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OBJECTIVES: This study evaluated the effectiveness of COVID-19 vaccines in preventing symptomatic and severe disease. STUDY DESIGN: This was an observational test-negative case-control study. METHODS: Study participants were adults with at least one symptom included in the World Health Organization COVID-19 definition who sought health care in a public emergency department between 1 November 2021 and 2 March 2022 (corresponding with the fifth pandemic wave in Portugal dominated by the Omicron variant). This study used multivariable logistic regression models to estimate and compare the odds ratio of vaccination between test-positive cases and test-negative controls to calculate the absolute and relative vaccine effectiveness. RESULTS: The study included 1059 individuals (522 cases and 537 controls) with a median age of 56 years and 58% were women. Compared with the effectiveness of the primary vaccination scheme that had been completed ≥180 days earlier, the relative effectiveness against symptomatic infection of a booster administered between 14 and 132 days earlier was 71% (95% confidence interval [CI]: 57%, 81%; P < 0.001). The effectiveness of the primary series against symptomatic infection peaked at 85% (95% CI: 56%, 95%) between 14 and 90 days after the last inoculation and decreased to 34% (95% CI: -43%, 50%) after ≥180 days. CONCLUSIONS: Despite the known immunological evasion characteristics of the Omicron variant, results from this study show that vaccine effectiveness increases after booster administration. COVID-19 vaccine effectiveness decreases to less than 50% between 3 and 6 months after completion of the primary cycle; therefore, this would be an appropriate time to administer a booster to restore immunity.
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Vacinas contra COVID-19 , COVID-19 , Adulto , Humanos , Feminino , Pessoa de Meia-Idade , Masculino , Vacinas contra COVID-19/uso terapêutico , COVID-19/epidemiologia , COVID-19/prevenção & controle , Estudos de Casos e Controles , SARS-CoV-2RESUMO
This randomised study aimed to assess and compare the efficacy of treatment protocols containing levamisole, ivermectin, or moxidectin against Capillaria spp. in naturally infected European hedgehogs (Erinaceus europaeus) presented to a British wildlife rehabilitation centre. Faecal analysis, consisting of wet mount and flotation, was performed for 229 hedgehogs weighing ≥200g. Animals testing positive for Capillaria spp. (81%), excluding pregnant females, were randomly allocated a treatment protocol. Initially, hedgehogs (n = 50) received one of six 'pilot' protocols, whereas the remaining animals (n = 97) received one of three 'main' protocols. Faecal analysis was repeated on day 8 and day 12 after treatment initiation. Efficacy of each treatment was assessed based on Capillaria reduction rate (CRR), weight gain, presence of respiratory clinical signs, and outcome. Pilot protocols containing only moxidectin had a significantly lower CRR (≥28.1%) compared to those with levamisole or ivermectin (≥86.6%), whereas the main protocols containing levamisole had a significantly higher CRR (≥93.0%) compared to those containing only ivermectin (≥69.3%). Clinical parameters did not differ significantly between treatments, but animals with respiratory clinical signs at the end of the trial were significantly more likely to have lower CRR and test positive for Crenosoma striatum. C. striatum often appeared refractory to treatment, and managing these infections requires additional anthelmintic therapy. Based on the formulations and dosages trialled, moxidectin is not recommended for treating capillariosis in European hedgehogs, whereas levamisole given orally for two consecutive days at 25-35 mg/kg is suggested as the treatment of choice.
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Anti-Helmínticos , Ivermectina , Feminino , Animais , Ivermectina/farmacologia , Ivermectina/uso terapêutico , Levamisol/uso terapêutico , Capillaria , Macrolídeos/farmacologia , Macrolídeos/uso terapêutico , Anti-Helmínticos/uso terapêutico , Ouriços , Fezes , Contagem de Ovos de ParasitasRESUMO
AIM: Physiological race determination of 143 Fusarium oxysporum f. sp. lycopersici (FOL) isolates collected along 30 years in major tomato-producing regions of Brazil. MATERIALS AND RESULTS: Physiological races were determined via root-dipping inoculation of differential tomato accessions and by the PCR-based marker system of Hirano and Arie (2006). According to pathogenicity/virulence assays, five race 1, 23 race 2 and 115 race 3 isolates were identified. FOL race 1 and 2 isolates prevailed up to early 2000s. Afterwards, the large majority of the isolates was classified as the invasive race 3. Novel reports of race 3 were done in five states, thus expanding its geographical distribution. Using this PCR-based marker system, a precise discrimination was observed for all race 3 isolates. However, all race 1 and 2 isolates displayed only the cosmopolitan race 1-specific amplicon pattern. CONCLUSION: The development and/or validation of novel race-specific marker systems are necessary to allow a precise discrimination of the potentially endemic Brazilian FOL race 2. SIGNIFICANCE AND IMPACT OF THE STUDY: The present characterization of isolates indicates that distinct evolutionary mechanisms are acting to select new FOL races and/or genetic variants across agroecosystems around the globe.
