Zebrafish Foxl2l functions in proliferating germ cells for female meiotic entry.

Zebrafish sex differentiation is a complicated process and the detailed mechanism has not been fully understood. Here we characterized a transcription factor, Foxl2l, which participates in female oogenesis. We show that it is expressed specifically in proliferating germ cells in juvenile gonads and mature ovaries. We have used CRISPR-Cas9 to generate zebrafish deficient in foxl2l expression. Zebrafish with foxl2l-/- are all males, and this female-to-male sex reversal cannot be reversed by tp53 mutation, indicating this sex reversal is unrelated to cell death. We have generated transgenic fish expressing GFP under the control of foxl2l promoter to track the development of foxl2l + -germ cells; these cells failed to enter meiosis and accumulated as cystic cells in the foxl2l-/- mutant. Our RNA-seq analysis also showed the reduced expression of genes in meiosis and oogenesis among other affected pathways. All together, we show that zebrafish Foxl2l is a nuclear factor controlling the expression of meiotic and oogenic genes, and its deficiency leads to defective meiotic entry and the accumulation of premeiotic germ cells.

Case series of ovarian neuroendocrine carcinoma: overview of clinicopathological features.

BACKGROUND: Ovarian neuroendocrine carcinoma (O-NEC) is a relatively uncommon neoplasm, and the current knowledge regarding its diagnosis and management is limited. In this series, our objective was to provide an overview of the clinicopathological characteristics of the disease by analyzing clinical case data to establish a theoretical foundation for the diagnosis and management of O-NEC. CASE PRESENTATION: We included three patients in the present case series, all of whom were diagnosed with primary O-NEC based on pathomorphological observation and immunohistochemistry. Patient 1 was a 62-year-old patient diagnosed with small cell carcinoma (SCC) of the pulmonary type. Post-surgery, the patient was diagnosed with stage II SCC of the ovary and underwent standardized chemotherapy; however, imaging examinations conducted at the 16-month follow-up revealed the existence of lymph node metastasis. Unfortunately, she passed away 21 months after the surgery. The other two patients were diagnosed with carcinoid tumors, one at age 39 and the other at age 71. Post-surgery, patient 2 was diagnosed with a carcinoid in the left ovary, whereas patient 3 was diagnosed with a carcinoid in her right ovary based on clinical evaluation. Neither of the cases received adjuvant therapy following surgery; however, they have both survived for 9 and 10 years, respectively, as of date. CONCLUSION: Primary O-NECs are rare and of diverse histological types, each of which has its own unique biological features and prognosis. SCC is a neoplasm characterized by high malignancy and a poor prognosis, whereas carcinoid tumors are of lesser malignancy and have a more favorable prognosis.

The potential use of Zymomonas mobilis for the food industry.

Zymomonas mobilis is a gram-negative facultative anaerobic spore, which is generally recognized as a safe. As a promising ethanologenic organism for large-scale bio-ethanol production, Z. mobilis has also shown a good application prospect in food processing and food additive synthesis for its unique physiological characteristics and excellent industrial characteristics. It not only has obvious advantages in food processing and becomes the biorefinery chassis cell for food additives, but also has a certain healthcare effect on human health. Until to now, most of the research is still in theory and laboratory scale, and further research is also needed to achieve industrial production. This review summarized the physiological characteristics and advantages of Z. mobilis in food industry for the first time and further expounds its research status in food industry from three aspects of food additive synthesis, fermentation applications, and prebiotic efficacy, it will provide a theoretical basis for its development and applications in food industry. This review also discussed the shortcomings of its practical applications in the current food industry, and explored other ways to broaden the applications of Z. mobilis in the food industry, to promote its applications in food processing.

Potential applications of Zymomonas mobilis in food industry summarized for the first time.Research status of Z. mobilis in food additive synthesis, fermentation applications, and probiotics are discussed in details.Future research perspectives of Z. mobilis in food industry further proposed.

Differentiation Between Solitary Pulmonary Inflammatory Lesions and Solitary Cancer Using Gemstone Spectral Imaging.

