RESUMEN
Fleece rot and dermatophilosis reduce health and production of sheep and predispose them to blow fly strike. This paper reviews aetiology, prevalence, pathogenesis, resistance, attempts to develop vaccines and prospects for new control strategies to these important skin diseases. Although the severity of fleece rot is associated with the abundance of Pseudomonas aeruginosa on skin, microbial ecology studies are providing new insights into the contribution of other bacteria to the disease. Wool traits and body conformation traits that predispose sheep to fleece rot and dermatophilosis are heritable and have been used as indirect selection criteria for resistance for many years. Selection against BoLA-DRB3-DQB class II haplotype in cattle can substantially reduce the prevalence of dermatophilosis and holds promise for identification of gene markers for resistance to these bacterial diseases in sheep. Immune responses in skin and systemic antibody responses to bacterial antigens are acquired through natural infection and contribute to resistance; however, prototype antibacterial vaccines have to date failed to provide protection against the diversity of isolates of Dermatophilus congolensis and Pseudomonas species present in the field. Opportunities for future control through breeding for resistance, vaccines and non-vaccine strategies for controlling the microbial ecology of fleece are discussed. In combination, control strategies need to reduce the risk of transmission, minimise exposure of animals to stressors that enhance the risk of infection, and enhance resistance though genetics or vaccines.
Asunto(s)
Infecciones por Actinomycetales/veterinaria , Vacunas Bacterianas , Dermatitis/veterinaria , Infecciones por Pseudomonas/veterinaria , Enfermedades de las Ovejas/microbiología , Lana/microbiología , Actinomycetales/inmunología , Infecciones por Actinomycetales/microbiología , Infecciones por Actinomycetales/patología , Infecciones por Actinomycetales/prevención & control , Animales , Dermatitis/microbiología , Dermatitis/patología , Dermatitis/prevención & control , Susceptibilidad a Enfermedades/inmunología , Susceptibilidad a Enfermedades/veterinaria , Predisposición Genética a la Enfermedad , Inmunidad Innata/genética , Inmunidad Innata/inmunología , Infecciones por Pseudomonas/microbiología , Infecciones por Pseudomonas/patología , Infecciones por Pseudomonas/prevención & control , Pseudomonas aeruginosa/inmunología , Ovinos , Enfermedades de las Ovejas/patología , Enfermedades de las Ovejas/prevención & controlRESUMEN
There is an increasing prevalence for work to be analysed through naturalistic study, especially using ethnographically derived methods of enquiry and qualitative field research. The relatively unexplored domain of railway control (in comparison to signalling) in the UK is described in terms of features derived from observations and semi-structured interviews. In addition, task diagrams (a technique taken from the Applied Cognitive Task Analysis toolkit) are used to represent controllers' core elements of work, i.e. to manage events or incidents, and to identify the challenging steps in the process. The work features identified, the task diagrams, and the steps identified as challenging form a basis from which future ergonomics studies on railway controllers in the UK will be carried out.
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Cognición , Eficiencia , Ergonomía , Vías Férreas , Interfaz Usuario-Computador , Recolección de Datos , Humanos , Entrevistas como Asunto , Análisis de Sistemas , Análisis y Desempeño de Tareas , Reino UnidoRESUMEN
Distinct motor patterns are selected from a multifunctional neuronal network by activation of different modulatory projection neurons. Subsets of these projection neurons can contain the same neuromodulator(s), yet little is known about the relative influence of such neurons on network activity. We have addressed this issue in the stomatogastric nervous system of the crab Cancer borealis. Within this system, there is a neuronal network in the stomatogastric ganglion (STG) that produces many versions of the pyloric and gastric mill rhythms. These different rhythms result from activation of different projection neurons that innervate the STG from neighboring ganglia and modulate STG network activity. Three pairs of these projection neurons contain the neuropeptide proctolin. These include the previously identified modulatory proctolin neuron and modulatory commissural neuron 1 (MCN1) and the newly identified modulatory commissural neuron 7 (MCN7). We document here that each of these neurons contains a unique complement of cotransmitters and that each of these neurons elicits a distinct version of the pyloric motor pattern. Moreover, only one of them (MCN1) also elicits a gastric mill rhythm. The MCN7-elicited pyloric rhythm includes a pivotal switch by one STG network neuron from playing a minor to a major role in motor pattern generation. Therefore, modulatory neurons that share a peptide transmitter can elicit distinct motor patterns from a common target network.
