RESUMO
Recently published near full-length KSHV genomes from a Cameroon Kaposi sarcoma case-control study showed strong evidence of viral recombination and mixed infections, but no sequence variations associated with disease. Using the same methodology, an additional 102 KSHV genomes from 76 individuals with KSHV-associated diseases have been sequenced. Diagnoses comprise all KSHV-associated diseases (KAD): Kaposi sarcoma (KS), primary effusion lymphoma (PEL), KSHV-associated large cell lymphoma (KSHV-LCL), a type of multicentric Castleman disease (KSHV-MCD), and KSHV inflammatory cytokine syndrome (KICS). Participants originated from 22 different countries, providing the opportunity to obtain new near full-length sequences of a wide diversity of KSHV genomes. These include near full-length sequence of genomes with KSHV K1 subtypes A, B, C, and F as well as subtype E, for which no full sequence was previously available. High levels of recombination were observed. Fourteen individuals (18%) showed evidence of infection with multiple KSHV variants (from two to four unique genomes). Twenty-six comparisons of sequences, obtained from various sampling sites including PBMC, tissue biopsies, oral fluids, and effusions in the same participants, identified near complete genome conservation between different biological compartments. Polymorphisms were identified in coding and non-coding regions, including indels in the K3 and K15 genes and sequence inversions here reported for the first time. One such polymorphism in KSHV ORF46, specific to the KSHV K1 subtype E2, encoded a mutation in the leucine loop extension of the uracil DNA glycosylase that results in alteration of biochemical functions of this protein. This confirms that KSHV sequence variations can have functional consequences warranting further investigation. This study represents the largest and most diverse analysis of KSHV genome sequences to date among individuals with KAD and provides important new information on global KSHV genomics.
Assuntos
Genoma Viral , Herpesvirus Humano 8 , Sarcoma de Kaposi , Humanos , Herpesvirus Humano 8/genética , Sarcoma de Kaposi/virologia , Sarcoma de Kaposi/genética , Masculino , Feminino , Pessoa de Meia-Idade , Adulto , Polimorfismo Genético , Idoso , Infecções por Herpesviridae/genética , Infecções por Herpesviridae/virologia , Etnicidade/genética , Hiperplasia do Linfonodo Gigante/virologia , Hiperplasia do Linfonodo Gigante/genética , FilogeniaRESUMO
Per- and polyfluoroalkyl substances (PFAS) enter the marine food web, accumulate in organisms, and potentially have adverse effects on predators and consumers of seafood. However, evaluations of PFAS in meso-to-apex predators, like sharks, are scarce. This study investigated PFAS occurrence in five shark species from two marine ecosystems with contrasting relative human population densities, the New York Bight (NYB) and the coastal waters of The Bahamas archipelago. The total detected PFAS (∑PFAS) concentrations in muscle tissue ranged from 1.10 to 58.5 ng g-1 wet weight, and perfluorocarboxylic acids (PFCAs) were dominant. Fewer PFAS were detected in Caribbean reef sharks (Carcharhinus perezi) from The Bahamas, and concentrations of those detected were, on average, â¼79% lower than in the NYB sharks. In the NYB, ∑PFAS concentrations followed: common thresher (Alopias vulpinus) > shortfin mako (Isurus oxyrinchus) > sandbar (Carcharhinus plumbeus) > smooth dogfish (Mustelus canis). PFAS precursors/intermediates, such as 2H,2H,3H,3H-perfluorodecanoic acid and perfluorooctanesulfonamide, were only detected in the NYB sharks, suggesting higher ambient concentrations and diversity of PFAS sources in this region. Ultralong-chain PFAS (C ≥ 10) were positively correlated with nitrogen isotope values (δ15N) and total mercury in some species. Our results provide some of the first baseline information on PFAS concentrations in shark species from the northwest Atlantic Ocean, and correlations between PFAS, stable isotopes, and mercury further contextualize the drivers of PFAS occurrence.
