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Interleukin-17A (IL-17A) is a pro-inflammatory cytokine implicated in diverse autoimmune and inflammatory disorders such as psoriasis and Kawasaki disease. Mature IL-17A is a homodimer that binds to the extracellular type-III fibronectin D1:D2-dual domain of its cognate IL-17 receptor A (IL-17RA). In this study, we systematically examined the structural basis, thermodynamics property, and dynamics behavior of IL-17RA/IL-17A interaction and computationally identified two continuous hotspot regions separately from different monomers of IL-17A homodimer that contribute significantly to the interaction, namely I-shaped and U-shaped segments, thus rendered as a peptide-mediated protein-protein interaction (PmPPI). Self-inhibitory peptides (SIPs) are derived from the two segments to disrupt IL-17RA/IL-17A interaction by competitively rebinding to the IL-17A-binding pocket on IL-17RA surface, which, however, only have a weak affinity and low specificity for IL-17RA due to lack of the context support of intact IL-17A protein, thus exhibiting a large flexibility and intrinsic disorder when splitting from the protein context and incurring a considerable entropy penalty when rebinding to IL-17RA. The U-shaped segment is further extended, mutated and stapled by a disulfide bridge across its two strands to obtain a number of double-stranded cyclic SIPs, which are partially ordered and conformationally similar to their native status at IL-17RA/IL-17A complex interface. Experimental fluorescence polarization assays substantiate that the stapling can moderately or considerably improve the binding affinity of U-shaped segment-derived peptides by 2-5-fold. In addition, computational structural modeling also reveals that the stapled peptides can bind in a similar mode with the native crystal conformation of U-shaped segment in IL-17RA pocket, where the disulfide bridge is out of the pocket for avoiding intervene of the peptide binding.
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Interleucina-17 , Receptores de Interleucina-17 , Interleucina-17/química , Interleucina-17/metabolismo , Interleucina-17/farmacologia , Receptores de Interleucina-17/química , Receptores de Interleucina-17/metabolismo , Peptídeos/química , Modelos Moleculares , Ligação ProteicaRESUMO
RATIONALE: Neuromyelitis optica spectrum disorder (NMOSD) involves autoimmune and inflammatory responses in the central nervous system, primarily affecting the optic nerves and spinal cord. Atypical presentations such as ataxia and syncope complicate the diagnosis, and lesions in the medulla are easily mistaken for cerebral infarction. This case report emphasizes the need to recognize such manifestations to avoid misdiagnosis and ensure timely treatment. PATIENT CONCERNS: This case report presents an NMOSD female patient who experienced ataxia, syncope, and neuropathic pain during her illness. DIAGNOSIS: NMOSD. INTERVENTIONS: The patient managed her blood sugar with insulin, controlled neuropathic pain with pregabalin, and underwent 5 plasma exchanges. OUTCOMES: Significant improvement was noted 1 week post-plasma exchange, with complete resolution of neuropathic pain and no symptom recurrence reported at 6-month follow-up. LESSONS: Atypical manifestations of NMOSD, such as ataxia, syncope, and trigeminal neuralgia, increase diagnostic difficulty. Recognizing these symptoms is crucial to avoid misdiagnosis and ensure timely and appropriate treatment for patients.
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Ataxia , Neuralgia , Neuromielite Óptica , Síncope , Humanos , Feminino , Neuromielite Óptica/complicações , Neuromielite Óptica/diagnóstico , Ataxia/diagnóstico , Ataxia/etiologia , Síncope/etiologia , Síncope/diagnóstico , Neuralgia/etiologia , Neuralgia/diagnóstico , Progressão da Doença , Adulto , Pessoa de Meia-Idade , Troca Plasmática/métodosRESUMO
Valproate encephalopathy is one of the unusual and severe but treatable side effect. This research focuses on four female patients who had valproate medication for epilepsy and developed an increased frequency of seizures, exacerbated disruption of consciousness, gastrointestinal problems, cognitive dysfunction, ataxia, and psychobehavioral abnormalities. The patient's symptoms improved over time once sodium valproate was stopped. As a result, when using sodium valproate, one should be aware of the risk of sodium valproate encephalopathy and cease using the medication right once if any of the above symptoms of unknown etiology manifest clinically. We also go over the potential pathogenesis that lead to valproate encephalopathy and the heightened risk of encephalopathy from taking antiepileptic medications together. It was stressed how crucial it is to identify, diagnose, and treat sodium valproate encephalopathy as soon as possible.
