RESUMO
Dynamics of endocytic clathrin-coated structures can be remarkably divergent across different cell types, cells within the same culture, or even distinct surfaces of the same cell. The origin of this astounding heterogeneity remains to be elucidated. Here we show that cellular processes associated with changes in effective plasma membrane tension induce significant spatiotemporal alterations in endocytic clathrin coat dynamics. Spatiotemporal heterogeneity of clathrin coat dynamics is also observed during morphological changes taking place within developing multicellular organisms. These findings suggest that tension gradients can lead to patterning and differentiation of tissues through mechanoregulation of clathrin-mediated endocytosis.
Assuntos
Vesículas Revestidas por Clatrina/fisiologia , Animais , Fenômenos Biomecânicos , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Membrana Celular/fisiologia , Células Cultivadas , Chlorocebus aethiops , Clatrina/metabolismo , Vesículas Revestidas por Clatrina/metabolismo , Invaginações Revestidas da Membrana Celular/metabolismo , Invaginações Revestidas da Membrana Celular/fisiologia , Citoplasma/metabolismo , Drosophila , Endocitose/fisiologia , Humanos , Análise Espaço-TemporalRESUMO
Frequency-dependent molar electrical conductivities for aqueous solutions of potassium borate, and sodium borate have been measured from ambient to near-critical temperatures and pressures to an accuracy of ±3 percent, using a unique high-precision flow-through AC conductance instrument. The concentration dependence of these conductivities was analyzed with the Turq-Blum-Bernard-Kunz ("TBBK") theoretical model to yield (i) limiting conductivities of the borate ion, λ(0)[B(OH)4(-)], and (ii) ion-pair formation constants, KA, for the species NaB(OH) and KB(OH) from T = 298 K to T = 623 K at a constant pressure p â¼ 20 MPa. The ion-pair formation constants for both borate salts were found to be consistent with previous literature studies at temperatures below 473 K. No significant difference in KA was observed between the species NaB(OH) and KB(OH). As temperature was increased from 473 up to 623 K, the degree of association increased significantly, and was found to be considerably higher than for any other 1-1 electrolyte previously studied. For instance, at 623 K, the association constant log KA[NaB(OH)] = 2.75 ± 0.21 was an order of magnitude higher than log KA[NaCl(0)] = 1.53 ± 0.03, and approximately equal to that of a 2 : 1 electrolyte, log KA[SrCF3SO3(+)] = 2.58 ± 0.06. Deviations in the limiting conductivities from Stokes Law show that the borate ion's unusual "structure making" effect, observed by other workers at sub-ambient conditions, persists up to temperatures above 500 K. The temperature dependence of the Walden product ratio is very different from that observed for other monovalent anions for which experimental data are available over this wide range of temperatures.
RESUMO
Immuno-biosensor inhibition assays for the detection of streptomycin and dihydrostreptomycin residues in whole cows' milk, honey, pig kidney and pig muscle are reported. The antibody showed high cross-reactivity with dihydrostreptomycin in various foodstuffs (buffer 103%, milk 96%, honey 84%, kidney extract 129% and muscle extract 98%). There was no significant cross-reaction with other aminoglycosides or commonly used antibiotics. A streptomycin derivative was used to prepare a stable, reusable sensor chip surface. The assay allowed the direct analysis of bovine whole milk (fat content approximately 3.5%). Honey samples required dilution with buffer, while kidney and muscle samples from pigs were homogenized in an aqueous extraction buffer and clarified by centrifugation. The limit of detection for each assay was determined from known streptomycin-free samples (n = 20; mean - (3 x standard deviation)) and the results were as follows: milk 30 microg kg(-1), honey 15 microg kg(-1), kidney 50 microg kg(-1) and muscle 70 microg kg(-1). Repeatability (or relative standard deviation) between runs were calculated (n = 3) at the respective Community maximum residue limits (MRL) and 0.5 x MRL with the exception of honey since no European MRL exists at present. Results were determined as 4.3% (200 microg kg(-1)) and 2.8% (100 microg kg(-1)) in milk, 13.3% (40 microg kg(-1)) and 9.5% (20 microg kg(-1)) in honey, 7.1% (1000 microg kg(-1)) and 7.6% (500 microg kg(-1)) in kidney and 7.1% (500 microg kg(-1)) and 11% (250 microg kg(-1)) in muscle.