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Fusarium , Doenças das Plantas/microbiologia , Solanum lycopersicum , Brasil , Fusarium/genética , Fusarium/patogenicidade , Solanum lycopersicum/microbiologia , VirulênciaRESUMO
Currently, the lack of reliable strategies for the diagnosis and treatment of cancer makes the identification and characterization of new therapeutic targets a pressing matter. Several studies have proposed the Six Transmembrane Epithelial Antigen of the Prostate 1 (STEAP1) as a promising therapeutic target for prostate cancer. Although structural and functional studies may provide deeper insights on the role of STEAP1 in cancer, such techniques require high amounts of purified protein through biotechnological processes. Based on the results presented, this work proposes the application, for the first time, of a fed-batch profile to improve STEAP1 biosynthesis in mini-bioreactor Komagataella pastoris X-33 Mut+ methanol-induced cultures, by evaluating three glycerol feeding profiles-constant, exponential, and gradient-during the pre-induction phase. Interestingly, different glycerol feeding profiles produced differently processed STEAP1. This platform was optimized using a combination of chemical chaperones for ensuring the structural stabilization and appropriate processing of the target protein. The supplementation of culture medium with 6 % (v/v) DMSO and 1 M proline onto a gradient glycerol/constant methanol feeding promoted increased biosynthesis levels of STEAP1 and minimized aggregation events. Deglycosylation assays with peptide N-glycosidase F showed that glycerol constant feed is associated with an N-glycosylated pattern of STEAP1. The biological activity of recombinant STEAP1 was also validated, once the protein enhanced the proliferation of LNCaP and PC3 cancer cells, in comparison with non-tumoral cell cultures. This methodology could be a crucial starting point for large-scale production of active and stable conformation of recombinant human STEAP1. Thus, it could open up new strategies to unveil the structural rearrangement of STEAP1 and to better understand the biological role of the protein in cancer onset and progression.
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Antígenos de Neoplasias/biossíntese , Glicerol , Metanol , Oxirredutases/biossíntese , Proteínas Recombinantes/biossíntese , Humanos , Pichia , Regiões Promotoras Genéticas , SaccharomycetalesRESUMO
Childhood-onset systemic lupus erythematosus (cSLE) is a chronic inflammatory multisystem autoimmune disease that requires multiple differential diagnoses. Munchausen by proxy syndrome (MBPS) is a form of child abuse, where a caregiver intentionally creates a medical history and induces or fabricates signs or disease in a patient. To our knowledge, there is no case report of MBPS mimicking cSLE diagnosis. We reported herein a 9-year-old male patient, with a history of multiple hospitalizations due to seizures with altered levels of consciousness. The mother reported malar rash, photosensitivity, alopecia, arthralgia, arterial hypertension, macroscopic hematuria, seizure and positive antinuclear antibodies. In the other service, he was treated with intravenous methylprednisolone, prednisone and mycophenolate mofetil. At 8 years and 8 months, he was admitted to our tertiary center with history of fever and macroscopic hematuria. Laboratory examinations were normal, including negative for antinuclear antibodies, anti-double stranded DNA, anticardiolipin, anti-Ro/SSA, anti-La/SSB, anti-RNP and anti-Sm antibodies. Multiple urine cultures revealed the presence of Enterococcus faecium, Acinetobacter sp., Stenotrophomonas maltophilia and Serratia marcescens, without any association with pyuria. At 8 years and 9 months, he was readmitted at emergency room with history of severe fever, headache, vomiting, photophobia, phonophobia and dizziness. The physical examination showed agitation, confusion, ataxic gait, slurred speech, horizontal nystagmus, painful facial expressions, tachycardia and weight loss. Brain magnetic resonance angiography and cerebrospinal fluid analysis were normal. During hospitalization, he had an acute episode of epistaxis and otalgia with excoriation in the auditory canal. At that moment, the suspicion of MBPS mimicking cSLE was raised and phenytoin intoxication was confirmed (peak phenytoin concentration was 45.4 mcg/mL, therapeutic range 10-20 mcg/mL). The mother and the patient were immediately separated, and she was replaced by another legal guardian. One week later, the neurological and other signs and symptoms were completely resolved. The child was placed under paternal custody with a court order and moved to another state. After that, the mother reported phenytoin use for her child and was referred to psychiatric follow-up. In conclusion, the first case of MBPS mimicking cSLE, resulting in multiple unnecessary examinations and treatments with delayed diagnosis was reported.
