Transcription factor Tox2 is required for metabolic adaptation and tissue residency of ILC3 in the gut.

Group 3 innate lymphoid cells (ILC3) are the major subset of gut-resident ILC with essential roles in infections and tissue repair, but how they adapt to the gut environment to maintain tissue residency is unclear. We report that Tox2 is critical for gut ILC3 maintenance and function. Gut ILC3 highly expressed Tox2, and depletion of Tox2 markedly decreased ILC3 in gut but not at central sites, resulting in defective control of Citrobacter rodentium infection. Single-cell transcriptional profiling revealed decreased expression of Hexokinase-2 in Tox2-deficient gut ILC3. Consistent with the requirement for hexokinases in glycolysis, Tox2-/- ILC3 displayed decreased ability to utilize glycolysis for protein translation. Ectopic expression of Hexokinase-2 rescued Tox2-/- gut ILC3 defects. Hypoxia and interleukin (IL)-17A each induced Tox2 expression in ILC3, suggesting a mechanism by which ILC3 adjusts to fluctuating environments by programming glycolytic metabolism. Our results reveal the requirement for Tox2 to support the metabolic adaptation of ILC3 within the gastrointestinal tract.

[Analysis and clinical application of preimplantation genetic testing for monogenic disorders in a case with Spinal muscular atrophy "2+0" genotype].

OBJECTIVE: To explore the clinical application of preimplantation genetic testing for monogenic disorders (PGT-M) in an unique case with Spinal muscular atrophy (SMA) type 2+0. METHODS: A special SMA family presented at the Third Affiliated Hospital of Guangzhou Medical University on October 19, 2020 was selected as the study subject. Multiple ligation-dependent probe amplification (MLPA) and molecular tagging linkage analysis were carried out to identify the SMN1 genotype of the couple and their fetus. Subsequently, next-generation sequencing (NGS), molecular tagging linkage analysis, and chromosomal microarray analysis were employed to determine the haplotypes and validate the result of PGT-M on the 11 embryos derived for the couple. RESULTS: The female partner was identified as a carrier of the rare SMN1[2+0] variant, and prenatal diagnosis confirmed the fetus to be affected by SMA. Ultimately, PGT-M has successfully selected four embryos free from the pathogenic SMN1 variants and X chromosome deletion. CONCLUSION: PGT-M can effectively prevent the transmission of rare genetic variants such as the SMA 2+0 subtype in the families. Above finding has provided guidance for genetic counseling and family planning for the couple.

Effect of noninvasive embryo viability testing versus conventional IVF on the live birth rate in IVF/ICSI patients: a study protocol for a double-blind, multicenter, randomized controlled trial.

BACKGROUND: Preimplantation genetic testing for aneuploidy (PGT-A) was demonstrated to be superior to conventional IVF in reducing the incidence of miscarriage and abnormal offspring after the first embryo transfer (ET). PGT-A requires several embryo trophectoderm cells, but its negative impacts on embryo development and long-term influence on the health conditions of conceived children have always been a concern. As an alternative, noninvasive PGT-A (niPGT-A) approaches using spent blastocyst culture medium (SBCM) achieved comparable accuracy with PGT-A in several pilot studies. The main objective of this study is to determine whether noninvasive embryo viability testing (niEVT) results in better clinical outcomes than conventional IVF after the first embryo transfer. Furthermore, we further investigated whether niEVT results in higher the live birth rate between women with advanced maternal age (AMA, > 35 years old) and young women or among patients for whom different fertilization protocols are adopted. METHODS: This study will be a double-blind, multicenter, randomized controlled trial (RCT) studying patients of different ages (20-43 years) undergoing different fertilization protocols (in vitro fertilization [IVF] or intracytoplasmic sperm injection [ICSI]). We will enroll 1140 patients at eight reproductive medical centers over 24 months. Eligible patients should have at least two good-quality blastocysts (better than grade 4 CB). The primary outcome will be the live birth rate of the first embryo transfer (ET). Secondary outcomes will include the clinical pregnancy rate, ongoing pregnancy rate, miscarriage rate, cumulative live birth rate, ectopic pregnancy rate, and time to pregnancy. DISCUSSION: In this study, patients who undergo noninvasive embryo viability testing (niEVT) will be compared to women treated by conventional IVF. We will determine the effects on the pregnancy rate, miscarriage rate, and live birth rate and adverse events. We will also investigate whether there is any difference in clinical outcomes among patients with different ages and fertilization protocols (IVF/ICSI). This trial will provide clinical evidence of the effect of noninvasive embryo viability testing on the clinical outcomes of the first embryo transfer. TRIAL REGISTRATION: Chinese Clinical Trial Registry (ChiCTR) Identifier: ChiCTR2100051408. 9 September 2021.