Assuntos
Tubarões , Poluentes Químicos da Água , Animais , Tubarões/metabolismo , Monitoramento Ambiental , Bahamas , Fluorocarbonos/análise , New York , Cadeia AlimentarRESUMO
Mercury (Hg) inputs have particularly impacted the northeastern United States due to its proximity to anthropogenic emissions sources and abundant habitats that efficiently convert inorganic Hg into methylmercury. Intensive research and monitoring efforts over the past 50 years in New York State, USA, have informed the assessment of the extent and impacts of Hg exposure on fishes and wildlife. By synthesizing Hg data statewide, this study quantified temporal trends of Hg exposure, spatiotemporal patterns of risk, the role that habitat and Hg deposition play in producing spatial patterns of Hg exposure in fish and other wildlife, and the effectiveness of current monitoring approaches in describing Hg trends. Most temporal trends were stable, but we found significant declines in Hg exposure over time in some long-sampled fish. The Adirondack Mountains and Long Island showed the greatest number of aquatic and terrestrial species with elevated Hg concentrations, reflecting an unequal distribution of exposure risk to fauna across the state. Persistent hotspots were detected for aquatic species in central New York and the Adirondack Mountains. Elevated Hg concentrations were associated with open water, forests, and rural, developed habitats for aquatic species, and open water and forested habitats for terrestrial species. Areas of consistently elevated Hg were found in areas driven by atmospheric and local Hg inputs, and habitat played a significant role in translating those inputs into biotic exposure. Continued long-term monitoring will be important in evaluating how these patterns continue to change in the face of changing land cover, climate, and Hg emissions.
Assuntos
Mercúrio , Compostos de Metilmercúrio , Poluentes Químicos da Água , Animais , Mercúrio/análise , New York , Monitoramento Ambiental , Peixes , Biota , Animais Selvagens , ÁguaRESUMO
In sub-Saharan Africa, Kaposi's sarcoma-associated herpesvirus (KSHV) is endemic, and Kaposi's sarcoma (KS) is a significant public health problem. Until recently, KSHV genotype analysis was performed using variable gene regions, representing a small fraction of the genome, and thus the contribution of sequence variation to viral transmission or pathogenesis are understudied. We performed near full-length KSHV genome sequence analysis on samples from 43 individuals selected from a large Cameroonian KS case-control study. KSHV genomes were obtained from 21 KS patients and 22 control participants. Phylogenetic analysis of the K1 region indicated the majority of sequences were A5 or B1 subtypes and all three K15 alleles were represented. Unique polymorphisms in the KSHV genome were observed including large gene deletions. We found evidence of multiple distinct KSHV genotypes in three individuals. Additionally, our analyses indicate that recombination is prevalent suggesting that multiple KSHV infections may not be uncommon overall. Most importantly, a detailed analysis of KSHV genomes from KS patients and control participants did not find a correlation between viral sequence variations and disease. Our study is the first to systematically compare near full-length KSHV genome sequences between KS cases and controls in the same endemic region to identify possible sequence variations associated with disease risk.
Assuntos
Herpesvirus Humano 8 , Sarcoma de Kaposi , Camarões/epidemiologia , Estudos de Casos e Controles , Herpesvirus Humano 8/genética , Humanos , Filogenia , Sarcoma de Kaposi/epidemiologia , Sarcoma de Kaposi/genéticaRESUMO
The respiratory cytochrome bc1 complex functions as a protonmotive ubiquinol:cytochrome c oxidoreductase. Lysine 228 (K228) located within the quinol reduction (Qi) site of the bc1 complex, has been reported as a key residue for proton transfer during the redox chemistry cycle to substrate quinone at Qi. In yeast, while single mutations had no effect, the combination of K228L and F225L resulted in a severe respiratory growth defect and inhibition of O2 consumption in intact cells. The inhibition was overcome by uncoupling the mitochondrial membrane or by suppressor mutations in the region of K228L-F225L. We propose that the K228L mutation introduces energetic (and kinetic) barriers into normal electron- and proton transfer chemistry at Qi, which are relieved by dissipation of the opposing protonmotive force or through the restoration of favourable intraprotein proton transfer networks via suppressor mutation.
