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1.
Rev. esp. anestesiol. reanim ; 70(10): 593-595, Dic. 2023.
Artigo em Espanhol | IBECS | ID: ibc-228136

RESUMO

La granulomatosis eosinofílica con poliangitis, también denominada síndrome de Churg-Strauss, es una vasculitis rara con compromiso multisistémico. Existe poca literatura en cuanto al enfoque anestésico de los pacientes con esta enfermedad. A nuestro saber, presentamos aquí el primer informe sobre cirugía ambulatoria en un paciente con granulomatosis eosinofílica con poliangitis. Este caso subraya la preocupación por la seguridad de la cirugía ambulatoria y el manejo anestésico exitoso de un paciente con granulomatosis eosinofílica con poliangitis con anestesia regional.(AU)


Eosinophilic granulomatosis with polyangiitis, also known as Churg-Strauss syndrome, is a rare type of vasculitis with multisystemic involvement. Very few authors have described the anaesthesia technique in these patients. We present the first report on ambulatory surgery in a patient with eosinophilic granulomatosis with polyangiitis. This case dispels concerns about the safety of day surgery and reports successful regional anaesthesia management in a patient with eosinophilic granulomatosis with polyangiitis.(AU)


Assuntos
Humanos , Masculino , Pessoa de Meia-Idade , Síndrome de Churg-Strauss/cirurgia , Procedimentos Cirúrgicos Ambulatórios/métodos , Anestesia por Condução , Vasculite/classificação , Eosinofilia/diagnóstico , Broncoscopia , Anestesiologia , Pacientes Internados , Exame Físico , Avaliação de Sintomas , Síndrome de Churg-Strauss/complicações , Consentimento Livre e Esclarecido , Síndrome de Churg-Strauss/diagnóstico
2.
Vet Parasitol ; 324: 110069, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37984155

RESUMO

Although ivermectin (IVM) has a wide spectrum and long half-life, its frequent use as an anthelmintic for the last 42 years led to its worldwide tolerance by Haemonchus contortus. We evaluated the combination of limonene (LIM), a P-glycoprotein (Pgp) modulator, with IVM in lambs infected with a multidrug-resistant H. contortus. Twenty-four male Dorper lambs were artificially infected with two doses (seven days apart) of 8000 infective larvae of a multidrug-resistant isolate of H. contortus. The infection was patent 25 days later. Fifteen days before treatment with IVM (DAY -15), animals were divided into 4 groups: Infected-untreated control (CTL), IVM, LIM, and LIM+IVM. From DAY -15 to DAY + 14, groups LIM and LIM+IVM received 200 mg/kg of body weight/day of LIM via oral. On DAY 0, a single dose of IVM at 200 µg/kg of body weight was administered orally to groups IVM and LIM+IVM. On DAY + 7 and DAY + 14, fecal egg counts (FEC) were performed and on DAY + 14 animals were euthanized for total worm count (TWC), worm length, fecundity of females, and Pgp-9 gene expression. On DAY + 7, group LIM+IVM had 96.29% efficacy based on Fecal Egg Count Reduction TEST (FECRT) and a highly significant reduction in FEC (P = 0.0005) when compared to CTL. On DAY + 14, the efficacy of LIM+IVM was 82.87% on FECRT, although no differences were found among groups for FEC, TWC, worm length, or Pgp-9 gene expression. Female worms from the CTL group had higher egg counts in their uterus when compared to LIM. No differences were found for hematological or biochemical parameters, body weight, or weight gain among groups. Thus, LIM given daily at 200 mg/kg was safe for animals and, when combined with IVM, decreased egg shedding and could reduce pasture contamination, although it was unable to kill multidrug-resistant H. contortus.


