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OBJECTIVE OF THE STUDY: The objective of the present study is to assess the mechanical behavior of trabecular bone based on microCT imaging and micro-finite-element analysis. In this way two methods are detailed: (i) direct determination of macroscopic elastic property of trabecular bone; (ii) inverse approach to assess mechanical properties of trabecular bone tissue. PATIENTS: Thirty-five females and seven males (forty-two subjects) mean aged (±SD) 80±11.7 years from hospitals of Assistance publique-Hôpitaux de Paris (AP-HP) diagnosed with osteoporosis following a femoral neck fracture due to a fall from standing were included in this study. MATERIALS AND METHODS: Fractured heads were collected during hip replacement surgery. Standardized bone cores were removed from the femoral head's equator by a trephine in a water bath. MicroCT images acquisition and analysis were performed with CTan® software and bone volume fraction was then determined. Micro-finite-element simulations were per-formed using Abaqus 6.9-2® software in order to determine the macroscopic mechanical behaviour of the trabecular bone. After microCT acquisition, a longitudinal compression test was performed and the experimental macroscopic Young's Modulus was extracted. An inverse approach based on the whole trabecular bone's mechanical response and micro-finite-element analysis was performed to determine microscopic mechanical properties of trabecular bone. RESULTS: In the present study, elasticity of the tissue was shown to be similar to that of healthy tissue but with a lower yield stress. CONCLUSION: Classical histomorphometric analysis form microCT imaging associated with an inverse micro-finite-element method allowed to assess microscopic mechanical trabecular bone parameters.
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Osso Esponjoso/fisiopatologia , Elasticidade , Fraturas do Fêmur/fisiopatologia , Osteoporose/fisiopatologia , Fraturas por Osteoporose/fisiopatologia , Acidentes por Quedas , Idoso , Idoso de 80 Anos ou mais , Fenômenos Biomecânicos , Osso Esponjoso/ultraestrutura , Módulo de Elasticidade , Feminino , Cabeça do Fêmur/fisiopatologia , Cabeça do Fêmur/ultraestrutura , Análise de Elementos Finitos , Humanos , Masculino , Paris , Estresse Mecânico , Microtomografia por Raio-XRESUMO
BACKGROUND: To investigate the effect of intra-articular injection of Chitosan (Cs) added to hyaluronic acid (HA) on subchondral bone during osteoarthritis (OA), microarchitectural parameters and mineral density were measured in a rabbit model of early OA. A novel hybrid hydrogel adding reacetylated Cs of fungal origin to HA was compared to high molecular weight HA commercial formulation. METHOD: Eighteen rabbits underwent unilateral anterior cruciate ligament transection (ACLT) and were divided into three groups (Saline-group, HA-group and Hybrid-group) depending on the intra-articular injection compound. Eight contralateral knees were used as non-operated controls (Contralateral-group). Micro-computed tomography was performed six weeks post-ACLT to study subchondral bone microarchitectural parameters and mineral density at an early stage of OA development. RESULTS: Cartilage thickness mean value was reduced only in Saline-group compared to Contralateral-group. When the Hybrid-group was compared to Saline-group, subchondral bone microarchitectural parameters (trabecular thickness and trabecular bone volume fraction) were significantly changed; subchondral bone plate and trabecular bone mineral densities (bone mineral density and tissue mineral density) were reduced. When the Hybrid-group was compared to HA-group, subchondral bone microarchitectural parameters (subchondral plate thickness and trabecular thickness) and trabecular bone mineral densities (bone mineral density and tissue mineral density) were significantly decreased. CONCLUSION: Conclusion: Compared to HA alone, the novel hybrid hydrogel, constituted of Cs added to HA, enhanced microarchitectural parameters and mineral density changes, leading to subchondral bone loss in a rabbit model of early experimental OA.