BACKGROUND: The distinction between solitary inflammatory lesion and solitary lung cancer remains a challenge because of their considerable overlapping computed tomography (CT) imaging features. PURPOSE: This study aimed to verify whether spectral CT parameters can differentiate solitary lung cancer from solitary inflammatory lesions and to find their correlations with lesion size. METHODS: A total of 78 patients with solitary lung lesions were included in our study. All of them underwent enhanced CT scans with Gemstone Spectral Imaging (GSI) mode, which was one of the dual-energy imaging technologies. According to maximum diameter (Dmax) of the lesion, regions of interest were collected and divided into inflammatory (group I: <3 cm [IA], n = 17; ≥3 cm [IB], n = 14) and cancer groups (group II: <3 cm [IIA], n = 20; ≥3 cm [IIB], n = 27). Computed tomography values (HU40keV, HU70keV), effective atomic number (Zeff), iodine concentration (IC), normalized IC (NIC), and spectral curve slopes (λ30, λ40) of each region of interest were calculated. The NIC was defined as the IC ratio of the lesion to the descending aorta. Mann-Whitney U test was used for intergroup (I vs II, IA vs IIA, IB vs IIB) and intragroup (IA vs IB, IIA vs IIB) comparisons, and receiver operating characteristic curve analysis was performed. Correlation analysis was applied to find the relationship between Dmax and GSI parameters. RESULTS: No significant correlation was found between GSI parameters and Dmax in the inflammatory group, whereas inverse correlations were found in the cancer group. Gemstone spectral imaging parameters (except HU70keV) of group IIA were significantly higher than those of group IIB. There were significant differences in HU40keV, IC, NIC, λ30, and λ40 between groups IB and IIB under both arterial and venous phase (P values < 0.05), whereas the area under the curve for λ30 under venous phase was largest, and sensitivity and specificity were 96.32% and 85.71%, respectively. However, only HU40keV and HU70keV values under the arterial phase of IIA were significantly higher than those of IA. CONCLUSIONS: Quantitative parameters of GSI demonstrated an inverse correlation with the lesion size of solitary lung cancer, and GSI parameters can be new ways to differentiate solitary lung cancer from solitary inflammatory lesions.

Pararcticibacter amylolyticus gen. nov., sp. nov., Isolated from a Rotten Hemp Rope, and Reclassification of Pedobacter tournemirensis as Pararcticibacter tournemirensis comb. nov.

A Gram-stain-negative, rod-shaped, non-motile, facultatively anaerobic bacterium, designated FJ4-8T, was isolated from a rotten hemp rope in Chongqing City, PR China. Phylogenetic analysis of 16S rRNA gene sequences indicated that the isolate was closely related to members of the family Sphingobacteriaceae, with the highest similarity to Pedobacter tournemirensis TF5-37.2-LB10T (95.3%) and low similarities to all other species of the genus Pedobacter (90.4-93.9%). Phylogenetic analyses demonstrated that strain FJ4-8T formed a stable subclade with Pedobacter tournemirensis TF5-37.2-LB10T. The clade with these two strains branched adjacent to a clade containing three species of the genus Arcticibacter. MK-7 was detected as the only respiratory quinone. The major fatty acids composed iso-C15:0, iso-C17:0 3-OH and summed feature three. Phosphatidylethanolamine, three aminophospholipids and one unidentified lipid were found as the major polar lipids. The major polyamine was identified as sym-homospermidine. The DNA-DNA hybridization value between strain FJ4-8T and Pedobacter tournemirensis TF5-37.2-LB10T was 42.0 ± 2.5%. Based on its phylogenetic, chemotaxonomic and phenotypic characteristics, the novel strain and TF5-37.2-LB10T were found to be different from members of genera Pedobacter and Arcticibacter. FJ4-8T and TF5-37.2-LB10T represented different species. Therefore, FJ4-8T should be classified as a novel species of a novel genus in the family Sphingobacteriaceae, for which the name Pararcticibacter amylolyticus gen. nov., sp. nov. is proposed. The type strain is FJ4-8T (= KCTC 62640T = CCTCC AB 2018052T). The draft genome sequence is 6290, 449 bp in length, the genomic DNA G+C content was 44.4 mol%. Pedobacter tournemirensis TF5-37.2-LB10T should be transferred to the novel genus as Pararcticibacter tournemirensis comb. nov. (The type strain is TF5-37.2-LB10T (= DSM 23085T = CIP 110085T = MOLA 820T).

