Oxidative Damage and Antioxidants as Markers for the Selection of Emersion Hardening Treatments in GreenshellTM Mussel Juveniles (Perna canaliculus).

Transport out of the water is one of the most challenging events for juvenile Perna canaliculus and can be a highly inefficient process, with many juveniles subsequently being lost following extended periods of emersion. Hardening techniques offer a possible method for reducing transport-related stress. In this study, different hardening treatments (short, long and intermittent sub-lethal emersion) were used to prepare ~1.2 mm P.canaliculus for transport (20 h) and subsequent reoxygenation stress during re-immersion (i.e., recovery). The oxidative stress responses, resettlement behaviour, respiration rates and survival of the mussels after transport and during recovery were all assessed. Short emersion (1 h) as a hardening treatment prior to transport did not cause major stress to the mussels, which maintained respiration at control levels, showed significantly stimulated antioxidant defences during recovery, showed greater resettlement behaviour and remained viable after 24 h of recovery. In comparison, the long and intermittent emersion treatments negatively impacted oxidative stress responses and affected the viability of the mussels after 24 h of recovery. This study showed that exposing juvenile P.canaliculus to a mild stress prior to transport may stimulate protective mechanisms, therefore eliciting a hardening response, but care must be taken to avoid overstressing the mussels. Improving the management of stress during the transport of juvenile mussels may be key to minimising mussel losses and increasing harvest production, and biomarkers associated with oxidative stress/antioxidant metabolism could be valuable tools to ensure emersion hardening does not overstress the mussels and reduce survival.

Physiological responses of juvenile New Zealand geoduck (Panopea zelandica) following emersion and recovery.

The New Zealand geoduck clam is a unique seafood delicacy, with animals selling for up to $US 220-330/kg. Stress accumulated during transport of juveniles to grow-out sites represent a bottleneck in the aquaculture process. In this study, the physiological responses of juvenile geoducks following emersion (3- and 8-h), and recovery (1- and 5-days) were investigated. An integrated approach of flow cytometry, osmolality and metabolomics, along with behavioural assessments was used. Both cellular and chemical haemolymph parameters and metabolite profiles were recorded for P. zelandica juveniles and are reported herein for the first time. An increase in haemolymph osmolality was experienced with an increase in emersion period, with significant differences seen between the 3- and 8-h emersion groups after 5 days of recovery. Viability measures of haemocytes varied insignificantly between experimental groups, creating baseline ranges. The proportion of haemocytes undergoing respiratory burst activity did not appear to be affected by emersion and re-immersion. Haemocyte mitochondrial membrane potential was highest following 1-day of recovery, likely linked to metabolic readjustment, and increased glycolysis, taking place following emersion. Metabolomics analyses suggest that protein, lipid and carbohydrate metabolite classes assist with energy production in geoducks. Activation of anaerobic metabolic pathways, with a high dependence on succinate, were prominent in the 8-h exposure group, with metabolic recovery still taking place following 5-days of immersion, mainly due to proteins restoring energy reserves. Elucidating the physiological responses of juvenile geoduck subjected to transport stress can aid cultivation methods already underway to develop a novel, high value aquaculture industry.

A new method to localise and quantify oxidative stress in live juvenile mussels.

Stress and survival of the juvenile New Zealand green-lipped mussel, Perna canaliculus, is a poorly understood bottleneck in the ecological and economic performance of a significant aquaculture crop. This species was therefore selected as a model organism for the development of a new method to quantify oxidative stress in whole individuals. An in vivo ROS-activated stain (CellROX™) was administered to anaesthetised, translucent juveniles that were subsequently formaldehyde fixed and then visualised using confocal microscopy. Subsequent application of image analysis to quantifying ROS-positive tissue areas was successfully used to detect stress differences in juvenile mussels exposed to varying levels of emersion. This integrated method can be used to localise and quantify ROS production in individual translucent bivalve life stages (larval and juvenile), while relative stability following fixation greatly expands potential practical field applications. This article has an associated First Person interview with the first and third authors of the paper.

Biochemical composition of the Australasian sea cucumber, Australostichopus mollis, from a nutritional point of view

BACKGROUND: Sea cucumbers are highly prized seafood in Asia, where the demand for them has increased dramatically in recent years. However, nutritional information is only available for only a few of the commercially traded species. METHODS: In this study, the biochemical composition of the sea cucumber Australostichopus mollis was evaluated in terms of the major functional components: collagen, amino acid and fatty acid content using standard analytical procedures. RESULTS: The collagen, mostly type I, formed a homogeneously clustered network constituting 1.4% of the wet weight of adult A. mollis which is lower than for some other species of sea cucumber. The collagen consisted ofα1andα2 chains (around 116 kDa),αchain dimers,ßchains (around 212 kDa), and small amounts of γ components.The most abundant amino acids were glycine, alanine, threonine, serine and proline (lysine/arginine ratio of 0.1).Threonine was the most abundant essential amino acid, followed by methionine and valine, while glycine was the dominant non-essential amino acid. The sea cucumbers had a low lipid content (0.2 and 0.1% of wet weight forthe viscera and body wall, respectively) which is below the range reported for most other species of sea cucumber.The lipid contained high levels of PUFA (54%) compared to MUFA (23%) and SFA (24%). The dominant PUFA was arachidonic acid in both the body wall and viscera, followed by eicosapentaenoic acid. CONCLUSIONS: Overall, the Australasian sea cucumber has strong potential as a functional food due to its high levels of PUFA and essential amino acids, comparing favourably with the most commercially valuable sea cucumbers.