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Human respiratory syncytial virus (RSV) is a primary cause of lower respiratory tract infections and hospital visits during infancy and childhood. The RSV phosphoprotein (P) is a major polymerase cofactor that interacts with nucleoprotein (N) to promote the recognition of ribonucleoprotein complex (RNP) by viral RNA polymerase. The binding pocket of N protein is chemically diverse, in or around which a number of aromatic and charged amino acid residues are observed. Previously, a nonapeptide segment (P peptide, 233DNDLSLEDF241) representing the C-terminal tail of P protein was identified to mediate the N-P interaction with a moderate affinity, in which the Phe241 at the end of P's C-terminus plays a critical role in the binding of P peptide to N protein. Here, we found that the side-chain aromatic phenyl moiety of P Phe241 residue can form short- and long-range cation-π interactions with N Arg132 and Arg150 residues, respectively, as well as T-shaped and parallel-displaced π-π stackings with N Tyr135 and His151 residues, respectively, which co-define a geometrically satisfactory π-stacking system at the complex interface of N protein with P peptide, thus largely stabilizing the complex architecture. The stacking effect was further optimized by systematically mutating the P Phe241 residue to other natural and non-natural aromatic amino acids with diverse chemical substitutions at the phenyl moiety to examine their structural and energetic effects on π-stacking system and on protein-peptide binding. The electron-donating mutations at the phenyl moiety of P Phe241 residue can effectively enhance the π-stacking system and then promote peptide binding, whereas the bulky and positively charged mutations would considerably impair the peptide potency by introducing steric hindrance and electrostatic repulsion. The [Tyr]P, [Thp]P and [Fph]P mutants were determined to have an increased affinity relative to wild-type P peptide, which could be used as self-inhibitory peptides to competitively disrupt the native interaction between N and P proteins.
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Vírus Sincicial Respiratório Humano , Humanos , Criança , Vírus Sincicial Respiratório Humano/genética , Vírus Sincicial Respiratório Humano/química , Vírus Sincicial Respiratório Humano/metabolismo , Fosfoproteínas/química , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Nucleocapsídeo/metabolismo , Nucleoproteínas , Peptídeos/químicaRESUMO
Sclerosing epithelioid fibrosarcoma (SEF) is a rare and poorly defined variant of fibrosarcoma, but generally insensitive to chemotherapy and progresses with poor prognosis. We report the marvelous effect of irinotecan hydrochloride (CPT-11) chemotherapy in rescuing a patient with atypical SEF from emergent condition, who underwent recurrences after several treatment methods. Small dose of CPT-11 was administered to the patient, with which, the size of superficial mass (cervical lymph node) gradually decreased observed by the naked eyes in 5 days. X-ray and CT image proved a marked reduction in the size of the tumor. CPT-11 is valuable for the treatment of this aggressive sarcoma. In condition of emergency caused by sarcoma oppression, administering a tolerable small dose of topoisomerase I-inhibiting drug could be a beneficial choice.
