RESUMO
BACKGROUND: Although there is growing demand for hospice care in China due to its aging population and increasing cancer rates, the sector remains slow to expand. Oncology nurses are the primary providers of hospice care, but little is known about their behaviors towards hospice care and related factors. METHODS: This cross-sectional study conveniently sampled 933 oncology nurses from six grade A tertiary hospitals in Hubei Province between January to March 2022. The questionnaire was composed of seven parts: general information (including sociodemographic and work-related information), hospice care behaviors, hospice care knowledge, hospice care attitudes, hospice care self-efficacy, hospice care outcome expectancy, and hospice care environment. Data were analyzed using descriptive analysis, independent sample t-tests, one-way ANOVA, Pearson's correlation, multiple linear regression, random forest regression, and BP neural network model analysis. RESULTS: A total of 852 questionnaires were valid. The mean score of hospice care behaviors was 50.47 ± 10.56, with a mean item score of 3.61 ± 0.75. The three highest scoring behaviors were "pain assessment of patients (4.21 ± 0.91)", "satisfying the physical and mental needs of dying patients (4.04 ± 0.92)", and "creating good relationships between the medical staff and family members (4.02 ± 0.87)". The two lowest-scoring behaviors were "proactively recommending medical institutions for hospice care to terminally ill patients and their families (2.55 ± 1.10)" and "proactively talking to patients and families about death-related topics for patients who are critically ill and cannot be reversed (2.87 ± 1.03)." Multiple linear regression, random forest regression, and BP neural network models all showed that the frequency of sharing hospice care experiences with colleagues, hospice care attitudes, hospice care self-efficacy, and hospice care environments were positively associated with hospice care behaviors. CONCLUSIONS: The frequency of hospice care behaviors among Chinese oncology nurses is generally at a moderate to high level. The results provide a basis for promoting hospice care behaviors among oncology nurses in order to improve the quality of life for terminally ill cancer patients.
Assuntos
Cuidados Paliativos na Terminalidade da Vida , Neoplasias , Enfermeiras e Enfermeiros , Recursos Humanos de Enfermagem Hospitalar , Humanos , Idoso , Cuidados Paliativos na Terminalidade da Vida/psicologia , Estudos Transversais , Qualidade de Vida , Recursos Humanos de Enfermagem Hospitalar/psicologia , Inquéritos e Questionários , Teoria Psicológica , Atitude do Pessoal de SaúdeRESUMO
BACKGROUND: WHO stated the environment is an important factor affecting the development of hospice care. The environment is the sum of factors affecting behavior besides the individual factors. Currently, a scale to comprehensively assess the hospice environment of nurse is still lacking. This study aimed to develop an instrument to investigate the environmental factors affecting hospice care of nurses. METHODS: Literature review and a semi-structured interview were conducted to form the items pool of the Hospice Care Environment Scale. Two rounds of Delphi expert consultation were conducted by 16 experts to revise the scale dimensions and entries to form the Hospice Care Environment Scale. A psychometric evaluation was then performed among 530 oncology nurses in a large tertiary oncology hospital in Hubei Province. The 500 valid questionnaires were randomly divided into two groups in a 1:1 ratio, sample 1 (n1 = 250) for item screening and sample 2 (n2 = 250) for quality evaluation of the resulting scale. Item analysis, reliability analysis, validity analysis and acceptability analysis were performed. RESULT: The Hospice Care Environment Scale consists of two dimensions and 13 entries. The Cronbach's α coefficient of the Hospice Care Environment Scale was 0.970, and the Cronbach's α coefficient of the two dimensions were 0.952 and 0.969, respectively, with the Item-content validity index and average Scale- content validity index of the scale was both 1.000. The validation factor analysis showed the standardized path coefficients of each item were basically above 0.5, and the factor structure model was stable and suitable. The average completion time of the scale was about 3 min, which had good feasibility. CONCLUSION: The Hospice Care Environment Scale to assess the environment of hospice care services, has good content and construct validity and reliability. This scale can provide guidance to evaluate the hospice care environment.
