Search

Secondary brain injury after aneurysmal subarachnoid hemorrhage (SAH-SBI) is a significant contributor to poor outcomes in patients after rupture of an intracranial aneurysm. The lack of diagnostic biomarkers and novel drug targets represent an unmet need. Prior experimental evidence has suggested cell-free hemoglobin in the cerebrospinal fluid (CSF-Hb) as a pathophysiological driver of SAH-SBI. The aim of this study was to investigate the clinical and pathophysiological association between CSF-Hb and SAH-SBI. We prospectively enrolled 47 consecutive patients and collected daily CSF samples within 14 days after aneurysm rupture. There was very strong evidence for a positive association between CSF-Hb and SAH-SBI. The diagnostic accuracy of CSF-Hb for SAH-SBI markedly exceeded that of established methods (area under the curve: 0.89 [0.85-0.92]). Temporal LC-MS/MS CSF proteomics demonstrated that erythrolysis accompanied by an adaptive macrophage response are the two dominant biological processes occurring in the CSF space after aneurysm rupture. To further investigate the pathophysiology between CSF-Hb and SAH-SBI, we explored the vasoconstrictive and lipid peroxidation activities of Hb ex-vivo. These experiments revealed critical inflection points overlapping CSF-Hb concentration thresholds in patients with SAH-SBI. Selective Hb depletion and in-solution neutralization by the Hb-scavenger haptoglobin or the heme-scavenger hemopexin efficiently attenuated the vasoconstrictive and lipid peroxidation activities of CSF-Hb in patient CSF. Collectively, the clinical association between high CSF-Hb levels and SAH-SBI, the underlying pathophysiological rationale, and the favorable effects of haptoglobin and hemopexin in ex-vivo experiments position CSF-Hb as a highly attractive biomarker and potential drug target.

The study of copper oxide nanoparticles green synthesis was carried out using facile, nontoxic, and low-cost approach using extract of Cedrus deodara as a safe and compatible reducing agent. Scanning electron microscope (SEM), X-ray diffraction (XRD), UV–visible spectroscopy (UV–Vis), Transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FTIR) machines were used to characterize the nanoparticles formed. The FTIR showed that different phytochemical present in cedrus extract are accountable for the synthesis of CuO (copper oxide) nanoparticles. The XRD result depicted that CuO nanoparticles were monoclinic in nature. The SEM and TEM analysis also confirmed that formed nanoparticles had spherical morphology with uniform symmetry. The CuO nanoparticles were tested to inhibit the growth human pathogenic strains with the help of disc diffusion method against E. coli and S. aureus. It is noted that the formed CuO nanoparticles showed efficient antibacterial activity especially against E. coli with maximum inhibition zone of 29 mm.

Funding: European and Developing Countries Clinical Trials Partnership (EDCTP) under Streamlining Health Research Ethics Review and Regulatory Framework in Tanzania (SMERT) project. Medicines with a stereogenic center (asymmetric carbon) are mainly present as racemates with a mixture of equal amounts of enantiomers. One enantiomer may be active while the other inactive, alternatively one may produce side-effects and even toxicity. However, there is lack of information on the chirality status (either racemates, single active enantiomer or achiral) of medicines circulated on the market particularly in African countries. We established the chirality status of registered medicines in Tanzania by conducting a retrospective cross-sectional study. Registration data for the past 15 years from 2003 to 2018 were extracted from TMDA-IMIS database to Microsoft excel for review and analysis. A total of 3,573 human medicines had valid registration. Out of which 2,150 (60%) were chiral and 1,423 (40%) achiral. Out of the chiral medicines, 1,591 (74%) and 559 (26%) were racemates and single active enantiomers, respectively. The proportion of racemates within chiral medicines was considerably higher than single enantiomer medicines. The use of racemates may cause harm to the public and may contribute to antimicrobial resistance due to potential existence of inactive and toxic enantiomers. In order to protect public health, regulatory bodies need to strengthen control of chiral medicines by conducting analysis of enantiomeric impurity. Publisher PDF

S100A8 and S100A9 are members of the Alarmin family; these proteins are abundantly expressed in neutrophils, form a heterodimer complex, and are secreted in plasma on pathogen infection or acute inflammatory diseases. Recently, both proteins were identified as novel biomarkers of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and were shown to play key roles in inducing an aggressive inflammatory response by mediating the release of large amounts of pro-inflammatory cytokines, called the “cytokine storm.” Although co-infection with SARS-CoV-2 in people living with HIV-1 may result in an immunocompromised status, the role of the S100A8/A9 complex in HIV-1 replication in primary T cells and macrophages is still unclear. Here, we evaluated the roles of the proteins in HIV replication to elucidate their functions. We found that the complex had no impact on virus replication in both cell types; however, the subunits of S100A8 and S100A9 inhibit HIV in macrophages. These findings provide important insights into the regulation of HIV viral loads during SARS-CoV-2 co-infection.

Increasing occurrence of harmful algal blooms across the land–water interface poses significant risks to coastal ecosystem structure and human health. Defining significant drivers and their interactive impacts on blooms allows for more effective analysis and identification of specific conditions supporting phytoplankton growth. A novel iterative Random Forests (iRF) machine-learning model was developed and applied to two example cases along the California coast to identify key stable interactions: (1) phytoplankton abundance in response to various drivers due to coastal conditions and land-sea nutrient fluxes, (2) microbial community structure during algal blooms. In Example 1, watershed derived nutrients were identified as the least significant interacting variable associated with Monterey Bay phytoplankton abundance. In Example 2, through iRF analysis of field-based 16S OTU bacterial community and algae datasets, we independently found stable interactions of prokaryote abundance patterns associated with phytoplankton abundance that have been previously identified in laboratory-based studies. Our study represents the first iRF application to marine algal blooms that helps to identify ocean, microbial, and terrestrial conditions that are considered dominant causal factors on bloom dynamics.

Ixora coccinea flower was utilized as source of natural dye to fabricate dye sensitized solar cells (DSSCs) using TiO2 electrodes. The electrodes were fabricated using Doctor Blade’s method and annealed at 600 °C, 450 °C, and 300 °C. The structural, morphological, and elemental composition analyses of the TiO2 electrodes were carried out. The dye extract and the TiO2 electrodes had low absorbance within the visible region of electromagnetic spectrum. The TiO2 photoanode annealed at 300 °C, 450 °C, and 600 °C had crystallite sizes of 26.52, 24.36, and 21.85 nm and average grain sizes of 85 nm, 169 nm, and 184 nm in that order. The DSSC fabricated based on the Ixora coccinea flower dye extract revealed conversion efficiency of 0.73%, 1.18%, and 1.01% for the TiO2 electrodes annealed at 300 °C, 450 °C, and 600 °C respectively.