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Background: A high-risk infant (HRI) is a child whose fetal, neonatal, and infant development is impacted by adverse factors that may cause cognitive, sensory, behavioral, or language defects. The complex situation in the treatment process is a continuous challenge and stressor for parents. If parents fail to take appropriate coping styles, it will have an adverse impact on the health of parents and the growth and development of children. The purpose of this study was to explore the impact of clinical characteristics, postpartum posttraumatic stress disorder (PTSD), and family coping on the neurodevelopment and language development of HRIs as a reference for targeted intervention. Methods: This study retrospectively recruited 211 children who were hospitalized in the neonatal intensive care unit (NICU) of Suzhou Kowloon Hospital from January 2018 to December 2021. HRI and their mother were interviewed by telephone with general information questionnaire, Perinatal Post-traumatic Stress Disorder Questionnaire, medical coping modes questionnaire, Bayley Scales of Infant Development the Early Language Milestone Scale to investigate HRI and their mothers; Logistic regression was used to analyze the relationship between HRI mothers' emotions and family coping with neurodevelopment and language development. Results: The neurodevelopmental scores of HRIs differed according to gestational week of delivery, birth weight, and disease diagnosis. The language development scores of HRIs differed according to gestational week of delivery, birth weight, disease diagnosis, and maternal education. Multiple stepwise regression analysis showed that the neurodevelopmental scores were affected by gestational week of delivery, postpartum PTSD score, and family coping. Logistic regression analysis showed that the language development scores were affected by maternal education and neurodevelopmental level. The correlation analysis showed that the postpartum PTSD scores were negatively correlated with family coping, neurodevelopment, and language development, and that family coping was positively correlated with the neurodevelopment and language development scores (P<0.05). Conclusions: The neurodevelopment and language development of HRIs were affected by the gestational week of delivery, maternal education, the child's birth weight, and disease diagnosis. NICU wards can promote the healthy development of HRIs by providing mothers with targeted health education concerning the child's condition, postpartum PTSD, and family coping strategies.
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Exploring the relationship between environmental air quality (EAQ) and climatic conditions on a large scale can help better understand the main distribution characteristics and the mechanisms of EAQ in China, which is significant for the implementation of policies of joint prevention and control of regional air pollution. In this study, we used the concentrations of six conventional air pollutants, i.e., carbon monoxide (CO), sulfur dioxide (SO2), nitrogen dioxide (NO2), fine particulate matter (PM2.5), coarse particulate matter (PM10), and ozone (O3), derived from about 1300 monitoring sites in eastern China (EC) from January 2015 to December 2018. Exploiting the grading concentration limit (GB3095-2012) of various pollutants in China, we also calculated the monthly average air quality index (AQI) in EC. The results show that, generally, the EAQ has improved in all seasons in EC from 2015 to 2018. In particular, the concentrations of conventional air pollutants, such as CO, SO2, and NO2, have been decreasing year by year. However, the concentrations of particulate matter, such as PM2.5 and PM10, have changed little, and the O3 concentration increased from 2015 to 2018. Empirical mode decomposition (EOF) was used to analyze the major patterns of AQI in EC. The first mode (EOF1) was characterized by a uniform structure in AQI over EC. These phenomena are due to the precipitation variability associated with the East Asian summer monsoon (EASM), referred to as the "summer-winter" pattern. The second EOF mode (EOF2) showed that the AQI over EC is a north-south dipole pattern, which is bound by the Qinling Mountains and Huaihe River (about 35° N). The EOF2 is mainly caused by seasonal variations of the mixed concentration of PM2.5 and O3. Associated with EOF2, the Mongolia-Siberian High influences the AQI variation over northern EC by dominating the low-level winds (10 m and 850 hPa) in autumn and winter, and precipitation affects the AQI variation over southern EC in spring and summer.
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Poluentes Atmosféricos , Poluição do Ar , Poluentes Atmosféricos/análise , Poluição do Ar/análise , China , Cidades , Monitoramento Ambiental , Mongólia , Material Particulado/análise , Estações do AnoRESUMO
The water-insoluble procedures in US Pharmacopeia (USP) General Chapter Residual Solvents <467>, which are based on European Pharmacopoeia procedures, were optimized and modified before their inclusion in the chapter to improve their scope, performance, and ruggedness. The optimized procedures use a static headspace introduction system with a gas chromatograph equipped with a flame ionization detector. This article describes some of the key changes made to the USP published procedures, including use of dimethyl sulfoxide (DMSO) or dimethylformamide (DMF) as the solvent, addition of 5 mL of water and 1 mL of sample (dissolved in DMSO or DMF) to the headspace vial, use of a 3:1 GC split ratio, and use of new matrix-matched system suitability solutions. These procedures were verified with two different active pharmaceutical ingredients--hydroxyzine pamoate and prednisone. In the investigation, the more polar material (hydroxyzine pamoate) showed greater recoveries for the optimized procedures when prepared in DMSO. The less polar material (prednisone) typically had greater recoveries in DMF for the optimized procedures. During experimentation, insights into sample preparation, additional types of headspace instrumentation, solvent purity, and other parameters were also gained.