Assuntos
Citocromos b/metabolismo , Citocromos c1/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Citocromos b/química , Citocromos b/genética , Citocromos c1/química , Citocromos c1/genética , Transporte de Elétrons , Hidroquinonas/metabolismo , Cinética , Modelos Moleculares , Mutação , Oxirredução , Oxigênio/metabolismo , Força Próton-Motriz , Prótons , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/genética , Ubiquinona/análogos & derivados , Ubiquinona/metabolismoRESUMO
Calanoid copepods, depending on feeding strategy, have different behavioral and biological controls on their movements, thereby responding differently to environmental conditions such as changes in seawater viscosity. To understand how copepod responses to environmental conditions are mediated through physical, physiological and/or behavioral pathways, we used high-speed microvideography to compare two copepod species, Acartia hudsonica and Parvocalanus crassirostris, under different temperature, viscosity and dietary conditions. Acartia hudsonica exhibited 'sink and wait' feeding behavior and typically responded to changes in seawater viscosity; increased seawater viscosity reduced particle-capture behavior and decreased the size of the feeding current. In contrast, P. crassirostris continuously swam and did not show any behavioral or physical responses to changes in viscosity. Both species showed a physiological response to temperature, with reduced appendage beating frequency at cold temperatures, but this did not generally translate into effects on swimming speed, feeding flux or active time. Both copepod species swam slower when feeding on diatom rather than dinoflagellate prey, showing that prey type mediates copepod behavior. These results differentiate species-specific behaviors and responses to environmental conditions, which may lead to better understanding of niche separation and latitudinal patterns in copepod feeding and movement strategies.
Assuntos
Copépodes , Animais , Comportamento Alimentar , Água do Mar , Temperatura , ViscosidadeRESUMO
Glucose-6-phosphate dehydrogenase (G6PDH) can initiate the glucose-6-phosphate (G6P) shunt around the Calvin-Benson cycle. To understand the regulation of flux through this pathway, we have characterized the biochemical parameters and redox regulation of the three functional plastidic isoforms of Arabidopsis G6PDH. When purified, recombinant proteins were measured, all three exhibited significant substrate inhibition by G6P but not NADP+, making the determination of enzyme kinetic parameters complex. We found that the half-saturation concentration of G6PDH isoform 1 is increased under reducing conditions. The other two isoforms exhibit less redox regulation, however, isoform 2 is strongly inhibited by NADPH. Redox regulation of G6PDH1 can be partially reversed by hydrogen peroxide or protected against by the presence of its substrate, G6P. Overall, our results support the conclusion that G6PDH can have significant activity throughout the day and can be dynamically regulated to allow or prevent flux through the glucose-6-phosphate shunt.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Glucosefosfato Desidrogenase/metabolismo , Luz , Plastídeos/enzimologia , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Glucosefosfato Desidrogenase/genética , Isoenzimas/genética , Isoenzimas/metabolismo , Oxirredução , Plastídeos/genéticaRESUMO
The importance of fish consumption as the primary pathway of human exposure to mercury and the establishment of fish consumption advisories to protect human health have led to large fish tissue monitoring programs worldwide. Data on fish tissue mercury concentrations collected by state, tribal, and provincial governments via contaminant monitoring programs have been compiled into large data bases by the U.S. Environmental Protection Agency's Great Lakes National Monitoring Program Office (GLNPO), the Ontario Ministry of the Environment's Fish Contaminants Monitoring and Surveillance Program (FMSP), and many others. These data have been used by a wide range of governmental and academic investigators worldwide to examine long-term and recent trends in fish tissue mercury concentrations. The largest component of the trend literature is for North American freshwater species important in recreational fisheries. This review of temporal trends in fish tissue mercury concentrations focused on published results from freshwater fisheries of North America as well as marine fisheries worldwide. Trends in fish tissue mercury concentrations in North American lakes with