Identification of QTLs and joint QTL segments of leaflet traits at different canopy layers in an interspecific RIL population of soybean.

KEY MESSAGE: A leaflet trait on different canopy layers may have different QTLs; leaflet trait QTLs may cluster to form joint QTL segments; all canopy layer QTLs form a complete QTL system for a leaflet trait. As the main part of the plant canopy structure, leaf/leaflet size and shape affect the plant architecture and yield. To explore the leaflet trait QTL system, a population composed of 199 recombinant inbred lines derived from Changling (annual wild, narrow leaflet) and Yiqianli (landrace, broad leaflet) with their parents was tested for leaflet length (LL), width (LW) and length to width (LLW). The population was genotyped with specific-locus amplified fragment sequencing (SLAF-seq) and applied for linkage mapping of the leaflet traits. The results showed that the leaflet traits varied greatly even within a plant, which supported a stratified leaflet sampling strategy to evaluate these traits at top, middle and bottom canopy layers. Altogether, 13 LL, 10 LW and 9 LLW in a total of 32 plus 3 duplicated QTLs were identified, in which, 17 QTLs were new ones, and 48.6%, 28.6% and 22.8% of QTLs were from the top, middle and bottom layers, respectively, indicating the genetic importance of the top layer leaves. Since a leaflet trait may have layer-specific QTLs, all layer QTLs form a complete QTL system. Five QTL clusters each with their QTL supporting intervals overlapped were designated as joint QTL segments (JQSs). In JQS-16, with its linkage map further validated using PCR markers, two QTLs, qLW-16-1 and qLLW-16-1 of the top layer leaflet, were identified six QTL·times. Six candidate genes were predicted, with Glyma.16G127900 as the most potential one for LW and LLW. Three PCR markers, Gm16PAV0653, BARCSOYSSR_16_0796 and YC-16-3, were suggested for marker-assisted selection for LW and LLW in JQS-16.

TOX and TOX2 transcription factors cooperate with NR4A transcription factors to impose CD8+ T cell exhaustion.

T cells expressing chimeric antigen receptors (CAR T cells) have shown impressive therapeutic efficacy against leukemias and lymphomas. However, they have not been as effective against solid tumors because they become hyporesponsive ("exhausted" or "dysfunctional") within the tumor microenvironment, with decreased cytokine production and increased expression of several inhibitory surface receptors. Here we define a transcriptional network that mediates CD8+ T cell exhaustion. We show that the high-mobility group (HMG)-box transcription factors TOX and TOX2, as well as members of the NR4A family of nuclear receptors, are targets of the calcium/calcineurin-regulated transcription factor NFAT, even in the absence of its partner AP-1 (FOS-JUN). Using a previously established CAR T cell model, we show that TOX and TOX2 are highly induced in CD8+ CAR+ PD-1high TIM3high ("exhausted") tumor-infiltrating lymphocytes (CAR TILs), and CAR TILs deficient in both TOX and TOX2 (Tox DKO) are more effective than wild-type (WT), TOX-deficient, or TOX2-deficient CAR TILs in suppressing tumor growth and prolonging survival of tumor-bearing mice. Like NR4A-deficient CAR TILs, Tox DKO CAR TILs show increased cytokine expression, decreased expression of inhibitory receptors, and increased accessibility of regions enriched for motifs that bind activation-associated nuclear factor κB (NFκB) and basic region-leucine zipper (bZIP) transcription factors. These data indicate that Tox and Nr4a transcription factors are critical for the transcriptional program of CD8+ T cell exhaustion downstream of NFAT. We provide evidence for positive regulation of NR4A by TOX and of TOX by NR4A, and suggest that disruption of TOX and NR4A expression or activity could be promising strategies for cancer immunotherapy.

