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BACKGROUND: Online dating apps are popular platforms for seeking romance and sexual relationships among young adults. As mobile apps can easily gain access to a pool of strangers ("new friends") at any time and place, it leads to heightened sexual health risks and privacy concerns. OBJECTIVE: This study aimed to evaluate the effectiveness of a peer-led web-based intervention for online dating apps to prepare Chinese college students so that they have better self-efficacy when using dating apps. METHODS: An open clustered randomized controlled trial was conducted among students from three colleges (The University of Hong Kong, Hang Seng University of Hong Kong, and Yijin Programme of Vocational Training College) in Hong Kong. Students aged 17 to 27 years who attended common core curriculum or general education were randomized into intervention and control groups. The intervention material, developed with high peer engagement, included four short videos, an interactive scenario game, and a risk assessment tool. An existing website promoting physical activities and healthy living was used as a control. Using the information, motivation, and behavioral skills (IMB) approach to design the evaluation, questionnaires covering participants' sociodemographics and dating app characteristics, as well as the general self-efficacy scale (GSE) as the primary outcome and the risk propensity scale (RPS) as the secondary outcome were administered before, immediately after, and at 1 month after the intervention. Intention-to-treat analysis was adopted, and between-group differences were assessed using the Mann-Whitney U test. A post-hoc multiple linear regression model was used to examine the correlates of the GSE and RPS. RESULTS: A total of 578 eligible participants (290 in the intervention group and 288 in the control group) participated in the study with 36 lost to follow-up. There were more female participants (318/542, 58.7%) than male participants in the sample, reflecting the distribution of college students. Over half of the participants (286/542, 52.8%) reported the following reasons for using dating apps: being curious (170/498, 34.1%), trying to make new friends (158/498, 31.7%), and finding friends with similar interests (121/498, 24.3%). Overall, the participants in the intervention group reported favorable experiences when compared with the finding in the control group. There was significant improvement in the GSE score and reduction in the RPS score (P<.001) in the intervention group. University of Hong Kong students were more susceptible to risk reduction after the intervention when compared with students from the other two institutions. CONCLUSIONS: The online intervention was effective in improving general self-efficacy and reducing risk tendency among young students. Future work is needed to determine if this approach is cost-effective and such behavioral change is sustainable. TRIAL REGISTRATION: ClinicalTrials.gov NCT03685643; https://clinicaltrials.gov/ct2/show/NCT03685643. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): RR2-10.1186/s13063-018-3167-5.
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Intervenção Baseada em Internet/tendências , Aplicativos Móveis/estatística & dados numéricos , Autoeficácia , Adolescente , Adulto , Exercício Físico , Feminino , Humanos , Masculino , Adulto JovemRESUMO
Severe acute respiratory syndrome (SARS) is caused by infection of a previously undescribed coronavirus (CoV). L-SIGN, encoded by CLEC4M (also known as CD209L), is a SARS-CoV binding receptor that has polymorphism in its extracellular neck region encoded by the tandem repeat domain in exon 4. Our genetic risk association study shows that individuals homozygous for CLEC4M tandem repeats are less susceptible to SARS infection. L-SIGN is expressed in both non-SARS and SARS-CoV-infected lung. Compared with cells heterozygous for L-SIGN, cells homozygous for L-SIGN show higher binding capacity for SARS-CoV, higher proteasome-dependent viral degradation and a lower capacity for trans infection. Thus, homozygosity for L-SIGN plays a protective role during SARS infection.