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Lesões do Ligamento Cruzado Anterior/diagnóstico por imagem , Lesões do Ligamento Cruzado Anterior/tratamento farmacológico , Quitosana/administração & dosagem , Viscossuplementação/métodos , Animais , Ligamento Cruzado Anterior , Injeções Intra-Articulares/métodos , Masculino , Coelhos , Distribuição Aleatória , Resultado do Tratamento , Microtomografia por Raio-X/métodosRESUMO
Camptocormia, a severe flexion deformity of the spine, presents challenges in monitoring its progression outside laboratory settings. This study introduces a customized method utilizing four inertial measurement unit (IMU) sensors for continuous recording of the camptocormia angle (CA), incorporating both the consensual malleolus and perpendicular assessment methods. The setup is wearable and mobile and allows measurements outside the laboratory environment. The practicality for measuring CA across various activities is evaluated for both the malleolus and perpendicular method in a mimicked Parkinson disease posture. Multiple activities are performed by a healthy volunteer. Measurements are compared against a camera-based reference system. Results show an overall root mean squared error (RMSE) of 4.13° for the malleolus method and 2.71° for the perpendicular method. Furthermore, patient-specific calibration during the standing still with forward lean activity significantly reduced the RMSE to 2.45° and 1.68° respectively. This study presents a novel approach to continuous CA monitoring outside the laboratory setting. The proposed system is suitable as a tool for monitoring the progression of camptocormia and for the first time implements the malleolus method with IMU. It holds promise for effectively monitoring camptocormia at home.
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The camptocormia angle has been established as a strong indicator for evaluating the progress of Parkinson's disease and the efficacy of therapeutical approaches. A wearable setup is proposed to measure the camptocormia angle with the perpendicular method using five inertial sensors. This study identifies suitable inertial measurement unit sensors for mobile long-term measurement. Moreover, a machine-learning approach is presented for segmenting the recorded data into periods with different dominant activities. An artificial neural network was the better classifier compared to a support vector machine to recognize certain common activities in patients with camptocormia. The artificial neural network's accuracy, sensitivity, and F1-score were 92.4 %, 82.9 %, and 82.1 %, respectively. Clinical Relevance- The presented approach is expected to lead to a wearable system for long-term monitoring of the progress of camptocormia, yielding improved parameters compared to the conventional static photo method.
Assuntos
Atrofia Muscular Espinal , Doença de Parkinson , Curvaturas da Coluna Vertebral , Humanos , Atrofia Muscular Espinal/diagnóstico , Redes Neurais de Computação , Doença de Parkinson/diagnóstico , Curvaturas da Coluna Vertebral/diagnósticoRESUMO
Prions are thought to consist of infectious proteins that cause transmissible spongiform encephalopathies. According to overwhelming evidence, the pathogenic prion protein PrPSc converts its host encoded isoform PrPC into insoluble aggregates of PrPSc, concomitant with pathological modifications (for review, see refs. 1-3). Although the physiological role of PrPC is poorly understood, studies with PrP knockout mice demonstrated that PrPC is required for the development of prion diseases. Using the yeast two-hybrid technology in Saccharomyces cerevisiae, we identified the 37-kDa laminin receptor precursor (LRP) as interacting with the cellular prion protein PrPC. Mapping analysis of the LRP-PrP interaction site in S. cerevisiae revealed that PrP and laminin share the same binding domain (amino acids 161 to 180) on LRP. The LRP-PrP interaction was confirmed in vivo in insect (Sf9) and mammalian cells (COS-7). The LRP level was increased in scrapie-infected murine N2a cells and in brain and spleen of scrapie-infected mice. In contrast, the LRP concentration was not significantly altered in these organs from mice infected with the bovine spongiform encephalopathic agent (BSE), which have a lower PrPSc accumulation. LRP levels, however, were dramatically increased in brain and pancreas, slightly increased in the spleen and not altered in the liver of crapie-infected hamsters. These data show that enhanced LRP concentrations are correlated with PrPSc accumulation in organs from mice and hamsters. The laminin receptor precursor, which is highly conserved among mammals and is located on the cell surface, may act as a receptor or co-receptor for the prion protein on mammalian cells.