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Dimetil Sulfóxido/química , Dimetilformamida/química , Avaliação Pré-Clínica de Medicamentos/métodos , Cromatografia Gasosa-Espectrometria de Massas/métodos , Solventes/química , Água/química , Sensibilidade e Especificidade , Solubilidade , VolatilizaçãoRESUMO
Only a month after the outbreak of pneumonia caused by 2019-nCoV, more than forty-thousand people were infected. This put enormous pressure on the Chinese government, medical healthcare provider, and the general public, but also made the international community deeply nervous. On the 25th day after the outbreak, the Chinese government implemented strict traffic restrictions on the area where the 2019-nCoV had originated-Hubei province, whose capital city is Wuhan. Ten days later, the rate of increase of cases in Hubei showed a significant difference (p = 0.0001) compared with the total rate of increase in other provinces of China. These preliminary data suggest the effectiveness of a traffic restriction policy for this pandemic thus far. At the same time, solid financial support and improved research ability, along with network communication technology, also greatly facilitated the application of epidemic prevention measures. These measures were motivated by the need to provide effective treatment of patients, and involved consultation with three major groups in policy formulation-public health experts, the government, and the general public. It was also aided by media and information technology, as well as international cooperation. This experience will provide China and other countries with valuable lessons for quickly coordinating and coping with future public health emergencies.
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Betacoronavirus/patogenicidade , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/prevenção & controle , Epidemias/prevenção & controle , Pneumonia Viral/epidemiologia , Pneumonia Viral/prevenção & controle , Animais , COVID-19 , China , Quirópteros/virologia , Comunicação , Governo , Humanos , Saúde Pública/métodos , SARS-CoV-2 , Viagem , Zoonoses/virologiaRESUMO
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to sweep the world, causing infection of millions and death of hundreds of thousands. The respiratory disease that it caused, COVID-19 (stands for coronavirus disease in 2019), has similar clinical symptoms with other two CoV diseases, severe acute respiratory syndrome and Middle East respiratory syndrome (SARS and MERS), of which causative viruses are SARS-CoV and MERS-CoV, respectively. These three CoVs resulting diseases also share many clinical symptoms with other respiratory diseases caused by influenza A viruses (IAVs). Since both CoVs and IAVs are general pathogens responsible for seasonal cold, in the next few months, during the changing of seasons, clinicians and public heath may have to distinguish COVID-19 pneumonia from other kinds of viral pneumonia. This is a discussion and comparison of the virus structures, transmission characteristics, clinical symptoms, diagnosis, pathological changes, treatment and prevention of the two kinds of viruses, CoVs and IAVs. It hopes to provide information for practitioners in the medical field during the epidemic season.
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Infecções por Coronavirus/diagnóstico , Influenza Humana/diagnóstico , Pneumonia Viral/diagnóstico , Infecções Respiratórias/virologia , Estações do Ano , Fatores Etários , Animais , Betacoronavirus , COVID-19 , Infecções por Coronavirus/complicações , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/transmissão , Humanos , Vírus da Influenza A/patogenicidade , Influenza Humana/complicações , Influenza Humana/prevenção & controle , Influenza Humana/transmissão , Coronavírus da Síndrome Respiratória do Oriente Médio/patogenicidade , Pandemias/prevenção & controle , Pneumonia Viral/complicações , Pneumonia Viral/prevenção & controle , Pneumonia Viral/transmissão , Saúde Pública , Infecções Respiratórias/transmissão , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/patogenicidade , SARS-CoV-2 , Síndrome Respiratória Aguda Grave/diagnóstico , Síndrome Respiratória Aguda Grave/virologiaRESUMO
Purpose: The prognostic role of programmed death-ligand 1 (PD-L1) in colorectal cancer remains unclear. We employed a meta-analysis to explore the prognostic value of PD-L1 and to ascertain the relationship between PD-L1 expression and clinicopathological characteristics in CRC patients. Methods: We systematically searched PubMed, Embase and the Cochrane Library until October 2018. Eligible studies about colorectal cancer that pay attention to PD-L1 expression and studies reporting survival information were included. In order to evaluate the prognostic role of PD-L1 for overall survival (OS) and recurrence-free survival (RFS)/disease-free survival (DFS), Hazard ratio (HR) with 95% confidence interval (CI) was used. Odds ratio (OR) with 95% CI was selected to appraise the correlation between PD-L1 with clinicopathological characteristics of colorectal cancer patients. Begg's funnel plot was used to assess publication bias. Results: Twelve studies involving 4344 patients published from 2013 to 2018 were included in this meta-analysis. Pooled results revealed that PD-L1 overexpression was relevant to shorter OS (HR 1.47, 95% CI =1.01-2.15, p=0.04) and shorter RFS/DFS (HR 1.47, 95% CI =1.01-2.15, p=0.04). Moreover, Patients with high expression of PD-L1 associated with inferior tumor stage (OR=0.57, 95% CI: 0.45, 0.74, p<0.0001) and Vascular invasion-negativity (OR=0.75, 95% CI: 0.6, 0.94, p=0.01). But the expression of PD-L1 is not related to age, sex, tumor location, tumor differentiation, pT stage, pN stage, MSI/MMR status. Conclusion: This meta-analysis revealed that PD-L1 can serve as a significant biomarker for negative prognosis and the adverse clinicopathological features of colorectal cancer and could facilitate the better management of individual patients.
