Egg-based influenza split virus vaccine with monoglycosylation induces cross-strain protection against influenza virus infections.

Each year influenza virus infections cause hundreds of thousands of deaths worldwide and a significant level of morbidity with major economic burden. At the present time, vaccination with inactivated virus vaccine produced from embryonated chicken eggs is the most prevalent method to prevent the infections. However, current influenza vaccines are only effective against closely matched circulating strains and must be updated and administered yearly. Therefore, generating a vaccine that can provide broad protection is greatly needed for influenza vaccine development. We have previously shown that vaccination of the major surface glycoprotein hemagglutinin (HA) of influenza virus with a single N-acetylglucosamine at each of the N-glycosylation sites [monoglycosylated HA (HAmg)] can elicit better cross-protection compared with the fully glycosylated HA (HAfg). In the current study, we produced monoglycosylated inactivated split H1N1 virus vaccine from chicken eggs by the N-glycosylation process inhibitor kifunensine and the endoglycosidase Endo H, and intramuscularly immunized mice to examine its efficacy. Compared with vaccination of the traditional influenza vaccine with complex glycosylations from eggs, the monoglycosylated split virus vaccine provided better cross-strain protection against a lethal dose of virus challenge in mice. The enhanced antibody responses induced by the monoglycosylated vaccine immunization include higher neutralization activity, higher hemagglutination inhibition, and more HA stem selectivity, as well as, interestingly, higher antibody-dependent cellular cytotoxicity. This study provides a simple and practical procedure to enhance the cross-strain protection of influenza vaccine by removing the outer part of glycans from the virus surface through modifications of the current egg-based process.

Actin cytoskeleton remodeling drives epithelial-mesenchymal transition for hepatoma invasion and metastasis in mice.

High invasiveness is a hallmark of human hepatocellular carcinoma (HCC). Large tumors predict invasion and metastasis. Epithelial-mesenchymal transition (EMT) is crucial for cancer invasion and metastasis. However, the mechanisms whereby large tumors tend to undergo EMT remain unclear. We conducted a subgenome-wide screen and identified KLHL23 as an HCC invasion suppressor by inhibiting EMT. KLHL23 binds to actin and suppresses actin polymerization. KLHL23 silencing induced filopodium and lamellipodium formation. Moreover, EMT was suppressed by KLHL23 through its action on actin dynamics. Traditionally, actin cytoskeleton remodeling is downstream of EMT reprogramming. It is therefore intriguing to ask why and how KLHL23 inversely regulates EMT. Activation of actin cytoskeleton remodeling by either KLHL23 silencing or treatment with actin cytoskeleton modulators augmented cellular hypoxic responses in a cell-density-dependent manner, resulting in hypoxia-inducible factor (HIF) and Notch signals and subsequent EMT. Environmental hypoxia did not induce EMT unless actin cytoskeleton remodeling was simultaneously activated and only when cells were at high density. The resulting EMT was reversed by either adenosine 5'-triphosphate supplementation or actin polymerization inhibitors. Down-regulation of KLHL23 was associated with invasion, metastasis, and poor prognosis of HCC and pancreatic cancer. Correlations of tumor size with EMT and inverse association of expression of KLHL23 with HIF/Notch signals were further validated in patient-derived xenograft HCCs in mice. CONCLUSION: Simultaneously activation of actin cytoskeleton remodeling by intrinsic (such as KLHL23 down-regulation) or microenvironment cues is crucial for cell-density-dependent and hypoxia-mediated EMT, providing a mechanistic link between large tumor size and invasion/metastasis. Our findings provide a means of developing the prevention and treatment strategies for tumor invasion and metastasis. (Hepatology 2018;67:2226-2243).

Vaccination of monoglycosylated hemagglutinin induces cross-strain protection against influenza virus infections.

The 2009 H1N1 pandemic and recent human cases of H5N1, H7N9, and H6N1 in Asia highlight the need for a universal influenza vaccine that can provide cross-strain or even cross-subtype protection. Here, we show that recombinant monoglycosylated hemagglutinin (HAmg) with an intact protein structure from either seasonal or pandemic H1N1 can be used as a vaccine for cross-strain protection against various H1N1 viruses in circulation from 1933 to 2009 in mice and ferrets. In the HAmg vaccine, highly conserved sequences that were originally covered by glycans in the fully glycosylated HA (HAfg) are exposed and thus, are better engulfed by dendritic cells (DCs), stimulated better DC maturation, and induced more CD8+ memory T cells and IgG-secreting plasma cells. Single B-cell RT-PCR followed by sequence analysis revealed that the HAmg vaccine activated more diverse B-cell repertoires than the HAfg vaccine and produced antibodies with cross-strain binding ability. In summary, the HAmg vaccine elicits cross-strain immune responses that may mitigate the current need for yearly reformulation of strain-specific inactivated vaccines. This strategy may also map a new direction for universal vaccine design.

