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Background and aims C-reactive protein (CRP) is a hepatocyte-produced marker of inflammation yet with undefined function in liver injury. We aimed to examine the role of CRP in acetaminophen-induced liver injury (AILI). Methods The effects of CRP in AILI were investigated using CRP knockout mice and rats combined with human CRP rescue. The mechanisms of CRP action were investigated in vitro and in mice with Fcγ receptor 2B knockout, C3 knockout, or hepatic expression of CRP mutants defective in complement interaction. The therapeutic potential of CRP was investigated by intraperitoneal administration at 2 or 6 hours post–AILI induction in wild-type mice. Results CRP knockout exacerbated AILI in mice and rats, which could be rescued by genetic knock-in, adeno-associated virus–mediated hepatic expression or direct administration of human CRP. Mechanistically, CRP does not act via its cellular receptor Fcγ receptor 2B to inhibit the early phase injury to hepatocytes induced by acetaminophen; instead, CRP acts via factor H to inhibit complement overactivation on already injured hepatocytes, thereby suppressing the late phase amplification of inflammation likely mediated by C3a-dependent actions of neutrophils. Importantly, CRP treatment effectively alleviated AILI with a significantly extended therapeutic time window than that of N-acetyl cysteine. Conclusion Our results thus identify CRP as a crucial checkpoint that limits destructive activation of complement in acute liver injury, and we argue that long-term suppression of CRP expression or function might increase the susceptibility to AILI.

The mud crab Scylla paramamosain is one of the economically important aquaculture species in China. The larval development of the mud crab is characterized by two significant morphological changes, from the 5th zoea (Z5) to the megalopa (M) stage and from the M to the first juvenile crab (C1) stage. In this study, we found that methyl farnesoate (MF) could prohibit the Z5 to M metamorphosis in a concentration-dependent manner, and that a concentration of 10 μM MF could completely prohibit the Z5 metamorphosis. Farnesoic acid (FA) could also prohibit the Z5 metamorphosis, but its effects seemed to be concentration-independent. In addition, MF could delay rather than prohibit the M to C1 metamorphosis, while FA had no effect on the M to C1 metamorphosis at all. To summarize, it is hypothesized that either absence of MF and FA, or at least very low levels of these substances, might be necessary for a successful Z5 to M metamorphosis.

To investigate the correlation between serum renin-angiotensin system (RAS) level and Symptoms of anxiety and depression in Parkinson disease patients (PD). A number of 90 PD patients (47 males and 43 females) were collected on an empty stomach 12 h after stopping taking anti-PD medicines. ELISA has been found in Serum RAS ((Ang) I, Ang II, Ang (1–7), Angiotensin converting enzyme (ACE), ACE2). Depression scale (HAMD) and Anxiety scale (HAMA) in Hamilton are used for the assessment of signs of depression and anxiety. The 90 patients were diagnosed with moderate depression (HAMD score 8 ~ 19); in 32 of those (35.56 percent), and 12 (13.33%) were diagnosed as moderate and severe depression (HAMD score ≥ 20). 20 cases (22.22%) were diagnosed as possible anxiety disorder (HAMA score 7 ~ 13) and 16 cases (17.78%) as definite anxiety disorder (HAMA score ≥ 14). The association of serum Ang I, Ang II and Ang (1–7) with HAMD (r= − 0.820, P 0.05) were not associated to the serum ACE and ACE2. The serum Ang I, Ang II, and Ang (1–7) were found to be adversely associated with HAMD score (r = 0.826, P >0,001; r = −0.876, P 0.001; The HAMD score and the HAMA score (all P > 0.05) were not correlated to the serum ACE and ACE2. Finally, in PD patients, non-engine signs, including depression and anxiety, are normal. Thus, Serum levels Ang I, Ang II and Ang (1–7) were substantially decreased in female and male patients and associated with symptoms of depression and anxiety, ACE and ACE2 levels have not been attributed to signs of depression and anxiety. Serum Ang I, Ang II, and Ang (1–7) are important markers of depression and anxiety prevention and diagnosis in patients with DP.

Large amounts of detrital organic matter and osmolytes accumulate in the sediments of hadal trenches (>6000 m depth) due to the funnelling effect. It is still unknown whether there are novel active microbes that depend on specific carbon sources in extreme and isolated environments. In this study, we present a novel active bacterial phylum, Candidatus Tianyabacteria in the FCB superphylum, which was enriched in sediments collected from the Challenger Deep. Genome binning resulted in high-quality Ca. Tianyabacteria genomes representing two Ca. Tianyabacteria lineages (L1 and L2) in sediments 0-21 cm below the surface (cmbsf); L1 tends to be abundant in the upper layers (0-9 cmbsf), and L2 seems to be more prevalent in the deeper layers (12-21 cmbsf). Gene annotation and transcriptomics results indicate that the two lineages might import and catalyse amino acids and myo-inositol into central carbon metabolism for a heterotrophic lifestyle. Probably due to differences in environmental oxygen levels, the L2 genomes harbour gene clusters responsible for denitrification and fermentation, while the L1 genomes encode octahaem cytochrome c and multicopper oxidase using unknown substrates. The Ca. Tianyabacteria are thus novel heterotrophic organisms that participate in processes of carbon, nitrogen and organic osmolyte cycling in hadal sediments.

Double-maize cropping system is an effective option for coping with climate change in the North China Plain. However, the effects of changes in climate on the growth and yield of maize in the two seasons are poorly understood. Forty-six cultivars of maize with different requirements for growing degree days (GDD), categorized as high (H), medium (M) or low (L), and three cultivar combinations for two seasons as LH (using JD27 and DMY1 from category L in the first season; and YD629 and XD22 from category H in the second season), MM (using JX1 and LC3 from category M in the first season; and ZD958 and JX1 from category M in the second season) and HL (using CD30 and QY9 from category H in the first season; and XK10 and DMY3 from category L in the second season) were tested to examine the eco-physiological determinants of maize yield from 2015 to 2017. The correlations between the combinations of cultivars and grain yield were examined. The combination LH produced the highest annual grain yield and total biomass, regardless of the year. It was followed, in decreasing order, by MM and HL. Higher grain yield and biomass in LH were mainly due to the greater grain yield and biomass in the second season, which were influenced mainly by the lengths of the pre- and post-silking periods and the rate of plant growth (PGR). Temperature was the primary factor that influenced dry matter accumulation. In the first season, low temperatures during pre-silking decreased both the duration and PGR in LH, whereas high temperatures during post-silking decreased the PGR in MM and HL, resulting in no significant differences in biomass being observed among the three combinations. In the second season, high temperatures decreased both the PGR and pre- and post-silking duration in MM and HL, and consequently, the biomass of those two combinations were lower than that in LH. Moreover, because of lower GDD and radiation in the first season and higher grain yield in the second season, production efficiency of temperature and radiation (Ra) was the highest in LH. More importantly, differences in temperature and radiation in the two seasons significantly affected the rate and duration of growth in maize, and thereby affecting both dry matter and grain yield. Our study indicated that the combination of LH is the best for optimizing the double-maize system under changing climatic conditions in the North China Plain.

The connection between diabetes and Alzheimer’s disease (AD) is not fully determined. Hyperphosphorylation of tau protein is mediated by binding and stabilization of truncated p25 with cyclin-dependent kinase-5 (CDK5) in AD. We recently showed that diabetes-associated hyperglycemia increased the CDK5 levels to promote development of AD. Here, we examined the underlying mechanisms. Hyperglycemia and glucose intolerance were induced in rats that had received a low dose of streptozotocin (STZ) and a high fat diet (HFD). Compared to the control rats that received no STZ and were fed a normal diet, the STZ + HFD rats exhibited poorer performance in the behavioral test and showed hyperacetylation of H3K9 histone on the CDK5 promoter, likely resulting from upregulation of a histone acetyltransferase, GCN5. Inhibition of acetylation of H3K9 histone by a specific GCN5 inhibitor, MB3, attenuated activation of CDK5, resulting in decreased tau phosphorylation in rat brain and improved performance of the rats in the behavior test. Thus, these data suggest that diabetes may promote future development of AD through hyperacetylation of H3K9 histone on CDK5 promoter.

Background & Aims Mucin 13 (MUC13) is reportedly overexpressed in human malignancies. However, the clinicopathological and biological significance of MUC13 in human intrahepatic cholangiocarcinoma (iCCA) remain unclear. The aim of this study was to define the role of MUC13 in the progression of iCCA. Methods Expression levels of MUC13 in human iCCA samples were evaluated by immunohistochemistry, western blot, and real-time PCR. In vitro and in vivo experiments were used to assess the effect of MUC13 on iCCA cell growth and metastasis. Crosstalk between MUC13 and EGFR/PI3K/AKT signaling was analyzed by molecular methods. The upstream regulatory effects of MUC13 were evaluated by Luciferase and DNA methylation assays. Results MUC13 was overexpressed in human iCCA specimens and iCCA cells. MUC13 overexpression positively correlated with clinicopathological characteristics of iCCA, such as vascular invasion and lymph node metastasis, and was independently associated with poor survival. Results from loss-of-function and gain-of-function experiments suggested that knockdown of MUC13 attenuated, while overexpression of MUC13 enhanced, the proliferation, motility, and invasiveness of iCCA cells in vitro and in vivo. Mechanistically, we found that the phosphatidylinositol 3-kinase-AKT signal pathway and its downstream effectors, such as tissue inhibitor of metalloproteinases 1 and matrix metallopeptidase 9, were required for MUC13-mediated tumor metastasis of iCCA. MUC13 interacted with epidermal growth factor receptor (EGFR) and subsequently activated the EGFR/PI3K/AKT signaling pathway by promoting EGFR dimerization and preventing EGFR internalization. We also found that MUC13 was directly regulated by miR-212-3p, whose downregulation was related to aberrant CpG hypermethylation in the promoter area. Conclusions These findings suggest that aberrant hypermethylation-induced downregulation of miR-212-3p results in overexpression of MUC13 in iCCA, leading to metastasis via activation of the EGFR/PI3K/AKT signaling pathway. Lay summary Mucin 13 overexpression has been implicated in the development of malignancies, although its role in intrahepatic cholangiocarcinoma has not been studied. Herein, we show that mucin 13 plays a critical role in intrahepatic cholangiocarcinoma. Mucin 13 could have therapeutic value both as a prognostic marker and as a treatment target.

Background Osteosarcoma (OS) is a highly aggressive bone cancer. Patients with OS frequently develop drug resistance in clinical treatment, and the prognosis has not been improved significantly. There is an urgent need to identify novel markers and therapeutic targets. Methods In this study, we focused on the highly expressed noncoding circular RNA circPDSS1 in OS, and studied its functional roles and downstream targets in OS cells by CCK-8, clone formation assay, transwell assays. Additionally, we performed luciferase reporter assay, RNA pull-down experiment and qRT-PCR to validate the micoRNA targets of circPDSS1. The involvement of circPDSS1 in tumorigenesis was also investigated in mouse xenografts model. Results The expression of circPDSS1 was significantly upregulated in OS tissues and cell lines. Patients with high circPDSS1 expression were associated with poorer progression-free survival (PFS) and overall survival (OS) as compared to those with low circPDSS1 expression. CircPDSS1 knockdown significantly inhibited the viability, clone formation ability and invasion ability of OS cells, and induced cell apoptosis, which were associated with the upregulation of proapoptotic proteins and the impairment of prosurvival signaling. Molecular mechanism study further demonstrated that circPDSS1 modulates OS cell functions by regulating the expression of miR-502-3p and miR-4436a. Conclusion Our data suggest that circPDSS1 acts as a molecular sponge of miR-502-3p and miR-4436a regulates the proliferation and invasion of OS cells and promote the malignant progression of OS.