Background: Motor and cognitive decline as part of the normal aging process is linked to alterations in synaptic plasticity and reduction of adult neurogenesis in the dorsal striatum. Neuroinflammation, particularly in the form of microglial activation, is suggested to contribute to these age-associated changes. Objective and Methods: To explore the molecular basis of alterations in striatal function during aging we analyzed RNA-Seq data for 117 postmortem human dorsal caudate samples and 97 putamen samples acquired through GTEx. Results: Increased expression of neuroinflammatory transcripts including TREM2, MHC II molecules HLA-DMB, HLA-DQA2, HLA-DPA1, HLA-DPB1, HLA-DMA and HLA-DRA, complement genes C1QA, C1QB, CIQC and C3AR1, and MHCI molecules HLA-B and HLA-F was identified. We also identified down-regulation of transcripts involved in neurogenesis, synaptogenesis, and synaptic pruning, including DCX, CX3CL1, and CD200, and the canonical WNTs WNT7A, WNT7B, and WNT8A. The canonical WNT signaling pathway has previously been shown to mediate adult neurogenesis and synapse formation and growth. Recent findings also highlight the link between WNT/β-catenin signaling and inflammation pathways. Conclusions: These findings suggest that age-dependent attenuation of canonical WNT signaling plays a pivotal role in regulating striatal plasticity during aging. Dysregulation of WNT/β-catenin signaling via astrocyte–microglial interactions is suggested to be a novel mechanism that drives the decline of striatal neurogenesis and altered synaptic connectivity and plasticity, leading to a subsequent decrease in motor and cognitive performance with age. These findings may aid in the development of therapies targeting WNT/β-catenin signaling to combat cognitive and motor impairments associated with aging.

Traditional medicines, derived from plants, could present alternative treatment strategy for cancer therapy. One such plant is Momordica charantia (MC) which possesses anti-carcinogenic properties. This study investigated the anticancer effect of an ethanol extract of MC fruit, Kuguacin-J (K-J), an isolated compound from the leaves of MC and cisplatin, either alone or combination on healthy MCF-10A mammary cells and compared with breast cancer MCF-7 and MDAMB-231 cell lines. Cell viability was tested using 8 μg/mL and 80 μg/mL doses of MC extract, K-J and cisplatin individually or combined for 24 and 48 hours. Caspase-3- activity was measured in MCF-7 and MDA-MB-231 cells using established methods. The results revealed that MC extract and K-J had no effect on healthy MCF-10A cell viability as compared to cisplatin which induced dose and time-dependent cell death. Similarly, treatment of MCF-7 cells with cisplatin induced cell death at high concentration at both the time points, while MC extract and K-J only induce MCF-7 cell death at high dose after 48 hours only. During combination therapy, both doses of cisplatin enhanced MCF-7 cell death when combined with MC extract or K-J after 24 and 48 hours. In MDAMB-231 cells, the three drugs, either alone or combined, evoked significant cell death at both the doses and time points. All three drugs, at high dose, elicited significant increase in caspase-3- activity in MCF-7 and MDA-MB-231 cells as compared to untreated cells. The results revealed that either MC extract or K-J alone or combined with cisplatin, can elicit significant increase in cell death and caspase–3-activity in MCF-7 and MDA-MB-231cells as compared to untreated cells.

Traditional medicines, derived from plants, could present alternative treatment strategy for cancer therapy. One such plant is Momordica charantia (MC) which possesses anti-carcinogenic properties. This study investigated the anticancer effect of an ethanol extract of MC fruit, Kuguacin-J (K-J), an isolated compound from the leaves of MC and cisplatin, either alone or combination on healthy MCF-10A mammary cells and compared with breast cancer MCF-7 and MDAMB-231 cell lines. Cell viability was tested using 8 μg/mL and 80 μg/mL doses of MC extract, K-J and cisplatin individually or combined for 24 and 48 hours. Caspase-3- activity was measured in MCF-7 and MDA-MB-231 cells using established methods. The results revealed that MC extract and K-J had no effect on healthy MCF-10A cell viability as compared to cisplatin which induced dose and time-dependent cell death. Similarly, treatment of MCF-7 cells with cisplatin induced cell death at high concentration at both the time points, while MC extract and K-J only induce MCF-7 cell death at high dose after 48 hours only. During combination therapy, both doses of cisplatin enhanced MCF-7 cell death when combined with MC extract or K-J after 24 and 48 hours. In MDAMB-231 cells, the three drugs, either alone or combined, evoked significant cell death at both the doses and time points. All three drugs, at high dose, elicited significant increase in caspase-3- activity in MCF-7 and MDA-MB-231 cells as compared to untreated cells. The results revealed that either MC extract or K-J alone or combined with cisplatin, can elicit significant increase in cell death and caspase–3-activity in MCF-7 and MDA-MB-231cells as compared to untreated cells.

Niche-related processes (e.g., density or niche-breadth compensation and competition) are fundamental to a broad understanding of community ecology and ecosystem functioning. Most evidences of competition are from controlled indoor trials with few species, and it remains a challenge to estimate competition among multiple species in the field. Here, we analyze stable isotopes and distributional data from 51 fish taxa in six locations in the southwestern Atlantic to predict intraspecific trophic pressure (ITP) and the potential competitive strength among species in a trophic-based framework. We used two proxies built upon 2-dimensional isotopic space (δ13C vs. δ15N), its predicted overlap, and fish density to calculate winner and loser taxa in potential paired interspecific competitive interactions. The intraspecific proxy indicated that cryptobenthic fishes are under high among-individual trophic pressure (high densities and small niche sizes). Also, cryptobenthic behavior together with feeding specialization and extremely small-sizes were the most important traits related to low success in interspecific simulations. Although cryptobenthic fishes face strong competitive pressures, there are some known inherent trade-offs to cryptobenthic life such as trophic and habitat use specializations. These seem to compensate and ensure coexistence among cryptobenthic fishes and non-cryptobenthic species. Habitat loss/degradation via urbanization, invasive species and climate-change-driven sea-level rise can reduce the suitability of habitat and increase competition on cryptobenthic species, especially in shallow reefs and intertidal shores.

Field studies were conducted on organic soils in Belle Glade, FL, in 2016 to 2017 to evaluate sugarcane tolerance and fall panicum control with topramezone applied alone or in combination with triazine herbicides (atrazine, metribuzin, ametryn). Treatments included topramezone (25 and 50 g ai ha−1) applied alone or in combination with atrazine (2,240 g ai ha−1), metribuzin (2,240 g ai ha−1), and ametryn (440 g ha−1) on four plant cane varieties to evaluate tolerance, and on second ratoon fields to determine efficacy on fall panicum control. Topramezone applied alone had no effect on sugarcane chlorophyll fluorescence (i.e., the ratio of variable fluorescence to maximum fluorescence), total chlorophyll, and carotenoid 7 to 28 d after treatment (DAT), suggesting sugarcane tolerance. Significant reduction of these parameters occured 7 to 14 DAT when topramezone (50 g ai ha−1) was applied with ametryn or metribuzin; however, reductions were not detected thereafter, indicating recovery. Sugarcane yield was not affected by topramezone applied alone or in combination with the triazine herbicides. Topramezone (50 g ai ha−1) plus metribuzin resulted in acceptable control of fall panicum (84%) with limited to no regrowth of meristematic tissue at sugarcane canopy closure, equivalent to 56 to 70 DAT. These results indicate that when sequential applications of topramezone, applied alone or in combination with these triazine herbicides, are required for efficacious weed control, topramezone applications alone can be made after 7 d, whereas the combinations can be made after 14 or 21 d, depending on sugarcane sensitivity.