Assuntos
Técnica Delphi , Cuidados Paliativos na Terminalidade da Vida , Psicometria , Humanos , Reprodutibilidade dos Testes , Psicometria/instrumentação , Psicometria/métodos , Inquéritos e Questionários , Cuidados Paliativos na Terminalidade da Vida/normas , Cuidados Paliativos na Terminalidade da Vida/métodos , Feminino , Masculino , Adulto , Pessoa de Meia-Idade , ChinaRESUMO
BACKGROUND: Ambient air pollutant exposure can change the composition of gut microbiota at 6-months of age, but there is no epidemiological evidence on the impacts of exposure to particulate matter with an aerodynamic diameter ≤1 µm (PM1) during pregnancy on gut microbiota in mothers and neonates. We aimed to determine if gestational PM1 exposure is associated with the gut microbiota of mothers and neonates. METHODS: Leveraging a mother-infant cohort from the central region of China, we estimated the exposure concentrations of PM1 during pregnancy based on residential address records. The gut microbiota of mothers and neonates was analyzed using 16 S rRNA V3-V4 gene sequences. Functional pathway analyses of 16 S rRNA V3-V4 bacterial communities were conducted using Tax4fun. The impact of PM1 exposure on α-diversity, composition, and function of gut microbiota in mothers and neonates was evaluated using multiple linear regression, controlling for nitrogen dioxide (NO2) and ozone (O3). Permutation multivariate analysis of variance (PERMANOVA) was used to analyze the interpretation degree of PM1 on the sample differences at the OTU level using the Bray-Curtis distance algorithm. RESULTS: Gestational PM1 exposure was positively associated with the α-diversity of gut microbiota in neonates and explained 14.8% (adj. P = 0.026) of the differences in community composition among neonatal samples. In contrast, gestational PM1 exposure had no impact on the α- and ß-diversity of gut microbiota in mothers. Gestational PM1 exposure was positively associated with phylum Actinobacteria of gut microbiota in mothers, and genera Clostridium_sensu_stricto_1, Streptococcus, Faecalibacterium of gut microbiota in neonates. At Kyoto Encyclopedia of Genes and Genomes pathway level 3, the functional analysis results showed that gestational PM1 exposure significantly down-regulated Nitrogen metabolism in mothers, as well as Two-component system and Pyruvate metabolism in neonates. While Purine metabolism, Aminoacyl-tRNA biosynthesis, Pyrimidine metabolism, and Ribosome in neonates were significantly up-regulated. CONCLUSIONS: Our study provides the first evidence that exposure to PM1 has a significant impact on the gut microbiota of mothers and neonates, especially on the diversity, composition, and function of neonatal meconium microbiota, which may have important significance for maternal health management in the future.
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Poluentes Atmosféricos , Microbioma Gastrointestinal , Gravidez , Recém-Nascido , Lactente , Feminino , Humanos , Mães , Poluentes Atmosféricos/toxicidade , Material Particulado/toxicidade , Mecônio , BactériasRESUMO
AIMS: To evaluate the effectiveness and safety of Xin Huang Pian skin patches for patients with acute gouty arthritis. BACKGROUND: In China, patients with acute gouty arthritis benefit from skin patcheses with herbal medicines. But the clinical effects of skin patches with Xin Huang Pian are rarely reported. DESIGN: A Randomized, Double-Blind, Active-Controlled Trial. METHODS: The trial was performed from January 2015-December 2018 at the First Affiliated Hospital of Sun Yat-sen University in China. It was conducted with one intervention group (skin patches of Xin Huang Pian, N = 30) and one active control group (skin patches of Diclofenac Diethylamine Emulgel, N = 31). Participants and study investigators were both blinded to the treatment assignments. The primary outcomes were the improvement of joints' symptoms. The secondary outcomes were changes in white blood cells, erythrocyte sedimentation rate and C-reactive protein. RESULTS: Skin patches of Xin Huang Pian showed quick effect on decreasing joint pain at 3rd day of treatment. Wherever only at 7th day, Diclofenac Diethylamine Emulgel markedly lowered joint pain. Xin Huang Pian also showed superior effect than Diclofenac Diethylamine Emulgel on improving joint swelling and range of motion and decreasing the levels of C-reactive protein and erythrocyte sedimentation rate. No adverse reactions were observed in skin patches of Xin Huang Pian treatment. CONCLUSION: Skin patches of Xin Huang Pian appeared to be safe and efficacious for relieving joint symptoms in patients with acute gouty arthritis. The mechanism might be associated with the decreased levels of C-reactive protein and erythrocyte sedimentation rate. IMPACT: Skin-patcheses with Xin Huang Pian are more effective than Diclofenac Diethylamine Emulgel on improving joint pain, swelling and range of motion. Xin Huang Pian treatment showed superior effects compared with Diclofenac Diethylamine Emulgel on decreasing levels of C-reactive protein and erythrocyte sedimentation rate. Patients with acute gouty arthritis may benefit from skin patches of Xin Huang Pian for effective relief from joint pain and swelling. Chinese Clinical Trial Registration: ChiCTR-TRC-1300 4122.
