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The potential usefulness of lopinavir-ritonavir on Covid 19 infection during the first wave of contamination in France had boosted Kaletra® syrup prescription to the point of causing its national shortage. In the intensive care units of Parisian hospitals in charge of patients with life-threatening viral contamination, caregivers had to resort to lopinavir-ritonavir-based tablets, crushing them and then dispersing the powder in milk to facilitate administration by nasogastric tube. The difficulties and poor control of this degraded mode, which does not always ensure control of the amount of the drug in the prepared dose and may induce insufficient antiviral exposure, led us to develop in a very short time, while ensuring quality control proportional to the risk, a liquid form as an alternative to Kaletra® oral solution shortage. For this purpose, we describe this compounding formulation and its preparation process, while justifying the quality control strategy adapted to the risk as well as its chemical and physical stability. Based on the chemical and physical studies, the preparation was showed to be stable during at least 2 months between +2°C and +8°C and 1 week at room temperature. This has resulted in the design of kits that include multi-dose packaging and a measuring device and contain the appropriate quantities of drugs to ensure at least one week's treatment for each patient, during which time the kit in use can be stored at room temperature. The intensive care team used this treatment under conditions that they considered well adapted until the imported specialty became available.
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Tratamento Farmacológico da COVID-19 , Ritonavir , Combinação de Medicamentos , Hospitais , Humanos , Lopinavir/farmacologia , Lopinavir/uso terapêutico , Ritonavir/uso terapêutico , SARS-CoV-2 , SuspensõesRESUMO
OBJECTIVES: Study of the impact of geriatricians' training on the improvement of their prescribing practices, and comparison of iatrogenesis between the two classes of oral anticoagulants. MATERIAL AND METHODS: Before/after and here/there comparison between a trained prescribers group and a control group, before and after the pharmacist intervention, with comparison of the iatrogenesis of two oral anticoagulant classes. Patients in the acute and post-acute geriatric departments treated with a vitamin K antagonist or a direct oral anticoagulant were included. Criteria for Good practice were rated according to a scale of severity: calculation of a score and a percentage of compliance per patient, and then an average of the percentage of compliance (main criterion) within the populations to be compared. The proportion of iatrogenic elements between the two classes was compared. We used statistical tests (significance threshold of 5%). RESULTS: Vitamin K antagonist: a decreasing trend in the control group (P=0.086) and an increasing trend in the trained group (P=0.183) was observed in prescription compliance before/after training. Direct oral anticoagulants: the compliance before/after decreased in the control group (P=0.005) and increased in the trained group (P=0.024). After training, compliance is higher among the group of trained prescribers for both vitamin K antagonist (P=0.018) and direct oral anticoagulant (P=0.003). The proportion of iatrogenic events in the two oral anticoagulants classes was not significantly different. CONCLUSIONS: Interest of good practice reminders in the quality of oral anticoagulants prescriptions with no difference in safety of use between the two classes.
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Fibrilação Atrial , Preparações Farmacêuticas , Acidente Vascular Cerebral , Administração Oral , Idoso , Anticoagulantes/efeitos adversos , Fibrilação Atrial/tratamento farmacológico , Humanos , Doença Iatrogênica/prevenção & controle , Acidente Vascular Cerebral/tratamento farmacológicoRESUMO
In this paper, a thermal-mechanical augmented finite-element method (TM-AFEM) has been proposed, implemented and validated for steady-state and transient, coupled thermal-mechanical analyses of complex materials with explicit consideration of arbitrary evolving cracks. The method permits the derivation of explicit, fully condensed thermal-mechanical equilibrium equations which are of mathematical exactness in the piece-wise linear sense. The method has been implemented with a 4-node quadrilateral two-dimensional (2D) element and a 4-node tetrahedron three-dimensional (3D) element. It has been demonstrated, through several numerical examples that the new TM-AFEM can provide significantly improved numerical accuracy and efficiency when dealing with crack propagation problems in 2D and 3D solids under coupled thermal-mechanical loading conditions. This article is part of the themed issue 'Multiscale modelling of the structural integrity of composite materials'.