Impacts on growth rates from exposure to ammonium (NH4+) and nitrate (NO3-), at non-limiting concentrations, in combination with irradiances varying from 25 to 600 µmol photons m-2 s-1 were investigated in the pelagic diatom Thalassiosira weisflogii and the benthic diatom Entomoneis paludosa recently isolated from Suisun Bay in northern San Francisco Bay. Growth rates were higher in T. weisflogii (0.76 ± 0.3 d-1) compared with E. paludosa (0.58 ± 0.2 d-1) across all irradiances and nitrogen (N) treatments. Differences in growth rates with N source were regulated by irradiance in both diatoms and were greatest at the intermediate irradiance due to saturation of rates at 85 µmol photons m-2 s-1 when growing on NH4+ and at 200 µmol photons m-2 s-1 when growing on NO3-. Notable physiological differences between these 2 diatoms included a larger range in the quantum yield of photosystem II (Fv/Fm) and in chlorophyll a per cell as a function of irradiance in T. weissflogii compared with E. paludosa. In addition, a negative interaction of high NH4+ concentration (1000 µmol l-1) and irradiance (≥200 µmol photons m-2 s-1) was observed on growth rates in E. paludosa that was not evident in T. weissflogii. Differences in physiological parameters of these diatoms are discussed in relation to their distributions and frequency of occurrence in Suisun Bay.

The marine biotoxin domoic acid (DA) is an analog of the neurotransmitter glutamate that exerts potent excitatory activity in the brain, heart, and other tissues. Produced by the diatomPseudo-nitzschiaspp., DA accumulates in marine invertebrates, fish, and sediment. Southern sea otters (Enhydra lutris nereis) feed primarily on invertebrates, including crabs and bivalves, that concentrate and slowly depurate DA. Due to their high prey consumption (25% of body weight/day), sea otters are commonly exposed to DA. A total of 823 necropsied southern sea otters were examined to complete this study; first we assessed 560 subadult, adult, and aged adult southern sea otters sampled from 1998 through 2012 for DA-associated pathology, focusing mainly on the central nervous system (CNS) and cardiovascular system. We applied what was learned to an additional cohort of necropsied sea otters of all demographics (including fetuses, pups, juveniles, and otters examined after 2012:n= 263 additional animals). Key findings derived from our initial efforts were consistently observed in this more demographically diverse cohort. Finally, we assessed the chronicity of DA-associated pathology in the CNS and heart independently for 54 adult and aged adult sea otters. Our goals were to compare the temporal consistency of DA-associated CNS and cardiovascular lesions and determine whether multiple episodes of DA toxicosis could be detected on histopathology. Sea otters with acute, fatal DA toxicosis typically presented with neurological signs and severe, diffuse congestion and multifocal microscopic hemorrhages (microhemorrhages) in the brain, spinal cord, cardiovascular system, and eyes. The congestion and microhemorrhages were associated with detection of high concentrations of DA in postmortem urine or gastrointestinal content and preceded histological detection of cellular necrosis or apoptosis. Cases of chronic DA toxicosis often presented with cardiovascular pathology that was more severe than the CNS pathology; however, the lesions at both sites were relatively quiescent, reflecting previous damage. Sea otters with fatal subacute DA toxicosis exhibited concurrent CNS and cardiovascular pathology that was characterized by progressive lesion expansion and host response to DA-associated tissue damage. Acute, subacute, and chronic cases had the same lesion distribution in the CNS and heart. CNS pathology was common in the hippocampus, olfactory, entorhinal and parahippocampal cortex, periventricular neuropil, and ventricles. The circumventricular organs were identified as important DA targets; microscopic examination of the pituitary gland, area postrema, other circumventricular organs, and both eyes facilitated confirmation of acute DA toxicosis in sea otters. DA-associated histopathology was also common in cardiomyocytes and coronary arterioles, especially in the left ventricular free wall, papillary muscles, cardiac apex, and atrial free walls. Progressive cardiomyocyte loss and arteriosclerosis occurred in the same areas, suggesting a common underlying mechanism. The temporal stage of DA-associated CNS pathology matched the DA-associated cardiac pathology in 87% (n= 47/54) of cases assessed for chronicity, suggesting that the same underlying process (e.g., DA toxicosis) was the cause of these lesions. This temporally matched pattern is also indicative of a single episode of DA toxicosis. The other 13% of examined otters (n= 7/54) exhibited overlapping acute, subacute, or chronic DA pathology in the CNS and heart, suggestive of recurrent DA toxicosis. This is the first rigorous case definition to facilitate diagnosis of DA toxicosis in sea otters. Diagnosing this common but often occult condition is important for improving clinical care and assessing population-level impacts of DA exposure in this federally listed threatened subspecies. Because the most likely source of toxin is through prey consumption, and because humans, sea otters, and other animals consume the same marine foods, our efforts to characterize health effects of DA exposure in southern sea otters can provide strong collateral benefits.

Given the effects of harmful algal blooms (HABs) on human and wildlife health, understanding how domoic acid (DA) is accumulated and transferred through food webs is critical for recognizing the most affected marine communities and predicting ecosystem effects. This study combines stable isotopes of carbon (δ13C) and nitrogen (δ15N) from bulk muscle tissue with DA measurements from viscera to identify the foraging strategies of important DA vectors and predators in Monterey Bay, CA. Tissue samples were collected from 27 species across three habitats in the summer of 2018 and 2019 (time periods without prominent HABs). Our results highlight an inshore-offshore variation in krill δ13C values and DA concentrations ([DA]; ppm) in anchovies indicating differences in coastal productivity and DA accumulation. The narrow overlapping isotopic niches between anchovies and sardines suggest similar diets and trophic positions, but striking differences in [DA] indicate a degree of specialization, thus, resource partitioning. In contrast, krill, market squid, and juvenile rockfish accumulated minimal DA and had comparatively broad isotopic niches, suggesting a lower capacity to serve as vectors because of potential differences in diet or feeding in isotopically distinct locations. Low [DA] in the liver of stranded sea lions and their generalist foraging tendencies limits our ability to use them as sentinels for DA outbreaks in a specific geographic area. Collectively, our results show that DA was produced a few kilometers from the coastline, and anchovies were the most powerful DA vector in coastal-pelagic zones (their DA loads exceeded the 20 ppm FDA regulatory limits for human consumption), while mussels did not contain detectable DA and only reflect in situ DA, δ13C, and δ15N values. Our study demonstrates the efficacy of combining multiple biogeochemical tracers to improve HAB monitoring efforts and identify the main routes of DA transfer across habitats and trophic levels.