Specific leaf area (SLA) is one of the most widely researched plant traits, integrating multiple functions and reflecting plant resource acquisition strategies. However, the underlying mechanisms that drive temporal and spatial variation in SLA are still not clear. We used a combination of transect investigation and short-term manipulative experiments to identify patterns of variation in SLA over naturally occurring and simulated environmental gradients. We measured SLA of 631 species from the Tibetan (TP), Loess (LP), and Mongolian Plateaus (MP), where plant growth is limited by temperature, soil nutrient availability, and precipitation, respectively. In each plateau, we established an east–west transect of 10 sites, including meadow steppe (meadow), typical steppe (steppe), and desert steppe (desert) habitats. We further measured SLA of 15 species from the TP and MP, using manipulative experiments that altered temperature, precipitation and soil nitrogen levels. SLA ranged from 0.9 to 58.90 m2 kg−1 on average, and was significantly larger in desert than in meadow and steppe, except in the LP. Plants from the TP had significantly lower SLA than those from the LP and MP, but variation in SLA across the environmental gradients was small and consistent across the three plateaus. Conversely, the short-term manipulative experiments revealed that SLA increased with resource availability. Shifts in SLA across the broad environmental gradients was relatively small, and SLA had high sensitivity to short-term resource enrichment, suggesting that grassland species in these plateaus express divergent short- (acclimation) and long-term responses (adaptation) to the environment, which may be an important strategy for coping with ongoing shifts in climate.

In this study, the complete mitochondrial genome of Ariosoma meeki was sequenced, assembled and annotated. The circular genome is 16,154 bp in length with nucleotide composition is 28.42% A, 26.53% T, 19.65% G, and 25.40% C and contains 13 protein-coding genes (PCGs), 21 transfer RNA genes (tRNAs), 2 ribosomal RNA unit genes and a large non-coding region (putative control region). To further explore the evolution relationship of the Anguilliformes, we constructed the phylogenetic tree and found that the A. meeki had closer relationship with Ariosoma shiroanago. This study provided the valuable evidence on phylogenetic relationship of the A. meeki at the molecular level and essential resource for further study the molecular phylogenetic, biogeography and adaptive evolution of this lineage.

Cyclin-dependent kinase-5 (CDK5) is activated by p35 and then binds to both p35 and its truncated form p25 to promote hyperphosphorylation of tau protein, thereby facilitating the pathological progression of Alzheimer's disease (AD). However, it is unknown whether a patient's diabetic status promotes the later onset of AD in a CDK5-dependent manner. Here, we induced pro-diabetic insulin resistance and glucose intolerance in rats using a combined high fat and high glucose diet. Compared to normal diet-fed rats, these pro-diabetic rats exhibited poorer behavioral performance in the Morris water maze test and the novel object recognition test. Increased phosphorylation of tau protein was detected in the hippocampal CA1 region of the rat brain, suggesting neurodegeneration. Moreover, CDK5 transcriptional activity was significantly increased in the HFGD-rat brain, likely resulting from an increase in acetylation and a decrease in methylation of the CDK5 promoter. Together, these data suggest that epigenetic control of the CDK5 promoter by acetylation and methylation may regulate the diabetes-associated development of AD.

It is largely unknown how living organisms—especially vertebrates—survive and thrive in the coldness, darkness and high pressures of the hadal zone. Here, we describe the unique morphology and genome of Pseudoliparis swirei—a recently described snailfish species living below a depth of 6,000 m in the Mariana Trench. Unlike closely related shallow sea species, P. swirei has transparent, unpigmented skin and scales, thin and incompletely ossified bones, an inflated stomach and a non-closed skull. Phylogenetic analyses show that P. swirei diverged from a close relative living near the sea surface about 20 million years ago and has abundant genetic diversity. Genomic analyses reveal that: (1) the bone Gla protein (bglap) gene has a frameshift mutation that may cause early termination of cartilage calcification; (2) cell membrane fluidity and transport protein activity in P. swirei may have been enhanced by changes in protein sequences and gene expansion; and (3) the stability of its proteins may have been increased by critical mutations in the trimethylamine N-oxide-synthesizing enzyme and hsp90 chaperone protein. Our results provide insights into the morphological, physiological and molecular evolution of hadal vertebrates.

The spike (S) protein receptor-binding domain (RBD) of SARS-CoV-2 is an attractive target for COVID-19 vaccine developments, which naturally exists in a trimeric form. Here, guided by structural and computational analyses, we present a mutation-integrated trimeric form of RBD (mutI tri-RBD) as a broadly protective vaccine candidate, in which three RBDs were individually grafted from three different circulating SARS-CoV-2 strains including the prototype, Beta (B.1.351) and Kappa (B.1.617). The three RBDs were then connected end-to-end and co-assembled to possibly mimic the native trimeric arrangements in the natural S protein trimer. The recombinant expression of the mutI tri-RBD, as well as the homo-tri-RBD where the three RBDs were all truncated from the prototype strain, by mammalian cell exhibited correct folding, strong bio-activities, and high stability. The immunization of both the mutI tri-RBD and homo-tri-RBD plus aluminum adjuvant induced high levels of specific IgG and neutralizing antibodies against the SARS-CoV-2 prototype strain in mice. Notably, regarding to the “immune-escape” Beta (B.1.351) variant, mutI tri-RBD elicited significantly higher neutralizing antibody titers than homo-tri-RBD. Furthermore, due to harboring the immune-resistant mutations as well as the evolutionarily convergent hotspots, the designed mutI tri-RBD also induced strong broadly neutralizing activities against various SARS-CoV-2 variants, especially the variants partially resistant to homo-tri-RBD. Homo-tri-RBD has been approved by the China National Medical Products Administration to enter clinical trial (No. NCT04869592), and the superior broad neutralization performances against SARS-CoV-2 support the mutI tri-RBD as a more promising vaccine candidate for further clinical developments.

Nine new compounds, including five natural rarely-occurring 2,3-dihydro-1H-indene derivatives named diaporindenes E−I ( 1 − 5 ), and four new benzophenone analogues named tenellones J−M ( 6 − 9 ) were isolated from the deep-sea sediment-derived fungus Phomopsis lithocarpus FS508. All the structures for these new compounds were fully characterized on the basis of spectroscopic data, NMR spectra, and ECD calculation and single-crystal X-ray diffraction analysis. The potential anti-tumor activities of compounds 1 − 9 against four tumor cell lines SF-268, MCF-7, HepG-2, and A549 were evaluated using the SRB method. Compound 7 exhibited cytotoxic activity against the SF-268 cell line with an IC50 value of 11.36 μmol·L−1.