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Moléculas de Adesão Celular/genética , Lectinas Tipo C/genética , Receptores de Superfície Celular/genética , Síndrome Respiratória Aguda Grave/genética , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/patogenicidade , Animais , Células CHO/virologia , Moléculas de Adesão Celular/metabolismo , Chlorocebus aethiops , Estudos de Coortes , Cricetinae , Cricetulus , Predisposição Genética para Doença , Homozigoto , Hong Kong/epidemiologia , Humanos , Intestino Delgado/fisiologia , Lectinas Tipo C/metabolismo , Pulmão/fisiologia , Pulmão/virologia , Dados de Sequência Molecular , Complexo de Endopeptidases do Proteassoma/metabolismo , Receptores de Superfície Celular/metabolismo , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/metabolismo , Síndrome Respiratória Aguda Grave/epidemiologia , Sequências de Repetição em Tandem , Células Vero/virologiaRESUMO
Laribacter hongkongensis is a newly discovered Gram-negative bacillus of the Neisseriaceae family associated with freshwater fish-borne gastroenteritis and traveler's diarrhea. The complete genome sequence of L. hongkongensis HLHK9, recovered from an immunocompetent patient with severe gastroenteritis, consists of a 3,169-kb chromosome with G+C content of 62.35%. Genome analysis reveals different mechanisms potentially important for its adaptation to diverse habitats of human and freshwater fish intestines and freshwater environments. The gene contents support its phenotypic properties and suggest that amino acids and fatty acids can be used as carbon sources. The extensive variety of transporters, including multidrug efflux and heavy metal transporters as well as genes involved in chemotaxis, may enable L. hongkongensis to survive in different environmental niches. Genes encoding urease, bile salts efflux pump, adhesin, catalase, superoxide dismutase, and other putative virulence factors-such as hemolysins, RTX toxins, patatin-like proteins, phospholipase A1, and collagenases-are present. Proteomes of L. hongkongensis HLHK9 cultured at 37 degrees C (human body temperature) and 20 degrees C (freshwater habitat temperature) showed differential gene expression, including two homologous copies of argB, argB-20, and argB-37, which encode two isoenzymes of N-acetyl-L-glutamate kinase (NAGK)-NAGK-20 and NAGK-37-in the arginine biosynthesis pathway. NAGK-20 showed higher expression at 20 degrees C, whereas NAGK-37 showed higher expression at 37 degrees C. NAGK-20 also had a lower optimal temperature for enzymatic activities and was inhibited by arginine probably as negative-feedback control. Similar duplicated copies of argB are also observed in bacteria from hot springs such as Thermus thermophilus, Deinococcus geothermalis, Deinococcus radiodurans, and Roseiflexus castenholzii, suggesting that similar mechanisms for temperature adaptation may be employed by other bacteria. Genome and proteome analysis of L. hongkongensis revealed novel mechanisms for adaptations to survival at different temperatures and habitats.
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Adaptação Fisiológica/genética , Genoma Bacteriano , Neisseriaceae/genética , Ecossistema , Proteoma , TemperaturaRESUMO
BACKGROUND: Prunella vulgaris (PV) is a low-growing perennial herb, which can be found in different parts of the world as Asia, Europe and North America. It is traditionally used for medicinal treatment in various cultures in India, China, Japan, Korea, Russia, and Eastern Europe for treating different ailments, such as fever, and healing wounds. In our previous article, we showed the anti-tumorous effect of the volatile organic compounds (VOCs) of PV and characterized the steam distillation process in the extraction of VOCs from PV. This has never been done before as we are aware of. To use the VOCs as drugs, there is a question of how much of the VOCs are lost before the prepared drugs reach the patients. Thus, the first aim of the present article is to try to explore the time depletion effect on the VOCs in the PV extracts. Then, the second aim is to extend the work in the previous paper and further understand the dynamics of the distillation process of PV by changing the steam flow rate in the extraction process. METHODS: To achieve the first aim to explore the aging effect of how much VOCs are depleted after they are extracted, the VOCs were first extracted by the same method as before, i.e., using steam distillation. Then, tubes of the aqueous solution containing the VOCs were then stored in a 5°C refrigerator. They were then taken out for GC-MS analysis according to a preplanned schedule up to 8 weeks after the VOCs were extracted. The chemical composition of the distillate could then be evaluated. This revealed the changes in the abundance of VOCs with aging. At the same time, the cell viability of SCC154 oral squamous cells treated by these herbal solutions, which were at different aging stages, was evaluated using a tetrazolium-based colorimetric reagent, Cell Counting Kit-8. To achieve the second aim of exploring the dynamics of the steam distillation process, the steam flow rate was adjusted by changing the temperature setting of the hot plate. GC-MS was again used to quantify the chemical constituents of the distillates. RESULTS: By using GC-MS to measure the abundance of volatile compounds at different time points after the distillation process, it was found that the volatile compounds persist for a very long time, or over 8 weeks, which was the longest period of our experiment. The aging of the distillates also did not depreciate much the cell cytotoxicity of the PV distillate on the cancer cells. With respect to the dynamics of the steam distillation process, it was found that, at a low steam flow rate, volatile compounds of lower molecular weight are more efficient to be extracted, while at a high steam flow rate, volatile compounds of higher molecular weight are more efficiently extracted. CONCLUSION: Our findings demonstrate that the VOC compounds extracted and present in aqueous form do not deplete much for at least 2 months after the extraction process, neither they exhibit cell cytotoxicity. The experiments on the dynamics of the steam distillation process demonstrate that the mass of herb present in the flow path of the steam has significant effects on the relative amounts of VOCs extracted.