Chemoresistance in gastric cancer is attributed to the overexpression of excision repair cross-complementing 1 (ERCC1) caused by microRNA-122 dysregulation.

MicroRNAs are deemed as key regulators of gene expression. In particular, the elevated expression of excision repair cross-complementing 1 (ERCC1) significantly reduced the effectiveness of gastric cancer treatment by cisplatin (CDDP)-based therapies. In this paper, qRT-PCR and western blot were adopted to measure miR-122 and ERCC1 messenger RNA (mRNA) expression in all samples. Luciferase assay was carried out to verify the role of ERCC1 as a target of miR-122. The CCK-8 assay was carried out to study the effect of ERCC1 and miR-122 on cell survival and apoptosis. The results demonstrated that miR-122 expression was reduced in cisplatin-resistant gastric cancer. Using bioinformatic analysis, miR-122 was shown to target the 3'-UTR of human ERCC1. A dual-luciferase assay demonstrated that miR-122 downregulated ERCC1 expression, while the mutations in ERCC1 3'-UTR abolished its interaction with miR-122. Transfection of miR-122 mimics decreased the levels of ERCC1 mRNA and protein expression, while the transfection of miR-122 inhibitors increased the levels of both ERCC1 mRNA and protein expression. Furthermore, we found that overexpressed miR-122 promoted the proliferation of MKN74 cells and reduced their apoptotic by targeting ERCC1. In addition, the levels of miR-122 and ERCC1 were negatively correlated in gastric cancer samples. In summary, the reduced miR-122 expression may play an essential role in the induction of cisplatin-resistance by increasing ERCC1 expression.

Chitinophaga salinisoli sp. nov., isolated from saline soil.

A novel aerobic bacterium, designated strain LAM9153T, was isolated from a saline soil sample collected from Lingxian County, Shandong Province, China. Cells of strain LAM9153T were observed to be Gram-stain negative, non-motile, non-spore-forming and rod-shaped. The new isolate grew optimally at 30-35 °C, pH 7.0 and 0.5% of NaCl concentration (w/v). According to the phylogenetic analysis based on the 16S rRNA gene sequence, strain LAM9153T shares high similarity with Chitinophaga terrae Gsoil 238T (96.9%) and Chitinophaga niabensis JS 13-10T (95.9%), forming a subcluster with C. terrae Gsoil 238T, Chitinophaga cymbidii R156-2T, C. niabensis JS 13-10T and Chitinophaga soli Gsoil 219T in the phylogenetic tree. The major cellular fatty acids (> 10%) were identified as iso-C15:0, iso-C17:0 3-OH and summed features 3 (C16:1 ω6c and/or C16:1 ω7c). The predominant respiratory quinone was identified as menaquinone MK-7. The polar lipids consisted of phosphatidylethanolamine, aminophospholipid, three unidentified aminolipids and five unidentified lipids. The genomic DNA G+C content was determined to be 53.2 ± 1.6 mol%. On the basis of phylogenetic, chemotaxonomic and phenotypic data, strain LAM9153T is concluded to represent a novel species of the genus Chitinophaga, for which the name Chitinophaga salinisoli sp. nov. is proposed. The type strain is LAM9153T (= ACCC 19960T = JCM 30847T).

Changes in the morphology and gene expression of developing zebrafish gonads.

