Partial denitrification is an alternative process to provide stable nitrite for anammox. In this study, based on full-scale and lab-scale experiments, achieving and control of partial denitrification and the microbial mechanism were studied for 17 months in municipal wastewater treatment plant (MWWTP). Using glucose (GLC) as sole carbon source, partial denitrification was successfully achieved with nitrite accumulation percentage (NAP) higher than 90%; whereas, using sodium acetate (NaAc) as sole carbon source, nitrite accumulation was effectively controlled with economic and efficient carbon usage. Candidatus Competibacter and Thaurea were the dominant communities for partial denitrification. Denitrifying glycogen accumulating organisms (DGAOs), Thauera, denitrifying phosphorus accumulating organisms (DPAOs), GAOs, PAOs and denitrifiers coexisted in MWWTP, resulting in COD specific removal rate (CODSRR) of 883.10 ~ 1188.92mgN/gMLVSS/h during partial denitrification. Through adjustment of Anoxic-Oxic (A/O) operation to anoxic operation, the growth of GAOs and PAOs could be limited.

Lipid synthesis is an important process in most organisms as well as in helminths. The present observation shows the variation of lipid and fatty acid uptake among cestode, Raillietina (Fuhrmannetta) echinobothrida; nematode, Ascaridia galli and their host, Gallus domesticus, the common country fowl. Total lipid (TL), neutral lipid (NL), glycolipid (GL), phospholipid (PL) and their fatty acid of cestode, nematode and liver and intestinal fluid of the host were analyzed by thin layer chromatography and gas liquid chromatography respectively. The result shows that liver take more TL, PL and GL except NL. Utilization of lipid from intestinal fluid when compare between the parasites, it is found that TL and PL content of cestode are higher than nematode, whereas, nematode absorbs more NL and GL than cestode. The percent of cholesterol is more in cestode than nematode. Palmitic, stearic, oleic and linoleic are the predominant fatty acids among all the samples. The present study reveals that the cestode having large surface area is more opportunistic in the resource utilization over the nematode as well as the host.

OBJECTIVE: To describe and analyse the prescription patterns and treatment outcomes of MDR-TB patients managed within Green Light Committee (GLC) and outside (non-GLC) the National TB programme in Viet Nam.
METHODS: Retrospective cohort study with two elements: (i) in-depth interviews and focus group discussions with clinical doctors, hospital pharmacists, and the non-GLC patients with MDR-TB; (ii) review of treatment cards and patients\' charts of all GLC and non-GLC patients with MDR-TB put on treatment during 2010.
RESULTS: Of 282 patients with MDR-TB, comprising 79 (28%) GLC patients MDR-TB and 203 (72%) non-GLC patients with MDR-TB, were enrolled in the study. Treatment delay was significantly higher in the GLC group (12.8 days) than the non-GLC group (2.3 days), (P = 0.004). The success rate was significantly better in GLC patients (84.8%) than in non-GLC patients (53.7%) (P < 0.001). The default rate was significantly higher in non-GLC patients than in GLC patients (25.6% vs. 6.3%), (P < 0.001). The risk of unsuccessful outcome was higher in non-GLC patients (Hazard ratio = 4.6, 95% CI: 1.8-11.8).
CONCLUSIONS: The treatment outcomes of patients with MDR-TB in the GLC group were significantly better than in the non-GLC group. Reasons for the high default rate in non-GLC patients with MDR-TB must be further investigated.