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PURPOSE: Immune suppression is common in patients with advanced breast cancer but the mechanisms underlying this phenomenon have not been sufficiently studied. In this study, we aimed to identify B7 family members that were able to predict the immune status of patients, and which may serve as potential targets for the treatment of breast cancer. We also aimed to identify microRNAs that may regulate the expression of B7 family members. METHODS: The Cancer Genome Atlas data from 1,092 patients with breast cancer, including gene expression, microRNA expression and survival data, were used for statistical and survival analyses. Polymerase chain reaction and Western blot were used to measure messenger RNA and protein expression, respectively. Luciferase assay was used to investigate direct microRNA target. RESULTS: Bioinformatic analysis predicted that microRNA (miR)-93, miR-195, miR-497, and miR-340 are potential regulators of the immune evasion of breast cancer cells, and that they exert this function by targeting CD274, PDCD1LG2, and NCR3LG1. We chose CD274 for further investigations. We found that miR-195, miR-497, and CD274 expression levels were inversely correlated in MDA-MB-231 cells, and miR-195 and miR-497 expressions mimic inhibited CD274 expression in vitro. Mechanistic investigations demonstrated that miR-195 and miR-497 directly target CD274 3' untranslated region. CONCLUSION: Our data indicated that the level of B7 family members can predict the prognosis of breast cancer patients, and miR-195/miR-497 regulate CD274 expression in triple negative breast cancer. This regulation may further influence tumor progression and the immune tolerance mechanism in breast cancer and may be able to predict the effect of immunotherapy on patients.
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The Raman spectra (3200-30 cm(-1)) of liquid and solid, and infrared spectra of gaseous and solid chloromethyl silyl dichloride, ClCH2SiHCl2, have been recorded. Variable temperature (-105 to -150 degrees C) studies of the infrared spectra of the sample dissolved in liquid krypton have been carried out. From these data, the enthalpy difference was determined to be 363 +/- 40 cm(-1) (4.34 +/- 0.48 kJ mol(-1)), with the more stable form being the gauche conformer, which is consistent with the prediction from ab initio calculations at both the Hartree-Fock level and with full electron correlation by the perturbation method to second order. It is estimated that 92% of the sample is in the gauche form at ambient temperature. A complete vibrational assignment is proposed for the gauche conformer and several of fundamentals of the trans conformer based on infrared band contours, relative intensities, depolarization values, and group frequencies, which is supported by normal coordinate calculations utilizing the force constants from the ab initio MP2/6-31G(d) calculations. The r0 SiH bond distances of 1.476 and 1.472 A have been obtained for the trans and gauche conformers, respectively, from the silicon-hydrogen stretching frequencies. The optimized geometries have also been obtained from ab initio calculations utilizing several different basis sets with full electron correlation by the perturbation method up to MP2/6-311 + G(2d,2p). The results are discussed and compared to some corresponding results for several related molecules.