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Methods for analyzing the full complement of a biomolecule type, e.g., proteomics or metabolomics, generate large amounts of complex data. The software tools used to analyze omics data have reshaped the landscape of modern biology and become an essential component of biomedical research. These tools are themselves quite complex and often require the installation of other supporting software, libraries and/or databases. A researcher may also be using multiple different tools that require different versions of the same supporting materials. The increasing dependence of biomedical scientists on these powerful tools creates a need for easier installation and greater usability. Packaging and containerization are different approaches to satisfy this need by delivering omics tools already wrapped in additional software that makes the tools easier to install and use. In this systematic review, we describe and compare the features of prominent packaging and containerization platforms. We outline the challenges, advantages and limitations of each approach and some of the most widely used platforms from the perspectives of users, software developers and system administrators. We also propose principles to make the distribution of omics software more sustainable and robust to increase the reproducibility of biomedical and life science research.
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ß-Lactoglobulin (ß-Lg) is the main protein in whey and is known for its allergenicity and resistance to the digestion of pepsin and trypsin. The UV-C photoinduced cleavage of disulfide bonds in ß-Lactoglobulin, as promoted by excitation of tryptophan residues (Trp), is shown to induce changes in the protein's secondary structure, significantly reducing the protein's resistance to pepsin digestion. The UV-C light-induced changes in the protein secondary structure are marked by an increase in the contribution of ß-sheet and α-helix structures with a concomitantly smaller contribution of the ß-turn structural motif. The photoinduced cleavage of disulfide bonds in ß-Lg has an apparent quantum yield of Ñ = 0.0015 ± 0.0003 and was shown by transient absorption laser flash photolysis to arise by two different pathways: a) the reduction of the disulfide bond Cys66Cys160 occurs by direct electron transfer from the triplet-excited 3Trp to the disulfide bond due to the existence of a CysCys/Trp triad (Cys66Cys160/Trp61) and b) the reduction of the buried Cys106Cys119 disulfide bond involves a reaction with a solvated electron originated by the photoejection of electrons from the triplet-excited 3Trp decay. The in vitro gastric digestion index for UV-C-treated ß-Lg is revealed to have increased significantly by 36 ± 4 % and 9 ± 2 % under simulated elderly and young adult digestive conditions, respectively. When compared to the native protein, the peptide mass fingerprint profile of digested UV-C-treated ß-Lg shows a higher content and variety of peptides, including the production of some exclusive bioactive peptides such as PMHIRL and EKFDKALKALPMH.
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Lactoglobulinas , Pepsina A , Humanos , Idoso , Lactoglobulinas/química , Pepsina A/metabolismo , Estômago , Digestão , Dissulfetos/químicaRESUMO
By generating phosphorylcholine (PC)-specific, wild-type (mu), and chimeric (mu-I-A alpha) antigen receptor transfectants of mature B cells, we have shown that the COOH terminus of the mu heavy chain is essential for three major functions: immediate signal transduction (measured as changes in intracellular Ca2+), antigen presentation, and induction of immunoglobulin M secretion. A more detailed analysis of structural requirements of the COOH-terminal domains contributing to these functions was achieved by systematically replacing the spacer, cytoplasmic, and transmembranal domains of the mu-I-A alpha chimeric chain with those of mu. Using this rescue approach, we show that the carboxyl two-thirds of the transmembranal domain (proximal to the cytoplasmic domain) is required for induction of intracellular Ca2+, whereas the complete transmembranal domain is required for the function of antigen presentation but is dispensable for induction of antibody secretion.
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Linfócitos B/imunologia , Fosforilcolina/imunologia , Receptores de Antígenos de Linfócitos B/fisiologia , Sequência de Aminoácidos , Animais , Cálcio/metabolismo , Linhagem Celular , Membrana Celular/imunologia , Imunoglobulina M/metabolismo , Linfoma de Células B , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Receptores de Antígenos de Linfócitos B/genética , Transdução de Sinais/imunologia , Relação Estrutura-Atividade , TransfecçãoRESUMO
OBJECTIVES: Nivolumab is an anti-programmed cell death-1 monoclonal antibody, approved for numerous indications in oncohaematological cancers. It is available as solution for infusion at 10 mg/ml. In accordance with the Summary of Product Characteristics (SmPCs), the product is stable for 24 h at 2-8 °C after dilution. However, to anticipate the needs and constraints related to the handling of the product, the aim was to obtain additional information that will contribute to the risk assessment in case of deviation. Potential changes in the stability of Opdivo® leftovers (10 mg/ml) and diluted nivolumab in normal saline solution (2 mg/ml) over a period exceeding 24 h, at different temperatures and after freezing/thawing cycles were studied. METHODS: Turbidimetry, Ultraviolet (UV)-spectroscopy, dynamic light scattering and chromatography were used to evaluate physicochemical stability. Potential pharmacological variations were monitored in vitro by a functional binding inhibition method. RESULTS: No change was detected after 1 month of storage at 2-8 °C neither after 7 days at 40 °C. Although slight changes were detected only after 3 months under 2-8 °C, major changes were found for the same period of time at 40 °C (variants in the distribution profile, slight increase in oligomers and fragments and UV spectral modifications). Physical instability was observed upon 3 freeze/thaw cycles, with the appearance of a new protein population associated with an increase in polydispersity index. CONCLUSION: In conclusion, our results provide additional rationale to the SmPCs, regarding the use of leftovers, reassignment of bags, pre-preparation or breaking the cold chain for Nivolumab.
