercial vaccines. rETXH106P/CTB-rETXY196E mixed with ISA201 induced the highest neutralizing antibody titer of 120 after the first immunization, suggesting that 0.1 ml of pooled sera could neutralize 120× mouse LD100 (100% lethal dose) of ETX. Following the second vaccination, rETXH106P/CTB-rETXY196E mixed with ISA201 or GR208 produced the highest neutralizing titer of 800. Rabbits from all vaccinated groups were completely protected from a 2× rabbit LD100 of ETX challenge. These results show that these novel recombinant proteins can induce a strong immune response and represent potential targets for the development of a commercial vaccine against the C. perfringens epsilon toxin.

Efficacy and safety of diazoxide for treating hyperinsulinemic hypoglycemia: A systematic review and meta-analysis.

Diazoxide is the first-line drug for treating hyperinsulinism and the only pharmacological agent approved for hyperinsulinism by the Federal Drug Administration. This systemic review and meta-analysis aimed to investigate the efficacy and safety of diazoxide for treating hyperinsulinemic hypoglycemia (HH). The meta-analysis of the efficacy and safety of diazoxide in treating HH was performed by searching relevant studies in the PubMed, Embase, and Cochrane databases. The findings were summarized, and the pooled effect size and its 95% confidence interval (CI) were calculated. A total of 6 cohort studies, involving 1142 participants, met the inclusion criteria. Among the cohort studies, the pooled estimate of the response rate of diazoxide therapy was 71% (95% CI 50%-93%, Pheterogeneity< 0.001, I2 = 98.3%, Peffect< 0.001). The common side effects were hypertrichosis (45%), fluid retention (20%), gastrointestinal reaction (13%), edema (11%), and neutropenia (9%). Other adverse events included pulmonary hypertension (2%) and thrombocytopenia (2%). This meta-analysis suggested that diazoxide was potentially useful in HH management; however, it had some side effects, which needed careful monitoring. Furthermore, well-designed large-scale studies, such as randomized controlled trials, might be necessary in the future to obtain more evidence.