marked overall decreases were reported over the period 1972-2016. These trends are consistent with reported mercury emission declines as well as trends in wet deposition across the U.S. and Canada. More recently, a leveling-off in the rate of decreases or increases in fish tissue mercury concentrations has been reported. Increased emissions of mercury from global sources beginning between 1990 and 1995, despite a decrease in North American emissions, have been advanced as an explanation for the observed changes in fish tissue trends. In addition to increased atmospheric deposition, the other factors identified to explain the observed mercury increases in the affected fish species include a systematic shift in the food-web structure with the introduction of non-native species, creating a new or expanding role for sediments as a net source for mercury. The influences of climate change have also been identified as contributing factors, including considerations such as increases in temperature (resulting in metabolic changes and higher uptake rates of methylmercury), increased rainfall intensity and runoff (hydrologic export of organic matter carrying HgII from watersheds to surface water), and water level fluctuations that alter either the methylation of mercury or the mobilization of monomethylmercury. The primary source of mercury exposure in the human diet in North America is from the commercial fish and seafood market which is dominated (>90%) by marine species. However, very little information is available on mercury trends in marine fisheries. Most of the data used in the published marine trend studies are assembled from earlier reports. The data collection efforts are generally intermittent, and the spatial and fish-size distribution of the target species vary widely. As a result, convincing evidence for the existence of fish tissue mercury trends in marine fish is generally lacking. However, there is some evidence from sampling of large, long-lived commercially-important fish showing both lower mercury concentrations in the North Atlantic in response to reduced anthropogenic mercury emission rates in North America and increases in fish tissue mercury concentrations over time in the North Pacific in response to increased mercury loading.
Assuntos
Monitoramento Ambiental , Peixes/metabolismo , Mercúrio/metabolismo , Poluentes Químicos da Água/metabolismo , Animais , Cadeia Alimentar , Lagos , Compostos de Metilmercúrio , OntárioRESUMO
BACKGROUND: Although measles was eliminated in the United States in 2000, importations of the virus continue to cause outbreaks. We describe the epidemiologic features of an outbreak of measles that originated from two unvaccinated Amish men in whom measles was incubating at the time of their return to the United States from the Philippines and explore the effect of public health responses on limiting the spread of measles. METHODS: We performed descriptive analyses of data on demographic characteristics, clinical and laboratory evaluations, and vaccination coverage. RESULTS: From March 24, 2014, through July 23, 2014, a total of 383 outbreak-related cases of measles were reported in nine counties in Ohio. The median age of case patients was 15 years (range, <1 to 53); a total of 178 of the case patients (46%) were female, and 340 (89%) were unvaccinated. Transmission took place primarily within households (68% of cases). The virus strain was genotype D9, which was circulating in the Philippines at the time of the reporting period. Measles-mumps-rubella (MMR) vaccination coverage with at least a single dose was estimated to be 14% in affected Amish households and more than 88% in the general (non-Amish) Ohio community. Containment efforts included isolation of case patients, quarantine of susceptible persons, and administration of the MMR vaccine to more than 10,000 persons. The spread of measles was limited almost exclusively to the Amish community (accounting for 99% of case patients) and affected only approximately 1% of the estimated 32,630 Amish persons in the settlement. CONCLUSIONS: The key epidemiologic features of a measles outbreak in the Amish community in Ohio were transmission primarily within households, the small proportion of Amish people affected, and the large number of people in the Amish community who sought vaccination. As a result of targeted containment efforts, and high baseline coverage in the general community, there was limited spread beyond the Amish community. (Funded by the Ohio Department of Health and the Centers for Disease Control and Prevention.).
