RESUMEN
Estuarine ecosystems are characterized by a wide physical-chemical variation that in the context of global change scenarios may be exacerbated in the future. The fitness of resident organisms is expected to be influenced by such variation and, hence, its study is a priority. Some of that variation relates to water vertical stratification, which may create "environmental refuges" or distinct layers of water with conditions favoring the fitness of some individuals and species. This study explored the performance of juvenile mussels (M. chilensis) settled in two distinctive water depths (1â¯m and 4â¯m) of the Reloncaví fjord (southern Chile) by conducting a reciprocal transplants experiment. Salinity, saturation state and the contents of CO3 in seawater were among the factors that best explained the differences between the two layers. In such environmental conditions, the mussel traits that responded to such variation were growth and calcification rates, with significantly higher values at 4â¯m deep, whereas the opposite, increased metabolic stress, was higher in mussels raised and transplanted to the surface waters (1â¯m). Such differences support the notion of an environmental refuge, where species like mussels can find better growth conditions and achieve higher performance levels. These results are relevant considering the importance of M. chilensis as a shellfish resource for aquaculture and a habitat forming species. In addition, these results shed light on the variable responses exhibited by estuarine organisms to small-scale changes in the characteristics of the water column, which in turn will help to better understand the responses of the organisms to the projected scenarios of climate global change.
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Mytilus , Animales , Chile , Ecosistema , Humanos , Agua de Mar , PlantonesRESUMEN
Biological interactions and environmental constraints alter life-history traits, modifying organismal performances. Trematode parasites often impact their hosts by inducing parasitic castration, frequently correlated with increased body size in the host (i.e., gigantism hypothesis), which is postulated to reflect the re-allocation of energy released by the reduction in the reproductive process. In this study, we compared the effect of a trematode species on shell size and morphology in adult individuals of the intertidal mussels Perumytilus purpuratus (>20â¯mm) collected from two local populations of contrasting environmental regimes experienced in central-southern Chile. Our field data indicates that in both study locations, parasitized mussels evidenced higher body sizes (shell length, total weight and volume) as compared with non-parasitized. In addition, parasitized mussels from the southern location evidenced thinner shells than non-parasitized ones and those collected from central Chile, suggesting geographical variation in shell carbonate precipitation across intertidal habitats of the Chilean coast. In laboratory conditions, mussels collected from a local population in central Chile were exposed to two temperature treatments (12 and 18⯰C). Parasitized mussels showed higher growth rates than non-parasitized, regardless of the seawater temperature treatments. However, the metabolic rate was not influenced by the parasite condition or the temperature treatments. Our field and laboratory results support the parasite-induced gigantism hypothesis, and suggest that both the thermal environment and geographic location explain only a portion of the increased body size, while the parasitic condition is the most plausible factor modulating the outcome of this host-parasite interaction.
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ABSTRACT In the last decade, the risk benefits ratio of MHT has been evaluated mainly in terms of cardiovascular risk. Present Consensus Statement is largely inspired by the Global Consensus on Menopausal Hormone Therapy in 2013 and 2016 by leading global menopause societies (The American Society for Reproductive Medicine, The Asia Pacific Menopause Federation, The Endocrine Society, The European Menopause and Andropause Society, The International Menopause Society, The International Osteoporosis Foundation and The North American Menopause Society). The aim of these Recommendations is to provide a simple and updated reference on postmenopausal MHT. The term MHT typically includes estrogen replacement therapy (ERT) and estrogen-progestogen therapy (EPT). EPT can be sequential (Seq) when progestogen is added to ERT for 10-14 days a month, or continuous combined (CC) when progestogen is administered continuously every day along with a fixed amount of estrogen. MHT also includes Tibolone and the Tissue Selective Estrogen Complex (TSEC).