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Compostos de Silício/química , Conformação Molecular , Espectrofotometria Infravermelho , Análise Espectral RamanRESUMO
The infrared (3100-40 cm(-1)) and Raman (3100-20 cm(-1)) spectra of gaseous and solid n-propylsilane, CH(3)CH(2)CH(2)SiH(3) and the Si-d(3) isotopomer, CH(3)CH(2)CH(2)SiD(3), have been recorded. Additionally, the Raman spectra of the liquids have been recorded and qualitative depolarization values obtained. Both the anti and gauche conformers have been identified in the fluid phases but only the anti conformer remains in the solid. Variable temperature (-105 to -150 degrees C) studies of the infrared spectra of n-propylsilane dissolved in liquid krypton have been recorded and the enthalpy difference has been determined to be 220+/-22 cm(-1) (2.63+/-0.26 kJ mol(-1)) with the anti conformer the more stable form. A similar value of 234+/-23 cm(-1) (2.80+/-0.28 kJ mol(-1)) was obtained for deltaH for the Si-d(3) isotopomer. At ambient temperature it is estimated that there is 30+/-2% of the gauche conformer present. The potential function governing the conformation interchange has been estimated from the far infrared spectral data, the enthalpy difference, and the dihedral angle of the gauche conformer, which is compared to the one predicted from ab initio MP2/6-31G(d) calculations. The barriers to conformational interchange are: 942, 970 and 716 cm(-1) for the anti to gauche, gauche to gauche, and gauche to anti conformers, respectively. Relatively complete vibrational assignments are proposed for both the n-propylsilane-d(0) and Si-d(3) molecules based on the relative infrared and Raman spectral intensities, infrared band contours, depolarization ratios, and normal coordinate calculations. The geometrical parameters, harmonic force constants, vibrational frequencies, infrared intensities, Raman activities and depolarization ratios, and energy differences have been obtained for the anti and gauche conformers from ab initio MP2/6-31G(d) calculations. Structural parameters and energy differences have also been obtained utilizing the larger 6-311 + G(d,p) and 6-311 + G(2d,2p) basis sets. From the isolated Si-H stretching frequency from the Si-d(2) isotopomer the r(0) distances of 1.484 and 1.485 A have been determined for the SiH(s) and SiH(a) bonds, respectively, for the anti conformer, and 1.486 A for the SiH bond for the gauche conformer. Utilizing previously reported microwave rotational constants for the anti conformer and the determined SiH distances along with ab initio predicted parameters 'adjusted r(0)' parameters have been obtained for the anti conformer. The results are discussed and compared to those obtained for some similar molecules.
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Compostos de Silício/química , Espectrofotometria Infravermelho , Análise Espectral RamanRESUMO
Variable temperature (-55 to -155 degrees C) infrared spectra of rare gas solutions of 2-chloro-3-fluoropropene, H2C=C(Cl)CH2F, have been recorded from 3500 to 400 cm(-1). The relative intensities of 16 conformer pairs at ten different temperatures of a krypton solution have been measured and from these data an enthalpy difference of 271+/-27 cm(-1) (3.17+/-0.32 kJ x mol(-1)) has been obtained with the cis conformer the more stable form. Similar studies were also carried out in xenon and an enthalpy difference of 334+/-33 cm(-1) (4.00+/-0.39 kJ x mol(-1)) was obtained again with the cis conformer the more stable form. It is estimated that there is 35+/-2% of the gauche conformer present at ambient temperature. However, in the crystalline solid the gauche conformer is the stable rotamer. Extensive ab initio calculations with full electron correlation by the perturbation method at the MP2 level with a variety of basis sets as well as density functional theory calculations (DFT) by the B3LYP method have been carried out. Several of these calculations predict an energy difference in the range of 400 cm(-1) with the cis form the more stable conformer but most of the predicted energy differences are significantly larger than the experimentally determined value. The spectroscopic and theoretical results are discussed and compared with the corresponding quantities for some similar molecules.