Study on Detection and Recognition of Traffic Lights Based on Improved YOLOv4.

To resolve the issues of a deep backbone network, a large model, slow reasoning speed on a mobile terminal, low detection accuracy for small targets and difficulties detecting and recognizing traffic lights in real time and accurately with YOLOv4, a traffic lights recognition method based on improved YOLOv4 is proposed. The lightweight ShuffleNetv2 network is utilized to replace the CSPDarkNet53 network of YOLOv4 to satisfy the requirements of a mobile terminal. The reformed k-means clustering algorithm is applied to generate anchor boxes for avoiding the sensitivity issue of outliers and initial values. A novel attention mechanism named CS2A is added to enhance the extraction capability of effective features. Multiple data augmentation methods are combined to improve the generalization ability of the model. Ultimately, the detection and recognition of traffic lights can be realized. The S2TLD dataset is selected for training and testing, and it can be proved that the recognition accuracy and model size are greatly optimized. Meanwhile, a self-made dataset is selected for training and testing. Compared with the conventional YOLOv4, the recognition accuracy of the proposed algorithm for traffic lights' state information increases by 1.79%, and the model size decreases by 81.97%. Appropriate scenes are selected for real-vehicle testing and the results demonstrate that the detection speed of the presented algorithm increases by 16%, and the recognition effect for small targets increases by 37% in comparison with conventional YOLOv4.

Identification of an Intronic Regulatory Element Necessary for Tissue-Specific Expression of Foxn1 in Thymic Epithelial Cells.

The thymus is critical for the establishment of the adaptive immune system and the development of a diverse T cell repertoire. T cell development depends upon cell-cell interactions with epithelial cells in the thymus. The thymus is composed of two different types of epithelial cells: cortical and medullary epithelial cells. Both of these express and critically depend on the transcription factor Foxn1 Foxn1 is also expressed in the hair follicle, and disruption of Foxn1 function in mice results in severe thymic developmental defects and the hairless (nude) phenotype. Despite its importance, little is known about the direct regulation of Foxn1 expression. In this study, we identify a cis-regulatory element (RE) critical for expression of Foxn1 in mouse thymic epithelial cells but dispensable for expression in hair follicles. Analysis of chromatin accessibility, histone modifications, and sequence conservation identified regions within the first intron of Foxn1 that possessed the characteristics of REs. Systematic knockout of candidate regions lead us to identify a 1.6 kb region that, when deleted, results in a near total disruption of thymus development. Interestingly, Foxn1 expression and function in the hair follicle were unaffected. RNA fluorescent in situ hybridization showed a near complete loss of Foxn1 mRNA expression in the embryonic thymic bud. Our studies have identified a genomic RE with thymic-specific control of Foxn1 gene expression.

CCDC114, DNAI2 and TOP2A involves in the effects of tibolone treatment on postmenopausal endometrium.