Assuntos
Anti-Helmínticos , Hemoncose , Haemonchus , Doenças dos Ovinos , Ovinos , Animais , Feminino , Masculino , Ivermectina/farmacologia , Ivermectina/uso terapêutico , Haemonchus/genética , Limoneno/farmacologia , Óvulo , Anti-Helmínticos/farmacologia , Anti-Helmínticos/uso terapêutico , Carneiro Doméstico , Peso Corporal , Subfamília B de Transportador de Cassetes de Ligação de ATP/genética , Expressão Gênica , Hemoncose/tratamento farmacológico , Hemoncose/veterinária , Doenças dos Ovinos/tratamento farmacológico , Fezes , Contagem de Ovos de Parasitas/veterinária
3.
Rev Esp Anestesiol Reanim (Engl Ed) ; 70(10): 593-595, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37666453

RESUMO

Eosinophilic granulomatosis with polyangiitis (EGPA), also known as Churg-Strauss syndrome, is a rare type of vasculitis with multisystemic involvement. Very few authors have described the anaesthesia technique in these patients. We present the first report on ambulatory surgery in a patient with EGPA. This case dispels concerns about the safety of day surgery and reports successful regional anaesthesia management in a patient with EGPA.


Assuntos
Síndrome de Churg-Strauss , Granulomatose com Poliangiite , Humanos , Síndrome de Churg-Strauss/complicações
5.
Rev. chil. endocrinol. diabetes ; 14(2): 74-76, 2021.
Artigo em Espanhol | LILACS | ID: biblio-1283555

RESUMO

La enfermedad de Gaucher (EG) es un trastorno genético lisosomal autosómico recesivo infrecuente, que conduce a la acumulación de lípidos y disfunción en múltiples órganos. La afectación del esqueleto es uno de los hallazgos más frecuentes de la EG y una de las principales causas de dolor y reducción de calidad de vida. El compromiso esquelético incluye anomalías en el remodelado óseo con pérdida mineral ósea, adelgazamiento cortical, lesiones líticas, fracturas por fragilidad y deformidades articulares. A continuación presentamos el caso de una paciente 61 años con osteoporosis grave secundaria a EG diagnosticada en la vida adulta, con antecedente de dos hermanas con EG. La paciente refería dolores óseos y lumbago crónico desde los 53 años. El 2012 fue evaluada en policlínico de hematología por trombocitopenia y debido a sus antecedentes familiares se le solicitaron exámenes que fueron compatibles con EG. El año 2016 la densitometría ósea (DXA) de columna lumbar y cuello femoral izquierdo, que mostró una osteoporosis. Se inició tratamiento con Alendronato, Calcio y Vitamina D, pero la paciente tuvo escasa adherencia. El 2018 se inició tratamiento de su EG con Taliglucerasa α. Al año siguiente se le realizó nueva DXA que evidenció persistencia de la osteoporosis y por mantención del lumbago se le solicitó una TAC de columna lumbar que mostró fracturas por aplastamiento de cuerpos vertebrales dorsales bajos. Se derivó a endocrinología para manejo de su osteoporosis grave. A su ingreso a endocrinología la paciente persitía con dolor lumbar alto y destacaba una marcada cifosis. Se decidió retomar tratamiento con Alendronato, calcio y vitamina D, además, se le solicitó una nueva evaluación densitométrica junto a una radiografía de columna total y evaluación dental. Durante el seguimiento la paciente mantuvo niveles de vitamina D adecuados con funciones renal, hepática y tiroidea normales.