Made available in DSpace on 2021-06-25T15:08:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-03-09 Wellcome Trust From 1985 to 2016, the prevalence of underweight decreased, and that of obesity and severe obesity increased, in most regions, with significant variation in the magnitude of these changes across regions. We investigated how much change in mean body mass index (BMI) explains changes in the prevalence of underweight, obesity, and severe obesity in different regions using data from 2896 population-based studies with 187 million participants. Changes in the prevalence of underweight and total obesity, and to a lesser extent severe obesity, are largely driven by shifts in the distribution of BMI, with smaller contributions from changes in the shape of the distribution. In East and Southeast Asia and sub-Saharan Africa, the underweight tail of the BMI distribution was left behind as the distribution shifted. There is a need for policies that address all forms of malnutrition by making healthy foods accessible and affordable, while restricting unhealthy foods through fiscal and regulatory restrictions. Imperial Coll London, London, England Harvard TH Chan Sch Publ Hlth, Boston, MA USA Middlesex Univ, London, England World Hlth Org, Geneva, Switzerland Minist Hlth, Victoria, Seychelles Univ Lausanne, Lausanne, Switzerland Eduardo Mondlane Univ, Maputo, Mozambique Victor Babes Univ Med & Pharm Timisoara, Timisoara, Romania Univ Sydney, Sydney, NSW, Australia Natl Inst Biomed Innovat Hlth & Nutr, Tokyo, Japan Univ Auckland, Auckland, New Zealand Seoul Natl Univ, Seoul, South Korea ICMR Natl Inst Nutr, Hyderabad, India Capital Med Univ, Beijing An Zhen Hosp, Beijing, Peoples R China Univ Peruana Cayetano Heredia, Lima, Peru Univ Tunis El Manar, Tunis, Tunisia Univ Oulu, Oulu, Finland Univ Zagreb, Zagreb, Croatia Univ Ljubljana, Ljubljana, Slovenia Caja Costarricense Seguro Social, San Jose, Costa Rica Al Quds Univ, East Jerusalem, State Of Palest, Palestine Natl Ctr Publ Healthcare, Nur Sultan, Kazakhstan Minist Hlth, Kuala Lumpur, Malaysia Qatar Univ, Doha, Qatar Birzeit Univ, Birzeit, State Of Palest, Israel Usmanu Danfodiyo Univ Teaching Hosp, Sokoto, Nigeria Inst Mexicano Seguro Social, Mexico City, DF, Mexico Flinders Univ S Australia, Adelaide, SA, Australia Mahidol Univ, Salaya, Nakhon Pathom, Thailand BRAC James P Grant Sch Publ Hlth, Dhaka, Bangladesh Univ Copenhagen, Copenhagen, Denmark Copenhagen Univ Hosp, Copenhagen, Denmark Food & Nutr Res Inst, Taguig, Philippines Urmia Univ Med Sci, Orumiyeh, Iran Inst Nacl Ciencias Med & Nut, Mexico City, DF, Mexico Univ Amsterdam, Amsterdam, Netherlands Shahrekord Univ Med Sci, Shahrekord, Iran Noncommunicable Dis Res Ctr, Tehran, Iran Univ Oslo, Oslo, Norway Univ Bremen, Bremen, Germany Republican Ctr Hlth Promot, Bishkek, Kyrgyzstan Natl Ctr Diabet Endocrinol & Genet, Amman, Jordan Princess Nourah Bint Abdulrahman Univ, Riyadh, Saudi Arabia Kuwait Inst Sci Res, Kuwait, Kuwait King Abdulaziz Univ, Jeddah, Saudi Arabia Dasman Diabet Inst, Kuwait, Kuwait Luxembourg Inst Hlth, Strassen, Luxembourg Aldara Hosp & Med Ctr, Riyadh, Saudi Arabia King Abdullah Int Med Res Ctr, Riyadh, Saudi Arabia Bispebjerg & Frederiksberg Hosp, Copenhagen, Denmark Barcelona Inst Global Hlth CIBERESP, Barcelona, Spain World Hlth Org Reg Off Eastern Mediterranean, Cairo, Egypt Bombay Hosp & Med Res Ctr, Mumbai, Maharashtra, India Res Ctr Social Determinants Hlth, Tehran, Iran UMR CNRS MNHN 7206 Ecoanthropologie, Paris, France Univ Lille, Lille, France Lille Univ Hosp, Lille, France Western Norway Univ Appl Sci, Sogndal, Norway Norwegian Sch Sport Sci, Oslo, Norway Madras Diabet Res Fdn, Chennai, Tamil Nadu, India Zahedan Univ Med Sci, Zahedan, Iran Natl Inst Publ Hlth, Tunis, Tunisia Univ Porto, Inst Publ Hlth, Porto, Portugal Norwegian Inst Publ Hlth, Oslo, Norway Univ Massachusetts, Amherst, MA 01003 USA Abt Associates Inc, Kathmandu, Nepal Univ Iceland, Reykjavik, Iceland Univ Yaounde I, Yaounde, Cameroon Univ Fed Pelotas, Pelotas, RS, Brazil Univ Med 1, Yangon, Myanmar Banska Bystrica Reg Author Publ Hlth, Banska Bystrica, Slovakia Tel Aviv Univ, Tel Aviv, Israel Hebrew Univ Jerusalem, Jerusalem, Israel Univ Porto, Med Sch, Porto, Portugal Neyshabur Univ Med Sci, Neyshabur, Iran Res Inst Endocrine Sci, Tehran, Iran Indian Council Med Res, New Delhi, India Natl Inst Publ Hlth, Copenhagen, Denmark Univ Sci & Technol, Sanaa, Yemen Med Univ Lodz, Lodz, Poland Med Univ Gdansk, Gdansk, Poland Univ Autonoma Madrid CIBERESP, Madrid, Spain Univ Rzeszow, Rzeszow, Poland Univ Palermo, Palermo, Italy Pan Amer Hlth Org, Washington, DC USA Mohammed V Univ Rabat, Rabat, Morocco Univ Pernambuco, Recife, PE, Brazil Baqai Inst Diabetol & Endocrinol, Karachi, Pakistan Univ Fed Santa Catarina, Florianopolis, SC, Brazil Dalhousie Univ, Halifax, NS, Canada Jordan Univ Sci & Technol, Irbid, Jordan Univ Fed Maranhao, Sao Luis, Brazil Farabi Kazakh Natl Univ, Alma Ata, Kazakhstan Univ Coll Dublin, Dublin, Ireland Christian Med Coll & Hosp, Vellore, Tamil Nadu, India Fed Minist Social Affairs Hlth Care & Consumer Pr, Vienna, Austria Cafam Univ Fdn, Bogota, Colombia Kazakh Natl Med Univ, Alma Ata, Kazakhstan Lithuanian Univ Hlth Sci, Kaunas, Lithuania Univ Sao Paulo, Sao Paulo, Brazil BJ Med Coll, Ahmadabad, Gujarat, India Chirayu Med Coll, New Delhi, India Sunder Lal Jain Hosp, Delhi, India Hosp Sick Children, Toronto, ON, Canada Shandong Univ Tradit Chinese Med, Jinan, Peoples R China Shanghai Jiao Tong Univ, Sch Med, Shanghai, Peoples R China Univ Republica, Montevideo, Uruguay Inst Med Res & Med Plant Studies, Yaounde, Cameroon Ufa Eye Res Inst, Ufa, Russia Nepal Hlth Res Council, Kathmandu, Nepal Univ Montenegro, Niksic, Montenegro Univ Southern Denmark, Copenhagen, Denmark Univ Gothenburg, Gothenburg, Sweden Univ Fed Rio de Janeiro, Rio De Janeiro, Brazil Natl Inst Publ Hlth & Environm, Bilthoven, Netherlands Univ Turin, Turin, Italy UCL, London, England Liverpool John Moores Univ, Liverpool, Merseyside, England Nanyang Technol Univ Singapore, Singapore, Singapore German Inst Human Nutr, Potsdam, Germany Endocrinol Res Ctr, Moscow, Russia Ctr Educ Med & Invest Clin, Buenos Aires, DF, Argentina IRCCS Neuromed, Pozzilli, Italy Toulouse Univ, Sch Med, Toulouse, France Univ Zurich, Zurich, Switzerland Univ Hosp KU Leuven, Leuven, Belgium Flemish Agcy Care & Hlth, Brussels, Belgium Univ Ghent, Ghent, Belgium FrieslandCampina, Amersfoort, Netherlands Univ Cent Venezuela, Caracas, Venezuela Bielefeld Univ, Bielefeld, Germany World Hlth Org Reg Off Europe, Moscow, Russia German Canc Res Ctr, Heidelberg, Germany Fred Hollows Fdn, Auckland, New Zealand Univ Med & Pharm Bucharest, Bucharest, Romania Swansea Univ, Swansea, W Glam, Wales Univ Coll Copenhagen, Copenhagen, Denmark Inst Publ Hlth, Tirana, Albania Munster Technol Univ, Cork, Ireland Univ La Laguna, Tenerife, Spain Univ Malta, Msida, Malta Vanderbilt Univ, 221 Kirkland Hall, Nashville, TN 37235 USA Minist Hlth, Tongatapu, Tonga Canadian Fitness & Lifestyle Res Inst, Ottawa, ON, Canada Hosp Santa Maria, Lisbon, Portugal Istanbul Univ Cerrahpasa, Istanbul, Turkey Univ Fed Juiz de Fora, Juiz De Fora, Brazil Minist Publ Hlth, Asuncion, Paraguay Natl Inst Publ Hlth, Prague, Czech Republic Gaetano Fucito Hosp, Mercato San Severino, Italy Univ Groningen, Groningen, Netherlands Karolinska Inst, Huddinge, Sweden Ctr Estudios Nutr Infantil, Buenos Aires, DF, Argentina Univ Porto, Porto, Portugal Univ Zaragoza, Zaragoza, Spain Santiago de Compostela Univ, Santiago De Compostela, Spain Minist Hlth, Ankara, Turkey Council Agr Res & Econ, Rome, Italy Fed Univ Rio Grande, Rio Grande, Brazil India Diabet Res Fdn, Chennai, Tamil Nadu, India Duke NUS Med Sch, Singapore, Singapore ICMR Natl Inst Med Stat, New Delhi, India Singapore Eye Res Inst, Singapore, Singapore Acad Sinica, Taipei, Taiwan Capital Inst Pediat, Beijing, Peoples R China Duke Univ, Durham, NH USA Kailuan Gen Hosp, Tangshan, Peoples R China Univ Oxford, Oxford, England Ahvaz Jundishapur Univ Med Sci, Ahvaz, Iran Gertner Inst Epidemiol & Hlth Policy Res, Ramat Gan, Israel Natl Ctr Publ Hlth & Anal, Sofia, Bulgaria Univ Fribourg, Fribourg, Switzerland Minist Hlth & Welf, Taipei, Taiwan CIBER Epidemiol & Salud Publ, Murcia, Spain Univ Southern Denmark, Odense, Denmark Univ Estadual Paulista, Presidente Prudente, Brazil Med Univ Silesia, Katowice, Poland Charles Univ Prague, Prague, Czech Republic Primary Hlth Care, Floriana, Malta Carol Davila Univ Med & Pharm, Bucharest, Romania Katholieke Univ Leuven, Leuven, Belgium STAT Canada, Ottawa, ON, Canada UMR CNRS MNHN 7206 Ecoanthropol, Marseille, France Agcy Prevent & Social Med, Bregenz, Austria Univ Southampton, Southampton, Hants, England Inst Pasteur, Lille, France Malawi Epidemiol & Intervent Res Unit, Lilongwe, Malawi CIBEROBN, Madrid, Spain Univ Debrecen, Debrecen, Hungary Univ Med & Pharm Carol Davila, Bucharest, Romania Univ Fed Rio Grande do Sul, Porto Alegre, RS, Brazil CNR, Reggio Di Calabria, Italy Eftimie Murgu Univ Resita, Resita, Romania Spanish Agcy Food Safety & Nutr, Madrid, Spain Indian Stat Inst, Kolkata, India Tabriz Hlth Serv Management Res Ctr, Tabriz, Iran Geneva Univ Hosp, Geneva, Switzerland Sciensano, Brussels, Belgium Univ Med Ctr Groningen, Groningen, Netherlands Natl Res Ctr Prevent Med, Moscow, Russia Innovating Hlth Int, Port Au Prince, Haiti Univ Montreal, Montreal, PQ, Canada French Natl Res Inst Sustainable Dev, Montpellier, France French Publ Hlth Agcy, St Maurice, France Mediterranea Cardioctr, Naples, Italy Univ Vale Rio dos Sinos, Sao Leopoldo, Brazil Natl Inst Hyg Epidemiol & Microbiol, Havana, Cuba Natl Council Sci & Tech Res, Buenos Aires, DF, Argentina Minist Hlth & Med Educ, Tehran, Iran Univ Novi Sad, Novi Sad, Serbia Natl Inst Nutr, Hanoi, Vietnam Univ Queensland, Brisbane, Qld, Australia Ist Super Sanita, Rome, Italy Univ Cuenca, Cuenca, Ecuador Helmholtz Zentrum Munchen, Munich, Germany Univ Tehran Med Sci, Tehran, Iran Minist Sante & Hyg Publ, Abidjan, Cote Ivoire Univ Hosp Dusseldorf, Dusseldorf, Germany Natl Inst Cardiol, Warsaw, Poland Beijing Ctr Dis Prevent & Control, Beijing, Peoples R China UMI 3189 ESS, Marseille, France Beth Israel Deaconess Med Ctr, Boston, MA 02215 USA Scuola Super Sant Anna, Pisa, Italy Univ Latvia, Riga, Latvia Minist Hlth & Med Serv, Gizo, Solomon Islands Hormozgan Univ Med Sci, Bandar Abbas, Iran Univ Benin, Benin, Nigeria Univ Skovde, Skovde, Sweden Natl Inst Nutr & Food Technol, Tunis, Tunisia Univ West Indies, Kingston, Jamaica Univ Calif Davis, Davis, CA 95616 USA Univ Stellenbosch, Cape Town, South Africa Univ Duisburg Essen, Duisburg, Germany Karadeniz Tech Univ, Trabzon, Turkey Univ Helsinki, Helsinki, Finland Mashhad Univ Med Sci, Mashhad, Razavi Khorasan, Iran Rafsanjan Univ Med Sci, Rafsanjan, Iran Queens Univ Belfast, Belfast, Antrim, North Ireland Tabriz Univ Med Sci, Tabriz, Iran Fasa Univ Med Sci, Fasa, Iran Shiraz Univ Med Sci, Shiraz, Iran Baqai Med Univ, Karachi, Pakistan Ctr Salud Villanueva Norte, Badajoz, Spain Hosp Don Benito Villanueva Serena, Badajoz, Spain Minist Hlth, Buenos Aires, DF, Argentina Univ Insubria, Varese, Italy Pontificia Univ Catolica Chile, Santiago, Chile Inst Mother & Child Hlth, Warsaw, Poland Swiss Trop & Publ Hlth Inst, Basel, Switzerland Univ Tartu, Tartu, Estonia Univ Sains Malaysia, Kota Baharu, Kelantan, Malaysia Umea Univ, Umea, Sweden Univ Fed Sao Paulo, Sao Paulo, Brazil Hosp Univ Son Espases, Palma De Mallorca, Spain Hosp Clin Porto Alegre, Porto Alegre, RS, Brazil Univ Sumatera Utara, Medan, Indonesia Kindai Univ, Osaka, Japan Kyoto Univ, Kyoto, Japan Med Univ Warsaw, Warsaw, Poland Jagiellonian Univ, Med Coll, Krakow, Poland Minist Salute DG Prevenz Sanit, Rome, Italy Univ Catania, Catania, Italy CIBER Epidemiol & Salud Publ, Alicante, Spain Africa Hlth Res Inst, Mtubatuba, South Africa Univ Geneva, Med Sch, Geneva, Switzerland CIBER Epidemiol & Salud Publ, Barcelona, Spain Univ Fed Minas Gerais, Belo Horizonte, MG, Brazil Univ Utrecht, Utrecht, Netherlands Wageningen Univ, Wageningen, Netherlands Univ Adelaide, Adelaide, SA, Australia Lund Univ, Lund, Sweden Aristotle Univ Thessaloniki, Thessaloniki, Greece Inst Natl Sante & Rech Med, Villejuif, France Minist Hlth, Jerusalem, Israel McGill Univ, Montreal, PQ, Canada Gasol Fdn, Barcelona, Spain PASs Hirszfeld Inst Immunol & Expt Therapy, Wroclaw, Poland Univ Politecn Madrid, Madrid, Spain St Annes Univ Hosp, Brno, Czech Republic Natl Inst Publ Hlth, Cuernavaca, Morelos, Mexico Ctr Estudios Diabet AC, Mexico City, DF, Mexico Univ Autonoma Santo Domingo, Santo Domingo, Dominican Rep Minist Hlth, Lisbon, Portugal Inst Clin & Expt Med, Prague, Czech Republic Childrens Mem Hlth Inst, Warsaw, Poland Natl Ctr Cardiovasc Dis, Beijing, Peoples R China Univ Ferrara, Ferrara, Italy Author Sanitaria San Marino, San Marino, CA USA Iceland Heart Assoc, Kopavogur, Iceland Univ Icesi, Cali, Colombia Univ Estadual Montes Claros, Montes Claros, MG, Brazil Kings Coll London, London, England Int Agcy Res Canc, Lyon, France Capital Med Univ, Beijing, Peoples R China Capital Med Univ, Beijing Tongren Hosp, Beijing, Peoples R China Healis Sekhsaria Inst Publ Hlth, Navi Mumbai, India Eternal Heart Care Ctr & Res Inst, Jaipur, Rajasthan, India Univ Ibadan, Ibadan, Nigeria Inst Clin Effectiveness & Hlth Policy, Buenos Aires, DF, Argentina Natl Hlth Insurance Serv, Wonju, South Korea Prevent Metab Disorders Res Ctr, Tehran, Iran Res & Educ Inst Child Hlth, Nicosia, Cyprus Danish Canc Soc Res Ctr, Copenhagen, Denmark Univ West Indies, Cave Hill, Barbados Kermanshah Univ Med Sci, Kermanshah, Iran Univ Eastern Finland, Kuopio, Finland Beijing Inst Ophthalmol, Beijing, Peoples R China Int Hellen Univ, Thessaloniki, Greece Kyushu Univ, Fukuoka, Japan Univ Bergen, Bergen, Norway Tulane Univ, New Orleans, LA USA Natl Res Inst Hlth & Family Planning, Beijing, Peoples R China Chinese