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Antineoplásicos Imunológicos/análise , Embalagem de Medicamentos , Inibidores de Checkpoint Imunológico/análise , Nivolumabe/análise , Antineoplásicos Imunológicos/administração & dosagem , Estabilidade de Medicamentos , Armazenamento de Medicamentos , Inibidores de Checkpoint Imunológico/administração & dosagem , Infusões Intravenosas , Nivolumabe/administração & dosagem , Solução Salina/administração & dosagem , Temperatura , Fatores de TempoRESUMO
BACKGROUND: When a test for diagnosis of infectious diseases is introduced in a resource-limited setting, monitoring quality is a major concern. An optimized design of experiment and statistical models are required for this assessment. METHODS: Interferon-gamma release assay to detect tuberculosis (TB) infection from whole blood was tested in Hanoi, Viet Nam. Balanced incomplete block design (BIBD) was planned and fixed-effect models with heterogeneous error variance were used for analysis. In the first trial, the whole blood from 12 donors was incubated with nil, TB-specific antigens or mitogen. In 72 measurements, two laboratory members exchanged their roles in harvesting plasma and testing for interferon-gamma release using enzyme linked immunosorbent assay (ELISA) technique. After intervention including checkup of all steps and standard operation procedures, the second trial was implemented in a similar manner. RESULTS: The lack of precision in the first trial was clearly demonstrated. Large within-individual error was significantly affected by both harvester and ELISA operator, indicating that both of the steps had problems. After the intervention, overall within-individual error was significantly reduced (P < 0.0001) and error variance was no longer affected by laboratory personnel in charge, indicating that a marked improvement could be objectively observed. CONCLUSION: BIBD and analysis of fixed-effect models with heterogeneous variance are suitable and useful for objective and individualized assessment of proficiency in a multistep diagnostic test for infectious diseases in a resource-constrained laboratory. The action plan based on our findings would be worth considering when monitoring for internal quality control is difficult on site.
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Ensaio de Imunoadsorção Enzimática/métodos , Interferon gama/sangue , Mycobacterium tuberculosis/isolamento & purificação , Tuberculose/diagnóstico , Análise de Variância , Antígenos de Bactérias , Humanos , Laboratórios Hospitalares/normas , Pessoal de Laboratório Médico/educação , Modelos Estatísticos , Controle de Qualidade , VietnãRESUMO
INTRODUCTION: Ear involvement by non-Hodgkin lymphoma is quite rare and can be mistaken for other common lesions encountered in otolaryngology. The literature on this subject is also limited. CASE SUMMARY: A 45-year-old man with bilateral ear nodules that progressed over two years. Biopsy of the right ear revealed a B-cell small lymphocytic lymphoma (SLL). The patient responded to radiotherapy well. He received an additional dose two months after the initial treatment because of a remaining nodularity on the right earlobe. After several months, he presented a new lesion on his nasal tip, for which a biopsy confirmed a lymphoma relapse. The patient was managed with oral prednisone and low-dose radiation with a favourable response. DISCUSSION: This case highlights the importance of including lymphoma in the differential diagnosis of ear lesions from an otolaryngology perspective. A biopsy of any lesion or nodule with an atypical course should be considered for appropriate diagnosis and management.