Gaucher disease (GD) is a rare autosomal recessive lysosomal genetic disorder, leading to the accumulation and dysfunction of lipids in multiple organs. Skeletal involvement is one of the most prevalent aspects of GD and one of the main causes of pain and reduced quality of life. Abnormalities of bones, which cause changes in the development and loss of bone mineral, cortical thinning, lytic lesions,fragility fractures and deformities. We present a case of a patient diagnosed with severe osteoporosis, secondary to GD diagnosed in adult life. The patient presents a disease pattern composed of bone pain and chronic low back pain since the age of 53. In 2012, she was evaluated at the hematology for thrombocytopenia and due to her family history, tests were performed to diagnose GD, which were compatible with it. In 2016 Bone Densitometry (DXA) of the lumbar spine and left femoral neck was requested, being consistent with osteoporosis. Treatment with Alendronate, Calcium and Vitamin D was started, however, there is little adherence. In 2018, treatment for Gaucher's disease was started with Taliglucerase α. The following year, DXA was performed with few changes and a CT scan of the lumbar spine was performed diagnosing crush fractures of the low dorsal vertebral bodies. She was referred to endocrinology. Upon admission to Endocrinology, it was decided to resume initial osteoporosis treatment and to perform skeletal evaluation with DXA of the lumbar spine and hips, total spine X-ray and dental evaluation. During follow-up, it maintains vitamin D at adequate levels and normal kidney, liver and thyroid functions.


Assuntos
Humanos , Feminino , Pessoa de Meia-Idade , Osteoporose/etiologia , Doença de Gaucher/complicações , Osteoporose/terapia , Dor Lombar/etiologia
6.
Nat Commun ; 5: 5199, 2014 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-25310906

RESUMO

Changing climatic conditions have led to a significant increase in the magnitude and frequency of extreme rainfall events in the Central Andes of South America. These events are spatially extensive and often result in substantial natural hazards for population, economy and ecology. Here we develop a general framework to predict extreme events by introducing the concept of network divergence on directed networks derived from a non-linear synchronization measure. We apply our method to real-time satellite-derived rainfall data and predict more than 60% (90% during El Niño conditions) of rainfall events above the 99th percentile in the Central Andes. In addition to the societal benefits of predicting natural hazards, our study reveals a linkage between polar and tropical regimes as the responsible mechanism: the interplay of northward migrating frontal systems and a low-level wind channel from the western Amazon to the subtropics.

7.
J Med Virol ; 85(11): 1983-9, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23926069

RESUMO

Since the 1980s, 2 antigenically distinct influenza B lineages have cocirculated in the world: B/Victoria/2/87 (first appeared in the 1980s) and B/Yamagata/16/88 (became predominant in the 1990s). B/Victoria/2/87 isolates were geographically restricted to eastern Asia during 1991-2000. During 2000-2001 and 2001-2002, B/Victoria/2/87 isolates reemerged in North America, Europe, and South America, and then spread globally. During influenza virus surveillance, season 2002, an outbreak of acute respiratory illness, which quickly spread among the population, has been notified by public health authorities living in Araraquara, São Paulo, Brazil. Instituto Adolfo Lutz and Secretariat of Health of São Paulo state teams initiate an investigation towards to describe the pattern of infection in this population temporally and by age and to characterize the strains by virus isolation and hemagglutination inhibition assay. The outbreak lasted approximately 10 weeks; many cases occurred between mid-August and mid-September. Children younger than 13 years were the most affected; the elderly were mostly immune to infection. Analysis of the clinical respiratory samples helped in identifying the B/Hong Kong/330/2001 and B/Brisbane/32/2002 subtypes-recent variants of B/Victoria/02/88, a lineage restricted to Southeast Asia until 2001. The Araraquara outbreak confirms the reemergence of the B/Victoria viruses in South America and highlights the importance of monitoring local circulating strains, especially in light of the absence of cross-protection between antigenically distinct influenza lineages. Based on influenza virus surveillance, public health authorities worldwide should decide whether trivalent vaccines or quadrivalent vaccines (containing both influenza virus B lineages) are to be used in each country.