Epidemiology of and risk factors associated with third-generation cephalosporin-resistant Escherichia coli carriage in children: A comparison between regions in Taiwan.

BACKGROUND: The increasing prevalence of antimicrobial-resistant Escherichia coli (E. coli) in the community is a global public health challenge. This study investigated the prevalence of third-generation cephalosporin-resistant (3GCR) E. coli fecal carriage in children, identified associated risk factors, and determined antimicrobial resistance patterns of E. coli across three regions of Taiwan. METHODS: Stool samples from children aged 0-18 years were collected in southern, northern, and eastern Taiwan from community or outpatient clinics between July 2022 and May 2023. E. coli colonies were selected and examined for antimicrobial susceptibility and multilocus sequence typing. Participant demographic data and potential risk factors for carrying resistant E. coli were surveyed using a questionnaire. RESULTS: Of the 246 children surveyed, 59.3% carried multidrug-resistant (MDR) E. coli, and 37.4% carried 3GCR E. coli. The prevalence of 3GCR E. coli carriage was highest in southern Taiwan (42.7%), followed by northern Taiwan (35.5%) and eastern Taiwan (28.4%). The study identified several risk factors which may be associated with the fecal carriage of 3GCR E. coli, such as having lower paternal education levels, being overweight or obese, having a nonvegetarian diet, and consuming eggs, with variations observed across regions. CONCLUSION: This study documented elevated fecal carriage rates of 3GCR and MDR E. coli across regions of Taiwan. The study also identified numerous demographic and environmental factors that require implementing comprehensive strategies to address this public health challenge.

Sugar Consumption Pattern among Cardiometabolic Risk Individuals: A Scoping Review.

BACKGROUND: The global prevalence of noncommunicable diseases has risen rapidly over the past decade. Research has focused on dietary management, particularly dietary sugar, to prevent and treat noncommunicable diseases. OBJECTIVE: This study undertakes a scoping review of research on the impacts of dietary sugar on cardiometabolic related health outcomes. METHODS: Ovid Medline, Scopus and Web of Science Core collection databases were used to identify papers published from January 1, 2010 onwards. The included studies had to be cross-sectional or cohort studies, peered review, published in English and in adults, aged 18 years old and above. Articles had to determine the impacts of sugar intake on cardiometabolic related health outcomes. Study quality was measured using the Quality Assessment Tool for Observational Cohort and Cross-sectional Studies. In addition, a narrative synthesis of extracted information was conducted. RESULTS: Thirty-one articles were included in this review. All studies had a large sample size, and the exposure measure was clearly defined, valid and applied consistently across all study participants. Exposure was measured using validated questionnaires. All data were statistically analysed and adjusted for critical potential confounding variables. Results showed that dietary sugar intake was significantly associated with metabolic syndrome, blood pressure, blood glucose, blood lipids, and body weight. CONCLUSION: Dietary sugar intake significantly increased cardiometabolic risks through mechanisms dependent and independent of weight gain. It is essential to create public awareness on the topics of cardiometabolic risk management and dietary sugars intake.

Effectiveness of education intervention to reduce sugar-sweetened beverages and 100% fruit juice in children and adolescents: a scoping review.

OBJECTIVE: To determine the impact of educational intervention in reducing the consumption of sugar-sweetened beverages (SSBs) and 100% fruit juice in children and adolescents. DESIGN: We conducted a comprehensive Ovid Medline and Scopus search. Articles had to be peer-reviewed, full-text and published in English. Studies had to be controlled intervention, published between 1 January 2010 to 7 February 2021, and in children and adolescents. Study quality was measured using the Quality Assessment Tool for Controlled Intervention Studies. RESULTS: Forty intervention articles were included in this review involving 25,069 children and adolescents. All studies employed a quantitative research method using the Randomized Controlled Trial designs. Twenty-eight out of forty interventions used psychosocial theories. Study quality ranged from 'fair' to 'good.' Results showed that education intervention effectively reduced SSBs, and reduced 100% fruit juice in children. EXPERT OPINION: Reducing the SSBs consumption in children should be initiated by conducting school-based programs consisting of interactive learning process, psychosocial theories and the involvement of parents or caregivers. Theories that focus on personal, behavior and environment factors improve the effectiveness of the intervention. More research is warranted to investigate the impacts of 100% fruit juice on obesity, dental caries and risk of co-morbidities in children.