Ctr Dis Control & Prevent, Beijing, Peoples R China Univ Pecs, Pecs, Hungary Danish Hlth Author, Copenhagen, Denmark Joep Lange Inst, Amsterdam, Netherlands Univ Fed Pernambuco, Recife, PE, Brazil Oulu Univ Hosp, Oulu, Finland Univ Autonoma Bucaramanga, Bucaramanga, Colombia Swiss Fed Inst Technol, Zurich, Switzerland Chron Dis Res Ctr, Tehran, Iran Univ Hong Kong, Hong Kong, Peoples R China Chinese Univ Hong Kong, Hong Kong, Peoples R China Univ Western Australia, Perth, WA, Australia Kingston Hlth Sci Ctr, Kingston, ON, Canada Fdn Oftalmol Santander, Bucaramanga, Colombia Univ Oran 1, Oran, Algeria Nay Pyi Taw, Independent Publ Hlth Specialist, Nay Pyi Taw, Myanmar Minist Hlth & Sports, Nay Pyi Taw, Myanmar Peking Univ, Beijing, Peoples R China Natl Inst Publ Hlth, Bucharest, Romania Vrije Univ Amsterdam Med Ctr, Amsterdam, Netherlands Amer Univ Beirut, Beirut, Lebanon CIBER Epidemiol & Salud Publ, San Sebastian, Spain Cairo Univ, Cairo, Egypt Erasmus MC, Rotterdam, Netherlands Med Univ Varna, Varna, Bulgaria Univ Tokyo, Tokyo, Japan Deakin Univ, Geelong, Vic, Australia Tokyo Metropolitan Inst Gerontol, Tokyo, Japan Hadassah Univ Med Ctr, Jerusalem, Israel Norwegian Univ Sci & Technol, Trondheim, Norway Univ Melbourne, Melbourne, Vic, Australia Sports Univ Tirana, Tirana, Albania Univ Hosp Ctr Zagreb, Zagreb, Croatia Univ Zagreb, Sch Med, Zagreb, Croatia Heart Fdn, Melbourne, Vic, Australia Guangzhou 12th Hosp, Guangzhou, Peoples R China Univ Eugenio Maria Hostos, Santo Domingo, Dominican Rep Simon Fraser Univ, Burnaby, BC, Canada Inst Mol & Clin Ophthalmol Basel, Basel, Switzerland Univ New South Wales, Sydney, NSW, Australia World Hlth Org Country Off, Delhi, India Guilan Univ Med Sci, Rasht, Iran Inst Publ Hlth, Belgrade, Serbia Univ Opole, Opole, Poland Finnish Inst Hlth & Welf, Helsinki, Finland Gulu Univ, Gulu, Uganda Univ Crete, Iraklion, Greece Sri Venkateswara Univ, Tirupati, Andhra Pradesh, India Sree Chitra Tirunal Inst Med Sci & Technol, Trivandrum, Kerala, India Hellen Med Assoc Obes, Athens, Greece Natl & Kapodistrian Univ Athens, Athens, Greece Maharajgunj Med Campus, Kathmandu, Nepal Aarhus Univ, Aarhus, Denmark Johns Hopkins Bloomberg Sch Publ Hlth, Baltimore, MD USA Pennington Biomed Res Ctr, Baton Rouge, LA USA Natl Inst Epidemiol, Chennai, Tamil Nadu, India Univ Munster, Munster, Germany Israel Ctr Dis Control, Ramat Gan, Israel Res Inst Primordial Prevent Noncommunicable Dis, Esfahan, Iran Amsterdam UMC Publ Hlth Res Inst, Amsterdam, Netherlands South African Med Res Council, Cape Town, South Africa Kyrgyz State Med Acad, Bishkek, Kyrgyzstan Res Inst Child Nutr, Dortmund, Germany Shahid Beheshti Univ Med Sci, Tehran, Iran Univ Cambridge, Cambridge, England Mazandaran Univ Med Sci, Sari, Iran Hypertens Res Ctr, Esfahan, Iran Med Univ Innsbruck, Innsbruck, Austria Muhimbili Univ Hlth & Allied Sci, Dar Es Salaam, Tanzania Yonsei Univ, Coll Med, Seoul, South Korea Natl Canc Ctr, Goyang Si, South Korea Univ Coll South Denmark, Haderslev, Denmark Stat Austria, Vienna, Austria BP Koirala Inst Hlth Sci, Dharan, Nepal Univ Vienna, Vienna, Austria Tartu Univ Clin, Tartu, Estonia Kansai Med Univ, Hirakata, Osaka, Japan Hungarian Sch Sport Federat, Budapest, Hungary Minist Hlth & Qual Life, Port Louis, Mauritius Univ Hosp Ulm, Ulm, Germany North West Univ, Potchefstroom, South Africa Univ Jyvaskyla, Jyvaskyla, Finland Inst Publ Hlth, Podgorica, Montenegro Amrita Inst Med Sci, Cochin, Kerala, India Inst Endocrinol, Prague, Czech Republic All India Inst Med Sci, New Delhi, India African Populat & Hlth Res Ctr, Nairobi, Kenya Hanoi Univ Publ Hlth, Hanoi, Vietnam Hassan First Univ Settat, Settat, Morocco Minist Hlth, Algiers, Algeria Harokopio Univ, Athens, Greece Sahlgrens Acad, Gothenburg, Sweden Endocrinol & Metab Res Ctr, Tehran, Iran Univ Publ Hlth, Yangon, Myanmar Int Food Policy Res Inst, Dakar, Senegal Natl Univ Singapore, Singapore, Singapore Hong Kong Polytech Univ, Hong Kong, Peoples R China Tampere Univ Hosp, Tampere, Finland Univ Douala, Douala, Cameroon Univ Cape Town, Cape Town, South Africa Natl Inst Hlth Dev, Tallinn, Estonia West Virginia Univ, Morgantown, WV 26506 USA Oswaldo Cruz Fdn Rene Rachou Res Inst, Belo Horizonte, MG, Brazil Natl Taiwan Univ, Taipei, Taiwan Univ Chinese Acad Sci, Shanghai, Peoples R China Uppsala Univ, Uppsala, Sweden Taipei Med Univ, Taipei, Taiwan Sports Med Ctr Minho, Braga, Portugal Univ San Martin Porres, Lima, Peru Consejeria Sanidad Junta Castilla & Leon, Valladolid, Spain Univ Fed Ciencias Saude Porto Alegre, Porto Alegre, RS, Brazil Norrbotten Cty Council, Lulea, Sweden Ilembula Lutheran Hosp, Ilembula, Iran Univ Fed Ouro Preto, Ouro Preto, Brazil Univ Coimbra, Coimbra, Portugal Coimbra Univ Hosp Ctr, Coimbra, Portugal Inst Neurosci Natl Res Council, Padua, Italy Baker Heart & Diabet Inst, Melbourne, Vic, Australia Agr Univ Athens, Athens, Greece Hosp Israelita Albert Einstein, Sao Paulo, Brazil SB RAS Fed Res Ctr Inst Cytol & Genet, Novosibirsk, Russia Univ Otago, Dunedin, New Zealand Univ Padua, Padua, Italy Hellen Mediterranean Univ, Siteia, Greece Loughborough Univ, Loughborough, Leics, England Minist Hlth, Nicosia, Cyprus Lausanne Univ Hosp, Lausanne, Switzerland Escola Nacl Saude Publ Sergio Arouca, Rio De Janeiro, Brazil CIBERCV, Madrid, Spain Mary Immaculate Coll, Limerick, Ireland Emory Univ, Atlanta, GA 30322 USA Hungarian Soc Sports Med, Budapest, Hungary Paracelsus Med Univ, Salzburg, Austria Univ Estadual Centro Oeste, Guarapuava, Brazil Sherikashmir Inst Med Sci, Srinagar, India UiT Arctic Univ Norway, Tromso, Norway ICMR Natl Ctr Dis Informat & Res, Bengaluru, India Cape Peninsula Univ Technol, Cape Town, South Africa Brown Univ, Providence, RI 02912 USA London Sch Hyg & Trop Med, London, England Univ Edinburgh, Edinburgh, Midlothian, Scotland Shahid Sadoughi Univ Med Sci, Tehran, Iran Inst Natl Sante & Rech Med, Lille, France Robert Koch Inst, Berlin, Germany Univ Lisbon, Lisbon, Portugal Womens Social & Hlth Studies Fdn, Trivandrum, Kerala, India Democritus Univ, Alexandroupolis, Greece Grigore T Popa Univ Med & Pharm, Iasi, Romania Univ Firenze, Florence, Italy Ain Shams Univ, Cairo, Egypt Isfahan Cardiovasc Res Ctr, Esfahan, Iran Univ Strasbourg, Strasbourg, France Mulago Hosp, Kampala, Uganda Univ San Carlos, Guatemala City, Guatemala Univ Limpopo, Sovenga, South Africa Univ Med Sci Cienfuegos, Cienfuegos, Cuba Royal Coll Surgeons Ireland, Dublin, Ireland Trobe Univ, Melbourne, Vic, Australia Int Inst Mol & Cell Biol, Warsaw, Poland Inst Conmemorat Gorgas Estudios Salud, Panama City, Panama World Hlth Org Country Off, Lilongwe, Malawi Dept Publ Hlth, Nay Pyi Taw, Myanmar Univ Brescia, Brescia, Italy Minist Hlth & Social Protect, Dushanbe, Tajikistan Univ Limerick, Limerick, Ireland Croatian Inst Publ Hlth, Zagreb, Croatia Univ Kebangsaan Malaysia, Kuala Lumpur, Malaysia Bushehr Univ Med Sci, Bushehr, Iran Ulm Univ, Ulm, Germany Dept Stat, Kuala Lumpur, Malaysia Kobe Univ, Kobe, Hyogo, Japan Suraj Eye Inst, Nagpur, Maharashtra, India UNICEF, Yaounde, Cameroon Univ Med Greifswald, Greifswald, Germany Karolinska Inst, Stockholm, Sweden Natl Inst Hyg & Epidemiol, Hanoi, Vietnam Univ Med & Pharm Ho Chi Minh City, Ho Chi Minh City, Vietnam Hanoi Med Univ, Hanoi, Vietnam LifeDoc Hlth, Memphis, TN USA Heartfile, Islamabad, Pakistan Eastern Mediterranean Publ Hlth Network, Amman, Jordan Univ Manchester, Manchester, Lancs, England State Univ Med & Pharm, Kishinev, Moldova Tachikawa Gen Hosp, Nagaoka, Niigata, Japan Korea Ctr Dis Control & Prevent, Cheongju, South Korea Japan Wildlife Res Ctr, Tokyo, Japan Istanbul Univ, Istanbul, Turkey Minist Hlth, Bandar Seri Begawan, Brunei Univ Fed Parana, Curitiba, Parana, Brazil Univ Madeira, Funchal, Portugal Univ Puerto Rico, San Juan, PR 00936 USA Ctr Clin Res & Prevent, Glostrup, Denmark MRC Lifecourse Epidemiol Unit, Southampton, Hants, England Kwame Nkrumah Univ Sci & Technol, Kumasi, Ghana UniSante, Lausanne, Switzerland Inst Canc Res, Prevent & Clin Network, Florence, Italy Univ Wisconsin, Madison, WI USA Heidelberg Univ, Heidelberg, Germany IRCCS Ente Osped Specializzato Gastroenterol S de, Bari, Italy Zayed Univ, Abu Dhabi, U Arab Emirates Catholic Univ Daegu, Daegu, South Korea Univ Med Pharm Sci & Technol Targu Mures, Targu Mures, Romania Jivandeep Hosp, Anand, Gujarat, India South African Med Res Council, Durban, South Africa Natl Dent Care Ctr Singapore, Singapore, Singapore Univ Malaga, Malaga, Spain Univ Hosp Varese, Varese, Italy Vietnam Natl Heart Inst, Hanoi, Vietnam Clin Med Avanzada Dr Abel Gonzalez, Santo Domingo, Dominican Rep Leibniz Inst Prevent Res & Epidemiol BIPS, Bremen, Germany Univ Sarajevo, Sarajevo, Bosnia & Herceg Cardiovasc Prevent Ctr Udine, Udine, Italy Pisa Univ Hosp, Pisa, Italy Minist Hlth & Med Serv, Honiara, Solomon Islands Publ Hlth Agcy Catalonia, Barcelona, Spain Inst Hosp del Mar Invest Med, Barcelona, Spain Ardabil Univ Med Sci, Ardebil, Iran Ctr Dis Prevent & Control, Riga, Latvia Alborz Univ Med Sci, Karaj, Iran Minist Hlth, Hanoi, Vietnam Pure Earth, Dhaka, Bangladesh Inst Epidemiol Dis Control & Res, Dhaka, Bangladesh Univ Turku, Turku, Finland Inst Univ Invest Atenc Primaria Jordi Gol, Girona, Spain Univ Putra Malaysia, Serdang, Malaysia Univ Malaya, Kuala Lumpur, Malaysia Sotiria Hosp, Athens, Greece Univ Valencia, Valencia, Spain Univ Philippines, Manila, Philippines Slovak Acad Sci, Bratislava, Slovakia Nutr Res Fdn, Barcelona, Spain Minas Gerais State Secretariat Hlth, Belo Horizonte, MG, Brazil CS S Agustin Ibsalut, Palma De Mallorca, Spain Amsterdam Inst Global Hlth & Dev, Amsterdam, Netherlands Natl Inst Hlth Doutor Ricardo Jorge, Lisbon, Portugal Univ Nove Julho, Sao Paulo, Brazil Publ Hlth Agcy Canada, Ottawa, ON, Canada Canarian Hlth Serv, Tenerife, Spain Univ Ind Santander, Bucaramanga, Colombia Assoc Calabrese Epatol, Reggio Di Calabria, Italy Univ Minho, Braga, Portugal Fiji Natl Univ, Suva, Fiji Spanish Nutr Fdn, Madrid, Spain Publ Hlth Agcy Sweden, Stockholm, Sweden Inst Food Sci Natl Res Council, Avellino, Italy Sitaram Bhartia Inst Sci & Res, New Delhi, India Kindergarten Avlonari, Evia, Greece Natl Inst Hlth, Lima, Peru Minist Hlth, Jakarta, Indonesia Catalan Dept Hlth, Barcelona, Spain Biodonostia Hlth Res Inst, San Sebastian, Spain Inst Saude Ambiental, Lisbon, Portugal South Karelia Social & Hlth Care Dist, Lappeenranta, Finland McMaster Univ, Hamilton, ON, Canada Natl Canc Ctr, Tokyo, Japan Univ Sao Paulo Clin Hosp, Sao Paulo, Brazil Hosp Italiano Buenos Aires, Buenos Aires, DF, Argentina Rigshosp, Copenhagen, Denmark Acad Med Ctr Univ Amsterdam, Amsterdam, Netherlands Ctr Oral Hlth Serv & Res Midnorway, Trondheim, Norway Lagos State Univ, Coll Med, Lagos, Nigeria Univ Las Palmas Gran Canaria, Las Palmas Gran Canaria, Spain Comenius Univ, Bratislava, Slovakia Natl Ctr Dis Control & Publ Hlth, Tbilisi, Georgia Nippon Med Sch, Tokyo, Japan Sungkyunkwan Univ, Seoul, South Korea Finnish Inst Occupat Hlth, Helsinki, Finland Publ Hlth Promot & Dev Org, Kathmandu, Nepal St Vincents Hosp, Sydney, NSW, Australia Nes Municipal, Aarnes, Norway Hlth Polytech Jakarta II Inst, Jakarta, Indonesia Diponegoro Univ, Semarang, Indonesia Univ Bari, Bari, Italy Inst Reg Sante Publ, Ouidah, Benin Univ Bordeaux, Bordeaux, France Oslo Metropolitan Univ, Oslo, Norway Inst Publ Hlth, Skopje, North Macedonia Univ Leuven, Leuven, Belgium Lamprecht & Stamm Sozialforsch & Beratung AG, Zurich, Switzerland Inst Natl Sante & Rech Med, Nancy, France Univ Bonn, Bonn, Germany Sotiria Hosp, Sotiria, Greece Kalina Malina Kindergarten, Pazardjik, Bulgaria Jikei Univ, Sch Med, Tokyo, Japan Fu Jen Catholic Univ, Taipei, Taiwan Natl Stat Off, Praia, Cape Verde Natl Inst Publ Hlth Natl Inst Hyg, Warsaw, Poland Deakin Univ, Sydney, NSW, Australia Sci Res Inst Maternal & Child Hlth, Ashkhabad, Turkmenistan Univ Lincoln, Lincoln, England Minist Hlth, Amman, Jordan UNICEF, Niamey, Niger Cent Univ Kerala, Kasaragod, India Amsterdam Publ Hlth Res Inst, Amsterdam, Netherlands Hlth Serv Murcia, Murcia, Spain Inst Invest Sanitaria Illes Balears, Menorca, Spain Univ Bologna, Bologna, Italy Hellen Hlth Fdn, Athens, Greece Govt Med Coll, Bhavnagar, Gujarat, India Sefako Makgatho Hlth Sci Univ, Ga Rankuwa, South Africa Addis Ababa Univ, Addis Ababa, Ethiopia Minist Hlth, Wellington, New Zealand Univ Ctr Occidental Lisandro Alvarado, Barquisimeto, Venezuela Meharry Med Coll, Nashville, TN 37208 USA Dokuz Eylul Univ, Izmir, Turkey Univ Tampere, Tays Eye Ctr, Tampere, Finland Sabiha Gokcen Ilkokulu, Ankara, Turkey Polytech Inst Porto, Porto, Portugal Icahn Sch Med Mt Sinai, New York, NY 10029 USA Univ CEU San Pablo, Madrid, Spain CNR, Pisa, Italy Univ Miguel Hernandez, Alicante, Spain Vrije Univ Amsterdam, Amsterdam, Netherlands Sunflower Nursery Sch, Craiova, Romania North Karelia Ctr Publ Hlth, Joensuu, Finland Univ Witwatersrand, Johannesburg, South Africa Med Univ Vienna, Vienna, Austria Cork Inst Technol, Cork, Ireland Inst Med Res, Kuala Lumpur, Malaysia Xinjiang Med Univ, Urumqi, Peoples R China Shanghai Educ Dev Co Ltd, Shanghai, Peoples R China Minist Hlth, Auckland, New Zealand Rebro Univ, Rebro, Sweden St Georges Univ London, London, England Univ Indonesia, Jakarta, Indonesia Minist Agr & Rural Affairs, Inst Food & Nutr Dev, Beijing, Peoples R China Fudan Univ, Childrens Hosp, Shanghai, Peoples R China Univ Cyprus, Nicosia, Cyprus Niigata Univ, Niigata, Japan Int Med Univ, Shah Alam, Selangor, Malaysia Hellen Mediterranean Univ, Iraklion, Greece Iran Univ Med Sci, Tehran, Iran Ctr Diabet & Endocrine Care, Srinagar, India Peking Univ First Hosp, Beijing, Peoples R China Jiangsu Prov Ctr Dis Control & Prevent, Nanjing, Peoples R China Sun Yat Sen Univ, Guangzhou, Peoples R China West Kazakhstan Med Univ, Aktobe, Kazakhstan Inner Mongolia Med Univ, Hohhot, Peoples R China Przedszkole 81, Warsaw, Poland Univ Estadual Paulista, Presidente Prudente, Brazil