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Pavilhão Auricular , Neoplasias da Orelha , Leucemia Linfocítica Crônica de Células B , Neoplasias Primárias Múltiplas , Neoplasias da Orelha/tratamento farmacológico , Neoplasias da Orelha/patologia , Neoplasias da Orelha/radioterapia , Humanos , Leucemia Linfocítica Crônica de Células B/tratamento farmacológico , Leucemia Linfocítica Crônica de Células B/patologia , Leucemia Linfocítica Crônica de Células B/radioterapia , Masculino , Pessoa de Meia-Idade , Neoplasias Primárias Múltiplas/tratamento farmacológico , Neoplasias Primárias Múltiplas/patologia , Neoplasias Primárias Múltiplas/radioterapia , Neoplasias Nasais/secundário , Doses de RadiaçãoRESUMO
Imatinib (IMA) is a highly potent tyrosine kinase inhibitor used as first-line anti-cancer drug in the treatment of chronic myeloid leukemia. Due to its universal mechanism of action, IMA also has endocrine and mutagenic disrupting effects in vivo and in vitro, which raises the question of its environmental impact. However, to date, very little information is available on its environmental fate and the potential role of its transformation products (TPs) on aquatic organisms. Given the IMA resistance to hydrolysis and direct photolysis according to the literature, we sought to generate TPs through oxidative and radical conditions using the AOPs pathway. Thus, the reactivity of the cytotoxic drug IMA in water in the presence of OH and h+ was investigated for the first time in the present work. In this regard, a non-targeted screening approach was applied in order to reveal its potential TPs. The tentative structural elucidation of the detected TPs was performed by LC-HRMSn. The proposed approach allowed detecting a total of twelve TPs, among which eleven are being described for the first time in this work. Although the structures of these TPs could not be positively confirmed due to lack of standards, their chemical formulas and product ions can be added to databases, which will allow their screening in future monitoring studies. Using the quantitative structure-activity relationship (QSAR) approach and rule-based software, we have shown that the detected TPs possess, like their parent molecule, comparable acute toxicity as well as mutagenic and estrogenic potential. In addition to the in silico studies, we also found that the samples obtained at different exposure times to oxidative conditions, including those where IMA is no longer detected, retained toxicity in vitro. Such results suggest further studies are needed to increase our knowledge of the impact of imatinib on the environment.
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Organismos Aquáticos/efeitos dos fármacos , Disruptores Endócrinos/toxicidade , Mesilato de Imatinib/toxicidade , Mutagênicos/toxicidade , Fotólise , Poluentes Químicos da Água/toxicidade , Adsorção , Aliivibrio fischeri/efeitos dos fármacos , Organismos Aquáticos/genética , Biodegradação Ambiental , Catálise , Simulação por Computador , Disruptores Endócrinos/química , Disruptores Endócrinos/efeitos da radiação , Mesilato de Imatinib/química , Mesilato de Imatinib/efeitos da radiação , Estrutura Molecular , Mutagênicos/química , Mutagênicos/efeitos da radiação , Relação Quantitativa Estrutura-Atividade , Titânio/química , Poluentes Químicos da Água/química , Poluentes Químicos da Água/efeitos da radiaçãoRESUMO
AIM: The aim of this case series was to report the use of 8% topical capsaicin patch (marketed under the trade name Qutenza®) a in the management of refractory neuropathic pain (NP) in adult patients with type 1 neurofibromatosis (NF1). METHODS: Capsaicin has been suggested for NF1 patients suffering from refractory peripheral NP despite several years of analgesic treatments. The patch was applied for 60 minutes on the painful area, with tolerability control (blood pressure, intensity of pain and dermal reaction). The evaluation was done at the beginning of treatment and during the 2 months following the first treatment (phone calls at weeks 1, 2, 4 and 8). The primary efficacy criterion was the response rate: a patient was considered to be responding if he or she reported an average relief ≥30% at the time of the follow-up calls. The secondary criteria were: interference scores (QCD), Patient Global Impression of Change (PGIC) and overall treatment satisfaction, self-reported by the patient. RESULTS: Eight patients (5 females/3 males, 41.8 ± 8.2 years of age) received a first treatment with capsaicin. Patients had pre-existing pain for 6.6 years (±6.0) and were currently receiving an average of 6.1 (±3.9) different analgesics. The response rate was 37.5%. The three responders felt globally improved and satisfied, with the improvement in overall condition as interference scores decreased. Apart from the expected local reactions, the treatment was not accompanied by systemic side effects. CONCLUSIONS: As suggested in this case series, capsaicin provided pain relief in certain NF1 patients with resistant NP. The response rate is that expected in multi-line refractory NP. A significant benefit on the overall condition of some patients was observed. In addition, this topical treatment is administered every 3 months without systemic effects. This study is limited by the small number of patients, but was intended to describe a new and well tolerated alternative treatment.