Zebrafish gonadal sexual differentiation is an important but poorly understood subject. The difficulty in investigating zebrafish sexual development lies in its sex determination plasticity, the lack of morphological tools to distinguish juvenile females from males, and the lack of sex chromosomes in laboratory strains. Zebrafish sexual differentiation starts at around 8 days post-fertilization when germ cells start to proliferate. The number of germ cells determines the future sex of the gonad. Gonads with more germ cells differentiate into ovaries, whereas a reduced germ cell number leads to male-biased sexual differentiation. Genes controlling sexual differentiation in pre-meiotic gonads encode proteins such as transcription factors, the transforming growth factor (TGF)-ß family of signaling proteins, and RNA-binding proteins. These proteins coordinately control germ cell proliferation/meiosis/maintenance and gonadal somatic cell differentiation, leading to stepwise differentiation of gonads. Morphological changes in differentiating gonads are characterized by the appearance of oocytes containing condensed chromatin, followed by incorporation of vitellogenin and oocyte maturation. Marker genes and morphological characteristics help distinguish the steps in zebrafish gonadal differentiation during this important sex-determining stage.

Arenimonas alkanexedens sp. nov., isolated from a frozen soil sample.

A novel facultatively anaerobic bacterium, designated strain LAM-WHM-D11T, was isolated from a frozen soil sample of China. Cells of strain LAM-WHM-D11T were observed to be Gram-stain negative, non-motile and rod-shaped. Colonies were yellowish, and circular with convex shape. Strain LAM-WHM-D11T was found to be able to grow at 4-40 °C (optimum 15 °C), pH 7.5-2.0 (optimum 9.5) and 0-2.5% NaCl (w/v) (optimum 1.5%). The 16S rRNA gene sequence similarity analysis showed that strain LAM-WHM-D11T is closely related to Arenimonas metalli CF5-1T (98.0%), Arenimonas aquaticum NA-09T (97.9%), Arenimonas donghaensis HO3-R19T (95.6%) and Arenimonas aestuarii S2-21T (95.3%). The DNA-DNA hybridization values between the isolate and A. metalli CGMCC 1.10787T, A. aquaticum KACC 14663T, A. donghaensis KACC 11381T were 41.0 ± 1.7, 44.7 ± 1.4 and 42.8 ± 1.2%, respectively. The genomic DNA G+C content was found to be 66.5 mol% as determined by the T m method. The major cellular fatty acids were identified as iso-C15:0 and iso-C16:0. The major isoprenoid quinone was identified as ubiquinone 8 (Q-8). The major polar lipids were found to be diphosphatidyglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmonomethylethanolamine, two phospholipids and five unidentified lipids. Based on the phylogenetic, phenotypic and chemotaxonomic characteristics, strain LAM-WHM-D11T is concluded to represent a novel species within the genus Arenimonas, for which the name Arenimonas alkanexedens sp. nov. is proposed. The type strain is LAM-WHM-D11T (ACCC 19750T = JCM 30464T).

Engineered Zymomonas mobilis for salt tolerance using EZ-Tn5-based transposon insertion mutagenesis system.

BACKGROUND: The cell growth and ethanol yield of Zymomonas mobilis may be detrimentally affected by salt stress frequently present in some biomass-based fermentation systems, leading to a decrease in the rate of sugar conversion to ethanol or other bioproducts. To address this problem, improving the salt tolerance of Z. mobilis is a desirable way. However, limited progress has been made in development of Z. mobilis with higher salt tolerance for some technical challenges in the past decades. Recently, transposon insertion mutant system has been widely used as a novel genetic tool in many organisms to develop mutant strains. In this study, Tn5-based transposon insertion mutagenesis system firstly used for construction of higher salt tolerance strain in Z. mobilis. RESULTS: Approximately 200 Z. mobilis ZM4 mutants were generated by using Tn5-based transposon mutagenesis system. The mutant strain ZMT2 with improved salt tolerance phenotype was obtained by screening on RM agar plates with additional 1 % NaCl. Strain ZMT2 was confirmed to exhibit better fermentation performance under NaCl stress than wild type of strain ZM4. The transposon insertion was located in ZMO1122 (himA) by genome walking. Discruption of himA gene showed that himA may play an important role in response to salt tolerance in Z. mobils. CONCLUSIONS: The mutant strain ZMT2 with a transposon insertion in himA gene of the genome showed obviously higher sugar conversion rate to ethonal under up to 2 % NaCl stress than did the wild ZM4 strain. Besides, ZMT2 exhibited shared fermentative capabilities with wild ZM4 strain under no or low NaCl stress. This report firstly showed that himA played a role in responding to NaCl stress. Furthermore, the result indicated that Tn5-based transposon mutagenesis system was a feasible tool not only for genetic engineering in Z. mobilis strain improvement, but also in tapping resistent genes.