Assuntos
Proteínas PrPSc/metabolismo , Precursores de Proteínas/metabolismo , Receptores de Laminina/metabolismo , Actinas/metabolismo , Animais , Sítios de Ligação , Células COS , Linhagem Celular , Cricetinae , Células Eucarióticas , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Precursores de Proteínas/química , Precursores de Proteínas/genética , Coelhos , Receptores de Laminina/química , Receptores de Laminina/genética , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Saccharomyces cerevisiae/metabolismo , Spodoptera/citologiaRESUMO
The importance of body sensor networks to monitor patients over a prolonged period of time has increased with an advance in home healthcare applications. Sensor nodes need to operate with very low-power consumption and under the constraint of limited memory capacity. Therefore, it is wasteful to digitize the sensor signal at a constant sample rate, given that the frequency contents of the signals vary with time. Adaptive sampling is established as a practical method to reduce the sample data volume. In this paper a low-power analog system is proposed, which adjusts the converter clock rate to perform a peak-picking algorithm on the second derivative of the input signal. The presented implementation does not require an analog-to-digital converter or a digital processor in the sample selection process. The criteria for selecting a suitable detection threshold are discussed, so that the maximum sampling error can be limited. A circuit level implementation is presented. Measured results exhibit a significant reduction in the average sample frequency and data rate of over 50% and 38% respectively.
RESUMO
The adsorption of the molecular acceptor hexaazatriphenylene-hexacarbonitrile on Ag(111) was investigated as function of layer density. We find that the orientation of the first molecular layer changes from a face-on to an edge-on conformation depending on layer density, facilitated through specific interactions of the peripheral molecular cyano groups with the metal. This is accompanied by a rehybridization of molecular and metal electronic states, which significantly modifies the interface and surface electronic properties, as rationalized by theoretical modeling.
RESUMO
Searching for new pi-conjugated charge-transfer systems, the electronic structure of a new acceptor-donor pair derived from coronene (C(24)H(12)) was investigated by ultraviolet photoelectron spectroscopy (UPS). The acceptor coronene-hexaone (C(24)H(6)O(6), in the following abbreviated as COHON) and the donor hexamethoxycoronene (C(30)H(24)O(6), abbreviated as HMC) were adsorbed as pure and mixed phases on gold substrates. At low coverage, COHON adsorption leads to the appearance of a charge-transfer induced interface state 1.75 eV below the Fermi energy. At multilayer coverage the photoemission intensity of the interface state drops and the valence spectrum of neutral COHON appears. The sample work function decreases from 5.3 eV (clean Au) to 4.8 eV (monolayer) followed by an increase to 5.6 eV (multilayer). The formation of a significant interface dipole due to charge-transfer at the metal-organic interface is possibly accompanied by a change in molecular orientation. HMC on Au exhibits no interface state and the sample work function decreases monotonically to ca. 4.8 eV (multilayer). The UPS spectra of individual donor and acceptor multilayers show good agreement with density functional theory modeling. In donor/acceptor mixed films the photoemission signal of the donor (acceptor) shifts to higher (lower) binding energy. This trend is predicted by the calculation and is anticipated when charge is transferred from donor to acceptor. We propose that mixed films of COHON and HMC constitute a weak charge-transfer system.