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Analgésicos/administração & dosagem , Capsaicina/administração & dosagem , Neuralgia/tratamento farmacológico , Neurofibromatose 1/tratamento farmacológico , Administração Tópica , Adulto , Capsaicina/uso terapêutico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neuralgia/etiologia , Manejo da Dor/métodosRESUMO
A simple detection system with a high-performance liquid chromatography (HPLC) with positive ionisation-tandem mass spectrometry (ESI-MS/MS) for determining diphemanil methylsulphate (DMS) levels in human plasma using 4-diphemanylmethylene,1-methylpiperidine as an internal standard (I.S.), is proposed. The acquisition was performed with the multiple reactional monitoring (MRM) mode, by monitoring the transitions: m/z 278>262 for DMS and m/z 263>247 for the I.S. The method involved a simple single-step deproteinisation with acetonitrile. The analyte was chromatographed on a Zorbax C18 reversed-phase chromatographic column by isocratic elution with 10(-3)M ammonium acetate and 10(-3)M hexafluorobutyric acid, adjusted to pH 7.0 with ammoniac/acetonitrile (40/60, v/v). The results were linear over the studied range (0.5-50.0 ng mL(-1)) and the total analysis time for each run was 10 min. The mean extraction apparent recoveries expressed at the 95% intervals of confidence were 94-104% for DMS and 92-106% for the I.S. The intra- and inter-assay precisions were 4.6-8.4% and 2.9-10.6%, respectively. The limit of quantification was 0.15 ng mL(-1). The devised assay was successfully applied to the residual concentrations monitoring in infant.
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Cromatografia Líquida de Alta Pressão/métodos , Piperidinas/sangue , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas em Tandem/métodos , Humanos , Lactente , Piperidinas/uso terapêutico , Reprodutibilidade dos TestesRESUMO
3,4-Diaminopyridine is used to treat some symptoms met in Lambert-Eaton myasthenia syndrome. It was shown efficient to reduce a form of variable muscle weakness and fatigability typical of the disease and correlated to a block of acetylcholine release. In France, 3,4-diaminopyridine is nowadays given to patients under capsules form and the status of hospital preparation. Whatever the diluant used in the formulation, the stability period could not exceed 12 months. Preliminary studies were made on a salt form in order to test the influence of various stress factors and determine if there is interaction between them. From this study, the most influent stress condition, presence of hydrogen peroxide, was selected and a comparative study was performed to compare the stability of molecular and salt species. Solutions of each species were exposed to 5 or 15% of hydrogen peroxide and analyzed at 8, 24, 72 and 216 h of degradation by HPLC-UV. Fractions of detected impurities were purified and collected by semi-preparative HPLC-UV and analyzed by HPLC-UV-ESI-MS and IR spectroscopy in order to determine their structure hypotheses. Theses experiments demonstrate that the salt species were more stable under oxidative stress condition than molecular species. The two main degradation products were collected and identified as 4-amino, 3-nitropyridine and 3,4-diaminopyridine-N-oxide when the molecular form was degraded whereas only 4-amino, 3-nitropyridine was found in less quantity in the salt solutions. Nitrogen pyridine and pyridine amine could not easily be oxidized by hydrogen peroxide in salt comparatively to molecular species due to the lone pair of electron engaged in a bound with hydrogen in the first case and by resonance change of the pyridine in the second case. This modification of structure promoted different pathways of degradation for the salt form which are more dependent of energy. Owing to the better stability of the salt species, a new pharmaceutical form containing it was developed to assess its stability under ICH standard conditions allowing an industrial manufacture of this drug.