Assuntos
Analgésicos/uso terapêutico , Anti-Inflamatórios/uso terapêutico , Artrite Gotosa/tratamento farmacológico , Diclofenaco/uso terapêutico , Dietilaminas/uso terapêutico , Medicamentos de Ervas Chinesas/uso terapêutico , Supressores da Gota/uso terapêutico , Administração Cutânea , Analgésicos/administração & dosagem , Anti-Inflamatórios/administração & dosagem , China , Diclofenaco/administração & dosagem , Método Duplo-Cego , Medicamentos de Ervas Chinesas/administração & dosagem , Feminino , Supressores da Gota/administração & dosagem , Humanos , Masculino , Pessoa de Meia-Idade , Fitoterapia , Distribuição AleatóriaRESUMO
Activation of mitogen-activated protein kinases (MAPKs) in neurons may underlie the pathogenesis of Alzheimer's disease (AD). Acrolein, a ubiquitous pollutant, has been reported to implicate in the etiology of AD. Our previous data showed that acrolein changed the levels of key AD-associated proteins, including advanced glycation end products (RAGE), A-disintegrin and metalloprotease (ADAM-10), and beta-site amyloid-beta peptide cleaving enzyme 1 (BACE-1). In this study, we investigated whether acrolein-induced alterations of AD-associated proteins are relevant to MAPKs activation, and strategies to inhibit MAPKs activation yield benefits to acrolein-induced neurotoxicity in HT22 mouse hippocampal cells. We found that acrolein activated MAPKs signaling pathways to mediate cells apoptosis, and then altered the levels of AD-associated proteins ADAM-10, BACE-1 and RAGE. Inhibitors of MAPKs signaling pathways attenuated the cells death and restored the proteins levels of ADAM-10, BACE-1 and RAGE in varying degrees induced by acrolein. Taken together, activated MAPKs signaling pathways should be underlying the pathology of acrolein-induced neuronal disorders. Inhibitors of MAPKs pathways might be promising agents for acrolein-related diseases, such as AD.
Assuntos
Acroleína/toxicidade , Hipocampo/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Neurônios/efeitos dos fármacos , Proteína ADAM10/metabolismo , Secretases da Proteína Precursora do Amiloide/metabolismo , Animais , Apoptose/efeitos dos fármacos , Ácido Aspártico Endopeptidases/metabolismo , Técnicas de Cultura de Células , Linhagem Celular , Inibidores Enzimáticos/farmacologia , Hipocampo/metabolismo , Hipocampo/patologia , Camundongos , Neurônios/metabolismo , Neurônios/patologia , Receptor para Produtos Finais de Glicação Avançada/metabolismoRESUMO
BACKGROUND: Arabinan is an important plant polysaccharide degraded mainly by two hydrolytic enzymes, endo-arabinanase and α-L-arabinofuranosidase. In this study, the characterization and application in arabinan degradation of an endo-arabinanase from Thermotoga thermarum were investigated. RESULTS: The recombinant endo-arabinanase was expressed in Escherichia coli BL21 (DE3) and purified by heat treatment followed by purification on a nickel affinity column chromatography. The purified endo-arabinanase exhibited optimal activity at pH 6.5 and 75°C and its residual activity retained more than 80% of its initial activity after being incubated at 80°C for 2 h. The results showed that the endo-arabinanase was very effective for arabinan degradation at higher temperature. When linear arabinan was used as the substrate, the apparent K(m) and V(max) values were determined to be 12.3 ± 0.15 mg ml⻹ and 1,052.1 ± 12.7 µmol ml⻹ min⻹, respectively (at pH 6.5, 75°C), and the calculated kcat value was 349.3 ± 4.2 s⻹. CONCLUSIONS: This work provides a useful endo-arabinanase with high thermostability andcatalytic efficiency, and these characteristics exhibit a great potential for enzymatic conversion of arabinan.
Assuntos
Regulação Bacteriana da Expressão Gênica , Glicosídeo Hidrolases/genética , Glicosídeo Hidrolases/metabolismo , Bacilos Gram-Negativos Anaeróbios Retos, Helicoidais e Curvos/enzimologia , Bacilos Gram-Negativos Anaeróbios Retos, Helicoidais e Curvos/genética , Sequência de Aminoácidos , Cromatografia de Afinidade , Escherichia coli/metabolismo , Glicosídeo Hidrolases/química , Concentração de Íons de Hidrogênio , Cinética , Dados de Sequência Molecular , Polissacarídeos/metabolismo , Estrutura Terciária de Proteína , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Especificidade por Substrato , TemperaturaRESUMO
BACKGROUND: Two-dimensional high-dose-rate brachytherapy (2D-HDR-BT) is an effective method of dose escalation for local tumor control in early T-stage nasopharyngeal carcinoma (NPC). Treatment outcomes for 3D-image-guided high-dose-rate brachytherapy (3D-image-guided-HDR-BT) after external beam radiotherapy (ERT) have not been examined in early T-stage NPC patients. The current study was designed to evaluate whether addition of 3D-HDR-BT to ERT showed further improvement in treatment outcomes in patients with early T-stage NPC when compared to 2D-HDR-BT after ERT. METHODS: The current study retrospectively analyzed and compared treatment outcomes for patients with nonmetastatic stage T1-2b NPC treated with 2D-HDR-BT (n =101) or 3D-HDR-BT (n =118) after ERT. Patients in both groups were treated with ERT at a mean dose of 60 Gy and a brachytherapy dose of 12Gy (8 ~ 20Gy), 2.5 ~ 5Gy per fraction under local anesthesia. RESULTS: Compared to patients treated with 2D-HDR-BT after ERT, patients treated with 3D-HDR-BT after ERT showed improvement in five-year actuarial local control survival rates (p = 0.024), local/regional relapse-free survival rates (p = 0.038), and disease-free survival rates (p = 0.021). Multivariate analysis showed that NPC patients treated with 3D-HDR-BT had improved local control survival (p = 0.042). The incidence rates of acute or chronic complications were similar between two groups. CONCLUSIONS: The current study showed that 3D-image-guided HDR-BT after ERT was an effective treatment modality for patients with stage T1-2 NPC with acceptable complications. The improvement in local tumor control and disease free survival is likely due to improved conformal dose distributions.