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Humanos , Femenino , Sociedades Médicas/tendencias , Menopausia , Terapia de Reemplazo de Estrógeno , Terapia de Reemplazo de Estrógeno/efectos adversos , Factores de Riesgo , Estrógenos/administración & dosificaciónRESUMEN
BACKGROUND: Neosaxitoxin (NeoSTX) and related paralytics shellfish toxins has been successfully used as local anesthetic and muscle relaxants to treat a variety of ailments. The primary mechanism of action of these toxins occurs by blocking voltage-gated sodium channels with compounds such as TTX, lidocaine, or derivatives. However, most of these non-classical sodium channel blockers act with a reduced time effect as well as ensuing neurotoxicity. NEW METHOD: In this report, we show that the use of local NeoSTX injections inactivates the hippocampal neuronal activity reversibly with a by long-term dynamics, without neuronal damage. RESULTS: A single 10 ng/µl injection of NeoSTX in the dorsal CA1 region abolished for up to 48 h memory capacities and neuronal activity measured by the neuronal marker c-fos. After 72 h of toxin injection, the animals fully recover their memory capacities and hippocampal neuronal activity. The histological inspection of NeoSTX injected brain regions revealed no damage to the tissue or reactive gliosis, similar to vehicle injection. Acute electrophysiological recording in vivo shows, also, minimal spreading of the NeoSTX in the cerebral tissue. COMPARISON WITH EXISTING METHODS: Intracerebral NeoSTX injection showed longer effects than other voltage sodium channel blocker, with minimal spreading and no neuronal damage. CONCLUSION: NeoSTX is a new useful tool that reversibly inactivates different brains region for a long time, with minimal diffusion and without neuronal damage. Moreover, NeoSTX can be used as a valuable sodium channel blocker for many studies in vivo and with potential therapeutic uses.
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Región CA1 Hipocampal/efectos de los fármacos , Neuronas/efectos de los fármacos , Saxitoxina/análogos & derivados , Bloqueadores de los Canales de Sodio/administración & dosificación , Memoria Espacial/efectos de los fármacos , Amnesia/inducido químicamente , Animales , Región CA1 Hipocampal/fisiología , Masculino , Neuronas/fisiología , Ratas Sprague-Dawley , Reconocimiento en Psicología/efectos de los fármacos , Saxitoxina/administración & dosificaciónRESUMEN
As a result of human activities, climate forecasts predict changes in the oceans pCO2 and salinity levels with unknown impacts on marine organisms. As a consequence, an increasing number of studies have begun to address the individual influence of pCO2 and salinity but much remains to be done to understand their combined effects on the physiology and ecology of marine species. Our study addressed this knowledge gap by measuring the influence of current and predicted levels of pCO2 (380 and 1200â¯ppm, respectively) and salinity (20, 25 and 30 psµ) on the energetic physiology of juvenile mussels (Mytilus chilensis) from the south-eastern Pacific region. Our results indicate that a reduced salinity caused a significant reduction in clearance rate, absorption efficiency and scope for growth of this species. Meanwhile, an increase in pCO2 levels caused a reduction in excretion rates and interacted significantly with salinity in the rate of oxygen uptake measured in the mussel. These results suggest that potential changes in salinity might have a direct role on the physiology of M. chilensis. The effect of pCO2, although less prevalent among the variables measured here, did interact with salinity and is also likely to alter the physiology of this species. Given the ecological and economic importance of M. chilensis, we call for further studies exploring the influence of pCO2 across a wider range of salinities.
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Dióxido de Carbono/análisis , Mytilus/fisiología , Salinidad , Agua de Mar/química , Animales , Océanos y Mares , Alimentos MarinosRESUMEN
Tide pools habitats are naturally exposed to a high degree of environmental variability. The consequences of living in these extreme habitats are not well established. In particular, little it is known about of the effects of hypercanic seawater (i.e. high pCO2 levels) on marine vertebrates such as intertidal pool fish. The aim of this study was to evaluate the effects of increased pCO2 on the physiology and behavior in juveniles of the intertidal pool fish Girella laevifrons. Two nominal pCO2 concentrations (400 and 1600µatm) were used. We found that exposure to hypercapnic conditions did not affect oxygen consumption and absorption efficiency. However, the lateralization and boldness behavior was significantly disrupted in high pCO2 conditions. In general, a predator-risk cost of boldness is assumed, thus the increased occurrence of shy personality in juvenile fishes may result in a change in the balance of this biological interaction, with significant ecological consequences.