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

Climate change is responsible for increased frequency, intensity, and duration of extreme events, such as marine heatwaves (MHWs). Within eastern boundary current systems, MHWs have profound impacts on temperature-nutrient dynamics that drive primary productivity. Bull kelp (Nereocystis luetkeana) forests, a vital nearshore habitat, experienced unprecedented losses along 350 km of coastline in northern California beginning in 2014 and continuing through 2019. These losses have had devastating consequences to northern California communities, economies, and fisheries. Using a suite of in situ and satellite-derived data, we demonstrate that the abrupt ecosystem shift initiated by a multi-year MHW was preceded by declines in keystone predator population densities. We show strong evidence that northern California kelp forests, while temporally dynamic, were historically resilient to fluctuating environmental conditions, even in the absence of key top predators, but that a series of coupled environmental and biological shifts between 2014 and 2016 resulted in the formation of a persistent, altered ecosystem state with low primary productivity. Based on our findings, we recommend the implementation of ecosystem-based and adaptive management strategies, such as (1) monitoring the status of key ecosystem attributes: kelp distribution and abundance, and densities of sea urchins and their predators, (2) developing management responses to threshold levels of these attributes, and (3) creating quantitative restoration suitability indices for informing kelp restoration efforts., Meredith McPherson et al. use a 34-year time series of satellite and in situ derived data to study bull kelp forests of northern California and demonstrate the ecosystem shifts following a marine heatwave event between 2014 and 2016. The results show that increased herbivory by sea urchins due to the loss of a predator reduced bull kelp forest resistance to fluctuating environmental conditions and point to the importance of ecosystem-based and adaptive management strategies.

Polymerase chain reaction (PCR) is a popular molecular tool for detection of bacteria. PCR allows millions of copies of a target segment of DNA to be produced. The DNA is extracted from overnight grown cultures of pure bacterial isolates using either the organo-solvent method or a commercial DNA extraction kit. The quality and purity of the DNA is determined by performing gel electrophoresis on 0.8% agarose gel. The DNA is amplified by performing PCR assay. Bands of approximately 1.5 kb in size are obtained from the amplified products of DNA. The PCR products run on 1.5% agarose gel are visualized with UV light and imaged by gel documentation system. This chapter outlines the protocol for isolation and amplification of DNA from cellulolytic bacteria. Cellulolytic bacteria are considered a potential source of cellulases for pretreatment of crop residues during biogas production. PCR is considered a very powerful, sensitive, specific, fast, and reliable tool in molecular detection and diagnostics.

BACKGROUND This study evaluated the effects of Bifidobacterium animalis subsp. lactis HN019 (B. lactis HN019) in the development of periodontitis (PE), associated or not with metabolic syndrome, (MS) in rats. METHODS Ninety-six rats were grouped according to a food protocol: high-fat diet for induction of MS or standard diet for the control groups (C). They were subdivided into groups with (+) and without (-) PE, receiving (*) or not (**) probiotic (PROB): C-**, CP-*, PE+**, PEP+*, MS- MSP-*, MSPE+**, and MSPEP+*. PROB administration started on the eighth week of the study and PE was induced on the 14th week by placing ligature on the animals' lower first molars. Euthanasia occurred in the 16th week. Biomolecular analyzes, immunoenzymatic assays, and microtomographic analyses were performed. The data obtained were analyzed statistically (P

Enzootic bovine leukosis (EBL) is an infectious disease caused by bovine leukemia virus (BLV) that affects cattle worldwide. Agar gel immunodiffusion (AGID) was the reference test for EBL diagnosis for many years, but enzyme-linked immunosorbent assay (ELISA) showed higher sensitivity, was faster to perform, and resulted in an objective reading. However, the importation of ELISA kits is lengthy and expensive, and currently, no AGID kits are available in Brazil. The aim of this work was to standardize an indirect ELISA (iELISA) for EBL diagnosis using BLV antigens produced in Tadarida brasiliensis lung (Tb1Lu) cells, which are Bovine viral diarrhea virus (BVDV) free, unlike fetal lamb kidney (FLK) cells, currently used for this purpose. Following standardization, iELISA results were compared with those obtained by AGID and the commercial Chekit Leucose-Serum ELISA. Compared to AGID, iELISA had 94,44% sensitivity, 75.68% specificity, 79.10% positive predictive value (PPV) and 93.30% negative predictive value (NPV), with 84% concordance and a Kappa index of 0.699. Compared to the Chekit Leucose-Serum ELISA, iELISA showed 92.60% sensitivity, 87.09% specificity, 90.27% PPV and 90,00% NPV, with 90.27% concordance and a Kappa index of 0.801. Taking into account the high agreement with the traditional tests and the absence of non-specific reactions with BVDV, the developed assay could be used as diagnostic method to control EBL in Brazil.

A heatwave that blanketed the northeast Pacific Ocean in 2013–2015 had severe impacts on the marine ecosystem through altered species composition and survival. A direct result of this marine heatwave was a sustained, record-setting harmful algal bloom (HAB), caused by the toxigenic diatom, Pseudo-nitzschia, that led to an unprecedented delay in harvest opportunity for commercial Dungeness crab (Metacarcinus magister) and closure of other recreational, commercial and tribal shellfish harvest, including razor clams. Samples collected during a cruise in summer 2015, showed the appearance of a highly toxic “hotspot” between Cape Mendocino, CA and Cape Blanco, OR that was observed again during cruises in the summers of 2016–2018. The transport of toxic cells from this retentive site northward during wind relaxations or reversals associated with storms resulted in economically debilitating delay or closure of Dungeness crab harvest in both northern California and Oregon in 2015–2019. Analyses of historic large-scale Pseudo-nitzschia HABs have shown that these events occur during warm periods such as El Niño, positive phases of the Pacific Decadal Oscillation, or the record-setting marine heatwave. In order to reduce the impacts of large-scale HABs along the west coast of North America, early warning systems have been developed to forewarn coastal managers. These early warning systems include the Pacific Northwest and California HAB Bulletins, both of which have documented elevated domoic acid and increased risk associated with the northern California hotspot. These early warnings enable mitigative actions such as selective opening of safe harvest zones, increased harvest limits during low risk periods, and early harvest in anticipation of impending HAB events. The aims of this study are to show trends in nearshore domoic acid along the US west coast in recent years, including the recent establishment of a new seed bed of highly-toxic Pseudo-nitzschia, and to explore how early warning systems are a useful tool to mitigate the human and environmental health and economic impacts associated with harmful algal blooms.