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Prunella , Compostos Orgânicos Voláteis , Destilação/métodos , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Extratos Vegetais/farmacologia , Vapor/análise , Compostos Orgânicos Voláteis/análise , Compostos Orgânicos Voláteis/química , Compostos Orgânicos Voláteis/farmacologiaRESUMO
We report a case of pulmonary tumoral thrombotic microangiopathy (PTTM) of pancreatobiliary origin who presented with fulminant right heart failure and was revived by extracorporeal membrane oxygenation-cardiopulmonary resuscitation (ECMO-CPR). Pathology of PTTM was illustrated. This case highlights the notorious difficulty of antemortem diagnosis of PTTM as well as the role of venoarterial-ECMO as a temporizing measure to diagnosis and treatment.
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Chromosome 3p was reported by previous studies as one of the regions showing strong evidence of linkage with schizophrenia. We performed a fine-mapping association study of a 6-Mb high-LD and gene-rich region on 3p in a Southern Chinese sample of 489 schizophrenia patients and 519 controls to search for susceptibility genes. In the initial screen, 4 SNPs out of the 144 tag SNPs genotyped were nominally significant (P < 0.05). One of the most significant SNPs (rs3732530, P = 0.0048) was a non-synonymous SNP in the neuroglycan C (NGC, also known as CSPG5) gene, which belongs to the neuregulin family. The gene prioritization program Endeavor ranked NGC 8th out of the 129 genes in the 6-Mb region and the highest among the genes within the same LD block. Further genotyping of NGC revealed 3 more SNPs to be nominally associated with schizophrenia. Three other genes (NRG1, ErbB3, ErbB4) involved in the neuregulin pathways were subsequently genotyped. Interaction analysis by multifactor dimensionality reduction (MDR) revealed a significant two-SNP interaction between NGC and NRG1 (P = 0.015) and three-SNP interactions between NRG1 and ErbB4 (P = 0.009). The gene NGC is exclusively expressed in the brain. It is implicated in neurodevelopment in rats and was previously shown to promote neurite outgrowth. Methamphetamine, a drug that may induce psychotic symptoms, was reported to alter the expression of NGC. Taken together, these results suggest that NGC may be a novel candidate gene, and neuregulin signaling pathways may play an important role in schizophrenia.
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Proteoglicanas de Sulfatos de Condroitina/genética , Predisposição Genética para Doença , Neurregulinas/genética , Esquizofrenia/terapia , Estudos de Casos e Controles , China , Humanos , Polimorfismo de Nucleotídeo ÚnicoRESUMO
BACKGROUND: Dating applications are a popular platform to meet new people. At the same time, they have been associated with risks such as unsafe sexual behavior and privacy concerns in young adults. This paper presents a study protocol of a cluster randomized controlled trial (RCT) to evaluate the effectiveness of a peer-led web-based intervention to promote its safe usage in young adults. METHODS: The study design is an open-labeled cluster RCT with an intervention and a placebo control arm. The intervention group will receive a web-based intervention developed through focus group discussions, a crowdsourcing contest, and a Peer-Vetted Creative Production (PVCP) workshop. The control group will receive a web-based resource on health and exercise. We aim to recruit approximately 338 young adults aged 17-27 years from three tertiary educational institutions in Hong Kong with the class as the cluster unit. Based on the Information, Motivation, and Behavioral Skills (IMB) model, the primary outcome of this study is self-efficacy in using dating applications measured by the General Self Efficacy Scale. Secondary outcomes include change in risk perception measured by the Risk Propensity Scale and a Risk Assessment Tool. Questionnaires will be administered before the intervention, after the intervention, and at one-month follow-up. Intention-to-treat analysis and multilevel regression modeling will be used to evaluate differences in outcomes between groups and the factors affecting these outcomes, respectively. DISCUSSION: Dating application usage presents opportunities as well as challenges to young adults meeting new friends. Innovative and relatable interventions are needed to promote the safe usage of dating applications to this population. Practical knowledge gained from the development process may be helpful for future intervention utilizing the peer-led approach. If effective, the intervention will be disseminated to non-governmental organizations and educational institutions to be used as a teaching resource. TRIAL REGISTRATION: ClinicalTrials.gov, NCT03685643 . Registered on 26 September 2018. University of Hong Kong Clinical Trials Registry, HKUCTR-2512.