Assuntos
Antibacterianos/análise , Resíduos de Drogas/análise , Leite/química , Estreptomicina/análise , Animais , Técnicas Biossensoriais , Sulfato de Di-Hidroestreptomicina , Mel/análise , Humanos , Rim/química , Carne/análise , Músculo Esquelético/química , Sensibilidade e Especificidade , SuínosRESUMO
The development of an assay for the detection of streptomycin residues in pasteurized whole milk using an optical biosensor (Biacore) is reported. Streptomycin-adipic hydrazide coupled to bovine thyroglobulin was used to produce a sheep polyclonal antibody. The antibody displayed excellent cross-reactivity with dihydrostreptomycin (106%). There was no significant cross-reaction with other aminoglycosides or common antibiotics. Streptomycin was also immobilized onto a CM5 sensor chip to provide a stable, reusable surface. The developed assay permitted the direct analysis of whole milk samples ( approximately 3.5% fat) without prior centrifugation and defatting. Results were available in 5 min. The limit of detection of the assay was determined as 4.1 ng/mL, well below the European maximum residue limit (MRL) of 200 ng/mL. Repeatability (or coefficient of variation) between runs was determined as 3.5% (100 ng/mL; 0.5 x MRL), 5.7% (200 ng/mL; MRL), and 7.6% (400 ng/mL; 2 x MRL).
Assuntos
Técnicas Biossensoriais , Resíduos de Drogas/isolamento & purificação , Leite/química , Estreptomicina/isolamento & purificação , Animais , Antibacterianos/isolamento & purificação , Sensibilidade e EspecificidadeRESUMO
A 33-year-old man with a history of severe asthma presented to the emergency department with a week-long history of severe unrelenting abdominal pain, nausea and decreased appetite. He was admitted to hospital, and routine gastrointestinal investigations were performed, which did not elucidate the cause of his abdominal pain. Exploratory laparotomy demonstrated patchy infarction of the entire small bowel, characteristic of Churg-Strauss syndrome. The patient subsequently underwent 12 separate laparotomies to salvage surviving small bowel. The patient is maintained on total parenteral nutrition.
Assuntos
Abdome Agudo/etiologia , Síndrome de Churg-Strauss/diagnóstico , Infarto/complicações , Infarto/etiologia , Intestino Delgado/irrigação sanguínea , Adulto , Síndrome de Churg-Strauss/complicações , Síndrome de Churg-Strauss/patologia , Diagnóstico Diferencial , Humanos , MasculinoRESUMO
The European Commission (EC) established the Standards, Measurements and Testing programme for the preparation of Reference Materials (RMs) as an aid to harmonise testing for veterinary drug residues throughout the European Union (EU). The production of chlortetracycline (CTC)-free and CTC-incurred pig tissues as candidate RMs is described. High performance liquid chromatography (HPLC) with fluorescence detection of CTC and 4-epi-CTC was used for all tissue analyses. A pilot study revealed that incurred CTC residues were stable in pig kidney, liver and muscle lyophilised powders during storage for 10 weeks at -70, -20 and +37 degrees C, obviating the need for addition of a stabiliser (thimerosal). In the main study, 500 vials each of CTC-free and CTC-incurred kidney, liver and muscle were produced. Target concentrations in the CTC-incurred lyophilised tissue powders were 750-1500, 500-1000 and 300-600 micrograms kg-1 for kidney, liver and muscle, respectively. Following lyophilisation, the mean +/- s concentrations of CTC in the incurred positive RMs were 1,315 +/- 56.9, 765 +/- 35.3 and 378 +/- 16.8 micrograms kg-1 for kidney, liver and muscle respectively. Residual moisture in the RMs ranged from 1.6 +/- 0.53% for muscle to 3.0 +/- 0.50% for liver. Between-vial homogeneity for incurred powders was determined for 20 vials of each material, which had been removed at regular intervals during the filling process. Relative standard deviations (RSDs) for kidney, liver and muscle were 4.3, 4.6 and 4.4% respectively, being within the interassay RSD of the method and indicating that mixing was effective. Stability of powders stored at -18, 4, 20 and 37 degrees C was assessed over a period of 79 weeks. No measurable degradation occurred over this time period at any of the storage temperatures. It is concluded that these candidate RMs are homogenous, stable and are suitable for certification.