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Conducta Animal , Dióxido de Carbono/química , Ecosistema , Perciformes/fisiología , Agua de Mar/química , Animales , HomeostasisRESUMEN
Zika virus (ZIKV) is an emerging mosquito-borne flavivirus circulating in Asia and Africa. In 2013, a large outbreak was reported on the archipelago of French Polynesia. In this study, we report the detection and molecular characterization of Zika virus for the first time in Chile from an outbreak among the inhabitants of Easter Island. A total of 89 samples from patients suspected of having ZIKV infection were collected between the period from January to May, 2014. Molecular diagnosis of the virus was performed by RT-PCR followed by the sequencing of the region containing the NS5 gene. A comparison of the viral nucleic acid sequence with those of other strains of ZIKA virus was performed using the MEGA software. Fifty-one samples were found positive for ZIKV by RT-PCR analysis. Further analysis of the NS5 gene revealed that the ZIKV strains identified in Easter Island were most closely related to those found in French Polynesia (99.8 to 99.9% nt and 100% aa sequence identity). These results strongly suggest that the transmission pathway leading to the introduction of Zika virus on Easter Island has its origin in French Polynesia.
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Brotes de Enfermedades , Infección por el Virus Zika/epidemiología , Virus Zika/aislamiento & purificación , Análisis por Conglomerados , Humanos , Epidemiología Molecular , Filogenia , Polinesia/epidemiología , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Homología de Secuencia , Virus Zika/clasificación , Virus Zika/genéticaRESUMEN
The impact of an endurance race on pulmonary pro-oxidative formation and lipoperoxidation was evaluated using exhaled breath condensate (EBC). 3 groups of 12, 12 and 17 healthy recreational runners of both sexes ran 10, 21.1 and 42.2 km, respectively. EBC samples were obtained before the run and at 20 and 80 min post run. Concentrations of H2O2, NO2 - , malondialdehyde and pH were determined. The 10 km group showed no post-run variations for H2O2 and NO2 - concentrations. The 21.1 km group showed significant increments for NO2 - , and H2O2 concentrations in 20 min and 80 min samples. The 42.2 km group, showed increased NO2 - concentration in 20 min and 80 min samples, while H2O2 concentration increased only in the 20 min sample. In the 10 and 42.2 km groups neither malondialdehyde concentration nor pH showed differences. The 42.2 km group exhibited ΔH2O2 and ΔNO2 - medians higher than the 10 km group. ΔpH median decreased in 21.1 and 42.2 km groups, exhibiting values significantly lower than the 10 km group. ΔH2O2 y ΔNO2 - correlated directly with race time, while ΔpH, correlated inversely. In conclusion, intense prolonged exercise favors the increase in pulmonary pro-oxidative levels, with no modifications on lipoperoxidation. Running time relates to the magnitude of acute post exercise pro-oxidative formation.
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Pruebas Respiratorias/métodos , Estrés Oxidativo/fisiología , Esfuerzo Físico/fisiología , Carrera/fisiología , Adulto , Femenino , Humanos , Peróxido de Hidrógeno/análisis , Concentración de Iones de Hidrógeno , Masculino , Malondialdehído/análisis , Nitritos/análisis , Adulto JovenRESUMEN
BACKGROUND: Gonyautoxin are phycotoxins, whose molecular mechanism of action is a reversible block of the voltage-gated sodium channels at axonal level, impeding nerve impulse propagation. OBJECTIVE: To evaluate clinical efficacy of gonyautoxin in the treatment of patients with chronic tensional-type headache. METHODS: Open trial from September 2004 to 2005 in Hospital Clínico Universidad de Chile. Twenty-seven patients with chronic tension-type headache were locally infiltrated with gonyautoxins (50 micrograms) in ten sites considered as pain trigger points in a fixed infiltration protocol. In each site, a volume of 200 microlitres was injected. EMG recording was performed before and immediately after infiltrations. Main outcome measures are where a significantly drop-off in acute headache pain score occurs and number of days without headache pain. RESULTS: No side effects were detected in the follow-up period. From base line of 2 weeks, 19 patients of 27 (70%) are the successfully responders to the treatment. They showed the remarkable immediate effect after infiltration demonstrated by trapezium EMG recording. Patients reported a fall in pain score 5 minutes post-injection from 5.0 +/- 2.8 to 1.6 +/- 1.6 (mean +/- SD). The responder showed an average of 8.1 +/- 9.9 weeks of headache pain-free, all of them without a second infiltration or use of any additional analgesic medication. DISCUSSION: The therapeutic properties of gonyautoxin local infiltration in chronic tension-type headache patients are shown to be safe and effective. This report describes a new therapy for chronic tension-type headache involving local infiltrations of gonyautoxins. The immediate headache pain relief effect shown only minutes after toxin infiltrations were the most remarkable feature of this protocol. This is the first gonyautoxins testing report in the treatment of chronic tension-type headache.