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Síndrome de Munchausen Causada por Terceiro/diagnóstico , Idade de Início , Criança , Diagnóstico Tardio , Diagnóstico Diferencial , Humanos , Lúpus Eritematoso Sistêmico/diagnóstico , Masculino , Procedimentos DesnecessáriosAssuntos
Ivermectina , Levamisol , Animais , Ivermectina/farmacologia , Levamisol/farmacologia , Capillaria , Macrolídeos/farmacologia , OuriçosRESUMO
Citrus canker, caused by the Gram-negative bacterium Xanthomonas citri subsp citri (Xac), severely affects most economically important citrus varieties worldwide. A previous study showed that disruption of the ORF XAC1201 from the Xac 306 strain by transposon Tn5 decreased bacterium virulence in the Rangpur lime host (Citrus limonia L. Osbeck). However, little is known regarding the possible function of the hypothetical protein XAC1201 and how it affects the virulence of Xac 306. Here, we confirmed that disruption of ORF XAC1201 reduces Xac 306 virulence in two different hosts, delaying the onset of typical symptoms. In silico analysis suggested that XAC1201 interacts with the flagellar proteins FliM and FliL, known to be an important factor for virulence. In fact, motility assays revealed that the XAC1201 mutant has a significant difference in motility compared to the wild-type Xac 306. Also, a 3-D structure model revealed modified cofactor binding sites and suggested that XAC1201 has a non-functional HD domain. This hypothesis was confirmed by enzymatic assays performed in purified, XAC1201 recombinant protein expressed in Escherichia coli, which revealed no significant activities previously associated with HD domains for the tested substrates. Thus, the role of the XAC1201 protein in Xac 306 virulence seems to be related to flagellar motility, although a non-classic role for the HD domain cannot be dismissed.
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Flagelos/metabolismo , Movimento , Fases de Leitura Aberta , Xanthomonas/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Flagelos/genética , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Mutação , Ligação Proteica , Domínios Proteicos , Virulência/genética , Xanthomonas/patogenicidade , Xanthomonas/fisiologiaRESUMO
Modeling and accurately forecasting trend and seasonal patterns of a time series is a crucial activity in economics. The main propose of this study is to evaluate and compare the performance of three traditional forecasting methods, namely the ARIMA models and their extensions, the classical decomposition time series associated with multiple linear regression models with correlated errors, and the Holt-Winters method. These methodologies are applied to retail time series from seven different European countries that present strong trend and seasonal fluctuations. In general, the results indicate that all the forecasting models somehow follow the seasonal pattern exhibited in the data. Based on mean squared error (MSE), root mean squared error (RMSE), mean absolute percentage error (MAPE), mean absolute scaled error (MASE) and U-Theil statistic, the results demonstrate the superiority of the ARIMA model over the other two forecasting approaches. Holt-Winters method also produces accurate forecasts, so it is considered a viable alternative to ARIMA. The performance of the forecasting methods in terms of coverage rates matches the results for accuracy measures.