BACKGROUND: This study aimed to explore the molecular mechanisms of tibolone treatment in postmenopausal women. METHODS: The gene set enrichment profile, GSE12446, which includes 9 human endometrial samples from postmenopausal women treated with tibolone (tibolone group) and 9 control samples (control group), was downloaded from GEO database for analysis. Differentially expressed genes (DEGs) in tibolone vs. control groups were identified and then used for function and pathway enrichment analysis. Protein-protein interaction (PPI) network and module analyses were also performed. Finally, drug-target interaction was predicted for genes in modules, and then were validated in Pubmed. RESULTS: A total of 238 up-regulated DEGs and 72 down-regulated DEGs were identified. These DEGs were mainly enriched in various biological processed and pathways, such as cilium movement (e.g., CCDC114 and DNAI2), calcium ion homeostasis, regulation of hormone levels and complement/coagulation cascades. PPI network contained 368 interactions and 166 genes, of which IGF1, DNALI1, CCDC114, TOP2A, DNAH5 and DNAI2 were the hue genes. A total of 96 drug-gene interactions were obtained, including 94 drugs and eight genes. TOP2A and HTR2B were found to be targets of 28 drugs and 38 drugs, respectively. Among the 94 obtained drugs, only 12 drugs were reported in studies, of which 7 drugs (e.g., epirubicin) were found to target TOP2A. CONCLUSIONS: CCDC114 and DNAI2 might play important roles in tibolone-treated postmenopausal women via cilium movement function. TOP2A might be a crucial target of tibolone in endometrium of postmenopausal women.

[Quercetin for the treatment of prostate cancer: Progress in studies].

Prostate cancer (PCa) is a common urinary malignancy, and advanced PCa has a poor prognosis and a high mortality. Drug therapies currently available for this malignancy often cause serious adverse reactions, and therefore new drugs with fewer adverse effects or the potential to reduce the adverse effects of traditional chemotherapeutic drugs are badly needed for the management of PCa. Quercetin, as a natural flavonoid, has been extensively studied in recent years for its anti-cancer effects, as in cell signal transduction, apoptosis promotion, anti-proliferation and -oxidation, and growth inhibition. In fact, quercetin has a variety of biological effects and can inhibit various enzymes involved in cell proliferation and signal transduction pathways. Besides, quercetin is also reported to have potential synergistic effects when used in combination with radiotherapy or chemotherapeutic drugs. This review summarizes the advances in the treatment of PCa with quercetin, focusing on its effects of promoting the apoptosis, inhibiting the proliferation and reducing the invasiveness and migration of tumor cells, and reversing drug resistance, aiming to provide a new theoretical basis and some new ideas for the studies of the treatment of PCa.

Gene expression changes in arterial and venous endothelial cells exposed to gestational diabetes mellitus.

We investigated the molecular changes in fetoplacental blood vessel endothelial cells in gestational diabetes mellitus (GDM). Raw gene expression profile data of arterial and venous endothelial cells from GDM complicated pregnancies and healthy controls were downloaded and used for bioinformatic analysis. There were two differentially expressed genes (DEGs) in venous endothelial cells and 178 DEGs in arterial endothelial cells induced by GDM. The altered genes were clustered to pathways associated with cell cycle, p53 signaling pathway, and cellular senescence. The disease associated gene-pathway network that was constructed comprised eight down-regulated genes (including FBXO5, CCNB1, and CDK1), one up-regulated gene (CCND2), hsa04068: FoxO signaling pathway and hsa04114: Oocyte mitosis pathway. CCND2 was a significant node in the microRNA (miRNA)-target network, which was regulated by seven miRNAs that included hsa-miR-1299, hsa-miR-1200, and hsa-miR-miR-593-5p. FBXO5 was a significant node regulated by two miRNAs. CCND2 and FBXO5 were also the significant nodes in the transcriptional factors-target network and integrated regulatory network. The cell cycle pathway was significantly altered in arterial endothelial cells during GDM, which was involved with the differential expression of CCND2 and FBXO5.

Transgenic Clustered Regularly Interspaced Short Palindromic Repeat/Cas9-Mediated Viral Gene Targeting for Antiviral Therapy of Bombyx mori Nucleopolyhedrovirus.

We developed a novel antiviral strategy by combining transposon-based transgenesis and the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system for the direct cleavage of Bombyx mori nucleopolyhedrovirus (BmNPV) genome DNA to promote virus clearance in silkworms. We demonstrate that transgenic silkworms constitutively expressing Cas9 and guide RNAs targeting the BmNPV immediate early-1 (ie-1) and me53 genes effectively induce target-specific cleavage and subsequent mutagenesis, especially large (∼7-kbp) segment deletions in BmNPV genomes, and thus exhibit robust suppression of BmNPV proliferation. Transgenic animals exhibited higher and inheritable resistance to BmNPV infection than wild-type animals. Our approach will not only contribute to modern sericulture but also shed light on future antiviral therapy.IMPORTANCE Pathogen genome targeting has shown its potential in antiviral research. However, transgenic CRISPR/Cas9 system-mediated viral genome targeting has not been reported as an antiviral strategy in a natural animal host of a virus. Our data provide an effective approach against BmNPV infection in a real-world biological system and demonstrate the potential of transgenic CRISPR/Cas9 systems in antiviral research in other species.