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Saxitoxina/análogos & derivados , Cefalea de Tipo Tensional/tratamiento farmacológico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Enfermedad Crónica , Electromiografía/efectos de los fármacos , Femenino , Humanos , Inyecciones Intramusculares/métodos , Masculino , Toxinas Marinas/administración & dosificación , Toxinas Marinas/efectos adversos , Persona de Mediana Edad , Dimensión del Dolor/efectos de los fármacos , Saxitoxina/administración & dosificación , Saxitoxina/efectos adversosRESUMEN
OBJECTIVE: The use of gonyautoxin has been reported to be safe and effective in healing acute and chronic anal fissures. This study was designed to show better efficacy in healing patients with chronic anal fissure by increasing the frequency of toxin injection. METHOD: Twenty-three chronic anal fissure patients were treated with doses of 100 units of gonyautoxin, which was intrasphincteric, infiltrated. The frequency of injection was every 4 days. Anorectal manometries were performed before and 4 min after infiltration. RESULTS: Total remissions were achieved within 7-14 days. The patients healed with a mean time of 8.2 +/- 2.4 days. No relapsed were observed during the 10-month follow up. Neither faecal incontinence nor other side effects were observed. All patients showed immediate sphincter relaxation detected by clinical examination. The maximum anal resting pressures, recorded 4 min after injections decreased to 62.9 +/- 27.7 mmHg, being 65.3 +/- 29.6% of baseline. Immediately after infiltration, patients reported anaesthetic effect with a fall down of the postdefecatory pain. CONCLUSION: Although, gonyautoxin anal fissures treatment recently published proved to be safe and effective, this study shows a better protocol for anal fissure treatment, showing better efficacy by shorting the healing time with better perception of healing by patients. Gonyautoxin anal sphincter infiltration proves to be safe and effective, reducing discomfort and healing time, advantageously comparing with alternative therapeutic approaches for chronic anal fissure.
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Fisura Anal/tratamiento farmacológico , Toxinas Marinas/uso terapéutico , Saxitoxina/análogos & derivados , Adulto , Anciano , Enfermedad Crónica , Femenino , Humanos , Masculino , Persona de Mediana Edad , Dolor , Inducción de Remisión , Saxitoxina/uso terapéutico , Factores de Tiempo , Resultado del TratamientoRESUMEN
Lung oxidative stress (OS) was explored in resting and in exercising subjects exposed to moderate and high altitude. Exhaled breath condensate (EBC) was collected under field conditions in male high-competition mountain bikers performing a maximal cycloergometric exercise at 670 m and at 2,160 m, as well as, in male soldiers climbing up to 6,125 m in Northern Chile. Malondialdehyde concentration [MDA] was measured by high-performance liquid chromatography in EBC and in serum samples. Hydrogen peroxide concentration [H(2)O(2)] was analysed in EBC according to the spectrophotometric FOX(2) assay. [MDA] in EBC of bikers did not change while exercising at 670 m, but increased from 30.0+/-8.0 to 50.0+/-11.0 nmol l(-1) (P<0.05) at 2,160 m. Concomitantly, [MDA] in serum and [H(2)O(2)] in EBC remained constant. On the other hand, in mountaineering soldiers, [H(2)O(2)] in EBC under resting conditions increased from 0.30+/-0.12 mumol l(-1) at 670 m to 1.14+/-0.29 mumol l(-1) immediately on return from the mountain. Three days later, [H(2)O(2)] in EBC (0.93 +/-0.23 mumol l(-1)) continued to be elevated (P<0.05). [MDA] in EBC increased from 71+/-16 nmol l(-1) at 670 m to 128+/-26 nmol l(-1) at 3,000 m (P<0.05). Changes of [H(2)O(2)] in EBC while ascending from 670 m up to 3,000 m inversely correlated with concomitant variations in HbO2 saturation (r=-0.48, P<0.05). AMS score evaluated at 5,000 m directly correlated with changes of [MDA] in EBC occurring while the subjects moved from 670 to 3,000 m (r=0.51, P<0.05). Lung OS may constitute a pathogenic factor in AMS.