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Compostos Alílicos/química , Clorofluorcarbonetos/química , Estabilidade de Medicamentos , Gases , Conformação Molecular , Soluções , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Temperatura , TermodinâmicaRESUMO
The infrared (3100-40 cm(-1)) spectra of gaseous and solid and Raman (3200-20 cm(-1)) spectra of liquid with qualitative depolarization values and solid n-propyltrifluorosilane, CH(3)CH(2)CH(2)SiF(3), have been recorded. Additionally the infrared spectra of the sample in nitrogen and argon matrices have been recorded. Both the anti and gauche conformers have been identified in the fluid phases but only the anti conformer remains in the solid. Variable temperature (-105 to -150 degrees C) studies of the infrared spectra of the sample dissolved in liquid krypton have been recorded and the enthalpy difference has been determined to be 135+/-14 cm(-1) (1.62+/-0.17 kJ mol(-1)) with the anti conformer the more stable form. At ambient temperature it is estimated that there is 51+/-2% of the gauche conformer present. Also the enthalpy difference in the liquid was obtained from variable temperature studies of the Raman spectra and from three conformer pairs an average value of 179+/-18 cm(-1) (2.14+/-0.22 kJ mol(-1)) was obtained again with the anti form the more stable conformer. Relatively complete vibrational assignments are proposed for both conformers based on the relative infrared and Raman spectral intensities, infrared band contours, depolarization ratios which are supported by normal coordinate calculations. The geometrical parameters, harmonic force constants, vibrational frequencies, infrared intensities, Raman activities, depolarization ratios, and energy differences have been obtained for the anti and gauche conformers from ab initio MP2/6-31G(d) calculations. Structural parameters and energy differences have also been obtained utilizing the larger 6-311+G(d, p) and 6-311+G(2d, 2p) basis sets. By utilizing the previously reported microwave rotational constants for five isotopomers of CH(3)SiF(3) along with ab initio predicted structural values, r(0) parameters have been obtained for methyltrifluorosilane. Similarly, from the ab initio predicted parameters "adjusted r(0)" parameters have been estimated for both conformers of n-propyltrifluorosilane. The results are discussed and compared with those obtained for some similar molecules.
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Compostos de Flúor/química , Silanos/química , Temperatura Baixa , Conformação Molecular , Espectrofotometria Infravermelho , Análise Espectral RamanRESUMO
OBJECTIVE: To study the effect of tumor necrosis factor-alpha (TNF-alpha) on the morphology and ultrastructure of dendritic cells (DCs). METHODS: Freshly isolated precursors of DCs were cultured in the medium consisting of RPMI 1640 medium and 10 % inactivated fetal bovine serum. In one experiment, the cells were pretreated by TNF-alpha (200 U/ml) for 4 h, followed by treatment with human granulocyte macrophage colony stimulating factor (rhGM-CSF, 800 U/ml) and interleukin-4 (IL-4, 500 U/ml). The above treatments were repeated again every 48 h. The cells in another experiment were treated with rhGM-CSF (800 U/ml) and IL-4 (500 U/ml) on the day of in vitro culture, which was repeated every 48 h with a treatment with TNF-alpha (200 U/ml) on day 5 of cell culture. On day 8, the cells in the 2 parallel experiments were collected with the former cells designated as T-DC and the latter DC. Morphological differences between the 2 group of cells were observed using light microscope and scanning electron microscope. RESULTS: The T-DC exhibited more typical dendric processes than DCs did. CONCLUSION: Pretreatment of the precursors of DCs with TNF-alpha for 4 h, in comparison with TNF-alpha treatment on day 5 of cell culture, produced DCs with more typical and conspicuous dendric morphologies.
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Células Dendríticas/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologia , Tamanho Celular/efeitos dos fármacos , Células Cultivadas , Células Dendríticas/citologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Humanos , Interleucina-4/farmacologiaRESUMO
OBJECTIVE: To explore a new method for in vitro culture of mature dendritic cells (DCs) by utilizing the maturation-inducing effect of tumor necrosis factor-alpha(TNF-alpha) on DC precursors. METHODS: Freshly isolated DC precursors (peripheral blood-derived monocytes) were initially cultivated for 4 h in RPMI 1640 supplemented with 10% fetal bovine serum and containing TNF-alpha (200 U/ml), followed by treatment with rhGM-CSF (800 U/ml) and interleukin (IL)-4 (500 U/ml) added into the medium. After 48 and 96 h, the 3 cytokines were simultaneously added to the medium for further culture. On day 8 of cell culture, the DCs were harvested and designated as T-DCs, which were subsequently subjected to examinations of flow cytometry analysis, assay by autologous and allogeneic mixed lymphocyte reaction (MLR) for estimating their maturation, and MTT assay for their specific killing activity. RESULTS: The T-DCs expressed specific surface markers (CD83,CD1a, CD54, CD11a, CD80, CD86, HLA-DR, CD83HLA-DR) of mature DCs, and stimulated strong T-cell proliferative responses in both autologous and autogeneic MLR. It also activated CD4+ T cells, which proliferated in response to autologous tumor, and specifically lysed tumor targets. CONCLUSION: The DCs thus cultured in vitro possess typical phenotype, and can present antigens to T cells, and this method provides an alternative in culturing mature and functionally competent DCs.