Asunto(s)
Ganglios de Invertebrados/citología , Neuronas Motoras/fisiología , Red Nerviosa/fisiología , Neuropéptidos , Oligopéptidos/fisiología , Transmisión Sináptica , Animales , Braquiuros , Electrofisiología , Ganglios de Invertebrados/anatomía & histología , Ganglios de Invertebrados/fisiología , Inmunohistoquímica , Técnicas In Vitro , Actividad Motora , Neuronas Motoras/metabolismo , Red Nerviosa/anatomía & histología , Red Nerviosa/metabolismo , Vías Nerviosas , Neurotransmisores/metabolismo , Neurotransmisores/fisiología , Oligopéptidos/metabolismo , Periodicidad , Estómago/inervación , Estómago/fisiología , Sinapsis/química , Sinapsis/fisiologíaRESUMEN
We have evaluated the antitumor activity of a murine antibody (IgG2a) against the leukocyte antigen CD48. CD48 is expressed on T and B lymphocytes, monocytes, and a wide range of lymphoid malignancies. To assess the therapeutic potential of an anti-CD48 antibody, we established a reproducible model of human B-cell (Raji) leukemia/lymphoma in C.B17/scid mice, where untreated mice develop hind leg paralysis due to tumor engraftment. Using this model, the murine anti-CD48 antibody HuLy-m3 was shown to mediate a strong in vivo antitumor effect. Long-term survival (>1 year) of scid mice was obtained after treatment with three 200-microg i.v. doses of anti-CD48 antibody on days 0, 2, and 4 after i.v. injection of tumor cells. In contrast, mice treated with an isotype control antibody developed hind leg paralysis after 34 +/- 3 days. A strong antitumor response was still observed when a dose of 20 microg of HuLy-m3 antibody was used. During preclinical investigations, we also examined a number of properties of the CD48 antigen. CD48 is present at high levels on the surface of T and B cells, but most (>95%) CD34-positive cells do not express CD48. Anti-CD48 antibodies are maintained on the surface of antigen-positive cells for extended periods (>24 h). These properties suggest that anti-CD48 antibodies may be useful in the treatment of a number of diseases including lymphoid leukemias and lymphomas.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Antígenos CD/inmunología , Antígenos de Neoplasias/inmunología , Linfoma de Células B/tratamiento farmacológico , Animales , Anticuerpos Monoclonales/administración & dosificación , Sitios de Unión , Antígeno CD48 , Humanos , Linfoma de Células B/patología , Ratones , Ratones SCID , Análisis de Supervivencia , Resultado del TratamientoRESUMEN
The serine proteinase inhibitor (serpin), plasminogen activator inhibitor type 2 (PAI-2), has been reported to inhibit tumor necrosis factor-alpha (TNF) induced apoptosis. In order to begin to understand the molecular basis for this protection, we have investigated the importance of a structural domain within the PAI-2 molecule, the C-D interhelical region, in mediating the protective effect. The C-D interhelical region is a 33 amino acid insertion which is unique among serpins and has been implicated in transglutaminase catalyzed cross-linking of PAI-2 to cell membranes. We have constructed a mutant of PAI-2 wherein 23 amino acids are deleted from the C-D interhelical region generating a structure predicted to be homologous to the closely related, but non-inhibitory serpin, chicken ovalbumin. The PAI-2Delta65/87 deletion mutant retained inhibitory activity against its known serine proteinase target, urokinase-type plasminogen activator (uPA); however expression of this mutant in HeLa cells failed to protect from TNF-induced apoptosis. Analyses of the cellular distribution of PAI-2 showed that intracellular PAI-2, and not secreted or cell-surface PAI-2, was likely responsible for the observed protection from TNF-induced apoptosis. No evidence was found for specific cross-linking of PAI-2 to the plasma membrane in either control or TNF/cycloheximide treated cells. The data demonstrate that the PAI-2 C-D interhelical domain is functionally important in PAI-2 protection from TNF induced apoptosis and suggest a novel function for the C-D interhelical domain in the protective mechanism.