Assuntos
Surtos de Doenças , Vírus da Influenza B/genética , Vírus da Influenza B/isolamento & purificação , Vacinas contra Influenza/imunologia , Influenza Humana/epidemiologia , Influenza Humana/virologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antivirais/sangue , Brasil/epidemiologia , Criança , Pré-Escolar , Reações Cruzadas , Feminino , Testes de Inibição da Hemaglutinação , Humanos , Lactente , Recém-Nascido , Vírus da Influenza B/classificação , Vacinas contra Influenza/administração & dosagem , Pessoa de Meia-Idade , Epidemiologia Molecular , Adulto Jovem
8.
Life Sci ; 92(10): 569-75, 2013 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-23352973

RESUMO

AIMS: We previously reported that mechanical atrial stretch (AS) by balloon distention increased gastric tonus in anesthetized rats. The present study evaluated the effect of AS on the gastric emptying of a liquid test meal in awake rats and its underlying neural mechanisms. MAIN METHODS: Anesthetized male rats received a balloon catheter into the right atrium and a gastrostomy cannula. The next day, mean arterial pressure (MAP), heart rate (HR), central venous pressure (CVP), and cardiac output (CO) were continuously monitored. After the first 20min of monitoring (basal interval), the balloon was either distended or not (control) with 30, 50, or 70µl saline for 5min. Fifteen minutes later, the rats received the test meal (glucose solution with phenol red), and fractional gastric dye retention was determined 10, 20, or 30min later. KEY FINDINGS: Heart rate and CVP values were transiently increased by 50 or 70µl AS but not 30µl AS, whereas gastric emptying was slower after 30, 50, or 70µl AS than after sham distention. Subdiaphragmatic vagotomy or splanchnicotomy+celiac ganglionectomy and capsaicin, ondansetron, hexamethonium, L-NAME, and glibenclamide treatment prevented the AS-induced delay in gastric emptying, whereas atropine and guanethidine treatment failed to prevent it. SIGNIFICANCE: Atrial stretch inhibited the gastric emptying of liquid via non-adrenergic and non-cholinergic pathways that activate nitric oxide-K(+)ATP channels.


Assuntos
Função do Átrio Direito/fisiologia , Esvaziamento Gástrico/fisiologia , Trânsito Gastrointestinal/fisiologia , Átrios do Coração/cirurgia , Hipovolemia/fisiopatologia , Análise de Variância , Animais , Atropina/farmacologia , Oclusão com Balão/métodos , Pressão Sanguínea , Capsaicina/farmacologia , Cateterismo Cardíaco/métodos , Cateterismo/métodos , Esvaziamento Gástrico/efeitos dos fármacos , Gastrostomia/métodos , Glibureto/farmacologia , Guanetidina/farmacologia , Frequência Cardíaca , Hexametônio/farmacologia , Masculino , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico/sangue , Ondansetron/farmacologia , Ratos
9.
Genet Mol Res ; 10(3): 1707-18, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21863565

RESUMO

The large number of microbial genomes deposited in databanks has opened the door for in-depth studies of organisms, including post-genomics investigations. Thanks to new generation sequencing technology, these studies have made advances that have lead to extraordinary discoveries in bacterial transcriptomics. In this review, we describe bacterial RNA sequencing studies that use these new techniques. We also examined the advantages and biases of these new generation technologies; advances in bioinformatics make it possible to overcome the biases, providing interesting and surprising results.


Assuntos
Bactérias/genética , Genoma Bacteriano , RNA Bacteriano/química , Transcriptoma , Sequência de Bases , RNA Bacteriano/genética , Análise de Sequência de RNA/métodos
10.
São Paulo; Secretaria da Saúde. Coordenação de Vigilância em Saúde. Gerência do Centro de Controle e Prevenção de Doenças; 2011. 1 p. ilus, graf.
Não convencional em Português | Coleciona SUS, COVISA-Producao, Sec. Munic. Saúde SP, Sec. Munic. Saúde SP | ID: biblio-937373
11.
São Paulo; Secretaria da Saúde. Coordenação de Vigilância em Saúde. Gerência do Centro de Controle e Prevenção de Doenças; 2011. 1 p. ilus, graf, tab.
Não convencional em Português | Coleciona SUS, COVISA-Producao, Sec. Munic. Saúde SP, Sec. Munic. Saúde SP | ID: biblio-937378
14.
Micron ; 41(6): 666-73, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20399104