RESUMO
BACKGROUND: The aim of this study was to evaluate the results of endoscopic lumbar sympathectomy for plantar hyperhidrosis. METHODS: A total of 178 endoscopic resections of the lower sympathetic lumbar trunk were carried out in 90 patients (59 men, 31 women) with severe plantar hyperhidrosis. The clinical results, including morbidity and satisfaction rates, were evaluated. Follow-up examination was carried out for all patients after a mean follow-up of 24 (range 3-45) months. RESULTS: All procedures were carried out endoscopically. There were no deaths and only three patients had a postoperative complication. All patients had evidence of postoperative sympathetic denervation of the feet. In 87 patients (97 per cent) hyperhidrosis was eliminated, but in three (3 per cent) it recurred. Compensatory sweating occurred in 40 patients (44 per cent), postsympathectomy neuralgia in 38 (42 per cent) and one man suffered temporary loss of ejaculation. A total of 86 patients (96 per cent) were very, or partly, satisfied with the result, and 83 (92 per cent) would have the procedure repeated if required. CONCLUSION: Endoscopic lumbar sympathectomy was a safe and effective option for patients with severe plantar hyperhidrosis.
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Endoscopia/métodos , Doenças do Pé/cirurgia , Hiperidrose/cirurgia , Plexo Lombossacral/cirurgia , Simpatectomia/métodos , Adolescente , Adulto , Idoso , Análise de Variância , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Dor Pós-Operatória/etiologia , Disfunções Sexuais Fisiológicas/etiologia , Resultado do Tratamento , Adulto JovemRESUMO
An ultrastructural study of Nemertoderma (Turbellaria, Nemertodermatida) has revealed that its spermatozoa have only a single falgellum. This is the first uniflagellate spermatozoon known in the Turbellaria; it is indicative of the primitiveness of Nemertoderma and is evidence in support of the view that the Turbellaria as a whole are among the most primitive living Bilateria.
Assuntos
Evolução Biológica , Filogenia , Cauda do Espermatozoide/ultraestrutura , Espermatozoides/ultraestrutura , Turbelários/ultraestrutura , Acrossomo/ultraestrutura , Animais , Núcleo Celular/ultraestrutura , Masculino , Microscopia Eletrônica , Microtúbulos/ultraestrutura , Turbelários/classificação , Turbelários/crescimento & desenvolvimentoRESUMO
BACKGROUND: Thoracic sympathectomy is a valuable treatment option for patients with primary hyperhidrosis. However, controversies exist about the optimal technique of sympathectomy and the association between localisation of the focal hyperhidrosis and postoperative results. PATIENTS AND METHODS: Retrospective analysis was performed on prospectively collected data of 402 thoracic sympathectomies performed in 204 consecutive patients with palmar-plantar (n=123), palmar-axillary (34), isolated axillary (35), and craniofacial (12) hyperhidrosis. The standard procedure was video-assisted thoracoscopic resection of the sympathetic chain from T2 to T4/5. RESULTS: All procedures were performed thoracoscopically without serious perioperative complications. Postoperative morbidity was 2.5% (10/402) including two cases of incomplete Horner's syndrome (0.5%). One hundred forty-three patients were followed for a mean of 21 months (6-86). Palmar hyperhidrosis was eliminated in 100% of cases and axillary hyperhidrosis in 98.5%. There were three axillary recurrences (1.5%). Of all patients, 60% suffered from transient postsympathectomy neuralgia which was mild in the majority of cases. Strong compensatory sweating occurred in 17% of patients with palmar-plantar and palmar-axillary hyperhidrosis and in 53% of patients with isolated axillary hyperhidrosis (P<0.001). In the palmar-plantar and palmar-axillary groups, 92% were very satisfied with the postoperative results, 90% reported increased quality of life, and 93% would repeat the operation. The corresponding numbers in patients with isolated axillary hyperhidrosis were 47%, 44%, and 66%, respectively (P<0.001). CONCLUSION: Video-assisted thoracoscopic resection of the sympathetic chain from T2 to T4-5 is safe and effective and leads in almost 100% of cases to the elimination of palmar and axillary hyperhidrosis. In contrast to the excellent results in patients with palmar-plantar and palmar-axillary hyperhidrosis, outcome in patients with isolated axillary hyperhidrosis was impaired by a high rate of disturbing compensatory sweating.