Impacts of a nutrition education intervention in individuals with cardiometabolic risk: Protocol of a randomized controlled trial.

BACKGROUND: Unhealthy dietary pattern including high dietary sugar intake is responsible for the cardiometabolic-related deaths. Nonetheless, adherence to the recommendation is low, and appropriate nutrition education to reduce sugar consumption is scarce. OBJECTIVE: This proposed project aims to evaluate the effectiveness of a nutrition education module in modifying sugar consumption and other cardiometabolic risk (CMR)-related outcomes in individuals with CMR. METHODS: This is an open-label, 3-group parallel-randomized controlled trial that will be conducted in Universiti Putra Malaysia, Serdang, Malaysia. Participants will be equally randomized into three groups. Participants in the Control Group (n = 35) would receive no intervention and undergo usual follow-up. In comparison, those participants in the Intervention Groups (n = 70) would receive nutrition education to modify sugars consumption. They will also be given either refined white sugar (n = 35), or Minimally Refined Brown Sugar (n = 35) for daily consumption throughout a six-month study period. Measurements include socio-demographic status, medical characteristics, nutritional status, lifestyle practices, Health Belief Model constructs, and knowledge of healthy diet will be measured at baseline, first- (V1), third- (V3) and sixth-month (V6) intervals. CONCLUSION: Participants of both Intervention Group aiming for sugar intake modification, would have lower sugar consumption and better CMR-related outcomes than the Control Group. Since Minimally Refined Brown Sugar contains fair amount of antioxidants, participants of Intervention Group (Minimally Refined Brown Sugar) will have better CMR-related outcomes than participants of Intervention Group (white sugar) at the end of the study.

Correlation between Visual Acuity and Optical Coherence Tomography Angiography Parameters in Unilateral Idiopathic Epiretinal Membrane.

BACKGROUND: The tangential traction by idiopathic epiretinal membrane (iERM) may alter the hemodynamics of the macula. We investigated the correlation between visual acuity and the optical coherence tomography angiography (OCTA) parameters in unilateral iERM. METHODS: We included 61 eyes of 61 consecutive patients with unilateral iERM between January 2018 and December 2018. The flow area of the retinal superficial capillary plexus (SCP), deep capillary plexus (DCP), and choroidal capillary plexus (CCP) were measured using OCTA. The normal fellow eyes were used for comparison. The iERM patients were divided into those with a presence of foveal concavity and those with a loss of foveal concavity. RESULTS: When compared with fellow eyes, the flow areas showed a statistically significant decrease in the SCP and CCP of those with iERM (p = 0.037 and p = 0.011, respectively). In the DCP, no significant reduction in flow area was found in iERM (p = 0.054). The flow area of the CCP was the only factor significantly associated with best vision (p = 0.012). No significant differences in the flow areas of the SCP, DCP, and CCP were found between the presence and loss of foveal concavity. CONCLUSIONS: The flow area of the CCP is an important determinant of vision, emphasizing the crucial role of choroidal circulation in iERM. Moreover, mechanical stretch by iERM is not the only mechanism affecting the flow area.

Elucidation of the stability and functional regions of the human coronavirus OC43 nucleocapsid protein.

Human coronavirus OC43 (HCoV-OC43) is one of the causes of the "common cold" in human during seasons of cold weather. The primary function of the HCoV-OC43 nucleocapsid protein (N protein) is to recognize viral genomic RNA, which leads to ribonucleocapsid formation. Here, we characterized the stability and identified the functional regions of the recombinant HCoV-OC43 N protein. Circular dichroism and fluorescence measurements revealed that the HCoV-OC43 N protein is more highly ordered and stabler than the SARS-CoV N protein previously studied. Surface plasmon resonance (SPR) experiments showed that the affinity of HCoV-OC43 N protein for RNA was approximately fivefold higher than that of N protein for DNA. Moreover, we found that the HCoV-OC43 N protein contains three RNA-binding regions in its N-terminal region (residues 1-173) and central-linker region (residues 174-232 and 233-300). The binding affinities of the truncated N proteins and RNA follow the order: residues 1-173-residues 233-300 > residues 174-232. SPR experiments demonstrated that the C-terminal region (residues 301-448) of HCoV-OC43 N protein lacks RNA-binding activity, while crosslinking and gel filtration analyses revealed that the C-terminal region is mainly involved in the oligomerization of the HCoV-OC43 N protein. This study may benefit the understanding of the mechanism of HCoV-OC43 nucleocapsid formation.