Background Huntington's disease is caused by a CAG repeat expansion in the huntingtin gene, HTT. Age at onset has been used as a quantitative phenotype in genetic analysis looking for Huntington's disease modifiers, but is hard to define and not always available. Therefore, we aimed to generate a novel measure of disease progression and to identify genetic markers associated with this progression measure. Methods We generated a progression score on the basis of principal component analysis of prospectively acquired longitudinal changes in motor, cognitive, and imaging measures in the 218 indivduals in the TRACK-HD cohort of Huntington's disease gene mutation carriers (data collected 2008–11). We generated a parallel progression score using data from 1773 previously genotyped participants from the European Huntington's Disease Network REGISTRY study of Huntington's disease mutation carriers (data collected 2003–13). We did a genome-wide association analyses in terms of progression for 216 TRACK-HD participants and 1773 REGISTRY participants, then a meta-analysis of these results was undertaken. Findings Longitudinal motor, cognitive, and imaging scores were correlated with each other in TRACK-HD participants, justifying use of a single, cross-domain measure of disease progression in both studies. The TRACK-HD and REGISTRY progression measures were correlated with each other (r=0·674), and with age at onset (TRACK-HD, r=0·315; REGISTRY, r=0·234). The meta-analysis of progression in TRACK-HD and REGISTRY gave a genome-wide significant signal (p=1·12 × 10−10) on chromosome 5 spanning three genes: MSH3, DHFR, and MTRNR2L2. The genes in this locus were associated with progression in TRACK-HD (MSH3 p=2·94 × 10−8 DHFR p=8·37 × 10−7 MTRNR2L2 p=2·15 × 10−9) and to a lesser extent in REGISTRY (MSH3 p=9·36 × 10−4 DHFR p=8·45 × 10−4 MTRNR2L2 p=1·20 × 10−3). The lead single nucleotide polymorphism (SNP) in TRACK-HD (rs557874766) was genome-wide significant in the meta-analysis (p=1·58 × 10−8), and encodes an aminoacid change (Pro67Ala) in MSH3. In TRACK-HD, each copy of the minor allele at this SNP was associated with a 0·4 units per year (95% CI 0·16–0·66) reduction in the rate of change of the Unified Huntington's Disease Rating Scale (UHDRS) Total Motor Score, and a reduction of 0·12 units per year (95% CI 0·06–0·18) in the rate of change of UHDRS Total Functional Capacity score. These associations remained significant after adjusting for age of onset. Interpretation The multidomain progression measure in TRACK-HD was associated with a functional variant that was genome-wide significant in our meta-analysis. The association in only 216 participants implies that the progression measure is a sensitive reflection of disease burden, that the effect size at this locus is large, or both. Knockout of Msh3 reduces somatic expansion in Huntington's disease mouse models, suggesting this mechanism as an area for future therapeutic investigation. Funding The European Commission FP7 NeurOmics project; CHDI Foundation; the Medical Research Council UK; the Brain Research Trust; and the Guarantors of Brain.

Harmful algal blooms produce toxins that bioaccumulate in the food web and adversely affect humans, animals, and entire marine ecosystems. Blooms of the diatom Pseudo-nitzschia can produce domoic acid (DA), a toxin that most commonly causes neurological disease in endothermic animals, with cardiovascular effects that were first recognized in southern sea otters. Over the last 20 years, DA toxicosis has caused significant morbidity and mortality in marine mammals and seabirds along the west coast of the USA. Identifying DA exposure has been limited to toxin detection in biological fluids using biochemical assays, yet measurement of systemic toxin levels is an unreliable indicator of exposure dose or timing. Furthermore, there is little information regarding repeated DA exposure in marine wildlife. Here, the association between long-term environmental DA exposure and fatal cardiac disease was investigated in a longitudinal study of 186 free-ranging sea otters in California from 2001 – 2017, highlighting the chronic health effects of a marine toxin. A novel Bayesian spatiotemporal approach was used to characterize environmental DA exposure by combining several DA surveillance datasets and integrating this with life history data from radio-tagged otters in a time-dependent survival model. In this study, a sea otter with high DA exposure had a 1.7-fold increased hazard of fatal cardiomyopathy compared to an otter with low exposure. Otters that consumed a high proportion of crab and clam had a 2.5- and 1.2-times greater hazard of death due to cardiomyopathy than otters that consumed low proportions. Increasing age is a well-established predictor of cardiac disease, but this study is the first to identify that DA exposure affects the risk of cardiomyopathy more substantially in prime-age adults than aged adults. A 4-year-old otter with high DA exposure had 2.3 times greater risk of fatal cardiomyopathy than an otter with low exposure, while a 10-year old otter with high DA exposure had just 1.2 times greater risk. High Toxoplasma gondii titers also increased the hazard of death due to heart disease 2.4-fold. Domoic acid exposure was most detrimental for prime-age adults, whose survival and reproduction are vital for population growth, suggesting that persistent DA exposure will likely impact long-term viability of this threatened species. These results offer insight into the pervasiveness of DA in the food web and raise awareness of under-recognized chronic health effects of DA for wildlife at a time when toxic blooms are on the rise.

The global proliferation of toxin producing cyanobacterial blooms has been attributed to a wide variety of environmental factors with nutrient pollution, increased temperatures, and drought being three of the most significant. The current study is the first formal assessment of cyanotoxins in two impaired lakes, Canyon Lake and Lake Elsinore, in southern California that have a history of cyanobacterial blooms producing high biomass as measured by chl-a. Cyanotoxins in Lake Elsinore were detected at concentrations that persistently exceeded California recreational health thresholds, whereas Canyon Lake experienced persistent concentrations that only occasionally exceeded health thresholds. The study results are the highest recorded concentrations of microcystins, anatoxin-a, and cylindrospermopsin detected in southern California lakes. Concentrations exceeded health thresholds that caused both lakes to be closed for recreational activities. Cyanobacterial identifications indicated a high risk for the presence of potentially toxic genera and agreed with the cyanotoxin results that indicated frequent detection of multiple cyanotoxins simultaneously. A statistically significant correlation was observed between chlorophyll-a (chl-a) and microcystin concentrations for Lake Elsinore but not Canyon Lake, and chl-a was not a good indicator of cylindrospermopsin, anatoxin-a, or nodularin. Therefore, chl-a was not a viable screening indicator of cyanotoxin risk in these lakes. The study results indicate potential acute and chronic risk of exposure to cyanotoxins in these lakes and supports the need for future monitoring efforts to help minimize human and domestic pet exposure and to better understand potential effects to wildlife. The frequent co-occurrence of complex cyanotoxin mixtures further complicates the risk assessment process for these lakes given uncertainty in the toxicology of mixtures.

Background Medicines with a stereogenic center (asymmetric carbon) mainly presents as racemates with a mixture of equal amounts of enantiomers. One enantiomer may be active while the other inactive, alternatively one may produce side-effects and even toxicity. Worldwide there is no mandatory regulatory requirement to enforce the development of new chiral medicines as exclusively pure active single enantiomers. This has contributed to the existence of both single enantiomers and racemates on the market. Moreover, literature shows the tendency of regulatory approval of medicines towards the development of pure single enantiomers 60%, only 5–10% racemates and 30–35% as achiral. This status has not been established particularly in African countries including Tanzania. Therefore, we aimed to establish the chirality status of registered medicines in Tanzania. Methodology: A retrospective cross-sectional study was conducted by reviewing all human medicines registered in Tanzania for the past 15 years from 2003 to 2018. Data were extracted from TMDA-IMIS database to Microsoft excel for analysis. Medicines were assessed to determine if they were chiral (racemates or single enantiomers) or achiral. Results A total of 3,573 human medicines had valid registration. Most of these medicines 2,150 (60%) are chiral and 1,423 (40%) are achiral. Out of the chiral medicines, 1,591 (74%) and 559 (26%) were racemates and single enantiomers, respectively. The percentage of chiral medicines for anti-infective (35%) was significantly higher (p

During hemolysis, macrophages in the liver phagocytose damaged erythrocytes to prevent the toxic effects of cell-free hemoglobin and heme. It remains unclear how this homeostatic process modulates phagocyte functions in inflammatory diseases. Using a genetic mouse model of spherocytosis and single-cell RNA sequencing, we found that erythrophagocytosis skewed liver macrophages into a unique anti-inflammatory phenotype that we defined as Marcohigh/Hmoxhigh/MHC-class IIlow erythrophagocytes. This phenotype transformation profoundly mitigated disease expression in a model of an anti-CD40-induced hyperinflammatory syndrome with necrotic hepatitis and in a non-alcoholic steatohepatitis model, representing two macrophage-driven sterile inflammatory diseases. We reproduced the anti-inflammatory erythrophagocyte transformation in vitro by heme-exposure of mouse and human macrophages, yielding a distinctive transcriptional signature that segregated heme-polarized from M1- and M2-polarized cells. Mapping transposase-accessible chromatin in single cells by sequencing (scATAC-seq) defined the transcription factor NFE2L2/NRF2 as a critical driver of erythrophagocytes, and Nfe2l2/Nrf2-deficiency restored heme-suppressed inflammation. Our findings point to a pathway that regulates macrophage functions to link erythrocyte homeostasis with innate immunity.