Lentibacillus amyloliquefaciens sp. nov., a halophilic bacterium isolated from saline sediment sample.

A Gram-stain positive, non-motile, non-sporogenous, aerobic, rod-shaped and halophilic bacterium, designated LAM0015(T), was isolated from a saline sediment sample collected from Yantai City in China. The isolate was found to be able to grow at NaCl concentrations of 5-25 % (w/v) (optimum: 7-12 %), 15-45 °C (optimum: 35 °C) and pH 5.0-9.0 (optimum: 7.0). The major fatty acids were determined to be anteiso-C15:0 and anteiso-C17:0. The predominant respiratory quinone was identified as MK-7. The cell wall peptidoglycan was determined to contain meso-diaminopimelic acid. The polar lipids were found to be diphosphatidyglycerol, phosphatidylglycerol, five phospholipids and one glycolipid. The DNA G+C content was 43.1 mol% as determined by the T m method. Analysis of the 16S rRNA gene sequence indicated that the isolate belongs within the genus Lentibacillus and is closely related to Lentibacillus persicus DSM 22530(T), Lentibacillus salicampi JCM 11462(T) and Lentibacillus jeotgali JCM 15795(T) with 97.3, 96.7 and 96.4 % sequence similarity, respectively. The DNA-DNA hybridization value between LAM0015(T) and L. persicus DSM 22530(T) was 51.2 ± 1.4 %. Based on its phenotypic, phylogenetic and chemotaxonomic characteristics, strain LAM0015(T) is concluded to represent a novel species of the genus Lentibacillus, for which the name Lentibacillus amyloliquefaciens sp. nov. is proposed. The type strain is LAM0015(T) (=ACCC 06401(T) = JCM 19838(T)).

Paenibacillus salinicaeni sp. nov., isolated from saline silt sample.

A novel facultatively anaerobic bacterium, designated strain LAM0A28(T), was isolated from a saline silt sample collected from the Chinese Sea of Death located in Suining city, Sichuan province, China. Cells of strain LAM0A28(T) were observed to be Gram-stain positive, motile, endospore-forming and straight-rod shaped. Strain LAM0A28(T) was found to be able to grow at 15-45 °C (optimum: 30-35 °C), pH 5.0-10.0 (optimum: 7.5) and 0-5 % NaCl (w/v) (optimum: 0.5 %). The 16S rRNA gene sequence similarity analysis showed that strain LAM0A28(T) is closely related to Paenibacillus jilunlii DSM 23019(T) (97.5 %) and Paenibacillus graminis DSM 15220(T) (97.2 %). The DNA-DNA hybridization values between the isolate and P. jilunlii DSM 23019(T), P. graminis DSM 15220(T) were 30.2 ± 1.6 % and 44.7 ± 2.1 %, respectively. The DNA G+C content was found to be 51.2 mol% as determined by the T m method. The major cellular fatty acids were identified as anteiso-C15:0, C16:0, iso-C16:0 and C14:0. The major isoprenoid quinone was identified as MK-7. The cell wall peptidoglycan was found to contain meso-diaminopimelic acid. The major polar lipids were found to be diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two aminophospholipids and six unidentified lipids. Based on the phylogenetic, phenotypic and chemotaxonomic characteristics, strain LAM0A28(T) is concluded to represent a novel species within the genus Paenibacillus, for which the name Paenibacillus salinicaeni sp. nov. is proposed. The type strain is LAM0A28(T) (=ACCC 00741(T) = JCM 30850(T)).