Mud crabs, found throughout the Indo-Pacific region, are coastal species that are important fisheries resources in many tropical and subtropical Asian countries. Here, we present a chromosome-level genome assembly of a mud crab (Scylla paramamosain). The genome is 1.55 Gb (contig N50 191 kb) in length and encodes 17,821 proteins. The heterozygosity of the assembled genome was estimated to be 0.47%. Effective population size analysis suggested that an initial large population size of this species was maintained until 200 thousand years ago. The contraction of cuticle protein and opsin genes compared with Litopenaeus vannamei is assumed to be correlated with shell hardness and light perception ability, respectively. Furthermore, the analysis of three chemoreceptor gene families, the odorant receptor (OR), gustatory receptor (GR) and ionotropic receptor (IR) families, suggested that the mud crab has no OR genes and shows a contraction of GR genes and expansion of IR genes. The numbers of the three gene families were similar to those in three other decapods but different from those in two nondecapods and insects. In addition, IRs were more diversified in decapods than in nondecapod crustaceans, and most of the expanded IRs in the mud crab genome were clustered with the antennal IR clades. These findings suggested that IRs might exhibit more diverse functions in decapods than in nondecapods, which may compensate for the smaller number of GR genes. Decoding the S. paramamosain genome not only provides insight into the genetic changes underpinning ecological traits but also provides valuable information for improving the breeding and aquaculture of this species.