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Mal de Altura/diagnóstico , Mal de Altura/metabolismo , Altitud , Pulmón/metabolismo , Estrés Oxidativo/fisiología , Adolescente , Adulto , Pruebas Respiratorias , Ejercicio Físico/fisiología , Prueba de Esfuerzo , Humanos , Peróxido de Hidrógeno/metabolismo , Peroxidación de Lípido , Masculino , Malondialdehído/metabolismo , Montañismo/fisiologíaRESUMEN
Copper mine tailings have been discharged around the city of Chanaral, in northern Chile, for more than 60 years. This report summarizes a 17-month long monitoring study of species richness and biodiversity at five intertidal sites around the point of the tailing discharge. Total dissolved copper in sites close to the point of discharge varied between 8.72 microg/l and 34.15 microg/l, showing that there has not been a significant reduction since 1994. However, species richness has increased, suggesting a possible recovery of the system. While diversity of sessile organisms correlates negatively with dissolved copper, diversity of mobile invertebrates did not correlate with the metal concentration. To explain the observed results we discuss the role of algal turf interference on the distribution of mobile invertebrates at reference sites, a top-down effect caused by the absence of carnivores at impacted sites, and an avoidance strategy by some species to reduce their contact with contaminated seawater.
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Biodiversidad , Monitoreo del Ambiente/estadística & datos numéricos , Contaminantes Ambientales/análisis , Eucariontes , Invertebrados , Agua de Mar/análisis , Animales , Chile , Análisis por Conglomerados , Cobre/análisis , Biología MarinaRESUMEN
This study reports the data recorded from four patients intoxicated with shellfish during the summer 2002, after consuming ribbed mussels (Aulacomya ater) with paralytic shellfish toxin contents of 8,066 n 61.37 mg/100 gr of tissue. Data associated with clinical variables and paralytic shellfish toxins analysis in plasma and urine of the intoxicated patients are shown. For this purpose, the evolution of respiratory frequency, arterial blood pressure and heart rate of the poisoned patients were followed and recorded. The clinical treatment to reach a clinically stable condition and return to normal physiological parameters was a combination of hydration with saline solution supplemented with Dobutamine (vasoactive drug), Furosemide (diuretic) and Ranitidine (inhibitor of acid secretion). The physiological condition of patients began to improve after four hours of clinical treatment, and a stable condition was reached between 12 to 24 hours. The HPLC-FLD analysis showed only the GTX3/GTX2 epimers in the blood and urine samples. Also, these epimers were the only paralytic shellfish toxins found in the shellfish extract sample.
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Humanos , Masculino , Animales , Persona de Mediana Edad , Mariscos/análisis , Mariscos/microbiología , Mariscos/toxicidad , Chile/epidemiología , /etiología , /microbiología , Parestesia/etiología , Parestesia/microbiología , Toxinas Marinas/aislamiento & purificación , Toxinas Marinas/análisis , Toxinas Marinas/efectos adversos , Toxinas Marinas/farmacología , Toxinas Marinas/toxicidadRESUMEN
Glycolytic metabolism in meiotic and post-meiotic spermatogenic cells shows differentiation-related changes. The developmental and physiological significance of these metabolic changes is not known. The aim of the present study was to test the hypothesis that glucose and lactate metabolism can modulate intracellular calcium [Ca2+](i) in spermatogenic cells in an opposing and dynamic manner. Fluorescent probes were used to measure [Ca2+](i) and pH(i), and HPLC was used to measure intracellular adenine nucleotides and mitochondrial sensing of ATP turnover. [Ca2+](i) in pachytene spermatocytes and round spermatids was modulated by changes in lactate and glucose concentrations in the media. The kinetics and magnitude of the [Ca2+](i) changes induced by lactate and glucose were different in meiotic and post-meiotic spermatogenic cells. The presence of glucose in the medium induced a decrease in pH(i) in spermatogenic cells. This glucose-induced pH(i) decrease occurred later than the changes in [Ca2+](i), which were also observed when the pH(i) decrease was inhibited, indicating that the glucose-induced [Ca2+](i) increase was not a consequence of pH(i) changes. Hexose phosphorylation in glycolysis was part of the mechanism by which glucose metabolism induced a [Ca2+](i) increase in spermatogenic cells. The sensitivity of [Ca2+](i) to carbohydrate metabolism was higher in round spermatids than in pachytene spermatocytes. Thus, differentiation-related changes in carbohydrate metabolism in spermatogenic cells determine a dynamic and differential modulation of their [Ca2+](i) by glucose and lactate, two substrates secreted by the Sertoli cells.