Asunto(s)
Apoptosis/fisiología , Secuencia Conservada , Inhibidor 2 de Activador Plasminogénico/genética , Serpinas/genética , Factor de Necrosis Tumoral alfa/farmacología , Apoptosis/efectos de los fármacos , Cartilla de ADN , Regulación Neoplásica de la Expresión Génica , Células HeLa , Humanos , Datos de Secuencia Molecular , Familia de Multigenes/fisiología , Mutagénesis/fisiología , Inhibidor 2 de Activador Plasminogénico/análisis , Inhibidor 2 de Activador Plasminogénico/química , Estructura Terciaria de Proteína , Homología de Secuencia de Aminoácido , Serpinas/análisis , Serpinas/química , TransfecciónRESUMEN
1. We have used multiple, simultaneous intra- and extracellular recordings as well as Lucifer yellow dye-fills to identify modulatory commissural neuron 5 (MCN5) and characterize its effects in the stomatogastric nervous system (STNS) of the crab, Cancer borealis. MCN5 has a soma and neuropilar arborization in the commissural ganglion (CoG; Figs. 1 and 2), and it projects through the inferior esophageal nerve (ion) and stomatogastric nerve (stn) to the stomatogastric ganglion (STG; Figs. 1-3). 2. Within the CoGs, MCN5 receives esophageal rhythm-timed excitation and pyloric rhythm-timed inhibition (Fig. 4). Additionally, during the lateral teeth protractor phase of the gastric mill rhythm, the pyloric-timed inhibition of MCN5 is reduced or eliminated. 3. Intracellular stimulation of MCN5 excites the pyloric pacemaker ensemble, including the anterior burster (AB), pyloric dilator (PD), and lateral posterior gastric (LPG) neurons. This produces a faster pyloric rhythm. MCN5 stimulation also inhibits all nonpacemaker pyloric neurons, reducing or eliminating their activity (Figs. 5 and 6A; Tables 1 and 2). After MCN5 stimulation, bursting is enhanced for several cycles in some pyloric neurons when compared with their prestimulus activity (Figs. 5 and 6A; Tables 1 and 2). 4. MCN5 evokes distinct responses from each pyloric pacemaker neuron (Figs. 6-8). The AB and LPG neurons respond with increased activity. The AB response includes the presence of large amplitude excitatory postsynaptic potentials (EPSPs) that contribute to a depolarization of the trough of its rhythmic oscillations (Fig. 6). LPG responds by exhibiting increased activity that prolongs the duration of its burst beyond that of AB and PD (Fig. 7). In contrast, MCN5 stimulation initially produces decreased PD neuron activity, followed by a slight enhancement of each PD burst (Figs. 7 and 8). PD activity is further enhanced after MCN5 stimulation (Figs. 7 and 8). 5. MCN5-elicited action potentials evoke discrete, constant latency inhibitory postsynaptic potentials (IPSPs) in all nonpacemaker pyloric neurons, including the inferior cardiac (IC), lateral pyloric (LP), pyloric (PY), and ventricular dilator (VD) neurons (Fig. 9). MCN5 activity also inhibits these neurons indirectly, via its excitation of the pacemaker neurons. The pyloric pacemaker neurons synaptically inhibit all four nonpacemaker neurons. 6. The increased activity in the VD neuron, after MCN5 stimulation, is not mimicked by either direct hyperpolarization or by synaptically inhibiting VD via another pathway (Fig. 10). The poststimulation increase in IC neuron activity is stronger than that after hyperpolarizing current injection but is comparable with that resulting from stimulation of another inhibitory pathway (Fig. 10). The enhanced PY neuron activity is comparable with that resulting from either direct current injection or synaptic inhibition from another pathway (Fig. 10). 7. MCN5 activity increases the pyloric cycle frequency of both slow (< 1 Hz) and fast (1-2 Hz) rhythms (Fig. 11), and it significantly alters the phase relationships that define this motor pattern (Fig. 12). These phase relationships change again after MCN5 stimulation (Fig. 12). 8. MCN5 acts in concert with the pyloric pacemaker ensemble to elicit a pyloric rhythm that exhibits enhanced pacemaker neuron activity and reduced activity in all nonpacemaker neurons. Additionally, despite their electrical coupling, the three types of pacemaker neurons exhibit distinct responses to MCN5 stimulation. This partially uncouples their normally coactive bursts. The resulting motor pattern is distinct from all previously characterized pyloric rhythms.