RESUMO

Human alpha2-macroglobulin (alpha 2M) is a 720 kDa glycoprotein that presents two ultrastructural conformations: slow (S-alpha 2M) and fast (F-alpha 2M). alpha 2M acts mainly as a proteinase scavenger, but an immunomodulatory role was also proposed. This work studies the effect of desialylation and deglycosylation on the structure patterns of alpha 2M by ultrastructural analysis of lectin-induced aggregates, which represents a new approach that had never been previously used. Transmission electron microscopy (TEM) analysis showed the loss of S-alpha 2M conformation after deglycosylation, indicating that glycosidic side-chains contribute to the molecular stability of S-alpha 2M. TEM proved to be an important tool to analyze the effect of biochemical changes on alpha 2M, yielding an objective qualitative control of its morphological state. Certain carbohydrate residues did not vary between the alpha 2M conformations, since both bound similarly ConA and WGA lectins. However, the binding of PNA and BSI-B(4) was slightly lower in F-alpha 2M than in S-alpha 2M. Among the neuraminidases used to desialylate both conformations of alpha 2M that from Arthrobacter ureafaciens was the most effective. Incubation with the lectins ConA or SNA, respectively specific for mannosyl and sialyl residues, led to dose-dependent patterns of aggregation of alpha 2M molecules, mediated by lectin binding and clearly visualized by TEM.


Assuntos
Glicoconjugados/análise , alfa-Macroglobulinas/química , Humanos , Lectinas/metabolismo , Microscopia Eletrônica de Transmissão/métodos , Ligação Proteica , Conformação Proteica , alfa-Macroglobulinas/ultraestrutura
15.
Diabetologia ; 53(7): 1428-37, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20361177

RESUMO

AIMS/HYPOTHESIS: Cell-cell coupling mediated by gap junctions formed from connexin (CX) contributes to the control of insulin secretion in the endocrine pancreas. We investigated the cellular production and localisation of CX36 and CX43, and gap junction-mediated beta cell coupling in pancreatic islets from rats of different ages, displaying different degrees of maturation of insulin secretion. METHODS: The presence and distribution of islet connexins were assessed by immunoblotting and immunofluorescence. The expression of connexin genes was evaluated by RT-PCR and quantitative real-time PCR. The ultrastructure of gap junctions and the function of connexin channels were assessed by freeze-fracture electron microscopy and tracer microinjection, respectively. RESULTS: Young and adult beta cells, which respond to glucose, expressed significantly higher levels of Cx36 (also known as Gjd2) than fetal and newborn beta cells, which respond poorly to the sugar. Accordingly, adult beta cells also showed a significantly higher membrane density of gap junctions and greater intercellular exchange of ethidium bromide than newborn beta cells. Cx43 (also known as Gja1) was not expressed by beta cells, but was located in various cell types at the periphery of fetal and newborn islets. CONCLUSIONS/INTERPRETATION: These findings show that the pattern of connexins, gap junction membrane density and coupling changes in islets during the functional maturation of beta cells.


Assuntos
Conexinas/metabolismo , Células Secretoras de Insulina/metabolismo , Ilhotas Pancreáticas/metabolismo , Animais , Animais Recém-Nascidos , Conexina 43/genética , Conexina 43/metabolismo , Conexinas/genética , Feminino , Imunofluorescência , Junções Comunicantes/metabolismo , Immunoblotting , Células Secretoras de Insulina/ultraestrutura , Ilhotas Pancreáticas/crescimento & desenvolvimento , Ilhotas Pancreáticas/ultraestrutura , Masculino , Microscopia Eletrônica , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteína delta-2 de Junções Comunicantes
16.
J Nanosci Nanotechnol ; 10(2): 1148-52, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20352770