Assuntos
Antibacterianos/normas , Clortetraciclina/normas , Resíduos de Drogas/análise , Carne/análise , Animais , Cromatografia Líquida de Alta Pressão , União Europeia , Rim/química , Fígado/química , Músculo Esquelético/química , Padrões de Referência , SuínosRESUMO
To determine the efficacy of single nighttime doses of arbaprostil [15(R)-15-methyl prostaglandin E2], 50 or 100 micrograms for 4 wk, a double-blind randomized placebo-controlled multiclinic trial was undertaken. Success was defined as complete healing of the ulcer documented by endoscopy. Fifty-one of 64 patients enrolled were considered evaluable. Ulcer healing was documented in 64.3%, 85.7%, and 31.2% of the 100-micrograms arbaprostil, 50-micrograms arbaprostil, and placebo treatment groups (p value vs. placebo = 0.003 and 0.002, respectively). No difference in side effects or changes in laboratory parameters were found between the treatment groups except that diarrhea, usually mild, was found more often in the 100-micrograms arbaprostil group (60.0%) than in the 50-micrograms arbaprostil (31.8%) or placebo groups (23.5%) (p value 100 micrograms arbaprostil vs. placebo = 0.02). A single nighttime administration of arbaprostil seems to be a safe and efficacious agent for the treatment of acute duodenal ulcer.
Assuntos
Arbaprostilo/uso terapêutico , Úlcera Duodenal/tratamento farmacológico , Prostaglandinas E Sintéticas/uso terapêutico , Adulto , Arbaprostilo/administração & dosagem , Arbaprostilo/efeitos adversos , Diarreia/induzido quimicamente , Relação Dose-Resposta a Droga , Método Duplo-Cego , Humanos , Masculino , Estudos Multicêntricos como Assunto , Distribuição AleatóriaRESUMO
The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or as reflecting the views of the Department of the Army or the Department of Defense. At the time this study was performed, LTs Ferguson, Blackley, Knight, and Sutlive were students in the US. Army-Baylor University Graduate Program in Physical Therapy. This research was performed in partial fulfillment of their requirements for the Master of Physical Therapy Degrees.The purpose of this study was to evaluate the effects of varying electrode placement on the torque output of an electrically stimulated involuntary quadriceps femoris muscle contraction. Twenty-two volunteer subjects (18 men, 4 women) with a mean age of 21.7 years received electrical stimulation according to a randomized treatment order which included: Femoral nerve and 1) vastus medialis (VM), 2) rectus femoris (RF), 3) vastus lateralis (VL), 4) opposite quadriceps (OQ), and 5) the ipsilateral vastus medialis and vastus lateralis (VM/VL). The subject's knee was placed in 60 degrees of flexion, and the isokinetic dynamometer set at 0 degrees /sec. The peak torque produced by the involuntary quadriceps contraction was measured as a percentage of maximum voluntary isometric contraction (MVIC). An analysis of variance with repeated measures was used to examine the data. The results indicated the mean percentages of MVIC produced by stimulating the VM, VL, and RF sites were significantly greater (p = 0.05) than the means of the OQ and VM/VL sites. Post hoc testing did not reveal a significant difference in the mean percent MVIC between the VM, VL, and RF sites. The VM, VL, or RF distal electrode site placement may be used clinically in conjunction with ipsilateral femoral nerve stimulation to produce a maximum involuntary isometric contraction of the quadriceps femoris muscle when stimulated electrically. J Orthop Sports Phys Ther 1989;11(1):24-29.
RESUMO
The seven papers in this section present individual viewpoints on the past 15 years, with a great variety in emphasis and perspective. Several authors take a nostalgic look back, one describes an overseas experience, and another provides a business outlook. Read and enjoy these contributions from your colleagues.