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Braquiterapia/métodos , Imageamento Tridimensional/métodos , Neoplasias Nasofaríngeas/patologia , Neoplasias Nasofaríngeas/radioterapia , Adolescente , Adulto , Idoso , Braquiterapia/efeitos adversos , Carcinoma , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/tratamento farmacológico , Dosagem Radioterapêutica , Estudos Retrospectivos , Taxa de Sobrevida , Resultado do Tratamento , Adulto JovemRESUMO
Acrolein is a highly electrophilic alpha, beta-unsaturated aldehyde to which humans are exposed in many situations and has been implicated in neurodegenerative diseases, such as Alzheimer's disease. Lithium is demonstrated to have neuroprotective and neurotrophic effects in brain ischemia, trauma, neurodegenerative disorders, and psychiatric disorders. Previously we have found that acrolein induced neuronal death in HT22 mouse hippocampal cells. In this study, the effects of lithium on the acrolein-induced neurotoxicity in HT22 cells as well as its mechanism(s) were investigated. We found that lithium protected HT22 cells against acrolein-induced damage by the attenuation of reactive oxygen species and the enhancement of the glutathione level. Lithium also attenuated the mitochondrial dysfunction caused by acrolein. Furthermore, lithium significantly increased the level of phospho-glycogen synthase kinase-3 beta (GSK-3ß), the non-activated GSK-3ß. Taken together, our findings suggest that lithium is a protective agent for acrolein-related neurotoxicity.
Assuntos
Acroleína/antagonistas & inibidores , Acroleína/toxicidade , Hipocampo/efeitos dos fármacos , Cloreto de Lítio/farmacologia , Fármacos Neuroprotetores/farmacologia , Animais , Linhagem Celular , Relação Dose-Resposta a Droga , Hipocampo/metabolismo , Hipocampo/patologia , Camundongos , Espécies Reativas de Oxigênio/metabolismoRESUMO
Oxidative stress is closely related to the pathogenesis of neurodegenerative disorders such as Parkinson's disease. Carvedilol, a nonselective ß-adrenergic receptor blocker with pleiotropic activity has been shown to exert neuroprotective effect due to its antioxidant property. However, the neuroprotective mechanism of carvedilol is still not fully uncovered. The phosphotidylinositol 3-kinase (PI3K)/Akt signaling pathway plays key role in cell survival and the nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) signaling pathway is the major cellular defense mechanism against oxidative stress. Here we investigated the effects of carvedilol on 6-hydroxydopamine (6-OHDA)-induced cell death as well as the Akt and Nrf2/ARE pathways in PC12 cells. We found that carvedilol significantly increased cell viability and decreased reactive oxygen species in PC12 cells exposed to 6-OHDA. Furthermore, carvedilol activated the Akt and Nrf2/ARE pathways in a concentration-dependent manner, and increased the protein levels of heme oxygenase-1(HO-1) and NAD(P)H quinone oxidoreductase-1(NQO-1), two downstream factors of the Nrf2/ARE pathway. In summary, our results indicate that carvedilol protects PC12 cells against 6-OHDA-induced neurotoxicity possibly through activating the Akt and Nrf2/ARE signaling pathways.