RESUMO

One of the factors that limit the efficiency of polymer-based optoelectronic devices, such as photovoltaic solar cells and light emitting diodes, is the exciton diffusion within the polymeric network. Due to the amorphous nature the of polymeric materials, the diffusion of excitons is limited by the energetic and spatial disorder in such systems, which is a consequence not only of the chemical structure of the polymer but also from its morphology at nanoscale. To get a deep understanding on how such effects influence exciton dynamics we performed a quantum molecular dynamics simulations to determine the energetic disorder within the polymer system, and Monte Carlo simulations to study exciton diffusion in three-dimensional (3D) polymer networks that present both spatial and energetic disorder at nanometre scale. Our results show clearly that exciton diffusion in poly(p-phenylenevinylene) (PPV) occurs preferentially in the direction parallel to the electrodes surface for a polymer-based optoelectronic devices with the orientation of the conjugated strands similar to those obtained by the spin-coating technique and the decay of such excitons occurs preferentially in longer strands which allow us to get insight on exciton behaviour in polymeric systems that are not possible to be obtained directly from the experiments.

17.
Antimicrob Agents Chemother ; 54(5): 2023-31, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20231395

RESUMO

From a series of 1,3,4-thiadiazole-2-arylhydrazone derivatives of megazol screened in vitro against Trypanosoma cruzi, eight (S1 to S8) were selected for in vivo screening by single-dose oral administration (200 mg/kg of body weight) to infected mice at 5 days postinfection (dpi). Based on significant decreases in both parasitemia levels and mortality rates, S2 and S3 were selected for further assays. Despite having no in vivo effect, S1 was included since it was 2-fold more potent against trypomastigotes than megazol in vitro. Trypomastigotes treated with S1, S2, or S3 showed alterations of the flagellar structure and of the nuclear envelope. When assayed on intracellular amastigotes, the selectivity index (SI) for macrophages was in the range of >27 to >63 and for cardiac cells was >32 for S1 and >48 for megazol. In noninfected mice, S1 did not alter the levels of glutamic oxalacetic transaminase (GOT), glutamate pyruvate transaminase (GPT), or urea. S2 led to an increase in GOT, S3 to increases in GOT and GPT, and megazol to an increase in GOT. Infected mice were treated with each derivative at 50 and 100 mg/kg from dpi 6 to 15: S1 did not interfere with the course of infection or reduce the number of inflammatory foci in the cardiac tissue, S2 led to a significant decrease of parasitemia, and S3 decreased mortality. There was no direct correlation between the in vitro effect on trypomastigotes and amastigotes and the results of the treatment in experimental models, as S1 showed a high potency in vitro while, in two different schemes of in vivo treatment, no decrease of parasitemia or mortality was observed.


Assuntos
Doença de Chagas/tratamento farmacológico , Hidrazonas/farmacologia , Tiadiazóis/farmacologia , Trypanosoma cruzi/efeitos dos fármacos , Alanina Transaminase/sangue , Animais , Antiprotozoários/química , Antiprotozoários/farmacologia , Aspartato Aminotransferases/sangue , Peso Corporal , Células Cultivadas , Doença de Chagas/mortalidade , Doença de Chagas/parasitologia , Hidrazonas/química , Técnicas In Vitro , Concentração Inibidora 50 , Macrófagos Peritoneais/citologia , Macrófagos Peritoneais/parasitologia , Masculino , Camundongos , Microscopia Eletrônica de Varredura , Miócitos Cardíacos/citologia , Miócitos Cardíacos/parasitologia , Parasitemia/tratamento farmacológico , Parasitemia/mortalidade , Parasitemia/parasitologia , Tiadiazóis/química , Trypanosoma cruzi/crescimento & desenvolvimento , Trypanosoma cruzi/ultraestrutura , Ureia/sangue
18.
J Proteomics ; 73(8): 1433-40, 2010 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-20123049

RESUMO

Considering the importance of bidimensional electrophoresis and image analysis in comparative proteomics, the parameters that influence the analysis of protein expression of transgenic and non-transgenic soybean seeds were evaluated. The loaded mass of the proteins (150-500 microg), the pH separation range (3-10 or 4-7), and manual/automatic image editing were evaluated. Additionally, after optimizing the conditions, histograms and matchings were obtained in order to accurately analyze the variations (90%) in protein expression. From this, 10 proteins displayed significant differences in expression, and eight of them were characterized and identified by mass spectrometry.


