Bacterial diversity in Arctic marine sediment determined by culture-dependent and -independent approaches

Bacterial diversity in surface sediment from the Arctic Ocean was investigated by culture-dependent and -independent approaches. Conventional culture-dependent techniques revealed 11 strains based on their distinct morphological characteristics on marine Zobell 2216E agar plates. Phylogenetic analysis showed that these isolates belonged to three major lineages of the Bacteria, γ-proteobacteria, Bacteroidetes and Actinobacteria, and that they included 10 genera. Most isolates were psychrotrophic, and NaCl was not necessary for their growth. Furthermore, they exhibited activity of at least one extracellular hydrolytic enzyme at 4°C and had various abilities to assimilate carbon sources. A total of 67 phylotypes were detected among 142 clones based on the 16S rRNA library of the total community DNA and grouped into nine major lineages of bacteria. Phylotypes affiliated with γ-, δ- and ε-proteobacteria accounted for 36.7%, 21.8% and 16.9% of the total clones, respectively. The rest of the clones belonged to Bacteroidetes, α-proteobacteria, Actinobacteria, Fusobacteria, Nitrospirae and an unclassified group

Multiplex glycan bead array for high throughput and high content analyses of glycan binding proteins

Glycan-binding proteins (GBPs) play critical roles in diverse cellular functions such as cell adhesion, signal transduction and immune response. Studies of the interaction between GBPs and glycans have been hampered by the availability of high throughput and high-content technologies. Here we report multiplex glycan bead array (MGBA) that allows simultaneous analyses of 384 samples and up to 500 glycans in a single assay. The specificity, sensitivity and reproducibility of MGBA are evaluated using 39 plant lectins, 13 recombinant anti-glycan antibodies, and mammalian GBPs. We demonstrate the utility of this platform by the analyses of natural anti-glycan IgM and IgG antibodies in 961 human serum samples and the discovery of anti-glycan antibody biomarkers for ovarian cancer. Our data indicate that the MGBA platform is particularly suited for large population-based studies that require the analyses of large numbers of samples and glycans

Evaluation of the national tobacco control mass media campaign in China

Background and challenges to implementation Tobacco use is the leading preventable cause of morbidity and mortality worldwide, yet the usage rate continues to increase in low- and middle-income countries such as China.Of over 310 million smokers lives in China, which accounts for 27.7% of the general population. Intervention or response In 2014, the BI collaborated with the Chinese Center for Health Education to implement the national tobacco control mass media campaign in China. The campaign was expected to improve knowledge about health effects of smoking and SHS exposure and shape attitudes and behaviors toward tobacco control policies. The campaign was adapted from materials which depicts a office environment with SHS to show how it affects everyone. It was broadcast on the national Television channels and six provincial satellite networks, meanwile on social media and mini-site for a four-week period. Results and lessons learnt To evaluate the effectiveness of this campaign, a total of 2400 participants were selected to complete interviews through a stratified random sampling. The participants consisted of 828(34.5%) smokers, and 1,572(65.5%) nonsmokers. The campaign reached about 24% of the Chinese population and increased smokers' knowledge about the harmful effects of SHS exposure(73% vs. 85%, p< 0.05). Both smokers and nonsmokers increased their knowledge that smoking lead to diseases other than lung cancer. Although smokers exposed to the campaign were more likely to take actions to reduce smoking than those not exposed(13% vs. 7%, p< 0.05), the percentage was still low. The support for bans on smoking in public places increased after the campaign, especially among nonsmokers and females. However, the support for ban on smoking in bars(48%) and restaurants(62%) were lower than that in hospitals(95%) and schools(94%). Conclusions and key recommendations The mass media campaign reinforced people's knowledge and attitudes about harmful health effects of smoking and SHS exposure, increased people's desire to quit, and improved people's support for smoking bans in public places

Novel Strategy to Release and Tag N‑Glycans for Functional Glycomics

Functional glycomics has been impeded by the lack of inexpensive enzymatic and mild chemical methods to acquire natural glycans in significant amounts. In this study, we have developed a new strategy we term “threshing and trimming” (TaT) to quickly obtain N-glycans from glycoproteins and animal tissues. TaT employs low-cost Pronase to degrade peptides and N-bromosuccinimide (NBS) to effect oxidative decarboxylation under very mild reaction conditions to generate homogeneous aglycon moieties as nitriles or aldehydes. These aglycons can be readily conjugated with fluorescent tags for profiling and functional study. TaT is an affordable alternative to expensive specialty enzymes and strong chemical treatment and unpleasant reagents, and should further drive the functional glycomics of N-glycans