Assuntos
Amish/estatística & dados numéricos , Surtos de Doenças , Vacina contra Sarampo-Caxumba-Rubéola/administração & dosagem , Sarampo/epidemiologia , Vacinação/estatística & dados numéricos , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Sarampo/transmissão , Pessoa de Meia-Idade , Ohio/epidemiologiaRESUMO
Elemental mercury (Hg0) formation from other mercury species in seawater results from photoreduction and microbial activity, leading to possible evasion from seawater to overlying air. Microbial conversion of monomethylmercury (MeHg) to Hg0 in seawater remains unquantified. A rapid radioassay method was developed using gamma-emitting 203Hg as a tracer to evaluate Hg0 production from Hg(II) and MeHg in the low pM range. Bacterioplankton assemblages in Atlantic surface seawater and Long Island Sound water were found to rapidly produce Hg0, with production rate constants being directly related to bacterial biomass and independent of dissolved Hg(II) and MeHg concentrations. About 32% of Hg(II) and 19% of MeHg were converted to Hg0 in 4 d in Atlantic surface seawater containing low bacterial biomass, and in Long Island Sound water with higher bacterial biomass, 54% of Hg(II) and 8% of MeHg were transformed to Hg0. Decreasing temperatures from 24°C to 4°C reduced Hg0 production rates cell-1 from Hg(II) 3.3 times as much as from a MeHg source. Because Hg0 production rates were linearly related to microbial biomass and temperature, and microbial mercuric reductase was detected in our field samples, we inferred that microbial metabolic activities and enzymatic reactions primarily govern Hg0 formation in subsurface waters where light penetration is diminished.
RESUMO
OBJECTIVES: Infanticide by males is common in mammals. According to the sexual selection hypothesis, the risk is inversely related to infant age because the older the infant, the less infanticide can shorten lactational amenorrhea; risk is also predicted to increase when an infanticidal male's chance of siring the replacement infant is high. Infanticide occurs in chimpanzees (Pan troglodytes), a species in which male dominance rank predicts paternity skew. Infanticidal male chimpanzees (if low-ranking) are unlikely to kill their own offspring, whereas those who are currently rising in rank, particularly when this rise is dramatic, have an increased likelihood of fathering potential future infants relative to any existing ones. Given that mothers should behave in ways that reduce infanticide risk, we predicted that female chimpanzees, and specifically those with younger, more vulnerable infants, would attempt to adjust the exposure of their infants to potentially infanticidal males. Specifically, mothers of young infants should reduce their association with adult males in general, and to a greater extent, with both low-ranking males and those rising in rank from a position where paternity of current infants was unlikely, to a rank where the probability of siring the next infant is significantly higher. We also investigated the alternative possibility that rather than avoiding all adult males, mothers would increase association with males of stable high rank on the basis that such males could offer protection against infanticide. MATERIALS AND METHODS: We examined data on female association patterns collected from the Budongo Forest, Uganda, during a period encompassing both relative stability in the male hierarchy and a period of instability with a mid-ranking male rising rapidly in rank. RESULTS: Using linear mixed models, we found that mothers reduced their association with the rank-rising male, contingent on infant age, during the period of instability. We also found evidence that females preferentially associated with a potential protector male during the high-risk period. DISCUSSION: Our results support the sexually selected hypothesis for infanticide and demonstrate that female chimpanzees are sensitive to the relative risks posed by adult males.
Assuntos
Agressão/fisiologia , Comportamento Animal/fisiologia , Pan troglodytes/fisiologia , Animais , Animais Recém-Nascidos , Feminino , Hierarquia Social , Masculino , MãesRESUMO
Cyclic electron flow around photosystem I (CEF) is critical for balancing the photosynthetic energy budget of the chloroplast by generating ATP without net production of NADPH. We demonstrate that the chloroplast NADPH dehydrogenase complex, a homolog to respiratory Complex I, pumps approximately two protons from the chloroplast stroma to the lumen per electron transferred from ferredoxin to plastoquinone, effectively increasing the efficiency of ATP production via CEF by 2-fold compared with CEF pathways involving non-proton-pumping plastoquinone reductases. By virtue of this proton-pumping stoichiometry, we hypothesize that NADPH dehydrogenase not only efficiently contributes to ATP production but operates near thermodynamic reversibility, with potentially important consequences for remediating mismatches in the thylakoid energy budget.