Salibacterium lacus sp. nov., a halophilic, non-spore-forming bacterium isolated from sediment of a saline lake.

A novel halophilic bacterium, strain GSS13T, capable of growing at salinities of 8-28 % (w/v) NaCl (optimally at 24 %, w/v) was isolated from Yuncheng Saline Lake in China. GSS13T was Gram-stain-positive, strictly aerobic, rod-shaped, motile and a non-spore-former. Growth occurred at pH 5.5-8.5 (optimum pH 7.0) and at 10-45 °C (optimum 30 °C). On the basis of the results of 16S rRNA gene sequences phylogenetic analyses, GSS13T represents a member of the genus Salibacterium and is closely related to Salibacterium halotolerans S7T, Salibacterium qingdaonense CM1T and Salibacterium halochares MSS4T, with 16S rRNA gene sequence similarities of 98.7, 98.4 and 97.9 %, respectively. The results of DNA-DNA pairing studies revealed that GSS13T displayed 52, 43 and 48 % relatedness to S. halotolerans S7T, S. qingdaonense CM1T and S. halochares MSS4T, respectively. The polar lipids of GSS13 consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol unidentified glycolipids, an unidentified phospholipid and an unidentified lipid. The predominant isoprenoid quinone was MK-7, and the major fatty acids were anteiso-C17 : 0 (32.0 %) and anteiso C15 : 0 (26.4 %). The DNA G+C content of the type strain was 52.1 mol%. On the basis of phylogenetic, chemotaxonomic and phenotypic data, a novel species of the genus Salibacterium is proposed, with the name Salibacterium lacus sp. nov. The type strain is GSS13T (=KCTC 33792T=MCCC 1K00567T).

Safety assessment of medium- and long-chain triacylglycerols containing 30% (w/w) medium-chain fatty acids in mice and rats.

A novel medium- and long-chain triacylglycerols (MLCT), with 30% (w/w) medium-chain fatty acids (MCFA) was evaluated for its safety as a dietary fat in mice and rats. The subacute oral toxicity study showed that the maximum tolerated dose exceeded 54.33 g/kg body weight (kg bw)/day. In the 90-day feeding study, no dose-related adverse effects were observed in rats administered diets formulated with different levels of MLCT (2.0, 4.0, and 8.0 g/kg bw/day) as compared to the rapeseed oil control diet. Further safety assessment in pregnant rats did not reveal any significant difference relative to the control at a treatment level up to 8.0 g MLCT/kg bw/day. The results from this study indicated the safe use of MLCT with high contents of MCFA in food products for improving human health.

Nitratireductor lacus sp. nov., isolated from Yuncheng Salt Lake, China.

This is the taxonomic study of a novel bacterial strain, designated GSS14T, isolated from a sediment sample of Yuncheng Salt Lake, China. Cells were Gram-negative, ovoid to rod-shaped and motile by means of flagella. The isolate could grow at 10-45 °C, at pH 6.5-11.0 and in the presence of 0-12 % (w/v) NaCl. The dominant fatty acids were summed feature 8 (consisting of C18 : 1ω7c and/or C18 : 1ω6c; 76.7 %) and the DNA G+C content was 61 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain GSS14T was affiliated with the genus Nitratireductor, and it was most closely related to Nitratireductor kimnyeongensis KY 101T (98.2 % 16S rRNA gene sequence similarity) and Nitratireductor aquibiodomus JCM 21793T (96.6 %). DNA-DNA hybridization between strains GSS14T and N. kimnyeongensis KY 101T revealed 52 % relatedness. Phenotypic, chemotaxonomic and phylogenetic data support assignment of this isolate to the genus Nitratireductor as a representative of a novel species. The name Nitratireductor lacus sp. nov. is proposed, with strain GSS14T (=KCTC 52186T=MCCC 1K02481) as the type strain.