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Comportamento do Adolescente , Internet , Relações Interpessoais , Aplicativos Móveis , Influência dos Pares , Assunção de Riscos , Comportamento Sexual , Adolescente , Adulto , Atitude Frente aos Computadores , Feminino , Hong Kong , Humanos , Masculino , Motivação , Estudos Multicêntricos como Assunto , Privacidade , Ensaios Clínicos Controlados Aleatórios como Assunto , Fatores de Risco , Autoeficácia , Fatores de Tempo , Sexo sem Proteção/prevenção & controle , Sexo sem Proteção/psicologia , Adulto JovemRESUMO
Loss of DNA copy number at the short arm of chromosome 3 is one of the most common genetic changes in human lung cancer, suggesting the existence of one or more tumor suppressor genes (TSG) at 3p. To identify most frequently deleted regions and candidate TSGs within these regions, a recently developed single-nucleotide polymorphism (SNP)-mass spectrometry-genotyping (SMSG) technology was applied to investigate the loss of heterozygosity (LOH) in 30 primary non-small-cell lung cancers. A total of 386 SNP markers that spanned a region of 70 Mb at 3p, from 3pter to 3p14.1, were selected for LOH analysis. The average intermarker distance in the present study is approximately 180 kb. Several frequently deleted regions, including 3p26.3, 3p25.3, 3p24.1, 3p23, and 3p21.1, were found. Several candidate TSGs within these frequently detected LOH regions have been found, including APG7L at 3p25.3, CLASP2 at 3p23, and CACNA2D3 at 3p21.1. This study also showed that SMSG technology is a very useful approach to rapidly define the minimal deleted region and to identify target TSGs in a given cancer.
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Carcinoma Pulmonar de Células não Pequenas/genética , Cromossomos Humanos Par 3/genética , Perda de Heterozigosidade , Neoplasias Pulmonares/genética , Genes Supressores de Tumor , Genótipo , Humanos , Hibridização de Ácido Nucleico , Polimorfismo de Nucleotídeo Único , Reprodutibilidade dos Testes , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Severe hemodilutional anemia may reduce cerebral oxygen delivery, resulting in cerebral tissue hypoxia. Increased nitric oxide synthase (NOS) expression has been identified following cerebral hypoxia and may contribute to the compensatory increase in cerebral blood flow (CBF) observed after hypoxia and anemia. However, changes in cerebral NOS gene expression have not been reported after acute anemia. This study tests the hypothesis that acute hemodilutional anemia causes cerebral tissue hypoxia, triggering changes in cerebral NOS gene expression. Anesthetized rats underwent hemodilution when 30 ml/kg of blood were exchanged with pentastarch, resulting in a final hemoglobin concentration of 51.0 +/- 1.2 g/l (n = 7 rats). Caudate tissue oxygen tension (Pbr(O(2))) decreased transiently from 17.3 +/- 4.1 to 14.4 +/- 4.1 Torr (P < 0.05), before returning to baseline after approximately 20 min. An increase in CBF may have contributed to restoring Pbr(O(2)) by improving cerebral tissue oxygen delivery. An increase in neuronal NOS (nNOS) mRNA was detected by RT-PCR in the cerebral cortex of anemic rats after 3 h (P < 0.05, n = 5). A similar response was observed after exposure to hypoxia. By contrast, no increases in mRNA for endothelial NOS or interleukin-1beta were observed after anemia or hypoxia. Hemodilutional anemia caused an acute reduction in Pbr(O(2)) and an increase in cerebral cortical nNOS mRNA, supporting a role for nNOS in the physiological response to acute anemia.