Kakkalide and irisolidone, the main isoflavones of Flos Puerariae, exhibit a wide spectrum of bioactivities. Intestinal bacteria biotransformation plays an important role in the metabolic pathways of flavones, and is directly related to the bioactivities of the prodrugs after oral administration. To the best of our knowledge, the metabolic pathways of kakkalide and irisolidone in vitro have not been comprehensively studied yet. This paper describes the strategy using ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOF MS) for the rapid analysis of the metabolic profiles of kakkalide and irisolidone after incubated with human and rat intestinal bacteria. Bacteria incubated samples were prepared and analyzed after incubated under anaerobic conditions for 48 h. A total of 17 metabolites, including parent compounds, were detected in human and rat intestinal bacteria incubated samples. The results obtained indicate that hydrolysis, dehydroxylation, demethoxylation, demethylation, hydroxylation, decarbonylation, and reduction were the detected metabolic pathways of kakkalide and irisolidone in vitro. The conversion rate of irisolidone in human and rat bacteria was 8.57% and 6.51%, respectively. Biochanin A was the relatively main metabolite of irisolidone, and the content of biochanin A in human and rat bacteria was 3.68% and 4.25%, respectively. The conversion rate of kakkalide in human and rat bacteria was 99.92% and 98.58%, respectively. Irisolidone was the main metabolite of kakkalide, and the content of irisolidone in human and rat bacteria was 89.58% and 89.38%, respectively. This work not only provides the evidence of kakkalide and irisolidone metabolites in vivo, but also demonstrates a simple, fast, sensitive, and inexpensive method for identification of metabolites of other compounds transformed by intestinal bacteria.

The bark of Parkia biglobosa is used in traditional medicine to cure a wide range of illnesses. Polysaccharides were extracted from the bark with 50% ethanol-water, 50°C and 100°C water, and seven active fractions obtained by anion exchange chromatography and gel filtration. The complement fixation and macrophage stimulating activities of the different fractions were determined. The acidic fractions PBEII-I and PBEII-IV were the most active in the complement fixation assay, but the other fractions were also potent compared to the positive control BPII from Biophytum petersianum. Fractions PBEII-I and PBEII-IV were also the most potent fractions in stimulating macrophages to release nitric oxide. Structural studies showed that PBEII-I and PBEII-IV were pectic type polysaccharides, containing arabinogalactan type II structures. The observed differences in biological activities among the seven purified polysaccharide sub-fractions are probably due to differences in monosaccharide compositions, linkage types and molecular sizes.

Cyanobacteria are known to be a rather diverse group of organisms regarding e.g. morphology, metabolism and composition of excreted exopolysaccharides (EPS). Considering the high number of known cyanobacterial species the EPS from only a small percentage are investigated in detail. This work examined EPS from the unicellular strains of Synechocystis aquatilis and S. pevalekii with various methods. The results emphasize the heterogeneity of cyanobacterial EPS. S. pevalekii secrets complex hetero-polysaccharides and acidic proteins as proteoglycan-complexes whereas the protein-free but highly sulfated EPS from S. aquatilis only consist of 4 dominant monosaccharides. Especially remarkable is the composition of these EPS: an arabinofucan with higher amounts of N-acetyl-fucosamine (FucNAc) and only minor quantities of glucose. Both EPS and the newly found component FucNAc in EPS from S. aquatilis extend the possible components of cyanobacterial EPS and the knowledge of heterogeneity of cyanobacterial metabolites.

Snails from the genus Pomacea lay conspicuous masses of brightly colored eggs above the water. Coloration is given by carotenoproteins that also which play important roles in protection against sun radiation, stabilizing and transporting antioxidant molecules and helping to protect embryos from desiccation and predators. They seem a key acquisition, but have been little studied. Here we report the characteristics of the major carotenoprotein from Pomacea maculata and the first comparison among these egg proteins. This particle, hereafter PmPV1, represents ~52% of perivitellin fluid protein. It is a glyco-lipo-carotenoprotein responsible for the bright reddish egg coloration. With VHDL characteristics, PmPV1 apparent molecular mass is 294kDa, composed of five non-covalently bound subunits of pI 4.7-9.8 and masses between 26 and 36kDa whose N-terminal sequences were obtained. It is a glyco-lipo-carotenoprotein scarcely lipidated (<1%) but highly glycosilated (13% by wt). Lipids include phospholipids, free fatty acids and carotenoids; mannose and galactose predominate over other monosaccharides. Main carotenoids are esterified and non-esterified astaxanthin (71 and 25%, respectively). Carotenoid removal does not seem to affect the structural characteristics of the oligomer, while deglycosilation reduces subunit number from five to a single one. The carotenoid-protein association protected the former against oxidation. PmPV1 cross reacts with polyclonal antibodies against the PcOvo, the major carotenoprotein from Pomacea canaliculata. The characterization of PmPV1 allows the first comparisons among snail carotenoproteins and further highlights the importance of these perivitellins in the reproductive strategy of Pomacea.