Assuntos
Antagonistas Adrenérgicos beta/farmacologia , Carbazóis/farmacologia , Morte Celular/efeitos dos fármacos , Fator 2 Relacionado a NF-E2/metabolismo , Propanolaminas/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Animais , Carvedilol , Oxidopamina/toxicidade , Células PC12 , RatosRESUMO
BACKGROUND: The present study aimed to determine that whether L-carnitine infusion could ameliorate fasting-induced adverse effects and improve outcomes. METHOD: In this 7-day, randomized, single-blind, placebo-controlled, pilot study, 15 metabolic syndrome (MetS) patients (11/4 F/M; age 46.9 ± 9.14 years; body mass index [BMI] 28.2 ± 1.8 kg/m2) were in the L-carnitine group (LC) and 15 (10/5 F/M; age 46.8 ± 10.9 years; BMI 27.1 ± 2.3 kg/m2) were in the control group (CT). All participants underwent a 5-day modified fasting therapy introduced with 2-day moderate calorie restriction. Patients in the LC group received 4 g/day of intravenous L-carnitine, while patients in the CT group were injected with saline. Blood pressure (BP), anthropometric characteristics, markers of liver function, metabolic indices (plasma glucose, lipid profiles, uric acid, free fatty acid and insulin) and hypersensitivity C-reactive protein were measured. Perceived hunger was recorded daily by self-rating visual analogue scales. Fatigue was evaluated by Wessely and Powell scores. RESULTS: In contrast to the CT group, total cholesterol, alanine aminotransferase, systolic and diastolic BP did not change significantly in the LC group after prolonged fasting. There were significant differences in weight loss (LC -4.6 ± 0.9 vs. CT -3.2 ± 1.1 kg, P = 0.03), and waist circumference (LC -5.0 ± 2.2 vs. CT -1.7 ± 1.16 cm, P < 0.001), waist hip ratio (LC -0.023 ± 0.017 vs. CT 0.012 ± 0.01, P < 0.001), insulin concentration (LC -9.9 ± 3.58 vs. CT -6.32 ± 3.44 µU/mL, P = 0.046), and γ-glutamyltransferase concentration (LC -7.07 ± 6.82 vs. CT -2.07 ± 4.18, P = 0.024). Perceived hunger scores were significantly increased (P < 0.05) in the CT group during starvation, which was alleviated with L-carnitine administration in the LC group. Physical fatigue (LC -3.2 ± 3.17 vs. CT 1.8 ± 2.04, P < 0.001) and fatigue severity (LC -11.6 ± 8.38 vs. CT 8.18 ± 7.32, P < 0.001) were significantly reduced in the LC group but were aggravated in the CT group. CONCLUSION: Intravenous L-carnitine can ameliorate fasting-induced hunger, fatigue, cholesterol abnormalities and hepatic metabolic changes and facilitate fasting-induced weight loss in MetS patients. TRIAL REGISTRATION: ChiCTR-TNRC-12002835.
Assuntos
Carnitina/administração & dosagem , Jejum/efeitos adversos , Fadiga/tratamento farmacológico , Fome/efeitos dos fármacos , Síndrome Metabólica/metabolismo , Administração Intravenosa , Adulto , Alanina Transaminase/sangue , Glicemia/metabolismo , Pressão Sanguínea/efeitos dos fármacos , Índice de Massa Corporal , Proteína C-Reativa/metabolismo , HDL-Colesterol/sangue , LDL-Colesterol/sangue , Dieta , Exercício Físico , Ácidos Graxos não Esterificados/sangue , Feminino , Humanos , Insulina/sangue , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Método Simples-Cego , Resultado do Tratamento , Ácido Úrico/sangue , Circunferência da Cintura , Redução de Peso/efeitos dos fármacos , gama-Glutamiltransferase/sangueRESUMO
The xyn10B gene, encoding the endo-1,4-ß-xylanase Xyn10B from Thermotoga thermarum, was cloned and expressed in Escherichia coli. The ORF of the xyn10B was 1,095 bp and encoded to mature peptide of 344 amino acids with a calculated MW of 40,531 Da. The recombinant xylanase was optimally active at 80 °C, pH 6.0 and retained approx. 60 % of its activity after 2 h at 75 °C. Apparent K m , k cat and k cat /K m values of the xylanase for beechwood xylan were 1.8 mg ml(-1), 520 s(-1) and 289 ml mg(-1) s(-1), respectively. The end products of the hydrolysis of beechwood xylan were mainly oligosaccharides but without xylose after 2 h hydrolysis.
Assuntos
Bactérias/enzimologia , Endo-1,4-beta-Xilanases/isolamento & purificação , Endo-1,4-beta-Xilanases/metabolismo , Xilanos/metabolismo , Bactérias/genética , Clonagem Molecular , Endo-1,4-beta-Xilanases/química , Endo-1,4-beta-Xilanases/genética , Estabilidade Enzimática , Escherichia coli/genética , Expressão Gênica , Concentração de Íons de Hidrogênio , Cinética , Peso Molecular , Fases de Leitura Aberta , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Temperatura , Fatores de TempoRESUMO
Microbial communities such as those residing in the human gut are highly diverse and complex, and many with important implications in health and diseases. The effects and functions of these microbial communities are determined not only by their species compositions and diversities but also by the dynamic intra- and inter-cellular states at the transcriptional level. Powerful and scalable technologies capable of acquiring single-microbe-resolution RNA sequencing information in order to achieve comprehensive understanding of complex microbial communities together with their hosts is therefore utterly needed. Here we report the development and utilization of a droplet-based smRNA-seq (single-microbe RNA sequencing) method capable of identifying large species varieties in human samples, which we name smRandom-seq2. Together with a triple-module computational pipeline designed for the bacteria and bacteriophage sequencing data by smRandom-seq2 in four human gut samples, we established a single-cell level bacterial transcriptional landscape of human gut microbiome, which included 29,742 single microbes and 329 unique species. Distinct adaptive responses states among species in Prevotella and Roseburia genus and intrinsic adaptive strategy heterogeneity in Phascolarctobacterium succinatutens were uncovered. Additionally, we identified hundreds of novel host-phage transcriptional activity associations in the human gut microbiome. Our results indicated the smRandom-seq2 is a high-throughput and high-resolution smRNA-seq technique that is highly adaptable to complex microbial communities in real-word situations and promises new perspectives in the understanding of human microbiomes.