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Anemia/enzimologia , Anemia/etiologia , Química Encefálica/genética , Regulação Enzimológica da Expressão Gênica/genética , Hemodiluição/efeitos adversos , Óxido Nítrico Sintase/biossíntese , Animais , Gasometria , Calibragem , Circulação Cerebrovascular/fisiologia , Eletroforese em Gel de Ágar , Hemoglobinas/metabolismo , Hipóxia/enzimologia , Fluxometria por Laser-Doppler , Microeletrodos , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase Tipo I , Óxido Nítrico Sintase Tipo III , Polarografia , RNA Mensageiro/biossíntese , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para CimaRESUMO
Epilepsy surgery is considered a treatment option for patients with intractable seizures. Relatively few studies of efficacy, safety, and long-term outcome are available for the pediatric age group. This study describes a 12-year experience with pediatric epilepsy surgery at the University of Alberta. Records of pediatric epilepsy surgery patients admitted to the Comprehensive Epilepsy Program at the University of Alberta between 1988 and 2000 were reviewed. All patients received preoperative and postoperative clinical evaluation, seizure charts, testing of drug levels, electroencephalogram, computed tomography/magnetic resonance imaging, neuropsychologic testing, and long-term video electroencephalogram monitoring. The patients were reassessed after surgery at 6 weeks, 6 months, and 1 year and then yearly. The duration of follow-up was 1 year to 12 years. Forty-two patients underwent temporal lobectomies; 35, extratemporal resection. The age at surgery ranged from 6 months to 16 years. Thirty-two (76%) of temporal lobe patients became seizure-free (Engel Class I) vs 24 (68%) for the extratemporal group (Engel Class I). One patient (2%) in the temporal group had an Engel Class II outcome and one patient (3%) in the extratemporal group had the same Engel Class II outcome. Three patients (4%) manifested postoperative complications, and there were no deaths. Patients reported improvement in cognitive abilities, behavior, and quality of life after the surgery. Epilepsy surgery in children is effective and safe. Many children are seizure-free after the operation and remain so, although the results of temporal lobectomy are better than for extratemporal resections. There are few complications, and children often have an improved quality of life.
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Epilepsia/cirurgia , Hospitais Universitários/estatística & dados numéricos , Adolescente , Adulto , Canadá , Criança , Pré-Escolar , Eletroencefalografia/métodos , Epilepsia/patologia , Epilepsia/fisiopatologia , Feminino , Seguimentos , Hospitais Universitários/tendências , Humanos , Lactente , Masculino , Estudos RetrospectivosRESUMO
Results of a systematic theoretical study of collisions between moving solitons in a fiber grating are presented. Various outcomes of the collision are identified, the most interesting one being merger of the solitons into a single zero-velocity pulse, which suggests a way to create pulses of "standing light." The merger occurs with the solitons whose energy takes values between 0.15 and 0.35 of the limit value, while their velocity is limited by approximately 0.2 of the limit light velocity in the fiber. If the energy is larger, another noteworthy outcome is acceleration of the solitons as a result of the collision. In the case of mutual passage of the solitons, inelasticity of the collision is quantified by the energy-loss share. Past the soliton's stability limit, the collision results in strong deformation and subsequent destruction of the solitons. Simulations of multiple collisions of two solitons in a fiber-loop configuration are performed too. In this case, the maximum velocity admitting the merger increases to approximately 0.4 of the limit velocity. The influence of an attractive local defect on the collision is also studied, with the conclusion that the defect does not alter the overall picture, although it traps a small-amplitude pulse. Related effects in single-soliton dynamics are considered too, the most important one being the possibility of slowing down the soliton (reducing its velocity to the above-mentioned values that admit fusion of colliding solitons) by passing it through an apodized fiber grating, i.e., one with a gradually increasing Bragg reflectivity. Additionally, transformation of an input sech signal into a gap soliton (which is quantified by the share of lost energy), and the rate of decay of a quiescent gap soliton in a finite fiber grating, due to energy leakage through loose edges, are also studied.