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Climate change further the world's human population increase is a mainstream political issue, and it's critical to search for solutions to produce enough food to feed everyone. Pesticides and fertilizers have been used as an easy solution to prevent pests and increase food production. Nevertheless, their overuse has dangerous effects on the ecosystems and communities. Oxyfluorfen (Oxy) and copper (Cu) based formulations are used as pesticides and widely applied on agricultural fields for crop protection. However, they have shown negative effects on non-target species. So, this work proposes to: a)determine the lethal concentration of Oxy and Cu to the zooplankton, Artemia franciscana, at different temperatures (15 °C, 20 °C and 25 °C); b)understand the biochemical impacts of these chemicals at the different temperatures scenarios, on A. franciscana and c)evaluate the impact of the climate changes, particularly the temperature increase, on this species sensitivity to the tested pesticides. Acute and sub-lethal bioassays with Oxy and Cu were performed at different temperatures to determine the lethal concentration of each chemical and to understand the effects of the compounds at different temperatures on the biochemical profiles of A. franciscana. Results showed an increase in chemicals toxicity with the temperature, and Oxy was revealed to be more noxious to A. franciscana than Cu; at a biochemical level, significant differences were observed among temperatures, with the biggest differences between the organisms exposed to 15 °C and 25 °C. Overall, a decrease in fatty acids (FA) and sugars was observed with the increase in Cu and oxyfluorfen concentrations. Different trends were observed with temperature increase, with FA increase in the organisms exposed to Cu and the opposite was observed in the ones exposed to oxyfluorfen. Sugar content decreases in the organisms exposed to oxyfluorfen with temperature increase and showed a non-linear behaviour in the ones exposed to Control and Cu treatments.
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Artemia , Cobre , Éteres Difenil Halogenados , Praguicidas , Temperatura , Animais , Cobre/toxicidade , Éteres Difenil Halogenados/toxicidade , Artemia/efeitos dos fármacos , Praguicidas/toxicidade , Praguicidas/análise , Poluentes Químicos da Água/toxicidade , Poluentes Químicos da Água/análise , Mudança ClimáticaRESUMO
The three races of Fusarium oxysporum f. sp. lycopersici (FOL) are important tomato pathogens throughout the world, causing severe economic losses (1). In Brazil, races 1 and 2 are widespread, but the current geographic distribution of race 3 is restricted to the mild climate areas of Espírito Santo and Rio de Janeiro States in the southeast region (2,3). Here we report the spread of FOL race 3 to the warm northeast region of Brazil. Plants in commercial fields of the hybrid 'Alambra' (resistant to FOL races 1 and 2) were found displaying chlorosis, vascular browning, and wilt symptoms in Jaguaquara County, Bahia State, Brazil. Disease incidence ranged from 10 to 50%. The virulence profile of six isolates obtained from three distinct tomato-producing fields was investigated by root-dipping inoculation (106 conidia/ml) of 21-day-old seedlings from a set of FOL race differential accessions: 'Ponderosa' (susceptible to all races), 'IPA-5' (FOL race 1 resistance; I-1 locus); 'Alambra' and 'Floradade' (FOL races 1 and 2 resistance; I-2 gene), and Solanum pennellii 'LA 716' (resistant to all three races; I-3 locus). All six isolates were able to induce severe wilt symptoms in 100% of the plants from all lines but S. pennellii 'LA 716'. FOL race 3 identity was confirmed via PCR assays employing a specific set of primers that are able to discriminate all the three FOL races as well as F. oxysporum f. sp. radicis-lycopersici isolates (1). Total DNA was extracted from pure fungal colonies growing in agar medium. The typical FOL race 3 amplicon profiles (i.e. positive for the primers uni, sp13, and sp23 and negative for the primer sprl) were observed only in the six FOL 3 isolates from Bahia as well as in five reference isolates of race 3 (previously obtained from tomato in Espírito Santo and Rio de Janeiro States), thus confirming their race identities. This recent, fast, and wide geographic expansion of the FOL race 3 in Brazil suggests that the pathogen has been introduced into new tomato producing areas via either contaminated seeds or seedlings. Because of the complexity of establishing effective chemical and cultural control strategies, these epidemics caused by FOL race 3 in distinct areas of Brazil might cause the replacement of the currently grown susceptible hybrids by resistant ones. References: (1) Y. Hirano and T. Arie. J. Gen. Plant Pathol. 72:273, 2006; (2) A. Reis et al. Fitopatol. Bras. 30:426, 2005; (3) A. Reis and L. S. Boiteux. Hort. Bras. 25:451, 2007.