Cyclosporin A Protects H9c2 Cells Against Chemical Hypoxia-Induced Injury via Inhibition of MAPK Signaling Pathway.

This study aimed to investigate the effects and molecular mechanism of cyclosporin A (CsA) on cobalt chloride (CoCl2)-induced injury in H9c2 embryonic rat cardiac cells. The results showed that CsA could protect H9c2 cells against CoCl2-induced hypoxic injury. CsA effectively improved cell viability, and decreased LDH leakage, cell apoptosis, MDA concentration, and ROS generation, and increased SOD activity, GSH production, and CAT activity in a dosedependent manner. In addition, CsA treatment blocked the CoCl2-induced increases in ROS production and mitochondrial dysfunction, including a decrease in membrane potential, cytochrome c (cyto-c) release, Bax/Bcl-2 imbalance, as well as the ratios of cl-casp-9/casp-9 and cl-casp-3/casp-3 ratios, via the inhibition of p38 and ERK MAPK signaling pathways. The results also suggested that CsA protected H9c2 cells against CoCl2-induced hypoxic injury, possibly by suppressing the MAPK signaling pathway. Thus, CsA is a potential therapeutic agent for cardiac hypoxic injury.

Porphyrin cosensitization for a photovoltaic efficiency of 11.5%: a record for non-ruthenium solar cells based on iodine electrolyte.

Dye-sensitized solar cells (DSSCs) are promising for utilizing solar energy. To achieve high efficiencies, it is vital to synergistically improve the photocurrent (Jsc) and the photovoltage (Voc). In this respect, conjugation framework extension and cosensitization are effective for improving the absorption and the Jsc, which, however, is usually accompanied by undesirably decreased Voc. Herein, based on a rationally optimized porphyrin dye, we develop a targeted coadsorption/cosensitization approach for systematically improving the Voc from 645 to 727, 746, and 760 mV, with synergistical Jsc enhancement from 18.83 to 20.33 mA cm(-2). Thus, the efficiency has been dramatically enhanced to 11.5%, which keeps the record for nonruthenium DSSCs using the I2/I3(-) electrolyte. These results compose an alternative approach for developing highly efficient DSSCs with relatively high Voc using traditional iodine electrolyte.

Bacillus dabaoshanensis sp. nov., a Cr(VI)-tolerant bacterium isolated from heavy-metal-contaminated soil.

A Cr(VI)-tolerant, Gram-staining-positive, rod-shaped, endospore-forming and facultative anaerobic bacterium, designated as GSS04(T), was isolated from a heavy-metal-contaminated soil. Strain GSS04(T) was Cr(VI)-tolerant with a minimum inhibitory concentration of 600 mg l(-1) and was capable of reducing Cr(VI) under both aerobic and anaerobic conditions. Growth occurred with presence of 0-3 % (w/v) NaCl (optimum 1 %), at pH 5.5-10.0 (optimum pH 7.0) and 15-50 °C (optimum 30-37 °C). The main respiratory quinone was MK-7 and the major fatty acids were anteiso-C15:0 and iso-C15:0. The DNA G+C content was 41.1 mol%. The predominant polar lipid was diphosphatidylglycerol. Based on 16S rRNA gene sequence similarity, the closest phylogenetic relative was Bacillus shackletonii DSM 18868(T) (97.6 %). The DNA-DNA hybridization between GSS04(T) and its closest relatives revealed low relatedness (<70 %). The results of phenotypic, chemotaxonomic and genotypic analyses clearly indicated that strain GSS04(T) represents a novel species of the genus Bacillus, for which the name Bacillus dabaoshanensis sp. nov. is proposed. The type strain is GSS04(T) (=CCTCC AB 2013260(T) = KCTC 33191(T)).