Antimicrobial Resistance Profile and Genotypic Characteristics of Streptococcus suis Capsular Type 2 Isolated from Clinical Carrier Sows and Diseased Pigs in China

Streptococcus suis serotype 2 is an important zoonotic pathogen. Antimicrobial resistance phenotypes and genotypic characterizations of S. suis 2 from carrier sows and diseased pigs remain largely unknown. In this study, 96 swine S. suis type 2, 62 from healthy sows and 34 from diseased pigs, were analyzed. High frequency of tetracycline resistance was observed, followed by sulfonamides. The lowest resistance of S. suis 2 for -lactams supports their use as the primary antibiotics to treat the infection of serotype 2. In contrast, 35 of 37 S. suis 2 with MLS B phenotypes were isolated from healthy sows, mostly encoded by the ermB and/or the mefA genes. Significantly lower frequency of mrp+/epf +/sly+ was observed among serotype 2 from healthy sows compared to those from diseased pigs. Furthermore, isolates from diseased pigs showed more homogeneously genetic patterns, with most of them clustered in pulsotypes A and E. The data indicate the genetic complexity of S. suis 2 between herds and a close linkage among isolates from healthy sows and diseased pigs. Moreover, many factors, such as extensive use of tetracycline or diffusion of Tn916 with tetM, might have favored for the pathogenicity and widespread dissemination of S. suis serotype 2

Fluorescent Glycosylamides Produced by Microscale Derivatization of Free Glycans for Natural Glycan Microarrays

A novel strategy for creating naturally derived glycan microarrays has been developed. Glycosylamines are prepared from free reducing glycans and stabilized by reaction with acryloyl chloride to generate a glycosylamide in which the reducing monosaccharide has a closed-ring structure. Ozonolysis of the protected glycan yields an active aldehyde, to which a bifunctional fluorescent linker is coupled by reductive amination. The fluorescent derivatives are easily coupled through a residual primary alkylamine to generate glycan microarrays. This strategy preserves structural features of glycans required for antibody recognition and allows development of natural arrays of fluorescent glycans in which the cyclic pyranose structure of the reducing-end sugar residue is retained

Glycan microarray analysis of P-type lectins reveals distinct phosphomannose glycan recognition

The specificity of the cation-independent and -dependent mannose 6-phosphate receptors (CI-MPR and CD-MPR) for high mannose-type N-glycans of defined structure containing zero, one, or two Man-P-GlcNAc phosphodiester or Man-6-P phosphomonoester residues was determined by analysis on a phosphorylated glycan microarray. Amine-activated glycans were covalently printed on N-hydroxysuccinimide-activated glass slides and interrogated with different concentrations of recombinant CD-MPR or soluble CI-MPR. Neither receptor bound to non-phosphorylated glycans. The CD-MPR bound weakly or undetectably to the phosphodiester derivatives, but strongly to the phosphomonoester-containing glycans with the exception of a single Man7GlcNAc2-R isomer that contained a single Man-6-P residue. By contrast, the CI-MPR bound with high affinity to glycans containing either phospho-mono- or -diesters although, like the CD-MPR, it differentially recognized isomers of phosphorylated Man7GlcNAc2-R. This differential recognition of phosphorylated glycans by the CI- and CD-MPRs has implications for understanding the biosynthesis and targeting of lysosomal hydrolases

Unique repertoire of anti-carbohydrate antibodies in individual human serum.

Humoral immunity to pathogens and other environmental challenges is paramount to maintain normal health, and individuals lacking or unable to make antibodies are at risk. Recent studies indicate that many human protective antibodies are against carbohydrate antigens; however, little is known about repertoires and individual variation of anti-carbohydrate antibodies in healthy individuals. Here we analyzed anti-carbohydrate antibody repertoires (ACARs) of 105 healthy individual adult donors, aged 20-60+ from different ethnic backgrounds to explore variations in antibodies, as defined by binding to glycan microarrays and by affinity purification. Using microarrays that contained > 1,000 glycans, including antigens from animal cells and microbes, we profiled the IgG and IgM ACARs from all donors. Each donor expressed many ACAs, but had a relatively unique ACAR, which included unanticipated antibodies to carbohydrate antigens not well studied, such as chitin oligosaccharides, Forssman-related antigens, globo-type antigens, and bacterial glycans. We also saw some expected antibodies to ABO(H) blood group and α-Gal-type antigens, although these also varied among individuals. Analysis suggests differences in ACARs are associated with ethnicity and age. Thus, each individual ACAR is relatively unique, suggesting that individualized information could be useful in precision medicine for predicting and monitoring immune health and resistance to disease