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Multilayer epitaxial graphene was obtained from a 6H-SiC(001) substrate subjected to a temperature gradient from 1250 to 1450 °C. Scanning tunneling microscopy and x-ray diffraction were used to identify the structure and morphology of the surface, from which the formation of a metastable phase was inferred. By a comparison between microscopy and diffraction data, we report the appearance of misoriented Si-doped graphene in cold regions (1250 °C) of the substrate. This metastable phase occurs in domains where silicon sublimation is incomplete and it coexists with small domains of epitaxial graphene. At 1350 °C this phase disappears and one observes complete graphene-like layers (although misoriented), where rotational registry between the underlying epitaxial graphene and additional layers is absent. At 1450 °C the stacking among layers is established and the formation of highly oriented single crystalline graphite is complete. The stability of this Si-rich metastable phase at 1250 °C was confirmed by first-principles calculations based on the density functional theory.
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A systematic study of the evolution of the electronic behavior and atomic structure of multilayer epitaxial graphene (MEG) as a function of growth time was performed. MEG was obtained by sublimation of a 4H-SiC(0001(-)) substrate in an argon atmosphere. Raman spectroscopy and x-ray diffraction were carried out in samples grown for different times. For 30 min of growth the sample Raman signal is similar to that of graphite, while for 60 min the spectrum becomes equivalent to that of exfoliated graphene. Conventional x-ray diffraction reveals that all the samples have two different (0001) lattice spacings. Grazing incidence x-ray diffraction shows that thin films are composed of rotated (commensurate) structures formed by adjacent graphene layers. Thick films are almost completely disordered. This result can be directly correlated to the single layer electronic behavior of the films as observed by Raman spectroscopy. Finally, to understand the change in lattice spacings as a result of layer rotation, we have carried out first principles calculations (using density functional theory) of the observed commensurate structures.
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The toxicity of single and combined formulated herbicides (Mikado and Viper) was assessed on several endpoints in species from two trophic levels: algae growth-Pseudokirchneriella subcapitata and Chlorella vulgaris-immobilization and life-history traits (only for single compound toxicity) of daphnids-Daphnia longispina and Daphnia magna. Viper was the most toxic formulated herbicide. It was hypothesized that the toxicity of both formulated herbicides could have been enhanced by adjuvants, especially for Viper. In most cases, the sublethal endpoints were the most sensitive and affected by both formulations, comparatively to their acute effects. Concentration addition (CA) and independent action (IA) models provided an accurate description of Mikado and Viper joint action on algae growth and immobilization of daphnids, although significant deviations were always detected. A low-dose antagonism and high-dose synergism were identified for P. subcapitata, whereas C. vulgaris response deviated antagonistically from CA and synergistically from IA. For both daphnids, however, synergistic effects were observed for higher mixture concentrations. Under a regulatory standpoint, CA provided the most conservative estimation either because the mixture effects were overestimated or less subestimated than IA. Overall, the great sensitivity differences observed within species did not allow the conclusion that one trophic level was more tolerant than the other. Instead, P. subcapitata was always the most sensitive species to both herbicide formulations, followed by D. longispina, while D. magna and C. vulgaris were the most tolerant species. On a whole, further studies are needed toward a comprehensive understanding of herbicides mode of action, their effects at lower biological-level endpoints, and under different mixture designs.