Asunto(s)
Braquiuros/fisiología , Ganglios de Invertebrados/fisiología , Neuronas Motoras/fisiología , Red Nerviosa/fisiología , Píloro/inervación , Animales , Vaciamiento Gástrico/fisiología , Potenciales de la Membrana/fisiología , Transmisión Sináptica/fisiologíaRESUMEN
1. In the isolated stomatogastric nervous system of the crab Cancer borealis (Fig. 1), the muscarinic agonist oxotremorine elicits several distinct gastric mill motor patterns from neurons in the stomatogastric ganglion (STG; Fig. 2). Selection of a particular gastric mill rhythm is determined by activation of distinct projection neurons that influence gastric mill neurons within the STG. In this paper we identify one such neuron, called commissural projection neuron 2 (CPN2), whose rhythmic activity is integral in producing one form of the gastric mill rhythm. 2. There is a CPN2 soma and neuropilar arborization in each commissural ganglion (CoG). The CPN2 axon projects through the superior esophageal nerve (son) and the stomatogastric nerve (stn) to influence neurons in the STG (Figs. 3 and 4A). 3. CPN2 activity influences most of the gastric mill neurons in the STG. Specifically, CPN2 excites gastric mill neurons GM and LG (gastric mill and lateral gastric, respectively) and inhibits the dorsal gastric (DG), anterior median (AM), medial gastric (MG), and inferior cardiac (IC) neurons (Figs. 5 and 6). CPN2 also indirectly inhibits gastric mill neurons Int1 and VD (interneuron 1 and ventricular dilator neuron, respectively) through its activation of LG. The CPN2 excitatory effects are mediated at least partly via discrete excitatory postsynaptic potentials (EPSPs; Fig. 4B), whereas its inhibitory effects are produced via smooth hyperpolarizations. 4. Within the CoG, CPN2 receives excitatory synaptic input from the anterior gastric receptor neuron (AGR), a gastric mill proprioceptive sensory neuron (Fig. 7) and inhibitory synaptic input from the gastric mill interneuron, Int1 (Fig. 8). 5. During one form of the gastric mill rhythm, CPN2 fires rhythmically in time with the gastric mill motor pattern, whereas it is silent or fires weakly during other gastric mill rhythms (Fig. 9). 6. When CPN2 rhythmic activity is suppressed during a CPN2-influenced gastric mill rhythm, the gastric mill rhythm continues, but the pattern is altered (Fig. 10). Moreover, transiently stimulating CPN2 during any ongoing gastric mill motor pattern can reset the timing of that rhythm (Fig. 11). 7. Tonic activity in CPN2 is insufficient to elicit a gastric mill rhythm (Fig. 12). Phasic activity in CPN2 can elicit a gastric mill rhythm only in preparations in which gastric mill neurons are already in an excited state (Figs. 12 and 13). 8. CPN2 recruitment plays a pivotal role in determining the final form of the gastric mill rhythm.(ABSTRACT TRUNCATED AT 400 WORDS)
Asunto(s)
Braquiuros/fisiología , Ganglios de Invertebrados/fisiología , Interneuronas/fisiología , Neuronas Motoras/fisiología , Boca/inervación , Reclutamiento Neurofisiológico/fisiología , Estómago/inervación , Transmisión Sináptica/fisiología , Animales , Contracción Muscular/fisiología , Red Nerviosa/fisiología , Inhibición Neural/fisiología , Receptores Muscarínicos/fisiologíaRESUMEN
mRNA from the dinoflagellate Symbiodinium sp. isolated from the staghorn coral Acropora formosa was used for the construction of cDNA libraries. A cDNA clone was identified which encoded the precursor of peridinin-chlorophyll a-binding protein (PCP), including a 52 amino acid transit peptide and the 313 amino acid mature protein. The deduced amino acid sequence clearly contains an internal duplication, implying that amongst dinoflagellates the M(r) 35,000 form of PCP has arisen by duplication and fusion of genes encoding the M(r) 15,000 form. This is the first reported sequence of a dinoflagellate light-harvesting protein. The anatomy of the mature protein and the transit peptide are discussed.