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Calcio/metabolismo , Glucosa/farmacología , Líquido Intracelular/metabolismo , Ácido Láctico/farmacología , Espermatozoides/metabolismo , Adenosina Trifosfato/metabolismo , Animales , Metabolismo de los Hidratos de Carbono , Diferenciación Celular , Concentración de Iones de Hidrógeno , Masculino , Consumo de Oxígeno , Ratas , Ratas Wistar , Espermátides/metabolismo , Espermatocitos/metabolismoRESUMEN
Se presenta un caso clínico de listeriosis diagnosticado a las 16 semanas de embarazo, con evolución perinatal satisfactoria y revisión del tema
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Humanos , Adulto , Femenino , Embarazo , Recién Nacido , Listeria monocytogenes , Listeriosis , Complicaciones Infecciosas del Embarazo , Listeria monocytogenes , Listeriosis , Intercambio Materno-Fetal , Complicaciones Infecciosas del Embarazo , Segundo Trimestre del EmbarazoRESUMEN
The hypothesis that intracellular [Ca2+] is a cell parameter responsive to extreme temperatures in rat meiotic and postmeiotic spermatogenic cells was tested using intracellular fluorescent probes for Ca2+ and pH. In agreement with this hypothesis, extreme temperatures induced a rapid increase of cytosolic [Ca2+] in rat pachytene spermatocytes and round spermatids. Oscillatory changes in temperature can induce oscillations in cytosolic [Ca2+] in these cells. Intracellular [Ca2+] homeostasis in round spermatids was more sensitive to high temperatures compared with pachytene spermatocytes. The calculated activation energies for SERCA ATPase-mediated fluxes in pachytene spermatocytes and round spermatids were 62 and 75 kJ mol(-1), respectively. The activation energies for leak fluxes from intracellular Ca2+ stores were 55 and 68 kJ mol(-1) for pachytene spermatocytes and round spermatids, respectively. Together with changes in cytosolic [Ca2+], round spermatids undergo a decrease in pH(i) at high temperatures. This temperature-induced decrease in pH(i) appears to be partially responsible for the increase in cytosolic [Ca2+] of round spermatids induced by high temperatures. This characteristic of rat meiotic and postmeiotic spermatogenic cells to undergo an increment in cytosolic Ca2+ at temperatures > 33 degrees C can be related to the induction of programmed cell death by high temperatures in these cells.
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Calcio/metabolismo , Citosol/metabolismo , Calor/efectos adversos , Espermatozoides/metabolismo , Animales , Apoptosis , Homeostasis , Concentración de Iones de Hidrógeno , Masculino , Meiosis , Ratas , Ratas Sprague-Dawley , Espermátides/metabolismo , Espermatocitos/metabolismoRESUMEN
The effects of parasitisation by Aphidius ervi on the thermoregulatory behaviour of the pea aphid Acyrtosiphon pisum were studied in alfalfa fields and in an experimental thermal gradient. In the field, mummies were found exclusively on the adaxial surface of the upper leaves, and aphids in the mid canopy. The adaxial surface of the upper leaves was ca. 2 degrees C hotter that the mid-canopy. In the thermal gradient, the thermal effect (selected minus exposure temperature) was higher in magnitude in non-parasitised than in parasitised aphids; the thermal effects of both types of aphids were linearly and negatively correlated with exposure temperature (i.e. aphids showed negative thermal sensitivity). The thermal sensitivity of parasitised aphids was lower than that of non-parasitised aphids. The results are discussed in relation to hypotheses on factors governing the host-parasite relationship.
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Protein phosphatases are involved in many cellular processes. One of the most abundant and best studied members of this class is protein phosphatase type-2A (PP2A). In this study, PP2A was purified from the mussel Mytilus chilensis. Using both SDS-PAGE and size exclusion gel filtration under denaturant conditions, it was confirmed that the PP2A fraction was essentially pure. The isolated enzyme is a heterodimer and the molecular estimated masses of the subunits are 62 and 28 kDa. The isolated PP2A fraction has a notably high p-NPP phosphatase activity, which is inhibited by NaCl. The hydrolytic p-NPP phosphatase activity is independent of the MgCl2 concentration. The time courses of the inhibition of the PP2A activity of p-NPP hydrolysis by increasing concentrations of three phycotoxins that are specific inhibitors of PP2A are shown. Inhibitions caused by Okadaic acid, dinophysistoxin-1 (DTX1, 35-methylokadiac acid) and Microcystine L-R are dose-dependent with inhibition constants (Ki) of 1.68, 0.40 and 0.27 nM respectively. Microcystine L-R, the most potent phycotoxin inhibitor of PP2A isolated from Mytilus chilensis with an IC50 = 0.25 ng/ml, showed the highest specific inhibition effect an the p-NPP hydrolisis. The calculated IC50 for DTX1 and OA was 0.75 ng/ml and 1.8 ng/ml respectively.