A new species of jawfish,Opistognathusthionyisp. n., is described from the Vitória-Trindade Chain and Fernando de Noronha Archipelago off Brazil, a disjunct distribution of ca. 1,800 km.Opistognathusthionyiand its allopatric Caribbean sister-species,Opistognathusmaxillosus, both have a wide, fan-like upper margin of the subopercular flap and mostly over-lapping meristic data. The new species differs fromO.maxillosusin having the darkest spot on the spinous dorsal fin, when present, between spines 2–5, versus always present between spines 6–9, the buccal area surrounding the esophageal opening pale versus very dark and fewer oblique scale rows in longitudinal series (45–52 vs. 69–85). A second new species,Opistognathusvicinussp. n., known from Brazil’s mainland, has completely over-lapping meristic values with its allopatric Caribbean sister-speciesO.whitehursti, but differs in lacking vomerine teeth and a supramaxilla and retaining the juvenile color pattern of the latter species in adults. Diagnoses, photographs, an identification key, and distributional maps are given for all Brazilian species ofOpistognathus.Molecular phylogenetic analysis of partial cytochrome c oxidase subunit-I sequences indicates that specimens of the two allopatric pairs,O.thionyi–O.maxillosusandO.vicinus–O.whitehursti, form reciprocally monophyletic groups that differ from each other on average by 9 to 11%, with less than 1% average pair-wise genetic distance within-species. Similar patterns of phylogenetic structure were observed between reciprocally monophyletic (predominately allopatric) groups within nominal species ofOpistognathusaurifrons, suggesting the possibility of at least two additional undescribed species from the Brazilian Province.

San Francisco Bay (SFB) is a eutrophic estuary that harbors both freshwater and marine toxigenic organisms that are responsible for harmful algal blooms. While there are few commercial fishery harvests within SFB, recreational and subsistence harvesting for shellfish is common. Coastal shellfish are monitored for domoic acid and paralytic shellfish toxins (PSTs), but within SFB there is no routine monitoring for either toxin. Dinophysis shellfish toxins (DSTs) and freshwater microcystins are also present within SFB, but not routinely monitored. Acute exposure to any of these toxin groups has severe consequences for marine organisms and humans, but chronic exposure to sub-lethal doses, or synergistic effects from multiple toxins, are poorly understood and rarely addressed. This study documents the occurrence of domoic acid and microcystins in SFB from 2011 to 2016, and identifies domoic acid, microcystins, DSTs, and PSTs in marine mussels within SFB in 2012, 2014, and 2015. At least one toxin was detected in 99% of mussel samples, and all four toxin suites were identified in 37% of mussels. The presence of these toxins in marine mussels indicates that wildlife and humans who consume them are exposed to toxins at both sub-lethal and acute levels. As such, there are potential deleterious impacts for marine organisms and humans and these effects are unlikely to be documented. These results demonstrate the need for regular monitoring of marine and freshwater toxins in SFB, and suggest that co-occurrence of multiple toxins is a potential threat in other ecosystems where freshwater and seawater mix.

Jatropha curcas L. has emerged in recent times as a leading energy crop in sub-Saharan Africa with over 32 countries in the region involved in its cultivation. By 2008, five countries in the region (Senegal, Nigeria, Mali, Ethiopia, and Zimbabwe) had policies promoting Jatropha cultivation. These policies were not informed by empirical evidence arising from the performance of Jatropha in the region. This paper analyses performance data of Jatropha in the region with a view to synthesizing information that is useful for evidence-based policy formulation. Production data in terms of agronomic issues and seed yields are analyzed. The results show that most of the attributes generically associated with Jatropha have not been achieved in the region and there has been inadequate research to support Jatropha cultivation. Policies supporting cultivation of Jatropha need to be informed by these observations in order to promote viable cultivation of the crop.

Length–weight relationships (LWRs) were determined for four fish species from two drainage areas, those of the Itapemirim and Jucu River, in the state of Espirito Santo, southeastern Brazil. Fishes were collected between May 2015 and November 2016, with gill nets, casting net, dip net, and sieve. Herein are presented LWRs for Astyanax lacustris, Astyanax taeniatus, Hyphessobrycon bifasciatus and Oligosarcus acutirostris.

Length‐weight and length‐length relationships were determined for nine fish species of Rocas Atoll, Fernando de Noronha and Trindade Island. Samples were conducted in 2014 (Rocas) and 2015 (Noronha and Trindade) in tidepools using anaesthetic clove oil and hand nets to collect fish fauna. Four species (Stegastes rocasensis, Scartella itajobi, Starksia multilepis and Bathygobius brasiliensis) are endemic from the Noronha‐Rocas ridge, two are endemic from the Trindade‐Martin Vaz insular complex (Scartella poiti and Malacoctenus brunoi), and three species have a widespread distribution. All relationships are novel for science.

Blooms of the diatom genus Pseudo-nitzschia occur annually in the Southern California Bight (SCB), and domoic acid (DA) associated with these events can contaminate fisheries, presenting both human and wildlife health risks. Recent studies have suggested that marine sediments may act as a reservoir for DA, extending the risk of food web contamination long after water column blooms have ended. In this study, we conducted a regional assessment of the extent and magnitude of DA in the benthic environment, and monthly observations of sediments and benthic infauna at multiple stations over a 16-month period. DA was widespread in continental shelf sediments of the SCB. The toxin was detected in 54% of all shelf habitats sampled. Detectable concentrations ranged from 0.11 ng/g to 1.36 ng/g. DA was consistently detected in benthic infauna tissues over the monthly timeseries, while the DA concentrations in sediments during the same period were commonly below detection or at low concentrations. The presence of DA in the benthic environment did not always have an apparent water column source, raising the possibility of lateral transport, retention/preservation in sediments or undetected blooms in subsurface waters. In most cases, DA was detected in tissues but not in the co-located surface sediments. Coarse taxonomic sorting of the infauna revealed that the accumulation of DA varied among taxa. We observed that DA was widespread among lower trophic level organisms in this study, potentially acting as a persistent source of DA to higher trophic levels in the benthos.