RESUMO
BACKGROUND: Mannan is one of the primary polysaccharides in hemicellulose and is widely distributed in plants. ß-Mannosidase is an important constituent of the mannan-degrading enzyme system and it plays an important role in many industrial applications, such as food, feed and pulp/paper industries as well as the production of second generation bio-fuel. Therefore, the mannose-tolerant ß-mannosidase with high catalytic efficiency for bioconversion of mannan has a great potential in the fields as above. RESULTS: A ß-mannosidase gene (Tth man5) of 1,827 bp was cloned from the extremely thermophilic bacterium Thermotoga thermarum DSM 5069 that encodes a protein containing 608 amino acid residues, and was over-expressed in Escherichia coli BL21 (DE3). The results of phylogenetic analysis, amino acid alignment and biochemical properties indicate that the Tth Man5 is a novel ß-mannosidase of glycoside hydrolase family 5. The optimal activity of the Tth Man5 ß-mannosidase was obtained at pH 5.5 and 85°C and was stable over a pH range of 5.0 to 8.5 and exhibited 2 h half-life at 90°C. The kinetic parameters K(m) and V(max) values for p-nitrophenyl-ß-D-mannopyranoside and 1,4-ß-D-mannan were 4.36±0.5 mM and 227.27±1.59 µmol min⻹ mg⻹, 58.34±1.75 mg mL⻹ and 285.71±10.86 µmol min⻹ mg⻹, respectively. The k(cat)/K(m) values for p-nitrophenyl-ß-D-mannopyranoside and 1,4-ß-D-mannan were 441.35±0.04 mM⻹ s⻹ and 41.47±1.58 s⻹ mg⻹ mL, respectively. It displayed high tolerance to mannose, with a K(i) value of approximately 900 mM. CONCLUSIONS: This work provides a novel and useful ß-mannosidase with high mannose tolerance, thermostability and catalytic efficiency, and these characteristics constitute a powerful tool for improving the enzymatic conversion of mannan through synergetic action with other mannan-degrading enzymes.
Assuntos
Proteínas de Bactérias/biossíntese , Escherichia coli/metabolismo , Bacilos Gram-Negativos Anaeróbios Retos, Helicoidais e Curvos/enzimologia , beta-Manosidase/biossíntese , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Clonagem Molecular , Biologia Computacional , DNA Bacteriano/genética , Escherichia coli/genética , Bacilos Gram-Negativos Anaeróbios Retos, Helicoidais e Curvos/genética , Meia-Vida , Concentração de Íons de Hidrogênio , Mananas/química , Manose/metabolismo , Dados de Sequência Molecular , Filogenia , Plasmídeos/genética , Polissacarídeos/química , Especificidade por Substrato , Temperatura , beta-Manosidase/genéticaRESUMO
Bacteria colonize almost all parts of the human body and can differ significantly. However, the population level transcriptomics measurements can only describe the average bacteria population behaviors, ignoring the heterogeneity among bacteria. Here, we report a droplet-based high-throughput single-microbe RNA-seq assay (smRandom-seq), using random primers for in situ cDNA generation, droplets for single-microbe barcoding, and CRISPR-based rRNA depletion for mRNA enrichment. smRandom-seq showed a high species specificity (99%), a minor doublet rate (1.6%), a reduced rRNA percentage (32%), and a sensitive gene detection (a median of ~1000 genes per single E. coli). Furthermore, smRandom-seq successfully captured transcriptome changes of thousands of individual E. coli and discovered a few antibiotic resistant subpopulations displaying distinct gene expression patterns of SOS response and metabolic pathways in E. coli population upon antibiotic stress. smRandom-seq provides a high-throughput single-microbe transcriptome profiling tool that will facilitate future discoveries in microbial resistance, persistence, microbe-host interaction, and microbiome research.
Assuntos
Escherichia coli , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Escherichia coli/genética , RNA-Seq , Antibacterianos/farmacologia , Primers do DNA , RNA Ribossômico/genéticaRESUMO
A novel series of tacrine-caffeic acid hybrids (5a-f) were designed and synthesized by combining caffeic acid (CA) with tacrine. The antioxidant study revealed that all the hybrids have much more antioxidant capacities compared to CA. Among these compounds, 5e showed the highest selectivity in inhibiting acetylcholinesterase (AChE) over butyrylcholinesterase (BuChE). Enzyme kinetic study had suggested that 5e binds to both catalytic (CAS) and peripheral anionic sites (PAS) of AChE. Moreover, compound 5e also inhibited self- or AChE-induced ß-amyloid(1-40) aggregation, as well as had potent neuroprotective effects against H(2)O(2)- and glutamate- induced cell death with low toxicity in HT22 cells.