Assuntos
Arabidopsis/enzimologia , Cloroplastos/enzimologia , Modelos Moleculares , NADPH Desidrogenase/metabolismo , Complexo de Proteína do Fotossistema I/metabolismo , Folhas de Planta/enzimologia , Spinacia oleracea/enzimologia , Trifosfato de Adenosina/metabolismo , Algoritmos , Biocatálise , Domínio Catalítico , Transporte de Elétrons , Ferredoxinas/química , Ferredoxinas/metabolismo , Cinética , NADPH Desidrogenase/química , NADPH Desidrogenase/isolamento & purificação , Complexo de Proteína do Fotossistema I/química , Complexo de Proteína do Fotossistema I/isolamento & purificação , Plastoquinona/química , Plastoquinona/metabolismo , Conformação Proteica , Domínios e Motivos de Interação entre Proteínas , Subunidades Proteicas/química , Subunidades Proteicas/isolamento & purificação , Subunidades Proteicas/metabolismo , Especificidade da Espécie , TermodinâmicaRESUMO
We quantified measles transmissibility during a measles outbreak in Ohio in 2014 to evaluate the impact of public health responses. Case incidence and the serial interval (time between symptom onset in primary cases and secondary cases) were used to assess trends in the effective reproduction number R (the average number of secondary cases generated per case). A mathematical model was parameterized using early R values to determine the size and duration of the outbreak that would have occurred if containment measures had not been initiated, as well as the impact of vaccination. As containment started, we found a 4-fold decline in R (from approximately 4 to 1) over the course of 2 weeks and maintenance of R < 1 as control measures continued. Under a conservative scenario, the model estimated 8,472 cases (90% confidence interval (CI): 8,447, 8,489) over 195 days (90% CI: 179, 223) without control efforts and 715 cases (90% CI: 103, 1,338) over 128 days (90% CI: 117, 139) when vaccination was included; 7,757 fewer cases (90% CI: 7,130, 8,365) and 67 fewer outbreak days (90% CI: 48, 98) were attributed to vaccination. Vaccination may not account entirely for transmission reductions, suggesting that changes in community behavior (social distancing) and other control efforts (isolation, quarantining) are important. Our findings highlight the benefits of measles outbreak response and of understanding behavior change dynamics.
Assuntos
Sarampo/transmissão , Modelos Biológicos , Prática de Saúde Pública , Surtos de Doenças , Humanos , Sarampo/epidemiologia , Ohio/epidemiologiaRESUMO
We analyzed publicly available data of Fukushima 137Cs concentrations in coastal fish, in surface and bottom waters, and in surface marine sediments and found that within the first year of the accident pelagic fish lost 137Cs at much faster rates (mean of â¼1.3% d-1) than benthic fish (mean of â¼0.1% d-1), with benthopelagic fish having intermediate loss rates (mean of â¼0.2% d-1). The loss rates of 137Cs in benthic fish in the first year were more comparable to the decline of 137Cs concentrations in sediments (0.03% d-1), and the declines in pelagic fish were more comparable to the declines in seawater. Retention patterns of 137Cs in pelagic fish were comparable to that in laboratory studies of fish in which there were no sustained 137Cs sources, whereas the benthopelagic and benthic fish species retained 137Cs to a greater extent, consistent with the idea that there is a sustained additional 137Cs source for these fish. These field data, based on 13â¯511 data points in which 137Cs was above the detection limit, are consistent with conclusions from laboratory experiments that demonstrate that benthic fish can acquire 137Cs from sediments, primarily through benthic invertebrates that contribute to the diet of these fish.
Assuntos
Acidente Nuclear de Fukushima , Monitoramento de Radiação , Poluentes Radioativos da Água , Animais , Radioisótopos de Césio , Sedimentos Geológicos , Japão , Água do MarRESUMO
Pelagic ecosystems are changing due to environmental and anthropogenic forces, with uncertain consequences for the ocean's top predators. Epipelagic and mesopelagic prey resources differ in quality and quantity, but their relative contribution to predator diets has been difficult to track. We measured mercury (Hg) stable isotopes in young (<2 years old) Pacific bluefin tuna (PBFT) and their prey species to explore the influence of foraging depth on growth and methylmercury (MeHg) exposure. PBFT total Hg (THg) in muscle ranged from 0.61 to 1.93 µg g-1 dw (1.31 µg g-1 dw ±0.37 SD; 99% ± 6% MeHg) and prey ranged from 0.01 to 1.76 µg g-1 dw (0.13 µg g-1 dw ±0.19 SD; 85% ± 18% MeHg). A systematic decrease in prey δ202Hg and Δ199Hg with increasing depth of occurrence and discrete isotopic signatures of epipelagic prey (δ202Hg: 0.74 to 1.49; Δ199Hg: 1.76-2.96) and mesopelagic prey (δ202Hg: 0.09 to 0.90; Δ199Hg: 0.62-1.95) allowed the use of Hg isotopes to track PBFT foraging depth. An isotopic mixing model was used to estimate the dietary proportion of mesopelagic prey in PBFT, which ranged from 17% to 55%. Increased mesopelagic foraging was significantly correlated with slower growth and higher MeHg concentrations in PBFT. The slower observed growth rates suggest that prey availability and quality could reduce the production of PBFT biomass.