Asunto(s)
Carotenoides/genética , Dinoflagelados/genética , Fotosíntesis/genética , Precursores de Proteínas/genética , Proteínas Protozoarias/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Carotenoides/química , Clonación Molecular , ADN Complementario/genética , Biblioteca de Genes , Datos de Secuencia Molecular , Peso Molecular , Precursores de Proteínas/química , Señales de Clasificación de Proteína/genética , Proteínas Protozoarias/química , ARN Mensajero/aislamiento & purificación , ARN Protozoario/aislamiento & purificación , Alineación de SecuenciaRESUMEN
We studied muscle fibers by quantitative biochemistry to determine whether metabolic capacity varied among fibers of a given type as a function of their anatomic location. Muscles were selected from both contiguous and diverse anatomic regions within the rats studied. The individual fibers, classified into myosin ATPase fiber types by histochemical means, were assessed for fiber diameters and analyzed for the activities of enzymes representing major energy pathways: malate dehydrogenase (MDH, oxidative), lactate dehydrogenase (LDH, glycolytic), and adenylokinase (AK, high-energy phosphate metabolism). We found that neither the average activities of each of the three enzymes nor the fiber diameters varied in Type I or Type IIa fibers selected from superficial to deep portions of the triceps surae of the hindlimb. However, the IIb fibers in the deep region of this muscle group had significantly greater oxidative capacity, less glycolytic capacity, and smaller diameters than the superficially situated IIb fibers. Type IIa fibers in lateral gastrocnemius, extensor digitorum longus, psoas, diaphragm, biceps brachii, superficial masseter, and superior rectus muscles were highly variable in both diameter and enzyme profiles, with a correlation between MDH activity and fiber diameter. Therefore, our results show that both intermuscular and intramuscular metabolic variations exist in muscle fibers of a given type.
Asunto(s)
Músculos/metabolismo , Adenilato Quinasa/metabolismo , Animales , Femenino , L-Lactato Deshidrogenasa/metabolismo , Malato Deshidrogenasa/metabolismo , Músculos/enzimología , Especificidad de Órganos , Oxidación-Reducción , Ratas , Ratas EndogámicasRESUMEN
The program of acquisition of adult metabolic phenotypes was studied in three jaw muscles in order to determine the link between muscle metabolism and functional development. During early postnatal stages, there were similar transitions in the masseter, anterior digastric, and internal pterygoid muscles with respect to fiber growth, fiber type composition, and whole muscle energy metabolism. Oxidative capacity, as judged by the activities of the enzymes succinate dehydrogenase (SDH), malate dehydrogenase (MDH), and beta-hydroxyacyl CoA dehydrogenase (beta OAC), rose sharply after birth to reach near maximal levels by 3 weeks. The capacities for glycolytic metabolism represented by lactate dehydrogenase (LDH), and for high-energy phosphate metabolism represented by adenylokinase (AK) and creatine kinase (CK) activities, rose gradually, not reaching peak values until 6 weeks or later. Thus, the maturation of oxidative metabolism preceded that of glycolytic metabolism in the developing jaw muscles. This was documented for individual fibers in the masseter muscle. Differential metabolic maturation among the jaw muscles was evident beyond 3 weeks. All three jaw muscles attained their specific adult fiber-type profile by about 6 weeks. This maturation program differed from that of hindlimb muscles [Nemeth et al., J Neurosci 9:2336-2343, 1989] and diaphragm muscle [Kelly et al., J Neurosci 11:1231-1242, 1991], reflecting their differential energy demands for contractile performance.
Asunto(s)
Maxilares/metabolismo , Músculo Masetero/metabolismo , Desarrollo Maxilofacial/fisiología , Músculos del Cuello/metabolismo , Músculos Pterigoideos/metabolismo , Animales , Animales Recién Nacidos , Femenino , Músculo Masetero/enzimología , Músculo Masetero/crecimiento & desarrollo , Desarrollo de Músculos , Músculos del Cuello/enzimología , Músculos del Cuello/crecimiento & desarrollo , Músculos Pterigoideos/enzimología , Músculos Pterigoideos/crecimiento & desarrollo , Ratas , Ratas EndogámicasRESUMEN
Neonatal rats were exposed to a high-fat low-carbohydrate diet to determine how substrate availability might affect the metabolic phenotype of muscle. Mixed-fiber homogenates of extensor digitorum longus, soleus, and diaphragm muscles were assayed for beta-hydroxyacyl-CoA dehydrogenase (beta-OAC), succinate dehydrogenase, malate dehydrogenase, lactate dehydrogenase, phosphofructokinase (PFK), adenylokinase, and creatine kinase. The three muscles showed significant increases in enzyme activity for fatty acid oxidation (beta-OAC) in weaned neonatal rats maintained on the high-fat diet compared with normal weaned controls. This effect persisted for 6 wk of the diet. The other consistent metabolic change was a decrease in PFK. Adult animals subjected to the same diet had similar increases in fatty acid oxidation and a fall in PFK after 1 wk, with most of these changes persisting for the 4 wk of the diet. Examination of individual fibers revealed enzyme changes in fibers of all types, but with the largest effect in type IIb fibers. The data indicate that both adult and neonatal muscles are similarly capable of adjusting their energy metabolism in response to dietary factors.