Three-dimensional (3D) brain organoids derived from human pluripotent stem cells (hPSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), appear to recapitulate the brain’s 3D cytoarchitectural arrangement and provide new opportunities to explore disease pathogenesis in the human brain. Human iPSC (hiPSC) reprogramming methods, combined with 3D brain organoid tools, may allow patient-derived organoids to serve as a preclinical platform to bridge the translational gap between animal models and human clinical trials. Studies using patient-derived brain organoids have already revealed novel insights into molecular and genetic mechanisms of certain complex human neurological disorders such as microcephaly, autism, and Alzheimer’s disease. Furthermore, the combination of hiPSC technology and small-molecule high-throughput screening (HTS) facilitates the development of novel pharmacotherapeutic strategies, while transcriptome sequencing enables the transcriptional profiling of patient-derived brain organoids. Finally, the addition of CRISPR/Cas9 genome editing provides incredible potential for personalized cell replacement therapy with genetically corrected hiPSCs. This review describes the history and current state of 3D brain organoid differentiation strategies, a survey of applications of organoids towards studies of neurodevelopmental and neurodegenerative disorders, and the challenges associated with their use as in vitro models of neurological disorders.

While 15N-labeled nitrogen (N) compounds have been used to quantify N uptake rates by plankton communities for decades, accurately ascribing those rates to individual populations or species has been a challenge. Here, we apply sorting flow cytometry combined with species-specific immuno-detection of a harmful alga, Aureococcus anophagefferens, to contrast the nutritional ecology of this alga with co-occurring picoplankton (picoeukaryotes, cyanobacteria, heterotrophic bacteria) during brown tides. The method was iteratively refined to yield close agreement (85–101%) between plankton community 15N uptake quantified via traditional filtration and this novel sorting method. Sorting of plankton revealed that the δ15N values of A. anophagefferens and phycocyanin-containing cyanobacteria were more enriched (∼ 10‰) than the values of other picoeukaryotes and heterotrophic bacteria that decreased to

An ecologically and economically disruptive harmful algal bloom (HAB) affected much of the northeast Pacific margin in 2015, during a prolonged oceanic warm anomaly. Caused by diatoms of the genus Pseudo-nitzschia, this HAB produced the highest particulate concentrations of the biotoxin domoic acid (DA) ever recorded in Monterey Bay, California. Bloom inception followed strong upwelling during the spring transition, which introduced nutrients and eliminated the warm anomaly locally. Subsequently, moderate and intermittent upwelling created favorable conditions for growth and accumulation of HAB biomass, which was dominated by a highly toxigenic species, P. australis. High cellular DA concentrations were associated with available nitrogen for DA synthesis coincident with silicate exhaustion. This nutrient influence resulted from two factors: (1) disproportionate depletion of silicate in upwelling source waters during the warm anomaly, the most severe depletion observed in 24 years, and (2) silicate uptake by the dense diatom bloom.

Processing of Jatropha seeds to produce oil leaves press-cake (JPC) as a by-product. Based on extraction efficiency, 80–85% of seed remains as press-cake. In places where large quantities of Jatropha oil are extracted, mainly for biodiesel production, huge quantities of press-cake would be produced. JPC constitutes a substantial by-product that has multiple applications. Most papers in literature have focused on a narrow range of product streams arising from valorisation of JPC. This paper presents data on nine possible product streams of JPC valorisation. These are organic fertiliser, protein, animal feed, biocomposites, briquettes, bioethanol, biogas, pyloritic products and syngas. This makes it easier for comparative purposes as it aggregates existing data. The objective of this paper is to review empirical data available in literature on the conversion of JPC into high-value products and map the various valorisation options. The product streams can be placed into three main groups; fertiliser, energy carriers and industrial chemical products. The energy carrier group dominates with five products (briquettes, bioethanol, biogas, pyrolytic products and syngas). Conversion technologies for these products are well established. Biogas and briquettes are the most promising and have wide application. Use of JPC as fertiliser is also widely practiced. Work on production of various industrial chemicals, such as protein and biocomposites is at different stages of maturity. The paper shows a lot of work that has and is still being conducted in order to broaden the product streams of JPC valorisation.

A 3-week diversion of the Orange County Sanitation District effluent discharge into nearshore waters off Newport Beach, CA constituted a considerable injection of secondarily-treated effluent into the coastal ecosystem. The location ≈1.6 km from shore, shallow water depth (≈16 m), volume and nutrient content of the discharge (≈5.3 × 10 8 L day −1 of effluent with inorganic nitrogen concentration >2 mM) during the diversion raised concerns regarding the potential for stimulating phytoplankton blooms and, in particular, blooms of toxic species. Remarkably, phytoplankton standing stocks during the event and shortly thereafter did not reach values associated even with minor blooms historically observed in the region (generally −1 ), although shifts in community composition were observed. Diatom abundances increased early during the diversion, dinoflagellates, phototrophic picoplanktonic eukaryotes and other algae increased mid-diversion, and cyanobacteria ( Synechococcus , Prochlorococcus ) increased near the end of the diversion. Concentrations of domoic acid (a phycotoxin commonly present in the area) remained near or below detection throughout the diversion, and abundances of potentially-harmful algal species were unresponsive. Bacterial biomass increased during the diversion, and equaled or exceeded total phytoplankton biomass in most samples. Abundances of microbial grazers were also elevated during the diversion. We speculate that nutrient uptake by the bacterial biomass, acting in concert with or a response to a negative effect of disinfection byproducts associated with chlorination on phytoplankton physiology, played a significant role in muting the response of the phytoplankton to nutrients released in the effluent.

DNA-encoded chemical libraries (DECLs) are pools of DNA-tagged small molecules that enable facile screening and identification of bio-macromolecule binders. The successful development of DECLs has led to their increasingly important role in drug development, and screening hits have entered clinical trials. In this review, we summarize the development and currently active research areas of DECLs with a focus on contributions from groups at academic institutes. We further look at opportunities and future directions of DECL research in medicinal chemistry and chemical biology based on the symbiotic relationship between academia and industry. Challenges associated with the application of DECLs in academic drug discovery are further discussed.

Most physical oceanographic data and some biological data used in this study are available through servers supported by federal and state agencies, as specified in the Methods section. Glider data are available from UCSD: https://spraydata.ucsd.edu/ projects/CUGN/. All other data presented have been deposited in a repository and are available at: https://doi.org/10.6084/m9.figshare.c.4584026.v1. The article of record as published may be found at https://doi.org/10.1371/journal.pone.0222456 This study examines the occurrence of humpback whale (Megaptera novaeangliae) song in the northeast Pacific from three years of continuous recordings off central California (36.713˚N, 122.186˚W). Song is prevalent in this feeding and migratory habitat, spanning nine months of the year (September–May), peaking in winter (November–January), and reaching a maximum of 86% temporal coverage (during November 2017). From the rise of song in fall through the end of peak occurrence in winter, song length increases significantly from month to month. The seasonal peak in song coincides with the seasonal trough in day length and sighting-based evidence of whales leaving Monterey Bay, consistent with seasonal migration. During the seasonal song peak, diel variation shows maximum occurrence at night (69% of the time), decreasing during dawn and dusk (52%), and further decreasing with increasing solar elevation during the day, reaching a minimum near solar noon (30%). Song occurrence increased 44% and 55% between successive years. Sighting data within the acoustic detection range of the hydrophone indicate that variation in local population density was an unlikely cause of this large interannual variation. Hydrographic data and modeling of acoustic transmission indicate that changes in neither habitat occupancy nor acoustic transmission were probable causes. Conversely, the positive interannual trend in song paralleled major ecosystem variations, including similarly large positive trends in winddriven upwelling, primary productivity, and krill abundance. Further, the lowest song occurrence during the first year coincided with anomalously warm ocean temperatures and an extremely toxic harmful algal bloom that affected whales and other marine mammals in the region. These major ecosystem variations may have influenced the health and behavior of humpback whales during the study period. David and Lucile Packard Foundation Central and Northern California Ocean Observing System Monterey Accelerated Research System (MARS) NOAA NOAA ECOHAB JPR DEC FPC JTP JTP CW RM KBB NOAA NA16NOS0120021 NOAA ECOHAB Program (NA11NOS4780030 NSF Award 1514756