Assuntos
Mercúrio , Compostos de Metilmercúrio , Poluentes Químicos da Água , Animais , Ecossistema , Monitoramento Ambiental , Isótopos , Isótopos de Mercúrio , AtumRESUMO
Cytochrome bc1 is a proven drug target in the prevention and treatment of malaria. The rise in drug-resistant strains of Plasmodium falciparum, the organism responsible for malaria, has generated a global effort in designing new classes of drugs. Much of the design/redesign work on overcoming this resistance has been focused on compounds that are presumed to bind the Q(o) site (one of two potential binding sites within cytochrome bc1 using the known crystal structure of this large membrane-bound macromolecular complex via in silico modeling. Cocrystallization of the cytochrome bc1 complex with the 4(1H)-pyridone class of inhibitors, GSK932121 and GW844520, that have been shown to be potent antimalarial agents in vivo, revealed that these inhibitors do not bind at the Q(o) site but bind at the Q(i )site. The discovery that these compounds bind at the Q(i) site may provide a molecular explanation for the cardiotoxicity and eventual failure of GSK932121 in phase-1 clinical trial and highlight the need for direct experimental observation of a compound bound to a target site before chemical optimization and development for clinical trials. The binding of the 4(1H)-pyridone class of inhibitors to Q(i) also explains the ability of this class to overcome parasite Q(o)-based atovaquone resistance and provides critical structural information for future design of new selective compounds with improved safety profiles.
Assuntos
Antimaláricos/metabolismo , Complexo III da Cadeia de Transporte de Elétrons/metabolismo , Piridonas/metabolismo , Sítios de Ligação , Complexo III da Cadeia de Transporte de Elétrons/química , Simulação de Acoplamento MolecularRESUMO
The chloroplast must regulate supply of reducing equivalents and ATP to meet rapid changes in downstream metabolic demands. Cyclic electron flow around photosystem I (CEF) is proposed to balance the ATP/NADPH budget by using reducing equivalents to drive plastoquinone reduction, leading to the generation of proton motive force and subsequent ATP synthesis. While high rates of CEF have been observed in vivo, isolated thylakoids show only very slow rates, suggesting that the activity of a key complex is lost or down-regulated upon isolation. We show that isolation of thylakoids while in the continuous presence of reduced thiol reductant dithiothreitol (DTT), but not oxidized DTT, maintains high CEF activity through an antimycin A sensitive ferredoxin:quinone reductase (FQR). Maintaining low concentrations (~2 mM) of reduced DTT while modulating the concentration of oxidized DTT leads to reversible activation/inactivation of CEF with an apparent midpoint potential of -306 mV (±10 mV) and n=2, consistent with redox modulation of a thiol/disulfide couple and thioredoxin-mediated regulation of the plastoquinone reductase involved in the antimycin A-sensitive pathway, possibly at the level of the PGRL1 protein. Based on proposed differences in regulatory modes, we propose that the FQR and NADPH:plastoquinone oxidoreductase (NDH) pathways for CEF are activated under different conditions and fulfill different roles in chloroplast energy balance.