[Teaching design and practice of human blood type traits in genetics comprehensive laboratory course].

Comprehensive laboratory courses, which enable students to aptly apply theoretic knowledge and master experiment skills, play an important role in the present educational reform of laboratory courses. We utilized human ABO blood type as the experimental subject, and designed the experiment--"Molecular Genotyping of Human ABO Blood Type and Analysis of Population Genetic Equilibrium". In the experiment, DNA in mucosal cells is extracted from students' saliva, and each student's genotype is identified using a series of molecular genetics technologies, including PCR amplification of target fragments, enzymatic digestion, and electrophoretic separation. Then, taking the whole class as an analogous Mendel population, a survey of genotype frequency of ABO blood type is conducted, followed with analyses of various population genetic parameters using Popgene. Through the open laboratory course, students can not only master molecular genetic experimental skills, but also improve their understanding of theoretic knowledge through independent design and optimization of molecular techniques. After five years of research and practice, a stable experimental system of molecular genetics has been established to identify six genotypes of ABO blood types, namely I(A)I(A), I(A)i, I(B)I(B), I(B)i, I(A)I(B) and ii. Laboratory courses of molecular and population genetics have been integrated by calculating the frequencies of the six genotypes and three multiple alleles and testing population genetic equilibrium. The goal of the open laboratory course with independent design and implementation by the students has been achieved. This laboratory course has proved effective and received good reviews from the students. It could be applied as a genetics laboratory course for the biology majors directly, and its ideas and methods could be promoted and applied to other biological laboratory courses.

[Discrimination of Plum Browning with Near Infrared Spectroscopy].

Flesh browning mostly happens in plum fruit during the post-harvest storage period, which is an important factor affecting the storage quality of plum fruits. Traditional methods used to discriminate plum browning involve the destruction of the intact fruit, which are highly subjective and error-prone. Therefore, the near-infrared (NIR) spectroscopy technique was applied to achieve rapid and non-destructive identification of plum browning and non-browning in this paper. The near infrared diffuse reflectance spectroscopy of 124 plum samples were collected in the band number of 4 000~12 500 cm-1. These samples were classified into two groups, browning (n=70) and non-browning (n=54). In order to find a new way to effectively discriminate plum fruits with flesh browning, three qualitative identification methods: the qualitative test, Mahalanobis distances discriminate analysis (DA) and Back Propagation-artificial neural networks (BP-ANN) were used to compare their capacity of recognizing browning plums and non-browning oneswhile the last two approaches were based on the principal component analysis (PCA) method. These results showed that DA and BP-ANN could be used to conctruct effective classification models for identifying plum browning, and the first ten principal components extracted from original spectra were applied as input variables to build DA and BP-ANN models. The optimal method was obtained with BP-ANN, which gained an accuracy of 100% for calibration set and 97.56% for prediction set, and the identification accuracy rate reached 100% and 98.57% for non-browning samples and browning ones, respectively. It could be concluded that NIR spectroscopy technique combined with chemometrics methods has great potential to recognize plums of browning and non-browning rapidly, non-destructively and effectively.

WITHDRAWN: Comparative proteome analysis of human testis from newborn, young adult, and aged men identified spermatogenesis-associated proteins.

Ahead of Print article withdrawn by publisher.

Aged men share the sperm protein PATE1 defect with young asthenozoospermia patients.