In this study, the first and second harvesting Pisang Awak in the field were sprayed with different dosage of paclobutrazol in order to analyze the mechanism of its induction of flowering. The results showed that there was no significant difference in the total number of newly extracted leaves between treatments. The first harvesting was 3.0 g/plant, and the second one was 2.0 g/plant. The pumping speed of leaves is the fastest and the accumulation of leaves number is the earliest. The 3.0 g/plant treatment of first harvesting Pisang Awak had the earliest bud extraction stage, and the bud extraction rate reached 62.5% at 170 days after treatment, about 40 days earlier than control group. Within the range of 1.0~5.0 g/plant, PP 333 promoted carbohydrate synthesis in leaves of Pisang Awak, significantly reduced the accumulation of nitrogen, significantly increased the carbon-nitrogen ratio (C/N), and significantly reduced the contents of GA 3 and IAA in the leaves. The results showed that the exogenous polyazole could accelerate the pumping speed of leaves, promote flowering and early bud extraction by regulating the distribution of carbon and nitrogen nutrients and the content of endogenous hormones, thus providing technical guidance for the management of early bud extraction culture of Pisang Awak.

Background: Genome-scale approaches have played a significant role in the analysis of evolutionary relationships. Because of rich polymorphisms, high evolutionary rate and rare recombination, mitochondrial DNA sequences are commonly considered as effective markers for estimating population genetics, evolutionary and phylogenetic relationships. Flying fishes are important components of epipelagic ecosystems. Up to now, only few complete mitochondrial genomes of flying fishes have been reported. In the present study, the complete mitochondrial DNA sequences of the Cheilopogon pinnatibarbatus japonicus and Hirundichthys rondeletii had been determined. Methods: Based on the published mitogenome of Cheilopogon atrisignis (GenBank: KU360729), fifteen pairs of primers were designed by the software Primer Premier 5.0 to get the complete mitochondrial genomes of two flying fishes. According to the reported data, the phylogenetic position of two flying fishes were detected using the conserved 12 protein-coding genes. Result: The complete mitochondrial genomes of Cheilopogon pinnatibarbatus japonicus and Hirundichthys rondeletii are determined. They are 16532bp and 16525bp in length, respectively. And they both consists of 13 protein-coding genes, 22 transfer RNA (tRNA) genes, two ribosomal RNA (rRNA) genes and a control region. The OL regions are conserved in these two flying fishes and might have no function. From the tree topologies, we found C.p. japonicus and H. rondeletii clustered in a group. The findings of the study would contribute to the phylogenetic classification and the genetic conservation management of C.p. japonicus and H. rondeletii.

Intrahepatic cholangiocarcinoma (iCCA) is a highly fatal malignant cancer worldwide. Elucidating the underlying molecular mechanism of iCCA progression is critical for the identification of new therapeutic targets. The present study explored the role of the miR-148a–GLUT1 axis in the progression of iCCA. The expression of GLUT1 was detected by using immunohistochemistry, western blot assays, and real-time polymerase chain reaction. The effects of GLUT1 on cell proliferation, invasion, and chemoresistance were investigated both in vitro and in vivo. A luciferase reporter assay was used to explore the effect of miR-148a on GLUT1 expression. GLUT1 was overexpressed in iCCA tissues. GLUT1 overexpression was associated with shorter overall and disease-free survival. Knockdown of GLUT1 reduced, while overexpression of GLUT1 promoted, the proliferation, motility, and invasiveness of iCCA cells in vitro and in vivo. Silencing GLUT1 significantly sensitized iCCA cells to gemcitabine in vitro and in vivo. GLUT1 was directly regulated by miR-148a, whose downregulation was associated with the proliferation, migration, and invasion of iCCA cells. WZB117, a GLUT1 inhibitor, inhibited tumor growth in an iCCA patient-derived xenograft model. These results indicate that downregulation of miR-148a levels results in GLUT1 overexpression in iCCA, leading to iCCA progression and gemcitabine resistance.

Ptinus japonicus Reitter is a subversive pest of stored-product throughout the world. Scanning electron microscopy (SEM) was used to observe the antennal morphology and sensilla ultrastructure in b...

Objective: We aimed to assess the possible relations between serum levels of macrophage migration inhibitory factor (MIF), a central cytokine of the innate immunity and inflammatory response, and benign paroxysmal positional vertigo (BPPV) risk and BPPV recurrence events. Methods: In the present study, 154 patients with BPPV, and 100 age-and sex-matched control subjects were enrolled in the study. All the patients and controls underwent a complete audio-vestibular test battery including the Dix–Hallpike maneuver and supine roll test. In the BPPV group, measurements of MIF levels were repeated 1 month after the vertigo attack. The patients were also divided into the recurrence group and the nonrecurrence group in the 1-year follow-up. Results: The serum levels of MIF in patients with BPPV were higher than in those controls (13.9[interquartile range {IQR}, 8.9–18.4] ng/ml vs. 9.8[7.8–11.8]; P Conclusions: Elevated MIF is positively correlated with BPPV risk and BPPV recurrence events, requiring further efforts to clarify the exact mechanism.

In subduction zones, serpentinization and biological processes may release alkanes to the deep waters, which would probably result in the rapid spread of Alcanivorax. However, the timing and area of the alkane distribution and associated enrichment of alkane-degrading microbes in the dark world of the deep ocean have not been explored. In this study, we report the richness (up to 17.8%) of alkane-degrading bacteria, represented by Alcanivorax jadensis, in deep water samples obtained at 3,000 to 6,000 m in the Mariana Trench in two cruises. The relative abundance of A. jadensis correlated with copy numbers of functional almA and alkB genes, which are involved in alkane degradation. In these water samples, we detected a high flux of alkanes, which probably resulted in the prevalence of A. jadensis in the deep waters. Contigs of A. jadensis were binned from the metagenomes for examination of alkane degradation pathways and deep sea-specific pathways, which revealed a lack of nitrate and nitrite dissimilatory reduction in our A. jadensis strains. Comparing the results for the two cruises conducted close to each other, we suggest periodic release of alkanes that may spread widely but periodically in the trench. Distribution of alkane-degrading bacteria in the world’s oceans suggests the periodic and remarkable contributions of Alcanivorax to the deep sea organic carbon and nitrogen sources. IMPORTANCE In the oligotrophic environment of the Mariana Trench, alkanes as carbohydrates are important for the ecosystem, but their spatial and periodic spreading in deep waters has never been reported. Alkane-degrading bacteria such as Alcanivorax spp. are biological signals of the alkane distribution. In the present study, Alcanivorax was abundant in some waters, at depths of up to 6,000 m, in the Mariana Trench. Genomic, transcriptomic, and chemical analyses provide evidence for the presence and activities of Alcanivorax jadensis in deep sea zones. The periodic spreading of alkanes, probably from the subductive plates, might have fundamentally modified the local microbial communities, as well as perhaps the deep sea microenvironment.

Platelet counts have been reported to be closely related to distant metastasis of many malignant tumors. Our previous study showed that elevated peripheral blood platelet counts may be an adverse prognostic factor of anaplastic thyroid carcinoma (ATC) patients, indicating that platelets may promote ATC progression. In the present study, we aimed to identify the role of platelets in ATC cell invasion and migration and to explore the underlying mechanisms. We found that platelets can promote the invasive and migratory of ATC cells, which may be related to the interaction between activated platelet-secreted chemokine (C-C motif) ligand 3 (CCL3) and its receptor CCR5. The interaction was shown to induce the upregulation of matrix metalloproteinase (MMP)-1 via NF-κB pathway. These findings could provide a new idea for the research of targeted platelets to inhibit tumor metastasis.

Bacteria produce metamorphosis-associated contractile (MAC) structures to induce larval metamorphosis in Hydroides elegans. The distribution and diversity of mac gene homologs in marine environments are largely unexplored. In the present study mac genes were examined in marine environments by analyzing 101 biofilm and 91 seawater metagenomes. There were more mac genes in biofilms than in seawater, and substratum type, location, or sampling time did not affect the mac genes in biofilms. The mac gene clusters were highly diverse and often incomplete while the three MAC components co-occurred with other genes of different functions. Genomic analysis of four Pseudoalteromonas and two Streptomyces strains revealed the mac genes transfers among different microbial taxa. It is proposed that mac genes are more specific to biofilms; gene transfer among different microbial taxa has led to highly diverse mac gene clusters; and in most cases, the three MAC components function individually rather than forming a complex.

Micromonospora craniellae LHW63014T is a novel marine Micromonospora, isolated from a Craniella species sponge collected in the South China Sea. In this study, we report the complete genome sequence of M. craniellae LHW63014T, which is comprised of a circular chromosome of 6,839,926 bp with the G + C content of 70.9 mol%. The complete genome contained 6572 protein-coding genes, 48 tRNA genes, and 9 rRNA genes. Genomic annotations revealed that 79.09% of the protein-coding genes were assigned to the COG database, among which, the abundant genes were predicted to be involved in transcription, replication, recombination and repair, and amino acid transport and metabolism. Secondary metabolites prediction using antiSMASH revealed that 22 biosynthetic gene clusters (BGC) of secondary metabolites were located in the genome of M. craniellae LHW63014T, 19 of which showed low similarity (

Asgard archaea, believed to be the ancestor of eukaryotes, constitute a new superphylum prevalent in diverse sediment environments. Their distribution and metabolic potentials are diversified and elusive. In the present study, the distribution and genomic characteristics of Asgard archaea in the Haima cold seep of the South China Sea (SCS) were investigated. In total, we obtained 48 high-quality metagenome-assembled genomes (MAGs), which fell into 7 separated lineages affiliated with the phyla Lokiarchaeota, Thorarchaeota, Heimdallarchaeota and Gerdarchaeota. These Asgard archaea were mainly distributed in the cold seep but rarely detected in the control marine sediments. We found that Asgard might encode complex CRISPR-Cas systems, which likely protected the archaea against abundant viruses in cold seep sediments. Phylogenomic tree indicated the novelty of cold-seep Asgard archaea, and highlighted that MAG HM7_B34 was affiliated with the newly proposed Gerdarchaeota phylum. Metabolic reconstruction indicated that Gerdarchaeota in the Haima cold seep were obligate anaerobic archaea and might undergo autotrophic lifestyle using the Wood-Ljungdahl pathway. Moreover, Gerdarchaeota might metabolize formaldehyde and likely had the ability to hydrolyze oligosaccharides including chitin. Altogether, our study expands the cognition of Asgard archaea in terms of spatial distribution, biodiversity and carbon cycle in cold seep ecosystems.

The mitochondrial genome (mitogenome) has been extensively used for studying phylogenetic relationships at different taxonomic levels. Several molecular analyses have been performed, but the phylogenetic relationships among infraorders in Polyphaga have not been well resolved. In this work, three nearly complete mitogenomes of Coleoptera, Sitophilus oryzae , Oryzaephilus surinamensis and Callosobruchus chinensis , were determined. The O. surinamensis and S. oryzae mitogenomes harbor gene content typical of other Polyphaga mitogenomes, while a gene rearrangement (trnQ) was found in the C. chinensis mitogenome. The mitogenomes of these three Coleoptera species each consist of approximately 13 protein-coding genes, 22 tRNA genes, two rRNA genes and one A + T-rich region. Phylogenetic analysis within Polyphaga was carried out based on mitochondrial data. The phylogenetic results within Polyphaga support the basal position of Cyphon sp., which belonged to Scirtoidea, Elateriformia. Within Cucujiformia, monophyletic Curculionoidea, Chrysomeloidea and Tenebrionoidea were confirmed.

Oxidative stress is a possible pathogenesis of hyperalgesia. Advanced oxidation protein products (AOPPs), a new family of oxidized protein compounds, have been considered as a novel marker of oxidative stress. However, the role of AOPPs in the mechanism of hyperalgesia remains unknown. Our study aims to investigate whether AOPPs have an effect on hyperalgesia and the possible underlying mechanisms. To identify the AOPPs involved, we induced hyperalgesia in rats by injecting complete Freund’s adjuvant (CFA) in hindpaw. The level of plasma AOPPs in CFA-induced rats was 1.6-fold in comparison with what in normal rats (P, Graphical abstract Schematic representation of AOPPs-induced TRPV1 sensitization through Nox1/Nox4-dependent ROS generation, intracellular Ca2+ increase and CGRP release.fx1, Highlights • Plasma AOPPs were first found to increase in CFA-induced hyperalgesia rats. • Intravenous injection of AOPPs in rats can induce hyperalgesia. • AOPPs sensitize Nox1/Nox4 to generate abundant ROS in DRG neurons. • AOPPs exacerbate hypersensitivity through Nox1/Nox4-ROS-TRPV1- CGRP/Ca2+ pathway.