Assuntos
Axila/inervação , Mãos/inervação , Hiperidrose/cirurgia , Simpatectomia/métodos , Cirurgia Torácica Vídeoassistida/métodos , Adolescente , Adulto , Idoso , Criança , Face/inervação , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Satisfação do Paciente , Complicações Pós-Operatórias/etiologia , Estudos Prospectivos , Qualidade de Vida , Recidiva , Estudos RetrospectivosRESUMO
BACKGROUND: Patients with plantar hyperhidrosis suffer physically and mentally from a disease that often cannot be treated sufficiently using conservative measures. This article reports the authors' experience with endoscopic lumbar sympathectomies for patients with plantar hyperhidrosis. METHODS: Bilateral lumbar sympathectomy was performed for eight patients (3 women and 5 men). A retroperitoneoscopic surgical technique was used, during which the lower lumbar sympathetic trunk was resected after radiologic localization of the lumbar spine, with care taken to protect the cranial lumbar ganglia. RESULTS: All 16 sympathectomies were conducted retroperitoneoscopically, with no intra- or postoperative complications. In all cases, pedal sweat secretion was completely suspended postoperatively. After a 3- to 8-month follow-up period, the anhidrosis persisted in all cases (100%). None of the patients experienced sexual dysfunction. Five patients (62%) reportedly experienced minor compensatory sweating of the torso, and four patients (50%) had postsympathectomy neuralgia. Seven of eight patients were very happy with the postoperative results and would agree to a repeat of the intervention any time. CONCLUSIONS: Retroperitoneoscopic resection of the lower lumbar sympathetic trunk is a safe and effective procedure for obtaining suspension of excessive sweat secretion in patients with plantar hyperhidrosis that cannot be treated with conservative methods.
Assuntos
Endoscopia , Pé , Hiperidrose/cirurgia , Espaço Retroperitoneal , Simpatectomia/métodos , Adaptação Fisiológica , Adulto , Feminino , Seguimentos , Humanos , Região Lombossacral , Neuralgia/etiologia , Satisfação do Paciente , Sudorese , Simpatectomia/efeitos adversos , Resultado do TratamentoRESUMO
The adsorption of molecular acceptors is a viable method for tuning the work function of metal electrodes. This, in turn, enables adjusting charge injection barriers between the electrode and organic semiconductors. Here, we demonstrate the potential of pyrene-tetraone (PyT) and its derivatives dibromopyrene-tetraone (Br-PyT) and dinitropyrene-tetraone (NO2-PyT) for modifying the electronic properties of Au(111) and Ag(111) surfaces. The systems are investigated by complementary theoretical and experimental approaches, including photoelectron spectroscopy, the X-ray standing wave technique, and density functional theory simulations. For some of the investigated interfaces the trends expected for Fermi-level pinning are observed, i.e., an increase of the metal work function along with increasing molecular electron affinity and the same work function for Au and Ag with monolayer acceptor coverage. Substantial deviations are, however, found for Br-PyT/Ag(111) and NO2-PyT/Ag(111), where in the latter case an adsorption-induced work function increase of as much as 1.6 eV is observed. This behavior is explained as arising from a face-on to edge-on reorientation of molecules in the monolayer. Our calculations show that for an edge-on orientation much larger work-function changes can be expected despite the prevalence of Fermi-level pinning. This is primarily ascribed to a change of the electron affinity of the adsorbate layer that results from a change of the molecular orientation. This work provides a comprehensive understanding of how changing the molecular electron affinity as well as the adsorbate structure impacts the electronic properties of electrodes.
RESUMO
3,N(4)-Ethano-2'-deoxycytidine (ethano-dC) may be incorporated successfully into synthetic oligodeoxynucleotides by omitting the capping procedure used in the automated DNA synthetic protocols immediately after inserting the lesion and in all iterations thereafter. Ethano-dC is sensitive to acetic anhydride found in the capping reagent, and multiple oligomeric products are formed. These products were identified by examining the reaction of ethano-dC with the capping reagent, and several acetylated, ring-opened products were characterized by electrospray mass spectrometry and collision induced dissociation experiments on a tandem quadrupole mass spectrometer. A scheme for the formation of the acetylated products is proposed. In addition, the mutagenic profile of ethano-dC was re-examined and compared to that for etheno-dC. Ethano-dC is principally a blocking lesion; however, when encountered by the exo(-)Klenow fragment of DNA polymerase, dAMP (22%), TMP (16%), dGMP (5.3%) and dCMP (1.2%) were all incorporated opposite ethano-dC, along with an oligomer containing a one-base deletion (0.6%).