STUDY QUESTION: Does a defect in the human sperm-located protein prostate and testis expressed 1 (PATE1) exist in both aged men and young asthenozoospermia patients? SUMMARY ANSWER: A defect in sperm PATE1 exists in both aged men and young asthenozoospermia patients, and an antibody against PATE1 can decrease human sperm motility and zona-free hamster oocyte penetration. WHAT IS KNOWN ALREADY: Both aged men and young asthenozoospermia patients have poor sperm quality. The PATE1 protein seems to mediate sperm-egg interactions; however, the mechanisms are still unknown. STUDY DESIGN, SIZE, DURATION: This was a case-control study including 60 young fathers (aged 28-32 years) and 60 aged fathers (68-72 years old) who donated semen by masturbation after 7 days of sexual abstinence. Comparative sperm proteome analysis from the young fathers and aged fathers was performed to discover key proteins. The target protein PATE1 was chosen and validated by western blotting and immunohistochemistry. Quantitative assessment of sperm PATE1 protein was performed on sperm from 60 young fathers, 60 aged fathers and 110 young asthenozoospermia patients. Furthermore, an antibody against PATE1 assay was used to test whether PATE1 participated in sperm motility and penetration of zona-free hamster egg. PARTICIPANTS/MATERIALS, SETTING, METHODS: Samples were pooled and separated by two-dimensional gel electrophoresis followed by identification by matrix-assisted laser desorption/ionization time of flight mass spectrometry. Western blotting and immunohistochemistry were used to validate the confidence of proteomic data. Sperm immunofluorescence quantification experiments disclosed whether the aged men indeed shared the same PATE1 defect with 110 young asthenozoospermia patients. The sperm motility test and penetration of zona-free hamster egg assay were performed for PATE1. MAIN RESULTS AND THE ROLE OF CHANCE: Twenty-two sperm proteins with significant differential expression between young adults and aged men were identified (P < 0.05, mean ratio >1.5), including 13 proteins with decreased expressions with aging. Based on bioinformatics, PATE1 was chosen for further study, and exhibited similar changes in expression level and localization on sperm from aged men and young asthenozoospermia patients. Antibody blocking revealed that PATE1 was involved in sperm-egg penetration and sperm motility. LIMITATIONS, REASONS FOR CAUTION: Before any clinical application of PATE1 as a biomarker for the diagnosis of male infertility, more cases should be used to evaluate confidence in this approach. WIDER IMPLICATIONS OF THE FINDINGS: This study revealed a common molecular basis underlying the decline in sperm quality in the natural aging process and in young men with asthenozoospermia. The data should greatly contribute to the development of molecular evaluation of sperm quality, and the diagnosis and treatment of asthenozoospermia. STUDY FUNDING/COMPETING INTERESTS: This work was supported by grants from the National Natural Science Foundation of China (NO. 81300533, 81370013 and 81000277) and Shandong Provincial Natural Science Foundation, China (ZR2013HQ002, ZR2014HQ068). The authors declare no competing financial interests.

In-depth Proteomic mapping of mouse (Mus musculus) epididymal constructive basis for sperm maturation.

BACKGROUND: The mouse epididymis performs an essential role in sperm maturation, but global protein expression data in mouse epididymis are still lacking. Here, we reported the first in-depth gel-based profiling of mouse epididymis proteome and established a 2-DE map. RESULTS: A total of 832 protein spots were detected in the reproducible gels, and 625 spots corresponding to 355 unique protein entries have been successfully identified by MALDI-TOF-MS. The confidence of proteome data was validated by Western blot. Functional annotations showed that these proteins were mainly related to general metabolism, antioxidant and structural molecule activity. Immunohistochemistry disclosed two structural proteins (myosin regulatory light polypeptide 9 and alpha-2 type I collagen) continuously expressed in the myoid cell since postpartum. CONCLUSION: This study provides a first-draft reference map of the mouse epididymis proteome, which will greatly expand the knowledge of the epididymal structural basis and contribute to the better understanding of those proteins in the process of mouse epididymal sperm maturation.

Paenibacillus populi sp. nov., a novel bacterium isolated from the rhizosphere of Populus alba.