Assuntos
Doença de Alzheimer/tratamento farmacológico , Peptídeos beta-Amiloides/antagonistas & inibidores , Ácidos Cafeicos/química , Ácidos Cafeicos/farmacologia , Tacrina/química , Tacrina/farmacologia , Acetilcolinesterase/metabolismo , Doença de Alzheimer/metabolismo , Peptídeos beta-Amiloides/metabolismo , Animais , Antioxidantes/síntese química , Antioxidantes/química , Antioxidantes/farmacologia , Butirilcolinesterase/metabolismo , Ácidos Cafeicos/síntese química , Morte Celular/efeitos dos fármacos , Linhagem Celular , Inibidores da Colinesterase/síntese química , Inibidores da Colinesterase/química , Inibidores da Colinesterase/farmacologia , Desenho de Fármacos , Peróxido de Hidrogênio/metabolismo , Camundongos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Fármacos Neuroprotetores/síntese química , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/farmacologia , Tacrina/síntese químicaRESUMO
BACKGROUND: An outbreak of Plasmodium malariae infection among forest goers in Sanya City of Hainan Island, China was reported in 2015. In response to this outbreak, an innovative three-layer strategy (TLS) targeted forest goers was adapted based on the 1-3-7 approach. MAIN TEXT: Key elements of TLS are: (i) The village with five malaria cases and adjacent villages were set as the first layer. All residents including forest goers were taken as the high-risk population (HRP). Active case detection (ACD) by blood smear microscopy and PCR was selected as the primary measure, and passive case detection (PCD) as complementary measure. One case was identified under TLS implementation. (ii) The township with cases (Gaofeng Town) and the nearby towns were chosen as the second layer. Only forest goers were screened by ACD, while PCD as a routine screening method. 7831 blood smears collected by ACD and PCD and tested with negative results. (iii) The city with cases (Sanya City) and others 12 counties/county-level cities were selected as the third layer. Malaria cases were monitored passively. A total of 77,555 blood slides were screened by PCD with zero positive sample. For each layer, the malaria vector mosquitoes were monitored using light traps, cattle-baited/human-bait traps. Anopheles minimus (dominant species), An. sinensis and An. dirus were captured. Vector control measures mainly include insecticide residual spraying and long-lasting insecticide nets. The capacity of clinicians, public health practitioners and laboratory technicians has been improved through training. During 2016â2018, TLS and chemoprophylaxis were implemented in the same areas. In the first layer, all residents were monitored by ACD, and malaria chemoprophylaxis were distributed, 89.5% of forest goers were using chemoprophylaxis against malaria. The blood smears (3126 by ACD plus 1516 by PCD) were with zero positive results. Chemoprophylaxis and ACD were offered to forest goers once a year, and PCD in residents as a complementary measure in the second and third layer, 77.8% and 95.1% of forest goers received chemoprophylaxis. In each layer, vector surveillance and control of malaria and trainings for medical staff were still in place. CONCLUSIONS: TLS was effective in blocking the outbreak by P. malariae among forest goers in Hainan in malaria elimination stage. However, whether it could prevent the malaria resurgence in the post-elimination phase needs to be further assessed.
Assuntos
Anopheles , Inseticidas , Malária , Animais , Anopheles/fisiologia , Bovinos , China/epidemiologia , Surtos de Doenças/prevenção & controle , Florestas , Humanos , Malária/epidemiologia , Malária/prevenção & controle , Mosquitos Vetores , Estudos RetrospectivosRESUMO
Hainan Province is in a tropical area of China and previously experienced serious P. falciparum and P. vivax malaria epidemics. After nearly 70 consecutive years of malaria prevention and control, malaria in Hainan has gradually been eliminated. To achieve the elimination of malaria, Hainan enacted six stages: investigative research and pilot prevention and control, large-scale antimalaria measures, adjustment of strategies for prevention and control, joint prevention and control measures, global funding of routine malaria control, and malaria elimination. Different strategies for malaria control were adopted at different stages. Malaria was most prevalent in the mountainous areas of central and southern Hainan, which contain a high-risk population (the forest goers) and two highly effective malaria vectors (An. dirus and An. minimus). Forest goers have been a high-risk population for malaria in Hainan since their identification in the 1990s. This paper summarizes malaria monitoring in forest goers and the response of forest goers to malaria control and elimination, distilling specific malaria control and elimination measures via case studies in Hainan Province. Two case studies in the malaria control stage demonstrated different measures for outbreaks and sporadic cases in forest goers. In view of the malaria outbreak in Sanya during the elimination stage, three-layered strategies (TLSs) were implemented to control outbreaks and improve control measures. Moreover, this paper also illustrates specific management measures to prevent malaria retransmission from sporadic imported malaria cases during the elimination phase. Hainan finally eliminated malaria in 2020. However, the risk of malaria retransmission is still high due to the prevalence of effective malaria vectors in Hainan, and forest goers are still a high-risk population for malaria retransmission.