Deep-sea isopod scavengers such as Bathynomus sp. are able to live in nutrient-poor environments, which is likely attributable to the presence of symbiotic microbes in their stomach. In this study we recovered two draft genomes of mycoplasmas, Bg1 and Bg2, from the metagenomes of the stomach contents and stomach sac of a Bathynomus sp. sample from the South China Sea (depth of 898 m). Phylogenetic trees revealed a considerable genetic distance to other mycoplasma species for Bg1 and Bg2. Compared with terrestrial symbiotic mycoplasmas, the Bg1 and Bg2 genomes were enriched with genes encoding phosphoenolpyruvate-dependent phosphotransferase systems (PTSs) and sodium-driven symporters responsible for the uptake of sugars, amino acids and other carbohydrates. The genome of mycoplasma Bg1 contained sialic acid lyase and transporter genes, potentially enabling the bacteria to attach to the stomach sac and obtain organic carbons from various cell walls. Both of the mycoplasma genomes contained multiple copies of genes related to proteolysis and oligosaccharide degradation, which may help the host survive in low-nutrient conditions. The discovery of the different types of mycoplasma bacteria in the stomach of this deep-sea isopod affords insights into symbiotic model of deep-sea animals and genomic plasticity of mycoplasma bacteria.

Aerophobetes (or CD12) is a recently defined bacterial phylum, of which the metabolic processes and ecological importance remain unclear. In the present study, we obtained the draft genome of an Aerophobetes bacterium TCS1 from saline sediment near the Thuwal cold seep in the Red Sea using a genome binning method. Analysis of 16S rRNA genes of TCS1 and close relatives revealed wide distribution of Aerophobetes in deep-sea sediments. Phylogenetic relationships showed affinity between Aerophobetes TCS1 and some thermophilic bacterial phyla. The genome of TCS1 (at least 1.27 Mbp) contains a full set of genes encoding core metabolic pathways, including glycolysis and pyruvate fermentation to produce acetyl-CoA and acetate. The identification of cross-membrane sugar transporter genes further indicates its potential ability to consume carbohydrates preserved in the sediment under the microbial mat. Aerophobetes bacterium TCS1 therefore probably carried out saccharolytic and fermentative metabolism. The genes responsible for autotrophic synthesis of acetyl-CoA via the Wood-Ljungdahl pathway were also found in the genome. Phylogenetic study of the essential genes for the Wood-Ljungdahl pathway implied relative independence of Aerophobetes bacterium from the known acetogens and methanogens. Compared with genomes of acetogenic bacteria, Aerophobetes bacterium TCS1 genome lacks the genes involved in nitrogen metabolism, sulfur metabolism, signal transduction and cell motility. The metabolic activities of TCS1 might depend on geochemical conditions such as supplies of CO2, hydrogen and sugars, and therefore the TCS1 might be a facultative bacterium in anaerobic saline sediments near cold seeps.

Autism is a neurodevelopmental disorder, and embryonic exposure to valproic acid (VPA) in rodents is the most frequently studied environmentally triggered autism models. Valproic acid can affect gene transcription as a histone deacetylase inhibitor, and thus may alter the expression of the most genes including reference genes. The aim of the current study is to validate suitable reference genes for quantitative real-time PCR (qPCR) quantification in prefrontal cortex and hippocampus of VPA rat models of autism. Female rats received a single intraperitoneal injection of 400 mg/kg sodium VPA at day 12.5 post-conception and controls were injected with saline. Male offspring were used to observe the expression of nine commonly used reference genes by qPCR, and the data were analyzed by four commonly used reference selection program including geNorm, BestKeeper, NormFinder and RefFinder. The results showed that VPA affected the expression of these commonly used reference genes in prefrontal cortex and hippocampus on postnatal 3, 5 weeks and 80 days, Gapdh and Actin, two very frequently used reference genes, were identified as the least stable genes in VPA group. Hprt1 was selected as the most stable gene, and Hmbs and Tbp were the optimum gene pair in prefrontal cortex and hippocampus across all VPA and controls. Problematically, the use of unstable reference genes results in calculation of different PGRN mRNA expression levels. The results suggest that selection of suitable references is critical for accurate mRNA quantification, and specifically in VPA induced rat models of autism.

β-thymosins, a family of highly conserved peptides, play a vital role in wound-healing, angiogenesis, antimicrobial process and antiviral immunity. Three novel β-thymosin-repeat proteins, named mjthm4, mjthm3 and mjthm2, were cloned from Marsupenaeus japonicus using expressed sequence tags (EST) from suppression subtractive hybridization. The mjthm4, mjthm3 and mjthm2 cDNAs possessed open reading frames that encoded 166, 128 and 90 amino acid residue polypeptides and contained four, three and two β-thymosin actin binding modules, respectively. Blast analysis demonstrated that mjthm4, mjthm3 and mjthm2 shared high homology with known invertebrate multi-repeat β-thymosins. These proteins are ubiquitously expressed in all of the examined tissues, and the transcriptional levels were highest in the intestine. Further investigation revealed that mjthm4, mjthm3 and mjthm2 were remarkably up-regulated 6 h after WSSV infection. Moreover, while mjthm4 transcriptional levels displayed no changes, mjthm3 and mjthm2 levels decreased in the virus-resistant shrimps. The results indicate that mjthm4, mjthm3 and mjthm2 are novel multi-repeat β-thymosin homologues, have a close relationship with WSSV infection, and might contribute to a better understanding of host defense and/or virus invasion interactions in shrimps.

The low biomass in environmental samples is a major challenge for microbial metagenomic studies. The amplification of a genomic DNA was frequently applied to meeting the minimum requirement of the DNA for a high-throughput next-generation-sequencing technology. Using a synthetic bacterial community, the amplification efficiency of the Multiple Annealing and Looping Based Amplification Cycles (MALBAC) kit that is originally developed to amplify the single-cell genomic DNA of mammalian organisms is examined. The DNA template of 10 pg in each reaction of the MALBAC amplification may generate enough DNA for Illumina sequencing. Using 10 pg and 100 pg templates for each reaction set, the MALBAC kit shows a stable and homogeneous amplification as indicated by the highly consistent coverage of the reads from the two amplified samples on the contigs assembled by the original unamplified sample. Although GenomePlex whole genome amplification kit allows one to generate enough DNA using 100 pg of template in each reaction, the minority of the mixed bacterial species is not linearly amplified. For both of the kits, the GC-rich regions of the genomic DNA are not efficiently amplified as suggested by the low coverage of the contigs with the high GC content. The high efficiency of the MALBAC kit is supported for the amplification of environmental microbial DNA samples, and the concerns on its application are also raised to bacterial species with the high GC content.

Background: Phosphatidylinositol binding clathrin assembly protein interacting mitotic regulator (PIMREG) is a protein associated with cell proliferation. Its aberrant expression was reported to be correlated with the development in multiple tumors. However, its role in cholangiocarcinoma (CAA) has not yet been evaluated in detail. Methods: Data were acquired from the public TCGA database for evaluating the expression pattern of PIMREG and assessing its clinical relevance as well as its correlation with overall survival. RBE and HUH28 cell lines were selected to perform loss- and gain-of-function of PIMREG assays respectively. Quantitative real-time PCR (RT-qPCR) and western blot analyses were used to measure the mRNA and protein levels of PIMREG. Cell Counting Kit-8, colony formation tests, and Transwell assays served to measure the effect of PIMREG on the proliferative, invasive and migratory capacities of CAA cells, appropriately. Gene set enrichment analysis (GSEA) was conducted to identify PIMREG associated gene set, which was further confirmed by western blot. Results: PIMREG was found to be highly expressed in CAA tissues and cell lines according to the public dataset and RT-qPCR analysis, and negatively related to the prognosis of patients with CAA. Moreover, knockdown of PIMREG suppressed and overexpression of PIMREG promoted the proliferation, invasion and migration of CAA cells. Furthermore, GSEA revealed that high PIMREG expression was positively associated with cell cycle signaling. And the next western blot analysis demonstrated that silencing PIMREG resulted in a reduction on the levels of p-CDK1, CCNE1, and CCNB1, whereas PIMREG overexpression led to an opposite result. Conclusion: The results suggested that PIMREG facilitates the growth, invasion and migration of CAA cells partly by regulating the cell cycle relative biomarkers, revealing that PIMREG may be a crucial molecule in the progression of CAA.

The low temperature and elevated hydrostatic pressure in hadal trenches at water depths below 6000 m render sample collection difficult. Here, in situ hadal water microbial samples were collected from the Mariana Trench and analysed. The hadal microbial communities at different depths were revealed to be consistent and were dominated by heterotrophic Marinimicrobia. Thirty high-quality metagenome-assembled genomes (MAGs) were retrieved to represent the major hadal microbes affiliated with 12 prokaryotic phyla. Most of the MAGs were newly reported and probably derived from novel hadal inhabitants as exemplified by a potentially new candidate archaeal phylum in the DPANN superphylum. Metabolic reconstruction indicated that a great number of the MAGs participated in nitrogen and sulfur cycling, in which the nitrification process was driven sequentially by Thaumarchaeota and Nitrospirae and sulfur oxidization by Rhodospirillales in the Alphaproteobacteria class. Moreover, several groups of hadal microbes were revealed to be potential carbon monoxide oxidizers. Metatranscriptomic result highlighted the contribution of Chloroflexi in degrading recalcitrant dissolved organic matter and Marinimicrobia in extracellular protein decomposition. The present work provides an in-depth view on the hadal microbial communities regarding their endemism and element cycles.

In deep-sea sediment, the microbes present in anhydrite crystals after mild hydrothermal activities are markers of the past environment. In this study, this hypothesis was tested by analyzing the metagenome of an anhydrite crystal sample from a hydrothermal and hypersaline sediment core sampled from the Atlantis II Deep in the Red Sea. The 16S/18S rRNA genes in the metagenome were assigned to Bacteria, Archaea, Fungi and even invertebrate species. The dominant species in the crystals was an alkane-degrading Alcanivorax bacterium, which was not detected in the adjacent sediment layer. Using a genome-binning method, a draft genome of the Alcanivorax bacterium was separated from the metagenome. Phylogenetic and genomic analyses revealed that this species was a close relative of Alcanivorax borkumensis Sk2. The draft genome contained all the functional genes for alkane utilization and the reduction of nitrogen oxides. Fluorescence microscopy using 16S rRNA and marker gene probes revealed intact cells of the Alcanivorax bacterium in the crystals. Moreover, the metagenomes of the anhydrites and control sediment contained aromatic degradation pathways, which were mostly derived from Ochrobactrum sp. The estimated age of the anhydrite layer was between 750–770 years, which might span the event of hydrothermal eruption into the benthic floor. Altogether, these results support the presence of an oxic, oil-spilling benthic environment in the Atlantis II basin of the Red Sea in approximately the 14th century. The original microbial inhabitants underwent a dramatic selection process via drastic environmental changes following the formation of an overlying anoxic brine pool in the basin due to hydrothermal activities.

Various lineages of ammonia-oxidizing archaea (AOA) are present in deep waters, but the mechanisms that determine ecotype formation are obscure. We studied 18 high-quality genomes of the marine group I AOA lineages (alpha, gamma and delta) from the Mariana and Ogasawara trenches. The genomes of alpha AOA resembled each other, while those of gamma and delta lineages were more divergent and had even undergone insertion of some phage genes. The instability of the gamma and delta AOA genomes could be partially due to the loss of DNA polymerase B (polB) and methyladenine DNA glycosylase (tag) genes responsible for the repair of point mutations. The alpha AOA genomes harbour genes encoding a thrombospondin-like outer membrane structure that probably serves as a barrier to gene flow. Moreover, the gamma and alpha AOA lineages rely on vitamin B12 -independent MetE and B12 -dependent MetH, respectively, for methionine synthesis. The delta AOA genome contains genes involved in uptake of sugar and peptide perhaps for heterotrophic lifestyle. Our study provides insights into co-occurrence of cladogenesis and anagenesis in the formation of AOA ecotypes that perform differently in nitrogen and carbon cycling in dark oceans.