A novel aerobic bacterium, designated strain LAM0705(T), was isolated from the rhizosphere of Populus alba in the Peking University Third Hospital. Cells of strain LAM0705(T) were observed to be Gram-stain positive, motile, spore-forming and rod-shaped. The optimal temperature and pH for growth were found to be 30 °C and pH 7.5, respectively. Strain LAM0705(T) was found to be able to grow in the presence 0-5 % NaCl (w/v) (optimum 1.0 %). The major fatty acids of strain LAM0705(T) were identified as anteiso-C15:0, C16:0 and iso-C16:0. The dominant polar lipids were found to consist of diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. The cell wall peptidoglycan of strain LAM0705(T) was found to contain meso-diaminopimelic acid. The predominant menaquinone was identified as MK-7. The G+C content of genomic DNA was found to be 48 mol% when determined by the T m method. The 16S rRNA gene sequence similarity analysis indicated that strain LAM0705(T) is closely related to Paenibacillus agaridevorans DSM 1355(T) and Paenibacillus thailandensis KCTC 13043(T) with 97.8 and 96.1 % sequence similarity, respectively. The DNA-DNA hybridization value between strain LAM0705(T) and P. agaridevorans DSM 1355(T) was 47 ± 0.8 %. On the basis of its phenotypic, phylogenetic and chemotaxonomic characteristics, strain LAM0705(T) is concluded to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus populi sp. nov. is proposed. The type strain is LAM0705(T) (=ACCC 06427(T) = JCM 19843(T)).

Paenibacillus vini sp. nov., isolated from alcohol fermentation pit mud in Sichuan Province, China.

A novel facultatively anaerobic bacterial strain, designated LAM0504(T), was isolated from a pit mud of Luzhou flavour liquor alcohol fermentation in Sichuan Province, China. Cells of strain LAM0504(T) were observed to be Gram-stain negative, spore-forming, rod shaped and motile by means of peritrichous flagella. Strain LAM0504(T) was found to be able to grow at 20-48 °C (optimum: 30 °C), pH 5.0-9.0 (optimum: 7.0) and 0-3 % NaCl (w/v) (optimum: 1.0 %). The 16S rRNA gene sequence similarity analysis showed that strain LAM0504(T) was most closely related to Paenibacillus konsisdensis JCM 14798(T), Fontibacillus phaseoli LMG 27589(T) and Paenibacillus motobuensis JCM 12774(T), with 97.0, 96.8 and 96.7 % sequence similarity, respectively. The DNA-DNA hybridization value between strain LAM0504(T) and P. konsisdensis JCM 14798(T) was 53.3 ± 1.2 %. The genomic DNA G+C content of strain LAM0504(T) was 43.0 mol% as determined by the Tm method. The major fatty acids of strain LAM0504(T) were identified as anteiso-C15:0, C16:0 and iso-C15:0. The cell-wall peptidoglycan was found to contain meso-diaminopimelic acid. The predominant menaquinone was identified as MK-7. The major polar lipids were found to be diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two unidentified phospholipids, two unidentified glycolipids and three unidentified lipids. On the basis of its physiological and phylogenetic characteristics, strain LAM0504(T) is concluded to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus vini sp. nov. is proposed. The type strain is LAM0504(T) (=ACCC 06420(T) = JCM 19842(T)).

Control the wettability of poly(n-isopropylacrylamide-co-1-adamantan-1-ylmethyl acrylate) modified surfaces: the more Ada, the bigger impact?

Surface-initiated SET-LRP was used to synthesize polymer brush containing N-isopropylacrylamide and adamantyl acrylate using Cu(I)Cl/Me6-TREN as precursor catalyst and isopropanol/H2O as solvent. Different reaction conditions were explored to investigate the influence of different parameters (reaction time, catalyst concentration, monomer concentration) on the polymerization. Copolymers with variable 1-adamantan-1-ylmethyl acrylate (Ada) content and comparable thickness were synthesized onto silicon surfaces. Furthermore, the hydrophilic and bioactive molecule ß-cyclodextrin-(mannose)7 (CDm) was synthesized and complexed with adamantane via host-guest interaction. The effect of adamantane alone and the effect of CDm together with adamantane on the wettability and thermoresponsive property of surface were investigated in detail. Experimental and molecular structure analysis showed that Ada at certain content together with CDm has the greatest impact on surface wettability. When Ada content was high (20%), copolymer-CDm surfaces showed almost no CDm complexed with Ada as the result of steric hindrance.