Asunto(s)
Envejecimiento/metabolismo , Animales Recién Nacidos/metabolismo , Grasas de la Dieta/farmacología , Músculos/enzimología , Animales , Animales Recién Nacidos/crecimiento & desarrollo , Diafragma , Miembro Posterior , Ratas , Ratas EndogámicasRESUMEN
The fluoroquinolones, temafloxacin, sarafloxacin, and difloxacin, are determined in the bulk drug substances and in a variety of dosage form using high-performance liquid chromatography (HPLC). The HPLC system used is also applicable for ciprofloxacin and norfloxacin. The procedure uses UV detection at 280 nm, which provides a linear response of the subject compounds to at least 20 micrograms/ml. Assay precision (RSD) values were +/- 1.2% or better for the bulk drugs and ranged from +/- 0.42 to +/- 2.3% for suspension, capsule, and tablet formulations. Drug recoveries were quantitative from the dosage forms tested. Sensitivity of the subject compounds is approximately 50 ng/ml (2.5 ng on column).
Asunto(s)
Antiinfecciosos/análisis , Fluoroquinolonas , Quinolonas , Cromatografía Líquida de Alta Presión , Ciprofloxacina/efectos adversos , Ciprofloxacina/análogos & derivados , Ciprofloxacina/análisis , Solubilidad , Espectrofotometría UltravioletaRESUMEN
Enzymes of energy production were measured in muscle homogenates and in individual muscle fibers from 5 patients with McArdle's disease. Individual fibers were investigated to determine whether fibers of all types were completely devoid of glycogen phosphorylase activity and whether the involved fibers might be biochemically altered in a fiber type dependent manner to enhance the energy-generating capabilities of the cells through other metabolic pathways. Using highly sensitive biochemical assays, a complete absence of glycogen phosphorylase, a and b, activity was found in fibers of all types in the McArdle's patients. Levels of enzymes representing glycolysis, the Krebs cycle, and high energy phosphate metabolism were essentially normal in each fiber type, indicating an apparent lack in metabolic adaptation of these energy pathways to the absence of glycogen utilization. However, a key enzyme in the beta-oxidation of fatty acids (beta-hydroxyacyl CoA dehydrogenase, beta OAC) was elevated in all patients, and substantially in 4 of the 5. This suggested that lipid substrates can provide support for oxidative endurance capacity in some patients. Individual fiber analyses indicated that the compensation involved fibers of all types.
Asunto(s)
3-Hidroxiacil-CoA Deshidrogenasas/metabolismo , Enfermedad del Almacenamiento de Glucógeno Tipo V/enzimología , Músculos/enzimología , Metabolismo Energético , Ácidos Grasos/metabolismo , Enfermedad del Almacenamiento de Glucógeno Tipo V/metabolismo , Humanos , Técnicas In Vitro , Músculos/metabolismo , Músculos/patología , Fosforilasas/deficienciaRESUMEN
1. Nerve terminals associated with longitudinal muscle in the leech show FMRFamide-like immunoreactivity. 2. Structure-activity studies using FMRFamide analogs show that the C-terminal RFamide portion of the molecule is crucial for biological activity on leech longitudinal muscle. 3. The putative protease inhibitor FA (Phe-Ala) increases the peak tension produced by longitudinal muscle in response to superfused FMRFamide and the majority of its analogs, suggesting the presence of peripheral proteases capable of degrading RFamide peptides. 4. FMRFamide decreases the relaxation rate of neurally evoked contractions of longitudinal muscle. FA also decreases the relaxation rate of neurally evoked contractions. 5. Intact and isolated muscle cells respond to superfused FMRFamide with a conductance increase, that leads to depolarization and often with a delayed conductance decrease as the membrane potential is restored to resting levels. 6. The depolarizing response of isolated muscle cells to FMRFamide is dependent on external calcium.