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A model of a lossy nonlinear fiber grating with a "hot spot," which combines a local gain and an attractive perturbation of the refractive index, is introduced. A family of exact solutions for pinned solitons is found in the absence of loss and gain. In the presence of the loss and localized gain, an instability threshold of the zero solution is found. If the loss and gain are small, it is predicted what soliton is selected by the energy-balance condition. Direct simulations demonstrate that only one pinned soliton is stable in the conservative model, and it is a semiattractor: solitons with a larger energy relax to it via emission of radiation, while those with a smaller energy decay. The same is found for solitons trapped by a pair of repulsive inhomogeneities. In the model with the loss and gain, stable pinned pulses demonstrate persistent internal vibrations and emission of radiation. If these solitons are nearly stationary, the prediction based on the energy balance underestimates the necessary gain by 10-15% (due to radiation loss). If the loss and gain are larger, the intrinsic vibrations of the pinned soliton become chaotic. The local gain alone, without the attractive perturbation of the local refractive index, cannot maintain a stable pinned soliton. For collisions of moving solitons with the "hot spot," passage and capture regimes are identified, the capture actually implying splitting of the soliton.
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We demonstrate that a symmetric system of two linearly coupled waveguides, with Kerr nonlinearity and resonant grating in both of them, gives rise to a family of symmetric and antisymmetric solitons in an exact analytical form, a part of which exists outside of the bandgap in the system's spectrum, i.e., they may be regarded as embedded solitons (ES's, i.e., the ones partly overlapping with the continuous spectrum). Parameters of the family are the soliton's amplitude and velocity. Asymmetric ES's, unlike the regular (nonembedded) gap solitons (GS's), do not exist in the system. Moreover, ES's exist even in the case when the system's spectrum contains no bandgap. The main issue is the stability of the solitons. We demonstrate that some symmetric ES's are stable, while all the antisymmetric solitons are unstable; an explanation is given to the latter property, based on the consideration of the system's Hamiltonian. We produce a full stability diagram, which comprises both embedded and regular solitons, quiescent and moving. A stability region for ES's is found around the point where the constant of the linear coupling between the two cores is equal to the Bragg-reflectivity coefficient accounting for the linear conversion between the right- and left-traveling waves in each core, i.e., the ES's are the "most endemic" solitary solitons in this system. The stability region quickly shrinks with the increase of the soliton's velocity c, and completely disappears when c exceeds half the maximum velocity. Collisions between stable moving solitons of various types are also considered, with a conclusion that the collisions are always quasielastic.
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OBJECTIVE: Hemodilution and endothelial nitric oxide synthase genetic polymorphism may contribute to cerebral and renal injury after cardiopulmonary bypass. This study tested the hypothesis that cardiopulmonary bypass and anemia stimulate an increase in cerebral and renal endothelial nitric oxide synthase gene expression in an experimental model of cardiopulmonary bypass. METHODS: Anesthetized rats underwent a sham procedure without cardiopulmonary bypass (sham, n = 5), normothermic bypass for 1 hour (CPB, n = 7), or bypass plus hemodilutional anemia (CPB anemia, n = 9). After 24 hours of recovery, RNA was extracted from the cerebral cortex, renal cortex, and renal medulla. Quantitative reverse transcriptase polymerase chain reaction was used to assess endothelial nitric oxide synthase messenger RNA levels in brain and kidney tissues. RESULTS: The hemoglobin concentration of anemic CPB rats was significantly lower than that of nonanemic rats on bypass (64 +/- 5 vs 99 +/- 8 g x L(-1), P < .001). Cerebral cortical endothelial nitric oxide synthase messenger RNA levels were increased after cardiopulmonary bypass relative to those of the sham group (11.2 +/- 4.2 vs 6.3 +/- 1.5 fg, P = .031), without a further increase in anemic rats. Renal medullary endothelial nitric oxide synthase messenger RNA levels were significantly higher in the CPB anemia group than in the sham and CPB groups (7.1 +/- 4.4 fg vs 1.8 +/- 0.4 fg vs 3.0 +/- 0.6 fg, P < .001). Renal cortical endothelial nitric oxide synthase messenger RNA levels did not change significantly. CONCLUSIONS: Normothermic cardiopulmonary bypass was associated with higher endothelial nitric oxide synthase messenger RNA levels in kidney and brain than was the sham procedure 24 hours after cardiopulmonary bypass. Anemia accentuated the increase in renal medullary, but not cerebral cortical, endothelial nitric oxide synthase expression. These data provide an approach for exploring potential mechanisms by which endothelial nitric oxide synthase may contribute to renal and cerebral dysfunction after cardiopulmonary bypass and anemia.