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4-Aminopiridina/análogos & derivados , 4-Aminopiridina/análise , Amifampridina , Cromatografia Líquida de Alta Pressão , Estabilidade de Medicamentos , Peróxido de Hidrogênio/química , Indicadores e Reagentes , Cinética , Padrões de Referência , Espectrometria de Massas por Ionização por Electrospray , Espectrofotometria Infravermelho , Espectrofotometria UltravioletaRESUMO
A new coccidian species (Protozoa: Apicomplexa: Isospora) is described parasitizing white-necked thrushes Turdus albicollis Vieillot, 1818; rufous-bellied thrushes Turdus rufiventris Vieillot, 1818; pale-breasted thrushes Turdus leucomelas Vieillot, 1818; and yellow-legged thrushes Turdus flavipes Vieillot, 1818 from 3 different localities in Brazil. Isospora sabiai n. sp. has oocysts that are subspherical to ellipsoidal, 20.9 × 18.6 µm, with smooth, delicate, bilayered wall, â¼1.1 µm thick. Micropyle inconspicuous or imperceptible. Oocyst residuum absent, but small polar granules rounded or comma-shaped are present. Sporocysts are elongate ellipsoidal to reniform, 16.5 × 9.2 µm. The Stieda body is knob-like. Sub-Stieda body rounded to conical, sometimes homogeneous with the Stieda body. Sporocyst residuum is present, usually as a cluster of numerous granules. Sporozoites are vermiform with 2 refractile bodies. The oocysts and sporocysts of I. sabiai n. sp. are uniform in the proportionality of width on length, but exhibited different patterns of size associated with each host species; therefore, an ecological discussion is introduced aimed at associating these morphometrical patterns of the oocysts with the habits of the different species of thrushes. This is the seventh isosporoid coccidian reported from New World turdids.
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Doenças das Aves/parasitologia , Isospora/classificação , Isosporíase/veterinária , Aves Canoras/parasitologia , Animais , Doenças das Aves/epidemiologia , Brasil/epidemiologia , Fezes/parasitologia , Ilhas/epidemiologia , Isospora/fisiologia , Isosporíase/epidemiologia , Isosporíase/parasitologia , Modelos Lineares , PrevalênciaRESUMO
We describe a patient with a history of recurrent squamous cell carcinoma of the tongue and abnormal FDG uptake in the left arm during a re-staging FDG PET/CT. After revision of the patient's clinical history, tests and physical exam, the abnormal FDG uptake was found to correspond to an extensive aseptic deep venous thrombosis of the upper extremity.
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Fluordesoxiglucose F18 , Tomografia por Emissão de Pósitrons/métodos , Tomografia Computadorizada por Raios X/métodos , Extremidade Superior/diagnóstico por imagem , Trombose Venosa/diagnóstico , Carcinoma de Células Escamosas/complicações , Carcinoma de Células Escamosas/diagnóstico por imagem , Humanos , Achados Incidentais , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/complicações , Neoplasias Bucais/diagnóstico por imagem , Recidiva Local de Neoplasia/complicações , Recidiva Local de Neoplasia/diagnóstico por imagem , Compostos Radiofarmacêuticos , Sepse/complicações , Sepse/diagnóstico , Trombose Venosa/complicaçõesRESUMO
The C-terminal domain of RNA polymerase II is an unusual series of repeated residues appended to the C-terminus of the largest subunit and serves as a flexible binding scaffold for numerous nuclear factors. The binding of these factors is determined by the phosphorylation patterns on the repeats in the domain. In this study, we generated a synthetic antibody library by replacing the third heavy chain complementarity-determining region of an anti-HER2 (human epidermal growth factor receptor 2) antibody (trastuzumab) with artificial sequences of 7-18 amino-acid residues. From this library, antibodies were selected that were specific to serine phosphopeptides that represent typical phosphorylation patterns on the functional unit (YSPTSPS)2 of the RNA polymerase II C-terminal domain (CTD). Antibody clones pCTD-1stS2 and pCTD-2ndS2 showed specificity for peptides with phosphoserine at the second residues of the first or second heptamer repeat, respectively. Additional clones specifically reacted to peptides with phosphoserine at the fifth serine of the first repeat (pCTD-1stS5), the seventh residue of the first repeat and fifth residue of the second repeat (pCTD-S7S5) or the seventh residue of either the first or second repeat (pCTD-S7). All of these antibody clones successfully reacted to RNA polymerase II in immunoblot analysis. Interestingly, pCTD-2ndS2 precipitated predominately RNA polymerase II from the exonic regions of genes in genome-wide chromatin immunoprecipitation sequencing analysis, which suggests that the phosphoserine at the second residue of the second repeat of the functional unit (YSPTSPS)2 is a mediator of exon definition.