Asunto(s)
Sanguijuelas/fisiología , Músculos/efectos de los fármacos , Neuropéptidos/farmacología , Neurotransmisores/farmacología , Animales , Calcio/farmacología , Electrofisiología , FMRFamida , Inmunohistoquímica , Contracción Muscular , Músculos/citología , Neuronas/fisiología , Inhibidores de Proteasas/farmacología , Relación Estructura-ActividadRESUMEN
Individual fibers of prospective fast (extensor digitorum longus; EDL) and slow (soleus) muscles of rats have been analyzed to determine the profiles of key energy-generating enzymes at successive stages of postnatal development. Mean activities of lactate dehydrogenase (LDH) and adenylokinase (AK), 2 enzymes associated with contractile function, are significantly different in the 2 fiber populations at birth; furthermore, wide variations in enzyme activities exist among the individual fibers. There is a progressive refinement of enzyme levels in the soleus into a more uniform fiber population, while the fibers in the EDL progressively diverge into 2 distinct phenotypes. Changes in EDL and soleus are punctuated by periods of rapid change, with the period between 10 and 21 d being most eventful. Generally, the maturation profiles of LDH and AK coincide with the transition from neonatal to adult fast myosins and closely reflect the timing of energy demands imposed by contractile activity patterns. In contrast, activities of the oxidative enzymes malate dehydrogenase and beta-hydroxyacyl CoA dehydrogenase are similar in both muscles at birth and steadily increase during the first 3 weeks, suggesting a progressive adaptation to the aerobic extrauterine environment. After 30 d, there are differential changes in the oxidative profiles of enzymes for fatty acid and glucose metabolism. The profiles follow dietary changes associated with weaning, which suggests a phenotypic dependence of neonatal muscle on the particular available energy substrate. All enzymes are low in all fibers of EDL and soleus at birth, indicating their modest metabolic capacity.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Músculos/enzimología , 3-Hidroxiacil-CoA Deshidrogenasas/metabolismo , Adenilato Quinasa/metabolismo , Animales , Animales Recién Nacidos , L-Lactato Deshidrogenasa/metabolismo , Malato Deshidrogenasa/metabolismo , Ratas , Ratas Endogámicas , Hormonas Tiroideas/farmacologíaRESUMEN
Motor units of the cat tibialis posterior muscle were selectively activated by prolonged electrical stimulation of functionally isolated motor axons in situ. During the activation, the glucose analog 2-deoxyglucose (DG) was administered systemically. Single muscle fibers were subsequently examined for accumulation of the metabolite 2-deoxyglucose-6-phosphate (DG6P) by an analytical assay and for depletion of glycogen by a PAS glycogen-specific staining reaction (periodic acid Schiff; PAS). In general, levels of DG6P were 20 times greater in unstained (PAS-negative) fibers compared with stained (PAS-positive) fibers. However, some glycogen-depleted fibers, particularly in putative ischemic fascicles of the muscle, did not have elevated DG6P, suggesting that depletion of glycogen is not always a reliable indicator of fiber activation. Furthermore, the PAS-staining reaction was not necessarily indicative of quantitative glycogen levels in single fibers. Thus, this report shows that DG6P accumulation enhances the identification of motor-unit fibers selectively activated via their common motor-nerve axon. Evidence is also presented for differential glucose uptake in muscle fibers of different phenotype, thereby indicating that the DG6P measurement in muscle has broad applicability to the investigation of cellular glucose utilization.