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DNA/síntese química , Oligodesoxirribonucleotídeos/química , Oligodesoxirribonucleotídeos/síntese química , Acetilação , Automação , Sequência de Bases , Cromatografia Líquida de Alta Pressão , DNA/química , DNA Polimerase I/metabolismo , Desoxicitidina/análogos & derivados , Desoxicitidina/farmacologia , Indicadores e Reagentes , Estrutura Molecular , Mutagênicos/química , Mutagênicos/farmacologia , Espectrometria de Massa de Íon SecundárioRESUMO
In the accompanying article, we describe the creation of novel cell lines derived from RIN 1046-38 rat insulinoma cells by stable transfection with combinations of genes encoding human insulin, GLUT2, and glucokinase. Herein we describe the regulation of insulin secretion and glucose metabolism in these new cell lines. A cell line (betaG I/17) expressing only the human proinsulin transgene exhibits a clear increase in basal insulin production (measured in the absence of secretagogues) relative to parental RIN 1046-38 cells. betaG I/17 cells engineered for high levels of GLUT2 expression and a twofold increase in glucokinase activity (betaG 49/206) or engineered for a 10-fold increase in glucokinase activity alone (betaG 40/110) exhibit a 66% and 80% suppression in basal insulin secretion relative to betaG I/17 cells, respectively. As a result, betaG 49/206 and betaG 40/110 cells exhibit potent insulin-secretory responses to glucose alone (6.1- and 7.6-fold, respectively) or to glucose plus isobutylmethylxanthine (10.8- and 15.1-fold, respectively) that are clearly larger than the corresponding responses of betaG I/17 or parental RIN 1046-38 cells. betaG 49/206 and betaG 40/110 cells also exhibit a rapid and sustained response to glucose plus isobutyl-methylxanthine in perifusion studies that is clearly larger in magnitude than that of the two control lines. Glucose dose-response studies show that both engineered and non-engineered lines respond maximally to submillimolar concentrations of glucose and that betaG 49/206 cells are the most sensitive to low concentrations of the hexose, consistent with their clearly elevated rate of [5-3H]glucose usage. Finally, 5-thioglucose, a potent inhibitor of low-K(m) hexokinases, most effectively normalizes glucose concentration dependence for insulin secretion in the cell line with highest glucokinase expression (betaG 40/110). We conclude that GLUT2 and/or glucokinase expression imposes tight regulation of basal insulin secretion in cell lines that overexpress human proinsulin, allowing a marked improvement in the range of secretagogue responsiveness in such cells.