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Anticorpos/metabolismo , Imunoprecipitação da Cromatina/métodos , Éxons , RNA Polimerase II/metabolismo , Anticorpos/imunologia , Células HEK293 , Células HeLa , Humanos , Fosforilação , Ligação Proteica , RNA Polimerase II/imunologiaRESUMO
In vivo pyruvate synthesis by malic enzyme (ME) and pyruvate kinase and in vivo malate synthesis by phosphoenolpyruvate carboxylase and the Krebs cycle were measured by 13C incorporation from [1-13C]glucose into glucose-6-phosphate, alanine, glutamate, aspartate, and malate. These metabolites were isolated from maize (Zea mays L.) root tips under aerobic and hypoxic conditions. 13C-Nuclear magnetic resonance spectroscopy and gas chromatography-mass spectrometry were used to discern the positional isotopic distribution within each metabolite. This information was applied to a simple precursor-product model that enabled calculation of specific metabolic fluxes. In respiring root tips, ME was found to contribute only approximately 3% of the pyruvate synthesized, whereas pyruvate kinase contributed the balance. The activity of ME increased greater than 6-fold early in hypoxia, and then declined coincident with depletion of cytosolic malate and aspartate. We found that in respiring root tips, anaplerotic phosphoenolpyruvate carboxylase activity was high relative to ME, and therefore did not limit synthesis of pyruvate by ME. The significance of in vivo pyruvate synthesis by ME is discussed with respect to malate and pyruvate utilization by isolated mitochondria and intracellular pH regulation under hypoxia.
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OBJECTIVE: Amplitude-integrated electroencephalography (aEEG) monitoring is increasing in the neonatal population, but the safety and feasibility of performing aEEG in extremely preterm infants have not been systematically evaluated. STUDY DESIGN: Inborn infants 23(0/7) to 28(6/7) weeks gestation or birth weight 401 to 1000 g were eligible. Serial, 6-h aEEG recordings were obtained from first week of life until 36 weeks postmenstrual age. Adverse events were documented, and surveys evaluated the impact of the aEEGs on routine care. Success of performing aEEGs according to protocol and aEEG quality were assessed. RESULT: A total of 102 infants were enrolled, with 755 recordings performed. 83% of recordings were performed according to schedule, and 96% were without adverse event. Bedside nurses reported no interference with routine care for 89% of recordings. 92% of recordings had acceptable signal quality. CONCLUSION: Serial aEEG monitoring is safe in preterm infants, with few adverse events and general acceptance by nursing staff.
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Eletroencefalografia/efeitos adversos , Eletroencefalografia/métodos , Recém-Nascido de Peso Extremamente Baixo ao Nascer , Lactente Extremamente Prematuro , Adulto , Encéfalo/fisiologia , Estudos de Coortes , Feminino , Idade Gestacional , Humanos , Lactente , Recém-Nascido , Masculino , Recursos Humanos de Enfermagem Hospitalar , Adulto JovemRESUMO
To determine the effect of nutritional agents on lipid peroxidation, 10 smokers were given 6 mg beta carotene, 200 IU vitamin E, and 250 mg vitamin C 4 times daily for 3 weeks. Lipid peroxidation was assessed by measuring baseline and postsupplementation levels of exhaled ethane. There was a 29% decrease in mean (+/-SD) exhaled ethane (4.06 +/- 1.49 vs 2.90 +/- 1.29 pmol.kg-1.min-1), with individual levels decreasing in 8 of the 10 smokers (p < 0.05, Wilcoxon sign rank test). Three nonsmokers had very low baseline levels of ethane that did not change with supplementation. Ethane production correlated with active (packs per day) and lifelong (pack-years) tobacco consumption. Also, a strong correlation was found between the decline in ethane output after micronutrient supplementation and the presupplement FEV1. Therefore, antioxidant vitamin supplementation resulted in attenuation of smoking-related lipid peroxidation, and the decreases in ethane production appears to be associated with preserved lung function.
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Antioxidantes/farmacologia , Etano/análise , Respiração , Fumar/metabolismo , Vitaminas/farmacologia , Ácido Ascórbico/farmacologia , Testes Respiratórios , Carotenoides/farmacologia , Feminino , Volume Expiratório Forçado , Humanos , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Fumar/fisiopatologia , Vitamina E/farmacologia , beta CarotenoRESUMO
[reaction: see text] This communication describes the first examples of tandem endo-regioselective and stereospecific oxacyclizations of 1, 5-diepoxides to oxepane products and a similar tandem oxacyclization of 1,5,9-triepoxides to fused bisoxepane cyclic carbonates. A mechanism for these biomimetic oxacyclizations is proposed in which the epoxides act as both electrophilic and nucleophilic reaction partners.