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Chlorella vulgaris/efeitos dos fármacos , Daphnia/efeitos dos fármacos , Herbicidas/toxicidade , Animais , Chlorella vulgaris/crescimento & desenvolvimento , Clorófitas/efeitos dos fármacos , Clorófitas/crescimento & desenvolvimento , Daphnia/crescimento & desenvolvimento , Sinergismo Farmacológico , Ecotoxicologia , Testes de ToxicidadeRESUMO
The Six-Transmembrane Epithelial Antigen of the Prostate 1 (STEAP1) is an integral membrane protein involved in cellular communications, in the stimulation of cell proliferation by increasing Reactive Oxygen Species levels, and in the transmembrane-electron transport and reduction of extracellular metal-ion complexes. The STEAP1 is particularly over-expressed in prostate cancer, in contrast with non-tumoral tissues and vital organs, contributing to tumor progression and aggressiveness. However, the current understanding of STEAP1 lacks experimental data on the respective molecular mechanisms, structural determinants, and chemical modifications. This scenario highlights the relevance of exploring the biosynthesis of STEAP1 and its purification for further bio-interaction and structural characterization studies. In this work, recombinant hexahistidine-tagged human STEAP1 (rhSTEAP1-His6) was expressed in Komagataella pastoris (K. pastoris) mini-bioreactor methanol-induced cultures and successfully solubilized with Nonidet P-40 (NP-40) and n-Decyl-ß-D-Maltopyranoside (DM) detergents. The fraction capacity of Phenyl-, Butyl-, and Octyl-Sepharose hydrophobic matrices were evaluated by manipulating the ionic strength of binding and elution steps. Alternatively, immobilized metal affinity chromatography packed with nickel or cobalt were also studied in the isolation of rhSTEAP1-His6 from lysate extracts. Overall, the Phenyl-Sepharose and Nickel-based resins provided the desired selectivity for rhSTEAP1-His6 capture from NP-40 and DM detergent-solubilized K. pastoris extracts, respectively. After a polishing step using the anion-exchanger Q-Sepharose, a highly pure, fully solubilized, and immunoreactive 35 kDa rhSTEAP1-His6 fraction was obtained. Altogether, the established reproducible strategy for the purification of rhSTEAP1-His6 paves the way to gather additional insights on structural, thermal, and environmental stability characterization significantly contributing for the elucidation of the functional role and oncogenic behavior of the STEAP1 in prostate cancer microenvironment.
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Detergentes , Próstata , Masculino , Humanos , Próstata/metabolismo , Próstata/patologia , Antígenos de Neoplasias/metabolismo , Níquel , Reatores Biológicos , Oxirredutases/metabolismoRESUMO
Hydrogen/sodium titanate nanotubes (TNTs) were investigated as hydrogen (H(2)) sensors. TNT films exhibit good sensing properties and a large response, in particular at room temperature. Electrical conductivity measurements performed under different atmospheres from 25 to 300 °C indicate that, for T > 100 °C, conduction is thermally activated and can be attributed to electronic transport, whereas for T < 100 °C conduction is dominated by protonic transport. The T dependence of the H(2) sensitivity was determined and related to this variation in the dominant transport mechanism. For low T, H(2) sensing originates from the modulation in protonic conduction. Such modulation was attributed to the creation/destruction of surface hydroxyl groups.
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The present work shows the application of nickel- and magnesium-crosslinked gellan microspheres in ionic and affinity capture strategies to directly extract hSCOMT from the complex Komagataella pastoris lysate through a simple batch method. Both formulations present similar morphology, but nickel-crosslinked microspheres present higher crosslinker content and smaller diameters. Four different capture strategies were established, by manipulating the ionic strength, pH, temperature and competing agents' presence. The most promising results for hSCOMT capture and clarification were obtained employing an ionic strategy with nickel-crosslinked microspheres and an affinity strategy with magnesium-crosslinked microspheres at 4 °C. The bioactivity results (200%) and purification degree (70%) of hSCOMT captured by the ionic strategy were more satisfactory probably due to the soft ionic conditions used (100 mM NaCl). For the first time, the gellan polysaccharide versatility was demonstrated in the microsphere application for the direct capture of hSCOMT from a complex lysate, simplifying isolation biotechnological procedures.