Assuntos
Regulação da Expressão Gênica/genética , Glucose/metabolismo , Insulina/metabolismo , Insulinoma/metabolismo , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/metabolismo , 1-Metil-3-Isobutilxantina/farmacologia , Animais , Relação Dose-Resposta a Droga , Glucoquinase/análise , Glucose/análise , Glucose/farmacologia , Transportador de Glucose Tipo 2 , Humanos , Immunoblotting , Insulina/análise , Insulina/genética , Secreção de Insulina , Insulinoma/genética , Insulinoma/patologia , Proteínas de Transporte de Monossacarídeos/análise , Neoplasias Pancreáticas/patologia , Inibidores de Fosfodiesterase/farmacologia , Fosforilação , Ratos , Trítio , Células Tumorais CultivadasRESUMO
The biochemical mechanisms involved in regulation of insulin secretion are not completely understood. The rat INS-1 cell line has been used to gain insight in this area because it secretes insulin in response to glucose concentrations in the physiological range. However, the magnitude of the response is far less than that seen in freshly isolated rat islets. In the current study, we have stably transfected INS-1 cells with a plasmid containing the human proinsulin gene. After antibiotic selection and clonal expansion, 67% of the resultant clones were found to be poorly responsive to glucose in terms of insulin secretion (< or =2-fold stimulation by 15 mmol/l compared with 3 mmol/l glucose), 17% of the clones were moderately responsive (2- to 5-fold stimulation), and 16% were strongly responsive (5- to 13-fold stimulation). The differences in responsiveness could not be ascribed to differences in insulin content. Detailed analysis of one of the strongly responsive lines (832/13) revealed that its potent response to glucose (average of 10-fold) was stable over 66 population doublings (approximately 7.5 months of tissue culture) with half-maximal stimulation at 6 mmol/l glucose. Furthermore, in the presence of 15 mmol/l glucose, insulin secretion was potentiated significantly by 100 pmol/l isobutylmethylxanthine (320%), 1 mmol/l oleate/palmitate (77%), and 50 nmol/l glucagon-like peptide 1 (60%), whereas carbachol had no effect. Glucose-stimulated insulin secretion was also potentiated by the sulfonylurea tolbutamide (threefold at 3 mmol/l glucose and 50% at 15 mmol/l glucose) and was abolished by diazoxide, which demonstrates the operation of the ATP-sensitive K+ channel (K(ATP)) in 832/13 cells. Moreover, when the K(ATP) channel was bypassed by incubation of cells in depolarizing K+ (35 mmol/l), insulin secretion was more effectively stimulated by glucose in 832/13 cells than in parental INS-1 cells, which demonstrates the presence of a K(ATP) channel-independent pathway of glucose sensing. We conclude that clonal selection of INS-1 cells allows isolation of cell lines that exhibit markedly enhanced and stable responsiveness to glucose and several of its known potentiators. These lines may be attractive new vehicles for studies of beta-cell function.
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Trifosfato de Adenosina/fisiologia , Glucose/farmacologia , Insulina/metabolismo , Ilhotas Pancreáticas/metabolismo , Canais de Potássio/fisiologia , Animais , Linhagem Celular , Células Clonais , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Humanos , Insulina/genética , Secreção de Insulina , Proinsulina/metabolismo , Processamento de Proteína Pós-Traducional , Ratos , TransfecçãoRESUMO
Using a selection based upon the ability of early Rec- mutations (e.g., rad50) to rescue the meiotic lethality of a rad52 spo13 strain, we have isolated 177 mutants. Analysis of 56 of these has generated alleles of the known Rec genes SPO11, ME14 and MER1, as well as defining five new genes: REC102, REC104, REC107, REC113 and REC114. Mutations in all of the new genes appear to specifically affect meiosis; they do not have any detectable mitotic phenotype. Mutations in REC102, REC104 and REC107 reduce meiotic recombination several hundred fold. No alleles of RED1 or HOP1 were isolated, consistent with the proposal that these genes may be primarily involved with chromosome pairing and not exchange.
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Genes Fúngicos , Meiose/genética , Mutação , Recombinação Genética , Saccharomyces cerevisiae/genética , Alelos , Clonagem Molecular , Diploide , Teste de Complementação Genética , Mitose/genética , Saccharomyces cerevisiae/citologia , Saccharomyces cerevisiae/crescimento & desenvolvimentoRESUMO
In the last decade there have been numerous reports of anthelmintic resistant cyathostomins in many parts of the world. The objective of the present study was to evaluate the efficacy of the commercially available anthelmintics against cyathostomin egg shedding in the Mid-Atlantic region of the United States. A total of 989 horses from 67 different farms located in southeastern Pennsylvania, northern Delaware, and northeastern Maryland were treated with fenbendazole, oxibendazole, pyrantel pamoate, ivermectin, or moxidectin at their recommended dosages. Fecal egg count reduction testing was used to determine the efficacy of each anthelmintic on those horses with fecal egg counts of ≥ 200 eggs per gram on the day of treatment (272 horses). Decreased efficacy (reduction of strongyle-type fecal egg counts by less than 90%) was found for fenbendazole, oxibendazole, and pyrantel pamoate, with only 6%, 21% and 43% of horses showing reductions of greater than 90%, respectively. The macrocyclic lactones showed high efficacy in all horses sampled in this study. The decreased anthelmintic efficacy detected in this study adds further evidence for the existence of resistant cyathostomins throughout much of the eastern United States. Findings from this study can be used to create a more sustainable approach for parasite control programs.