The complete mitochondrial genome DNA sequence of Istigobius campbelli was 16,527 bp in length. It consists of 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and 1 control region. 28 of the 37 genes were encoded on the heavy strand, and 9 of them were encoded on the light strand. The overall base composition of the genome is 27.84% A, 25.81% T, 29.68% C, and 16.67% G. The phylogenetic tree suggested I. campbelli was genetically closest to Acentrogobius pflaumii and Oxyurichthys formosanus. This study could provide valuable information for further studies on I. campbelli.

In this study, the complete mitochondrial genome of Dysomma anguillare was obtained, which was 16,898 bp in length. It consists of 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA ge...

3-Hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) reductases (HMGRs), which catalyze the conversion of HMG-CoA to mevalonate, may have an important role in the synthesis of methyl farnesoate (MF). In this study, we obtained two HMGR cDNA sequences termed Sp-HMGR1 (membrane-bound form) and Sp-HMGR2 (soluble form), which encode 967 and 654 amino acids, respectively. The two cDNAs possess entirely identical sequences except that Sp-HMGR1 is 1,382 bp, which encodes a sterol-sensed domain (SSD; a membrane-bound domain) and was first found in crustacean HMGR, larger than Sp-HMGR2. Thus, it was deduced that these cDNAs might be derived from a single genomic DNA sequence. Sp-HMGRs have the typical features of the HMGR class of proteins. However, residue 844 in Sp-HMGR1, which is usually occupied by a Ser residue in other species, has an unusual Ala substitution. This Ser is thought to be involved in enzyme activity regulation by reversible phosphorylation. A putative "PEST" sequence that, until now, has only been found in crustacean species was also identified in the C-terminus of both transcripts, and a sterol-sensing domain, which was first found in crustacean species, was identified in Sp-HMGR1; these findings suggest that Sp-HMGR might function in some special regulatory mechanism. Furthermore, the quantitative real-time polymerase chain reaction results showed that the two transcripts have different expression patterns; Sp-HMGR2 was mainly expressed in the mandibular organ (MO) of adult crabs, whereas Sp-HMGR1 was mainly expressed in other tissues and fertilized eggs up until the fourth juvenile crab stage. The fluctuating gene expression seemed to suggest a relationship between Sp-HMGRs and the development of the crab, especially during the larval stage. Besides, the fluctuation of Sp-HMGR1 in ovary, brain, and thoracic ganglia during the ovary development seemed to have some correlation with the nutrition accumulation of ovaries, whether the SSD domain evolved in this process deserve further investigation. Moreover, it remains unclear whether the significant variation in ovary, brain, and thoracic ganglia during ovary development suggests that other tissues in addition to the MO could synthesize MF.

Pro-inflammatory cytokine-induced chondrocyte apoptosis is a primary cause of cartilage destruction in the progression of rheumatoid arthritis (RA). Advanced oxidation protein products (AOPPs), a novel pro-inflammatory mediator, have been confirmed to accumulate in patients with RA. However, the effect of AOPPs accumulation on chondrocyte apoptosis and the associated cellular mechanisms remains unclear. The present study demonstrated that the plasma formation of AOPPs was enhanced in RA rats compared with normal. Then, chondrocyte were treated with AOPPs-modified rat serum albumin (AOPPs-RSA) in vitro. Exposure of chondrocyte to AOPPs activated nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and increased expression of NADPH oxidase subunits, which was mediated by receptor for advanced glycation end products (RAGE), but not scavenger receptor CD36. Moreover, AOPPs challenge triggered NADPH oxidase-dependent ROS generation which induced mitochondrial dysfunction and endoplasmic reticulum stress resulted in activation of caspase family that eventually lead to apoptosis. Lastly, blockade of RAGE, instead of CD36, largely attenuated these signals. Our study demonstrated first time that AOPPs induce chondrocyte apoptosis via RAGE-mediated and redox-dependent intrinsic apoptosis pathway in vitro. These data implicates that AOPPs may represent a novel pathogenic factor that contributes to RA progression. Targeting AOPPs-triggered cellular mechanisms might emerge as a promising therapeutic option for patients with RA.

Rhabdomyosarcoma (RMS) is the most common type of soft-tissue sarcoma in children. Immunotherapy has been proposed as a treatment for this deadly tumor. In the present study, the cytotoxicity of ex vivo expanded γδ T cells on RMS cell lines was evaluated and the molecular interactions involved were investigated. γδ T cells were expanded in vitro using peripheral blood mononuclear cells from 5 healthy donors and were stimulated with zoledronic acid (Zol) and interleukin 2. RMS cell lines RD and A-673 were used as target cells. The cytotoxicity of the γδ T cells against RMS was assessed in vitro and in vivo. γδ T cells were cytotoxic to RMS cells. Importantly, Zol markedly increased their cytotoxic potential. RMS cells treated with Zol-stimulated γδ T cells to produce interferon γ. γδ T cell-mediated cytotoxicity was primarily through the T cell receptor-dependent signaling pathway in blocking studies. Transfer of γδ T cells together with Zol into nude mice induced the regression of RD tumor xenotransplants. The results of the present study provide the rationale for the clinical evaluation of γδ T cells in RMS.

The complete sequence of the mitochondrial genome of Podagrion sp. (Hymenoptera: Torymidae) is described. The mitogenome was 15,845 bp in size, and contained typical sets of mitochondrial genes. The base composition of the Podagrion sp. mitogenome was also biased toward A + T bases (81.8%). The mitochondrial genome of Podagrion sp. has a weak AT skew (0.07) and a strong GC skew (-0.26). Podagrion sp. exhibits a novel rearrangement compared with the ancestral order, including six protein-coding genes (nad3, cox3, atp6, atp8, cox2 and cox1), which have inverted to the minor strand from the major strand. The A + T-rich region of Podagrion sp., which is located between trnN and trnI, have five tandem repeats. The apomorphic rearrangements, including the conserved block "cox3-atp6-atp8-cox2-cox1-nad5-nad4-nad4l-nad6-cob" and the special locations of trnV and trnA, were mapped onto the phylogeny of Proctotrupomorpha.

The South China Sea is a special reservoir of sponges of which prokaryotic communities are less studied. Here, a new record of the sponge Dactylospongia elegans is reported near the coast of Jinqing Island in the South China Sea, and its prokaryotic community is comprehensively investigated. Sponge specimens displayed lower microbial diversity compared with surrounding seawater. At the phylum level, prokaryotic communities were consistently dominated by Proteobacteria, followed by Cyanobacteria, Chloroflexi, Acidobacteria, Actinobacteria, Gemmatimonadetes, Thaumarchaeota, and Poribacteria. Operational taxonomic unit (OTU) analysis alternatively showed that the most abundant symbiont was the sponge-specific cyanobacterial species “Candidatus Synechococcus spongiarum,” followed by OTUs belonging to the unidentified Chloroflexi and Acidobacteria. Phylogenetic tree based on 16S-23S internal transcribed spacer regions indicated that the dominated cyanobacterial OTU represented a new clade of “Ca. Synechococcus spongiarum.” More reliable metagenomic data further revealed that poribacterial symbionts were highly abundant and only secondary to the cyanobacterial symbiont. One draft genome for each of the Cyanobacteria, Chloroflexi and Acidobacteria and three poribacterial genomes were extracted from the metagenomes. Among them, genomes affiliated with the Chloroflexi and Acidobacteria were reported for the first time in sponge symbionts. Eukaryotic-like domains were found in all the binned genomes, indicating their potential symbiotic roles with the sponge host. The high quality of the six recovered genomes of sponge symbionts from the sponge D. elegans makes it possible to understand their symbiotic roles and interactions with the sponge host as well as among one another.

Prophenoloxidase (PPO) is an essential enzyme in insect innate immunity because of its role in humoral defense. In this study, we have cloned a full-length cDNA of Antheraea pernyi prophenoloxidase (ApPPO) with an open-reading frame encoding 683 amino acids, and the deduced amino acid sequence of ApPPO exhibited a high similarity with those of lepidoptera. The expression of ApPPO was inducible so that the mRNA level was significantly upregulated in the microbial challenged tissues, including fat body, hemocytes, and midgut. To better investigate the enzymatic and immunological properties of ApPPO, recombinant ApPPO (rApPPO) was produced in Escherichia coli. Several functional verification experiments were performed after studying the enzymatic properties. It was found that rApPPO could be stimulated by the microbial challenged larvae hemolymph and then killed bacteria in the radial diffusion assay. Furthermore, rApPPO also induced the transcription of cecropins after injected into the larvae 24 h later.

This study evaluated the potential adverse effects of butenolide, a promising antifouling compound, using the marine medaka (Oryzias melastigma), a model fish for marine ecotoxicology. The active ingredient used in the commercial antifoulant SeaNine 211, 4,5-dichloro-2-n-octyl-4-isothiazolin-3-one (DCOIT) was employed as the positive control. Adult marine medaka (4-month-old) were exposed to various concentrations of butenolide or DCOIT for 28 days and then depurated in clean seawater for 14 days (recovery). A suite of sensitive biomarkers, including hepatic oxidative stress, neuronal signal transmission, endocrine disruption, and reproductive function, was used to measure significant biological effects induced by the chemicals. Compared to DCOIT, chronic exposure to butenolide induced a lower extent of oxidative stress in the liver of male and female medaka. Furthermore, butenolide-exposed fish could recover faster from oxidative stress than fish exposed to DCOIT. Regarding neurotransmission, DCOIT significantly inhibited acetylcholinesterase (AChE) activity in the brain of both male and female medaka, whereas this was not significant for butenolide. In addition, plasma estradiol (E2) level was elevated and testosterone (T) level was decreased in male medaka exposed to DCOIT. This greatly imbalanced sex hormones ratio (E2/T) in exposed males, indicating that DCOIT is a potent endocrine disruptive chemical. In contrast, butenolide induced only moderate effects on sex hormone levels in exposed males, which could be gradually recovered during depuration. Moreover, the endocrine disruptive effect induced by butenolide did not affect normal development of offspring. In contrast, DCOIT-exposed fish exhibited a decrease of egg production and impaired reproductive success. Overall, the above findings demonstrated that chronic exposure to butenolide induced transient, reversible biological effect on marine medaka, while DCOIT could impair reproductive success of fish, as evident by clear alterations of the E2/T ratio. The relatively low toxicity of butenolide on marine biota highlights its promising application in the antifouling industry. The present findings also emphasize gender difference in fish susceptibility to chemical treatment (male > female), which is an important consideration for ecological risk assessment.

Microbial communities in hadal sediments are least explored in hadal zone (>6,000 m), especially in the Challenger Deep with high pressure (∼110 M pa at the bottom). In this study, we investigated the microbial communities in the sediments of the slope and trench-axis bottom of the Challenger Deep in the Mariana Trench. Classification of the reads of the 16S rRNA gene amplicons showed vertical distribution of prokaryotic microbial inhabitants from the surface to up to 60 centimeter below surface floor (cmbsf). The most dominant phyla were Proteobacteria, Chloroflexi, Actinobacteria, Planctomycetes and candidate phyla Patescibacteria and Marinimicrobia. Distinct dominant groups in the microbial communities were observed in trench-axis sediment (water depth >8,600 m), compared to the slopes of the Challenger Deep. A sampling site at the northern slope was enriched with archaea from mesophilic Euryarchaeota Marine Group II (MGII) as a biomarker of specific geochemical setting. Among archaeal community, Thaumarchaeota represented byNitrosopumiluswere dominant in the upper layers and diminished drastically in the deeper layers. “Ca. Woesearchaeota”, however, became the dominant group in the deeper layers. Overall, our study provides a better understanding on the pattern of the microbial communities in the deepest hadal sediments on Earth, and highlights the extraordinary diversity still waiting to be discovered.