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Ponte Cardiopulmonar , Córtex Cerebral/metabolismo , Expressão Gênica , Rim/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Animais , Hemodiluição , Hemoglobinas/análise , Masculino , Óxido Nítrico Sintase Tipo III/genética , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Adiponectin is a multifunctional adipokine that circulates as several oligomeric complexes in the blood stream. However, the molecular basis that regulates the production of the adiponectin oligomers remains largely elusive. We have shown previously that several conserved lysine residues (positions 68, 71, 80, and 104) within the collagenous domain of adiponectin are modified by hydroxylation and glycosylation (Wang, Y., Xu, A., Knight, C., Xu, L. Y., and Cooper, G. J. (2002) J. Biol. Chem. 277, 19521-19529). Here, we investigated the potential roles of these post-translational modifications in oligomeric complex formation of adiponectin. Gel filtration chromatography revealed that adiponectin produced from mammalian cells formed trimeric, hexameric, and high molecular weight (HMW) oligomeric complexes. These three oligomeric forms were differentially glycosylated, with the HMW oligomer having the highest carbohydrate content. Disruption of hydroxylation and glycosylation by substitution of the four conserved lysines with arginines selectively abrogated the intracellular assembly of the HMW oligomers in vitro as well as in vivo. In type 2 diabetic patients, both the ratios of HMW to total adiponectin and the degree of adiponectin glycosylation were significantly decreased compared with healthy controls. Functional studies of adiponectin-null mice revealed that abrogation of lysine hydroxylation/glycosylation markedly decreased the ability of adiponectin to stimulate phosphorylation of AMP-activated protein kinase in liver tissue. Chronic treatment of db/db diabetic mice with wild-type adiponectin alleviated hyperglycemia, hypertriglyceridemia, hepatic steatosis, and insulin resistance, whereas full-length adiponectin without proper post-translational modifications and HMW oligomers showed substantially decreased activities. Taken together, these data suggest that hydroxylation and glycosylation of the lysine residues within the collagenous domain of adiponectin are critically involved in regulating the formation of its HMW oligomeric complex and consequently contribute to the insulin-sensitizing activity of adiponectin in hepatocytes.
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Adiponectina/química , Lisina/química , Processamento de Proteína Pós-Traducional , Adipócitos/metabolismo , Animais , Glicosilação , Hepatócitos/metabolismo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mutagênese Sítio-Dirigida , Estrutura Terciária de Proteína , RatosRESUMO
The fragmentation mechanisms of protonated triglycine and its first-generation dissociation products have been investigated using a combination of density functional theory calculations and threshold collision-induced dissociation experiments. The activation barrier measured for the fragmentation of protonated triglycine to the b(2) ion and glycine is in good agreement with a calculated barrier at the B3LYP/6-31++G(d,p) level of theory reported earlier [Rodriquez, C. F. et al. J. Am. Chem. Soc. 2001, 123, 3006-3012]. The b(2) ion fragments to the a(2) ion via a transition state structure that is best described as acylium-like. Contrary to what is commonly assumed, the lowest energy structure of the a(2) ion is not an iminium ion, but a cyclic, protonated 4-imidazolidone. Furthermore, fragmentation of the b(2) to the a(1) ion proceeds not via a mechanism that results in HNCO and H(2)C=C=O as byproducts, as have been postulated, but via a transition state that contains an incipient a(1) ion and an incipient carbene. The fragmentation of a(2) to a(1) proceeds via a transition state structure that contains the a(1) ion, CO and an imine as incipient components.