Assuntos
Antimicina A/farmacologia , Complexo de Proteína do Fotossistema I/metabolismo , Tilacoides/metabolismo , Elétrons , Ferredoxinas/metabolismo , Oxirredução , Quinona Redutases/metabolismoRESUMO
The 2011 release of Fukushima-derived radionuclides into the Pacific Ocean made migratory sharks, teleosts, and marine mammals a source of speculation and anxiety regarding radiocesium (134+137Cs) contamination, despite a lack of actual radiocesium measurements for these taxa. We measured radiocesium in a diverse suite of large predators from the North Pacific Ocean and report no detectable (i.e., ≥ 0.1 Bq kg-1 dry wt) Fukushima-derived 134Cs in all samples, except in one olive ridley sea turtle (Lepidochelys olivacea) with trace levels (0.1 Bq kg-1). Levels of 137Cs varied within and across taxa, but were generally consistent with pre-Fukushima levels and were lower than naturally occurring 40K by one to one to two orders of magnitude. Predator size had a weaker effect on 137Cs and 40K levels than tissue lipid content. Predator stable isotope values (δ13C and δ15N) were used to infer recent migration patterns, and showed that predators in the central, eastern, and western Pacific should not be assumed to accumulate detectable levels of radiocesium a priori. Nondetection of 134Cs and low levels of 137Cs in diverse marine megafauna far from Fukushima confirms negligible increases in radiocesium, with levels comparable to those prior to the release from Fukushima. Reported levels can inform recently developed models of cesium transport and bioaccumulation in marine species.
Assuntos
Acidente Nuclear de Fukushima , Monitoramento de Radiação , Poluentes Radioativos da Água , Animais , Radioisótopos de Césio , Cadeia Alimentar , Japão , Oceano Pacífico , Tubarões , TartarugasRESUMO
The largest bioconcentration step of most metals, including methylmercury (MeHg), in aquatic biota is from water to phytoplankton, but the extent to which dissolved organic matter (DOM) affects this process for MeHg largely remains unexplored in marine systems. This study investigated the influence of specific sulfur-containing organic compounds and naturally occurring DOM on the accumulation of MeHg in a marine diatom Thalassiosira pseudonana. Initial uptake rate constants and volume concentration factors (VCFs) of MeHg were calculated to evaluate MeHg enrichment in algal cells in the presence of a range of organic compound concentrations. At environmentally realistic and higher concentrations, the addition of glycine and methionine had no effect on algal MeHg uptake, but thiol-containing compounds such as cysteine and thioglycolic acid reduced MeHg accumulation in algal cells at high added concentrations (> 100 times higher than naturally occurring concentrations). However, environmentally realistic concentrations of glutathione, another thiol-containing compound as low as 10 nM, resulted in a decline of ~ 30% in VCFs, suggesting its possible importance in natural waters. Humic acid additions of 0.1 and 0.5 mg C/L also reduced MeHg VCFs by ~ 15% and ~ 25%, respectively. The bioaccumulation of MeHg for T. pseudonana in coastal waters with varying levels of dissolved organic carbon (DOC) was inversely correlated with bulk DOC concentrations. Generally, naturally occurring DOM, particularly certain thiol-containing compounds, can reduce MeHg uptake by phytoplankton.
RESUMO
The respiratory chain bc1 complex is central to mitochondrial bioenergetics and the target of antiprotozoals. We characterized a modified yeast bc1 complex that more closely resemble Plasmodium falciparum enzyme. The mutant version was generated by replacing ten cytochrome b Qo site residues by P. falciparum equivalents. The Plasmodium-like changes caused a major dysfunction of the catalytic mechanism of the bc1 complex resulting in superoxide overproduction and respiratory growth defect. The defect was corrected by substitution of the conserved residue Y279 by a phenylalanine, or by mutations in or in the vicinity of the hinge domain of the iron-sulphur protein. It thus appears that side-reactions can be prevented by the substitution Y279F or the modification of the iron-sulphur protein hinge region. Interestingly, P. falciparum - and all the apicomplexan - contains an unusual hinge region. We replaced the yeast hinge region by the Plasmodium version and combined it with the Plasmodium-like version of the Qo site. This combination restored the respiratory growth competence. It could be suggested that, in the apicomplexan, the hinge region and the cytochrome b Qo site have co-evolved to maintain catalytic efficiency of the bc1 complex Qo site.