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Catecol O-Metiltransferase/química , Polímeros/química , Polissacarídeos Bacterianos/química , Proteínas Recombinantes/química , Saccharomycetales/química , Humanos , Concentração de Íons de Hidrogênio , Microesferas , Concentração Osmolar , TemperaturaRESUMO
In order to better understand the ossification processes in anurans our study was carried out on tadpoles and adults of Lithobates catesbeianus. In this sense, we characterized the kinetic properties of alkaline phosphatase with p-nitrophenylphosphatase (pNPP) and pyrophosphate (PPi) and evaluated the activities of tartrate-resistant acid phosphatase and acid phosphatase. The enzyme extracts were obtained from tadpoles and adult femurs, which were divided into epiphysis and diaphysis. After homogenization, the samples were submitted to differential centrifugation to obtain cell membranes and, further, to phospholipase C (PIPLC) treatment, to remove membrane-bound proteins anchored by phosphatidylinositol. The average of specific activity for pNPP hydrolysis (at pH 10.5) by alkaline phosphatase released by phosphatidylinositol-specific phospholipase C (PIPLC) from Bacillus cereus among different bone regions at different animal ages was 1,142.57 U.mg-1, while for PPi hydrolysis (at pH 8.0), it was 1,433.82 U.mg-1. Among the compounds tested for enzymatic activity, the one that influenced the most was EDTA, with approximately 67% of inhibition for pNPPase activity and 77% for PPase activity. In the case of kinetic parameters, the enzyme showed a "Michaelian" behavior for pNPP and PPi hydrolysis. The Km value was around 0.6mM for pNPPase activity and ranged from 0.01 to 0.11mM for PPase activity, indicating that the enzyme has a higher affinity for this substrate. The study of pNPP and PPi hydrolysis by the enzyme revealed that the optimum pH of actuation for pNPP was 10.5, while for PPi, which is considered the true substrate of alkaline phosphatase, was 8.0, close to the physiological value. The results show that regardless of the ossification type that occurs, the same enzyme or isoenzymes act on the different bone regions and different life stages of anurans. The similarity of the results of studies with other vertebrates shows that anurans can be considered excellent animal models for the study of biological calcification.
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Fosfatase Alcalina , Osteogênese , Fosfatase Alcalina/metabolismo , Animais , Osso e Ossos/metabolismo , Cinética , Rana catesbeianaRESUMO
The methodology used to extract and quantify microplastics (MPs) in aquatic systems are still not standardized. Salt saturated solutions, such as sodium chloride (NaCl), zinc chloride (ZnCl2) and/or sodium iodide (NaI), are normally added to separate dense plastics from aquatic samples. However, the most effective reagents are also the most expensive (e.g. ZnCl2 and NaI). To decrease this cost, a reuse process of the salt solutions should be applied. The reuse process has been widely investigated for the NaI solution neglecting the ZnCl2. Hence, the aim of this study was to present a simple methodology to reuse the ZnCl2 solution ensuring the efficiency of the product. Results of the present study showed that ZnCl2 solution could be reused at least five times maintaining an efficiency above 95 %. â¢The ZnCl2 reuse decreases the cost of the methodology.â¢The efficiency of ZnCl2 solution after five filtrations remains above 95 % (all polymers are detected and recovered).â¢The use of this salt solution is the most cost-effective methodology to isolate MPs from aquatic samples.
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Nudix pyrophosphatases are a well represented protein family in the Deinococcus radiodurans genome. These hydrolases, which are known to be enzymatically active towards nucleoside diphosphate derivatives, play a role in cleansing the cell pool of potentially deleterious damage products. Here, the structure of DR2204, the only ADP-ribose pyrophosphatase in the D. radiodurans genome that is known to be active towards flavin adenosine dinucleotide (FAD), is presented at 2.0 angstrom resolution.
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Proteínas de Bactérias/química , Deinococcus/enzimologia , Estrutura Quaternária de Proteína , Estrutura Terciária de Proteína , Pirofosfatases/química , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Cristalografia por Raios X , Modelos Moleculares , Dados de Sequência Molecular , Pirofosfatases/genética , Alinhamento de Sequência , Nudix HidrolasesRESUMO
Huge efforts have been made both in adopting more environmental-friendly bleaching processes, and in developing advanced oxidation processes and more effective biological treatments for the reduction of deleterious impacts of paper mill effluents. Even so, the success of such treatments is frequently reported in terms of chemical parameters without a proper evaluation of the effluent's toxicity mitigation. This is the first study reporting an exhaustive evaluation of the toxicity of a secondary bleached kraft pulp mill effluent, after either tertiary treatment with the soft-rot fungi Rhizopus oryzae or with a photo-Fenton oxidation, using a battery of freshwater species. As it has been reported the photo-Fenton/UV treatment has proved to be the most effective in reducing the colour and the COD (chemical oxygen demand) of the effluent. Nevertheless, extremely low EC(50) values were reported for almost all species, after this tertiary treatment. The treatment with R. oryzae was less effective in terms of colour removal and COD reduction, but proved to be the most promising in reducing toxicity.