Asunto(s)
Glucosa-6-Fosfato/análogos & derivados , Glucofosfatos/metabolismo , Neuronas Motoras/fisiología , Músculos/fisiología , Animales , Gatos , Estimulación Eléctrica , Glucógeno/metabolismo , Músculos/inervación , Músculos/metabolismo , Concentración Osmolar , Reacción del Ácido Peryódico de SchiffRESUMEN
Excitatory motor neurons in the leech are cholinergic. By using a combination of intracellular Lucifer yellow injection and indirect immunofluorescence, we localized FMRFamidelike immunoreactivity to a number of the motor neurons innervating longitudinal and dorsoventral muscle in the leech. All excitatory motor neurons innervating longitudinal muscle (cells 3, 4, 5, 6, 8, L, 106, 107, 108) were labeled with an antiserum to FMRFamide, while the inhibitory motor neurons innervating longitudinal muscle (cells, 1, 2, 7, 9, 102) were not. The excitatory motor neuron innervating medial dorsoventral muscle (cell 117) was labeled, while the excitatory motor neuron innervating lateral dorsoventral muscle (cell 109) was not. The inhibitory motor neuron innervating dorsoventral muscle (cell 101) was also labeled. Nerve terminals along dorsoventral muscle were also labeled with the antiserum. FMRFamide was bath applied to strips of longitudinal muscle while recording tension, and the muscle's response was compared to its response to the previously identified neuromuscular transmitter ACh. Brief applications of FMRFamide caused a contraction approximately one-tenth as large as that caused by an equimolar amount of ACh. The muscle response to FMRFamide was unaffected by curare. During extended exposures, FMRFamide caused a maintained contraction in longitudinal muscle without any apparent desensitization of the FMRFamide receptors and occasionally triggered an irregular myogenic rhythm. This extended exposure to FMRFamide caused a post-exposure potentiation of the longitudinal muscle's response to ACh that shorter applications of FMRFamide did not. Thus FMRFamide may act as a transmitter or modulator in cholinergic motor neurons innervating longitudinal and dorsoventral muscles in the leech.
Asunto(s)
Sanguijuelas/citología , Neuronas Motoras/citología , Músculos/inervación , Neuropéptidos/análisis , Acetilcolina/farmacología , Animales , FMRFamida , Técnica del Anticuerpo Fluorescente , Colorantes Fluorescentes , Isoquinolinas , Neuronas Motoras/fisiología , Contracción Muscular/efectos de los fármacos , Músculos/citología , Músculos/fisiología , Neuropéptidos/farmacologíaRESUMEN
Methods for 2-deoxyglucose (2-DG) and 2-deoxyglucose 6-phosphate (DG6P) are described which are based on the fact that DG6P is oxidized by glucose-6-phosphate dehydrogenase (G6PDH), but at a rate 1000-fold slower than for glucose 6-phosphate, whereas hexokinase phosphorylates 2DG and glucose at comparable rates. Therefore, by adding the two enzymes in a suitable order, and in appropriate concentrations, 2DG, glucose, DG6P, and glucose 6-P can all be separately measured. To avoid a side reaction from the use of a high level of G6PDH, when measuring DG6P, glucose is first removed with glucose oxidase plus aldose reductase.
Asunto(s)
Desoxiazúcares/análisis , Desoxiglucosa/análisis , Glucosa-6-Fosfato/análogos & derivados , Glucosafosfato Deshidrogenasa , Glucofosfatos/análisis , Hexoquinasa , Química Encefálica , Glucosafosfato Deshidrogenasa/análisis , Hexoquinasa/análisis , Cinética , Saccharomyces cerevisiae/enzimologíaRESUMEN
Two sisters with progressive myopathy demonstrated microscopic and biochemical evidence of lipid storage in skeletal muscle. Their muscle biopsy specimens resembled those seen in Duchenne's muscular dystrophy and some of the biochemical features were similar, including increased muscle concentration of long-chain acyl-coenzyme A (a fatty oxidation intermediate) and decreased oxidation of radioactively labeled fatty acids by muscle homogenates in vitro. Although the site of the defect was not localized, the data suggested impairment of intramitochondrial beta-oxidation of fatty acids. These two patients may be important in understanding the pathogenesis of muscular dystrophy.
Asunto(s)
Metabolismo de los Lípidos , Distrofias Musculares/metabolismo , Niño , Preescolar , Coenzima A/metabolismo , Femenino , Humanos , Distrofias Musculares/enzimología , Distrofias Musculares/genética , Distrofias Musculares/patología , Oxidación-Reducción , Palmitatos/metabolismo , Piruvatos/metabolismoRESUMEN
Compared with normal skeletal muscle, muscle from patients with Duchenne dystrophy had decreased [U-14 C] palmitic acid oxidation. [1-14 C] palmitic acid oxidation was normal. These results may indicate a defect in intramitochondrial fatty acid oxidation.