The thermostable 1,3-1,4-β-glucanase PtLic16A from the fungus Paecilomyces thermophila catalyzes stringent hydrolysis of barley β-glucan and lichenan with an outstanding efficiency and has great potential for broad industrial applications. Here, we report the crystal structures of PtLic16A and an inactive mutant E113A in ligand-free form and in complex with the ligands cellobiose, cellotetraose and glucotriose at 1.80Å to 2.25Å resolution. PtLic16A adopts a typical β-jellyroll fold with a curved surface and the concave face forms an extended ligand binding cleft. These structures suggest that PtLic16A might carry out the hydrolysis via retaining mechanism with E113 and E118 serving as the nucleophile and general acid/base, respectively. Interestingly, in the structure of E113A/1,3-1,4-β-glucotriose complex, the sugar bound to the -1 subsite adopts an intermediate-like (α-anomeric) configuration. By combining all crystal structures solved here, a comprehensive binding mode for a substrate is proposed. These findings not only help understand the 1,3-1,4-β-glucanase catalytic mechanism but also provide a basis for further enzymatic engineering.

Glycogen synthesis is initiated by self-glucosylation of the glycosyltransferases glycogenin-1 and -2 that, in the presence of UDP-glucose, form both the first glucose-O-tyrosine linkage, and then stepwise add a series of α1,4-linked glucoses to a growing chain of variable length. Glycogen-1 and -2 coexist in liver glycogen preparations where the proteins are known to form homodimers, and they also have been shown to interact with each other. In order to study how glycogenin-1 and -2 interactions may influence each other\'s glucosylations we setup a cell-free expression system for in vitro production and glucosylation of glycogenin-1 and -2 in various combinations, and used a mass spectrometry based workflow for the characterization and quantitation of tryptic glycopeptides originating from glycogenin-1 and -2. The analysis revealed that the self-glucosylation endpoint was the incorporation of 4-8 glucose units on Tyr 195 of glycogenin-1, but only 0-4 glucose units on Tyr-228 of glycogenin-2. The glucosylation of glycogenin-2 was enhanced to 2-4 glucose units by the co-presence of enzymatically active glycogenin-1. Glycogenin-2 was, however, unable to glucosylate inactive glycogenin-1, at least not an enzymatically inactivated Thr83Met glycogenin-1 mutant, recently identified in a patient with severe glycogen depletion.

Congenital disorders of glycosylation (CDG) are a growing group of inherited metabolic disorders where enzymatic defects in the formation or processing of glycolipids and/or glycoproteins lead to variety of different diseases. The deficiency of GDP-Man:GlcNAc2-PP-dolichol mannosyltransferase, encoded by the human ortholog of ALG1 from yeast, is known as ALG1-CDG (CDG-Ik). The phenotypical, molecular and biochemical analysis of a severely affected ALG1-CDG patient is the focus of this paper. The patient\'s main symptoms were feeding problems and diarrhea, profound hypoproteinemia with massive ascites, muscular hypertonia, seizures refractory to treatment, recurrent episodes of apnoea, cardiac and hepatic involvement and coagulation anomalies. Compound heterozygosity for the mutations c.1145T>C (M382T) and c.1312C>T (R438W) was detected in the patient\'s ALG1-coding sequence. In contrast to a previously reported speculation on R438W we confirmed both mutations as disease-causing in ALG1-CDG.