Diarrhetic shellfish toxins (DSTs) are produced by the marine dinoflagellate, Dinophysis, as well as select species of benthic Prorocentrum. The DSTs can bioaccumulate in shellfish and cause gastrointestinal illness when humans consume high levels of this toxin. Although not routinely monitored throughout the U.S., recent studies in Washington, Texas, and New York suggest DSTs may be widespread throughout U.S. coastal waters. This study describes a four-year time series (2013-2016) of Dinophysis concentration and DST level in California mussels (Mytilus californianus) from Santa Cruz Municipal Wharf (SCMW) in Monterey Bay, California. Results show a maximum Dinophysis concentration of 9404 cells/L during this study and suggest Dinophysis persists as a member of the background phytoplankton community throughout the year. In California mussels, DSTs were found at persistent low levels throughout the course of this study, and exceeded the FDA guidance level of 160 ng/g 19 out of 192 weeks sampled. Concentrations of Dinophysis alone are a positive but weak predictor of DST level in California mussels, and basic environmental variables (temperature, salinity, and nutrients) do not sufficiently explain variation in Dinophysis concentration at SCMW. This study demonstrates that Dinophysis in Monterey Bay are producing DSTs that accumulate in local shellfish throughout the year, occasionally reaching levels of concern.

Occludin(OCLN) mutations cause human microcephaly and cortical malformation. A tight junction component thought absent in neuroepithelium after neural tube closure, OCLN isoform-specific expression extends into corticogenesis. Full-length and truncated isoforms localize to neuroprogenitor centrosomes, but full-length OCLN transiently localizes to plasma membranes while only truncated OCLN continues at centrosomes throughout neurogenesis. Mimicking human mutations, full-length OCLN depletion in mouse and in human CRISPR/Cas9-edited organoids produce early neuronal differentiation, reduced progenitor self-renewal and increased apoptosis. Human neural progenitors were more severely affected, especially outer radial glial cells, which mouse embryonic cortex lacks. Rodent and human mutant progenitors displayed reduced proliferation and prolonged M-phase. OCLN interacted with mitotic spindle regulators, NuMA and RAN, while full-length OCLN loss impaired spindle pole morphology, astral and mitotic microtubule integrity. Thus, early corticogenesis requires full-length OCLN to regulate centrosome organization and dynamics, revealing a novel role for this tight junction protein in early brain development.

The present study investigated the circulation of avian metapneumovirus (aMPV) in wild birds in Brazil. To do so, 131 samples from 366 oropharyngeal or cloacal swabs collected from 18 species of birds were tested individually or in pools by RT-PCR. Samples detected by RT-PCR were selected for DNA sequencing. Thirteen (9.9%) samples were detected by the RT-PCR targeting the N gene and four out of 13 samples were sequenced. Sequencing results showed a high identity with the aMPV subtype A. Our results confirm the circulation of the aMPV subtype A in wild birds in Brazil even five years after its last detection. RESUMO: O presente estudo investigou a circulação de metapneumovírus aviário em aves silvestres no Brasil. Para tanto, 131 amostras de 366 suabes orofaringeanos ou cloacais coletados de 18 espécies de aves foram testadas individualmente ou na forma de pools por RT-PCR. As amostras detectadas por RT-PCR foram selecionadas para sequenciamento. Treze (9,9%) das amostras foram detectadas por RT-PCR tendo o gene N como alvo; destas, quatro foram sequenciadas com sucesso. Resultados do sequenciamento mostraram alta identidade com o aMPV de subtipo A. Nossos resultados confirmam a circulação de aMPV subtipo A em aves silvestres no Brasil mesmo cinco anos após sua última detecção.

San Francisco Bay (SFB), California, USA is the largest estuary in the western United States and is home to more than 7 million people in nine counties and 101 cities. It is highly nutrient enriched and is directly connected to the Gulf of the Farallones and coastal Pacific ocean through the Golden Gate strait. The Gulf of the Farallones is one of several “hotspots” for the neurotoxin domoic acid, produced by members of the genus Pseudo-nitzschia. Despite the close proximity, SFB has few reports of harmful algal blooms and low concentrations of domoic acid, suggesting that SFB is somehow resistant to toxic blooms. Here we evaluate the potential growth and toxicity of the dominant toxigenic species in California coastal waters, P. australis and P. multiseries, to directly test the hypothesis that SFB waters confer resistance to blooms. We specifically evaluate the effect of varying temperature, salinity, and to a lesser extent, nutrients on growth and toxin production. Results show equivalent growth in SFB water (maximum growth rates of 0.71 and 1.35 d−1 for P. multiseries and P. australis) compared to open-coast water, and comparable or greater toxicity (0 to >100 pg DA cell−1). The historical resistance to blooms in SFB is hypothesized to be caused by a combination of insufficient acclimation time for advected Pseudo-nitzschia populations to become established and suppression of toxin production in warm waters.

Delayed ischemic neurological deficit (DIND) is a major driver of adverse outcomes in patients with aneurysmal subarachnoid hemorrhage (aSAH), defining an unmet need for therapeutic development. Cell-free hemoglobin that is released from erythrocytes into the cerebrospinal fluid (CSF) is suggested to cause vasoconstriction and neuronal toxicity, and correlates with the occurrence of DIND. Cell-free hemoglobin in the CSF of patients with aSAH disrupted dilatory NO signaling ex vivo in cerebral arteries, which shifted vascular tone balance from dilation to constriction. We found that selective removal of hemoglobin from patient CSF with a haptoglobin-affinity column or its sequestration in a soluble hemoglobin-haptoglobin complex was sufficient to restore physiological vascular responses. In a sheep model, administration of haptoglobin into the CSF inhibited hemoglobin-induced cerebral vasospasm and preserved vascular NO signaling. We identified 2 pathways of hemoglobin delocalization from CSF into the brain parenchyma and into the NO-sensitive compartment of small cerebral arteries. Both pathways were critical for hemoglobin toxicity and were interrupted by the large hemoglobin-haptoglobin complex that inhibited spatial requirements for hemoglobin reactions with NO in tissues. Collectively, our data show that compartmentalization of hemoglobin by haptoglobin provides a novel framework for innovation aimed at reducing hemoglobin-driven neurological damage after subarachnoid bleeding.