Coupling bioleaching and electrokinetics to remediate heavy metal contaminated soils.

In this study, bioleaching was coupled with electrokinetics (BE) to remove heavy metals (Cu, Zn, Cr and Pb) from contaminated soil. For comparison, bioleaching (BL), electrokinetics (EK), and the chemical extraction method were also applied alone to remove the metals. The results showed that the BE method removed more heavy metals from the contaminated soil than the BL method or the EK method alone. The BE method was able to achieve metal solubilization rates of more than 70 % for Cu, Zn and Cr and of more than 40 % for Pb. Within the range of low current densities (<1 mA cm(-2)), higher current density led to more metal removal. However, the metal solubilization rates did not increase with increasing current density when the current density was higher than 1 mA cm(-2). Therefore, it is suggested that bioleaching coupled with electrokinetics can effectively remediate heavy metal-contaminated soils and that preliminary tests should be conducted before field operation to detect the lowest current density for the greatest metal removal.

Enhanced dewaterability of waste-activated sludge by combined cationic polyacrylamide and magnetic field pretreatment.

The potential function of combining magnetic field (MF) pretreatment with cationic polyacrylamide (CPAM) additive on enhancing the dewaterability of waste-activated sludge was investigated in the present work. Two reactors were involved in a specially designed experimental apparatus, one of which was built with MF accessories. Several parameters were conducted, including CPAM dosages, MF strengths and processing times, respectively. Capillary suction time (CST) and specific resistance to filtration (SRF) were used to evaluate sludge dewaterability. Extracellular polymeric substances (EPS) concentration was also determined in an attempt to identify the observed changes in dewaterability. It was indicated by the results that both CPAM conditioning and MF pretreatment on sludge can lower CST and SRF values. However, subjecting to a combination of MF pretreatment and CPAM conditioning, sludge dewaterability was significantly enhanced beyond the level observed of CPAM addition alone. The lowest CST and SRF values of 36.5 s and 0.75×10(12) m kg(-1), respectively, were obtained when sludge was co-conditioned by CPAM (at a dosage of 40 mg L(-1)) and MF (at an induction of 40 mT) for 30 min, suggesting the optimal condition for enhancing sludge dewaterability. It is also shown from the significant correlations between EPS, protein, polysaccharide and CST/SRF that the increment of EPS concentration in sludge supernatant may be the major reason for the enhancement of dewaterability.

Oceanobacillus halophilum sp. nov. isolated from a mangrove forest soil.

A halophilic, aerobic bacterium, designated GD01(T), was isolated from a mangrove forest soil near the South China Sea. Cells of strain GD01(T) were Gram staining positive, oxidase positive, and catalase positive. The strain was rod shaped and motile by means of peritrichous flagella and produced ellipsoidal endospores. The strain was able to grow with NaCl at concentrations of 0.5-12 % (optimum 3-5 %, w/v), at temperatures of 20-50 °C (optimum 30 °C), and at pH 6.0-8.5 (optimum pH 7.0). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain GD01(T) formed a cluster with O. profundus DSM 18246(T) (96.4 % 16S rRNA gene sequence similarity), O. caeni KCTC 13061(T) (95.4 %), and O. oncorhynchi JCM 12661(T) (94.5 %). The G+C content of strain GD01(T) was 38.7 mol%. The major respiratory quinone was MK-7. The major cellular fatty acids (>5 %) were anteiso-C15:0, iso-C16:0 (13.7 %), anteiso-C17:0 (12.6 %), iso-C15:0 (9.9 %), iso-C14:0 (9.5 %), and C16:0 (5.0 %). The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, glycolipid, four unknown lipids, and four unknown phospholipids. Based on phenotypic characteristics, chemotaxonomic features, and phylogenetic analysis based on 16S rRNA gene sequences, the strain was identified to represent a distinct novel species in the genus Oceanobacillus, and the name proposed is Oceanobacillus halophilum sp. nov. with type train GD01(T) (=CCTCC AB 2012863(T) = KCTC 33101(T)).