Assuntos
Eletroforese em Gel Bidimensional/métodos , Proteômica/métodos , Proteínas de Soja/química , Concentração de Íons de Hidrogênio , Processamento de Imagem Assistida por Computador/métodos , Plantas Geneticamente Modificadas , Sementes/química , Glycine max/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos
19.
J Chromatogr A ; 1217(9): 1429-36, 2010 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-20083249

RESUMO

The DNA binding fusion protein, LacI-His6-GFP, together with the conjugate PEG-IDA-Cu(II) (10 kDa) was evaluated as a dual affinity system for the pUC19 plasmid extraction from an alkaline bacterial cell lysate in poly(ethylene glycol) (PEG)/dextran (DEX) aqueous two-phase systems (ATPS). In a PEG 600-DEX 40 ATPS containing 0.273 nmol of LacI fusion protein and 0.14% (w/w) of the functionalised PEG-IDA-Cu(II), more than 72% of the plasmid DNA partitioned to the PEG phase, without RNA or genomic DNA contamination as evaluated by agarose gel electrophoresis. In a second extraction stage, the elution of pDNA from the LacI binding complex proved difficult using either dextran or phosphate buffer as second phase, though more than 75% of the overall protein was removed in both systems. A maximum recovery of approximately 27% of the pCU19 plasmid was achieved using the PEG-dextran system as a second extraction system, with 80-90% of pDNA partitioning to the bottom phase. This represents about 7.4 microg of pDNA extracted per 1 mL of pUC19 desalted lysate.


Assuntos
Cromatografia de Afinidade/métodos , DNA/isolamento & purificação , Iminoácidos/química , Plasmídeos/isolamento & purificação , Polietilenoglicóis/química , Proteínas Recombinantes de Fusão/metabolismo , Eletroforese em Gel de Ágar , Escherichia coli/metabolismo , Proteínas de Escherichia coli/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Histidina/metabolismo , Repressores Lac/metabolismo , Lactoglobulinas/metabolismo , Oligopeptídeos/metabolismo
20.
Clin Exp Immunol ; 158(3): 294-9, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19778295

RESUMO

Human T lymphotropic virus type 1 (HTLV-1) infects 10-20 million people worldwide. The majority of infected individuals are asymptomatic; however, approximately 3% develop the debilitating neurological disease HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). There is also currently no cure, vaccine or effective therapy for HTLV-1 infection, and the mechanisms for progression to HAM/TSP remain unclear. NK T cells are an immunoregulatory T cell subset whose frequencies and effector functions are associated critically with immunity against infectious diseases. We hypothesized that NK T cells are associated with HAM/TSP progression. We measured NK T cell frequencies and absolute numbers in individuals with HAM/TSP infection from two cohorts on two continents: São Paulo, Brazil and San Francisco, CA, USA, and found significantly lower levels when compared with healthy subjects and/or asymptomatic carriers. Also, the circulating NK T cell compartment in HAM/TSP subjects is comprised of significantly more CD4(+) and fewer CD8(+) cells than healthy controls. These findings suggest that lower numbers of circulating NK T cells and enrichment of the CD4(+) NK T subset are associated with HTLV-1 disease progression.


Assuntos
Células T Matadoras Naturais/imunologia , Paraparesia Espástica Tropical/imunologia , Adulto , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Portador Sadio/imunologia , Progressão da Doença , Feminino , Infecções por HTLV-I/imunologia , Humanos , Imunidade Celular , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Subpopulações de Linfócitos T/imunologia , Adulto Jovem
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