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Anti-Helmínticos/uso terapêutico , Doenças dos Cavalos/parasitologia , Contagem de Ovos de Parasitas/veterinária , Animais , Resistência a Medicamentos , Fezes/parasitologia , Helmintos/efeitos dos fármacos , Doenças dos Cavalos/tratamento farmacológico , Cavalos , Óvulo/efeitos dos fármacos , Estados Unidos/epidemiologiaRESUMO
Electrospray mass spectrometry techniques were used to characterize components of the active site in Endonuclease VIII by identifying the amino acid sequence and the binding site for a tryptic peptide derived from Endo VIII in a cross-linked DNA-peptide complex. Endo VIII, a DNA repair enzyme with both glycosylase and lyase activities, was covalently bound to a thymidine glycol-containing oligodeoxynucleotide duplex by converting a transient Schiff base formed during the course of the glycosylase activity to a stable covalent bond by chemical reduction with sodium borohydride. After tryptic digestion of the initial product, the identification of the cross-linked peptide was deduced initially from the molecular mass of the tryptic product obtained by negative ion electrospray mass analysis. Nanospray tandem mass spectrometry (MS/MS) analysis of the tryptic product corroborated the molecular mass of the peptide fragment and verified the point of attachment to the oligomer, but failed to produce sufficient fragmentation to sequence the peptide completely. Direct evidence for the amino acid sequence of the peptide was obtained after enzymatic digestion of the DNA portion of the cross-linked DNA-peptide product and analysis by negative ion nanospray MS/MS. Examination of the ions from collision induced fragmentation disclosed that this substance was the N-terminal tryptic fragment of Endo VIII cross-linked to a portion of the oligomer, and that the N-terminal proline from Endo VIII was covalently bound to the residual deoxyribose moiety at the original location of the thymine glycol in the oligomer.
Assuntos
Reparo do DNA , Endodesoxirribonucleases/química , Sequência de Aminoácidos , Aminoácidos/análise , Sítios de Ligação , Reagentes de Ligações Cruzadas , DNA/química , Desoxirribonuclease (Dímero de Pirimidina) , Glicóis/química , Hidrólise , Indicadores e Reagentes , Espectrometria de Massas , Dados de Sequência Molecular , Oligonucleotídeos/química , Peptídeos/química , Timidina/química , TripsinaRESUMO
In a 17-year period 1848 patients with hyperthyroidism were operated on. Fourteen (0.76%) had a coexisting thyroid malignancy. Preoperative scintiscan and pathologic diagnoses were compared: 10 malignancies were in cold nodules, two were unidentifiable preoperatively due to small size, and two were in hot areas. Five patients had papillary cancer, four follicular, three anaplastic, and two medullary. Patients with uninodular toxic goiter had a low rate of associated malignancy (0.27%, 3/1108). In contrast, patients with multinodular toxic goiter had an incidence of 1.63% (11/676). No patient with Graves' disease (n = 64) had a carcinoma. Extensive use of fine-needle aspiration biopsy enabled preoperative diagnosis in a majority of the cases (9/14, 64%). We conclude that the incidence of coexisting thyroid malignancy and hyperthyroidism is rare in our endemic iodine-deficiency goiter area.