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Células Dendríticas/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologia , Células Dendríticas/imunologia , Humanos , Técnicas In Vitro , Leucócitos Mononucleares/citologia , Linfócitos T/imunologiaRESUMO
OBJECTIVE: To study the influence of albumin supplementation in the medium and the placement time outside the incubator on the viability of isolated cytokine-induced killer (CIK) cells. METHODS: CIK cells labeled with anti-CD3-FITC and anti-CD56-PE or with FQ-AE were observed under fluorescence microscope. The effect of albumin in the cell medium on the cell viability was analyzed using flow cytometry with Annexin V-FITC after different time lengths of placement. RESULTS: The clones of CIK cells from the patient with esophageal carcinoma were small and scattered in the medium, but the clones from the patients with pancreatic cancer were large and densely distributed. CD3(+)CD56(+) cells could be detected under fluorescence microscope. The addition of albumin in the medium did not obviously affect cell apoptosis and death of CIK cells. CIK cells placed outside the incubator for less than 90 min showed a significant lower apoptosis rate than the cells placed for 150 min, whereas the cell death rate did not vary significantly with the placement time. CONCLUSION: CIK cells from different cancer patients present with different growth pattern of the cells clones. Labeling with anti-CD3-FITC and anti-CD56-PE allows convenient counting of the newly generated CD3(+)CD56(+) CIK cells and FQ-AE labeling can be used for quantitative assessment of cell death. Albumin is not necessary in the medium of CIK cells. Prolonged placement (for over 90 min) of CIK cells outside the incubator should be avoided, and the placement time should be shorten as much as possible.
Assuntos
Albuminas/análise , Células Matadoras Induzidas por Citocinas/metabolismo , Citocinas/metabolismo , Idoso , Apoptose , Complexo CD3/imunologia , Antígeno CD56/imunologia , Sobrevivência Celular , Meios de Cultura/química , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Tempo , Células Tumorais CultivadasRESUMO
AIMS: The presence of genetic variants for autism spectrum disorders (ASDs) was investigated for the metabotropic glutamate receptor 7 (GRM7) gene in a case-control study. MAIN METHODS: Employing Affymetrix SNP microarrays, 297 single nucleotide polymorphisms (SNPs) covering the GRM7 gene were selected and analyzed in ASD patients (n=22), non-ASD patients [n=14, including seven patients with development delay (DD)/mental retardation (MR), four patients with language delay (LD), and three patients with attention deficit hyperactivity disorder (ADHD)] and normal control subjects (n=18). KEY FINDINGS: Twenty-one statistically significant SNPs with different inheritance models (recessive, dominant and allele) were demonstrated in three groups (ASDs vs. combined controls, ASDs vs. normal controls, ASDs vs. non-ASD patients). Associations of rs779867 and rs6782011 with ASDs were significant in all three groups and independent associations of rs779867 and rs6782011 with ASDs were found in the ASD vs. combined controls group, which are in modest linkage disequilibrium (D'>0.5). Further haplotype analysis showed that rs6782011/rs779867 (T-C) was statistically significantly related to ASDs in both the ASD vs. combined controls and ASD vs. normal controls groups (bootstrap P value=0.013, permutation P value=0.013 for the former group and bootstrap P value=0.002, permutation P value=0.020 for the latter). SIGNIFICANCE: These findings support a role for genetic variants within the GRM7 gene in 3p26.1 in ASDs.