Assuntos
Malária , China/epidemiologia , Florestas , Humanos , Malária/epidemiologia , Malária/prevenção & controle , Prevalência , Fatores de RiscoRESUMO
This communication reports a strategy for scale-up of an in situ polymerization technique for polyolefin-based nanocomposites preparation, taking layered silicate (clay) and multi-walled carbon nanotubes (MWCNTs) as examples of nanofillers. The strategy is realized by transforming the nanofillers into granular "nanosupports" for Ziegler-Natta catalysts. With a catalyst to polymer replication effect on particle morphology, the in situ prepared nanocomposites are of controlled granular particle morphology. With the polymer particle morphology controlled, the in situ polymerization technique becomes suitable for industrial olefin polymerization processes for mass production of polyolefin nanocomposites.
Assuntos
Química Orgânica/métodos , Nanocompostos/química , Polienos/química , Polímeros/química , Catálise , Polimerização , Polímeros/síntese químicaRESUMO
OBJECTIVE: To evaluate the effects of the modified Linggui Zhugan Decoction (see text) combined with short-term very low calorie diets (VLCDs) on glycemic control in newly diagnosed type 2 diabetes mellitus (T2DM) patients. METHODS: A total of 20 subjects with newly diagnosed T2DM were treated with the modified Linggui Zhugan Decoction (one-month administration) combined with short-term VLCDs (5 days), and 3-months follow-up. A standard 75-g oral-glucose-tolerance test (OGTT) indexes fasting plasma glucose (FPG), post-prandial 0.5 h and 2 h plasma glucose (P0.5hPG, P2hPG), glycated hemoglobin A(1c) (GHbA(1c)), body weight, body mass index (BMI), insulin function, insulin resistance index, incidence of hypoglycemia, and the liver and renal functions were evaluated before and after treatment. Correlations of BMI with insulin function and insulin resistance were also assessed. RESULTS: After the treatment, the patients' plasma glucose decreased steadily, FPG decreased from 5.8 +/- 0.9 mmol/L at pre-treatment to 5.0 +/- 0.6 mmol/L at 3-months follow-up (P < 0.05), and P2hPG decreased from 11.7 +/- 3.8 mmol/L at pre-treatment to 6.9 +/- 0.9 mmol/L at 3-months follow-up (P < 0.01). The level of GHbA(1c) declined from (6.47 +/- 1.24)% at pre-treatment to (6.14 +/- 0.99)% at 3-months follow-up (P < 0.01). Body weight and BMI also declined significantly. Insulin resistance index was improved obviously and no event of hypoglycemia occurred. Part of the patients companied with fatty liver had a transient increase in hepatic transaminase during the treatment, but it turned to normal after the treatment. CONCLUSIONS: The modified Linggui Zhugan Decoction combined with short-term VLCDs can be safely implemented for steady glycemic control in newly diagnosed T2DM patients.
Assuntos
Restrição Calórica , Diabetes Mellitus Tipo 2/dietoterapia , Diabetes Mellitus Tipo 2/tratamento farmacológico , Extratos Vegetais/uso terapêutico , Adulto , Idoso , Glicemia/efeitos dos fármacos , Índice de Massa Corporal , Peso Corporal/efeitos dos fármacos , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: There are conflicting reports on the outcomes of patients with metabolic liver disease after liver transplantation. We aimed to compare the outcomes of living donor liver transplantation (LDLT) for metabolic disease vs orthotopic liver transplantation (OLT) from deceased donation. METHODS: Clinical data of 89 patients undergoing liver transplantation for the treatment of metabolic disorders were reviewed. Pre- and peri-transplant demographics, survival rate, complications and laboratory test data were collected and analyzed. RESULTS: For the 89 patients, only 2 died by the end of the last follow-up. The post-transplant EAD rate and severe complications were higher for OLT than LDLT. No significant difference was found between LDLT and OLT for the incidence of EBV and CMV infections. In terms of laboratory indexes, the recovery time of PLT, AKP and AST levels were significantly longer for OLT than LDLT. Among different types of metabolic disease, no significant difference was found in viral infection, EAD, laboratory indexes, severe complications or duration of hospital stay. CONCLUSIONS: LDLT shows a lower incidence rate of EAD and complications, while it also shows a 1-year survival rate and incidence of viral infections compared similar to that of OLT. LDLT is the better treatment option of pediatric liver transplantation for metabolic liver disease compared with OLT.