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Bivalvos/enzimología , Inhibidores Enzimáticos/farmacología , Ácido Ocadaico/farmacología , Péptidos Cíclicos/farmacología , Fosfoproteínas Fosfatasas/antagonistas & inhibidores , Piranos/farmacología , Animales , Cromatografía Líquida de Alta Presión , Electroforesis en Gel de Poliacrilamida , Toxinas Marinas , Microcistinas , Fosfoproteínas Fosfatasas/química , Fosfoproteínas Fosfatasas/aislamiento & purificación , Proteína Fosfatasa 2RESUMEN
Energy metabolism and intracellular adenine nucleotides of meiotic and postmeiotic spermatogenic cells are highly dependent on external substrates for oxidative phosphorylation and glycolysis. Using fluorescent probes to measure the changes in cytosolic [Ca2+] ([Ca2+]i) and pH (pHi), we were able to demonstrate that changes in energy metabolism of meiotic and postmeiotic spermatogenic cells were rapidly translated into changes of pHi and [Ca2+]i in the absence or presence of external Ca2+. Under these conditions, mitochondria were gaining cytosolic calcium in these cells. Our results indicate that Ca2+ mobilised by changes in metabolic energy pathways originated in thapsigargin-sensitive intracellular Ca2+ stores. Changes in intracellular adenine nucleotides, measured by HPLC, and a likely colocalization of ATP-producing and ATP-consuming processes in the cells seemed to provide the linkage between metabolic fluxes and the changes in pHi and [Ca2+]i in pachytene spermatocytes and round spermatids. Glucose metabolism produced an increase of [Ca2+]i in round spermatids but not in pachytene spermatocytes, and a decrease in pHi in both cell types. Hence, glucose emerges as a molecule that can differentially modulate [Ca2+]i and pHi in pachytene spermatocytes and round spermatids in rats.
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Calcio/metabolismo , Metabolismo Energético , Concentración de Iones de Hidrógeno , Espermatocitos/metabolismo , Adenosina Trifosfato/metabolismo , Animales , Fluoresceínas , Colorantes Fluorescentes , Fura-2 , Cinética , Masculino , Mitocondrias/metabolismo , Fosforilación Oxidativa/efectos de los fármacos , Ratas , Ratas Wistar , Espectrometría de Fluorescencia , Espermátides/metabolismo , Espermátides/ultraestructura , Espermatocitos/ultraestructuraRESUMEN
Protein phosphatases are involved in many cellular processes. One of the most abundant and best studied members of this class is protein phosphatase type-2A (PP2A). In this study, PP2A was purified from the mussel Mytilus chilensis. Using both SDS-PAGE and size exclusion gel filtration under denaturant conditions, it was confirmed that the PP2A fraction was essentially pure. The isolated enzyme is a heterodimer and the molecular estimated masses of the subunits are 62 and 28 kDa. The isolated PP2A fraction has a notably high p-NPP phosphatase activity, which is inhibited by NaCl. The hydrolytic p-NPP phosphatase activity is independent of the MgCl2 concentration. The time courses of the inhibition of the PP2A activity of p-NPP hydrolysis by increasing concentrations of three phycotoxins that are specific inhibitors of PP2A are shown. Inhibitions caused by Okadaic acid, dinophysistoxin-1 (DTX1, 35-methylokadiac acid) and Microcystine L-R are dose-dependent with inhibition constants (Ki) of 1.68, 0.40 and 0.27 nM respectively. Microcystine L-R, the most potent phycotoxin inhibitor of PP2A isolated from Mytilus chilensis with an IC50 = 0.25 ng/ml, showed the highest specific inhibition effect an the p-NPP hydrolisis. The calculated IC50 for DTX1 and OA was 0.75 ng/ml and 1.8 ng/ml respectively.