(Pro)renin receptor activation increases profibrotic markers and fibroblast-like phenotype through MAPK-dependent ROS formation in mouse renal collecting duct cells.

Recent studies suggested that activation of the PRR upregulates profibrotic markers through reactive oxygen species (ROS) formation; however, the exact mechanisms have not been investigated in CD cells. We hypothesized that activation of the PRR increases the expression of profibrotic markers through MAPK-dependent ROS formation in CD cells. Mouse renal CD cell line (M-1) was treated with recombinant prorenin plus ROS or MAPK inhibitors and PRR-shRNA to evaluate their effect on the expression of profibrotic markers. PRR immunostaining revealed plasma membrane and intracellular localization. Recombinant prorenin increases ROS formation (6.0 ± 0.5 vs 3.9 ± 0.1 nmol/L DCF/µg total protein, P < .05) and expression of profibrotic markers CTGF (149 ± 12%, P < .05), α-SMA (160 ± 20%, P < .05), and PAI-I (153 ± 13%, P < .05) at 10-8  mol/L. Recombinant prorenin-induced phospho ERK 1/2 (p44 and p42) at 10-8 and 10-6  mol/L after 20 minutes. Prorenin-dependent ROS formation and augmentation of profibrotic factors were blunted by ROS scavengers (trolox, p-coumaric acid, ascorbic acid), the MEK inhibitor PD98059 and PRR transfections with PRR-shRNA. No effects were observed in the presence of antioxidants alone. Prorenin-induced upregulation of collagen I and fibronectin was blunted by ROS scavenging or MEK inhibition independently. PRR-shRNA partially prevented this induction. After 24 hours prorenin treatment M-1 cells undergo to epithelial-mesenchymal transition phenotype, however MEK inhibitor PD98059 and PRR knockdown prevented this effect. These results suggest that PRR might have a significant role in tubular damage during conditions of high prorenin-renin secretion in the CD.

Vasopressin/V2 receptor stimulates renin synthesis in the collecting duct.

Renin is synthesized in the principal cells of the collecting duct (CD), and its production is increased via cAMP in angiotensin (ANG) II-dependent hypertension, despite suppression of juxtaglomerular (JG) renin. Vasopressin, one of the effector hormones of the renin-angiotensin system (RAS) via the type 2-receptor (V2R), activates the cAMP/PKA/cAMP response element-binding protein (CREB) pathway and aquaporin-2 expression in principal cells of the CD. Accordingly, we hypothesized that activation of V2R increases renin synthesis via PKA/CREB, independently of ANG II type 1 (AT1) receptor activation in CD cells. Desmopressin (DDAVP; 10(-6) M), a selective V2R agonist, increased renin mRNA (∼3-fold), prorenin (∼1.5-fold), and renin (∼2-fold) in cell lysates and cell culture media in the M-1 CD cell line. Cotreatment with DDAVP+H89 (PKA inhibitor) or CREB short hairpin (sh) RNA prevented this response. H89 also blunted DDAVP-induced CREB phosphorylation and nuclear localization. In 48-h water-deprived (WD) mice, prorenin-renin protein levels were increased in the renal inner medulla (∼1.4- and 1.8-fold). In WD mice treated with an ACE inhibitor plus AT1 receptor blockade, renin mRNA and prorenin protein levels were still higher than controls, while renin protein content was not changed. In M-1 cells, ANG II or DDAVP increased prorenin-renin protein levels; however, there were no further increases by combined treatment. These results indicate that in the CD the activation of the V2R stimulates renin synthesis via the PKA/CREB pathway independently of RAS, suggesting a critical role for vasopressin in the regulation of renin in the CD.

Cordoma sacrococcígeo: presentación de un caso / Sacrococcigeal cordoma: a case report

Introducción: El cordoma sacrococcígeo representa más del 50 por ciento de los cordomas. La resección total en bloquecombinada con radioterapia en altas dosis ha mostrado prolongar la recurrencia.Caso clínico: Se presenta un paciente masculino de 58 años operado de cordoma sacrococcígeo mediante resección total en bloque por vía posterior. Los estudios de imagen postoperatorios confirmaron la remoción total. Como complicacionesposquirúrgicas mediatas el paciente presentó una fístula de líquido cefalorraquídeo y una infección local que remitieron con drenaje espinal y antibioticoterapia. Además, presentó trastornos transitorios del control del esfínter vesical que desaparecieron a los 6 meses. Posteriormente recibió radioterapia con cobalto. Se ha mantenido asintomático y sin evidencia de recidiva en los estudios de imágenes por resonancia magnética.Conclusiones: Las mejores opciones de tratamiento se obtienen con un enfoque multidisciplinario, en centrosespecializados y con avanzadas técnicas neuroquirúrgicas asociada a la radioterapia(AU)

Introduction: Sacrococcigeal cordoma represents more than 50 percent of the cordomas. The total resection in blockcombined with radiotherapy in high dose has shown to prolong the recurrence.Clinical case: Male, 58 years old operated of sacrococcigeal cordoma is presented by means of total resection in block for posterior approach. The postoperative image studies confirmed the total removal. It presented cerebrospinal fistulae and local infection treated with spinal drainage and antibiotics. Also, transient urinary incontinence occurred for 6 months. After that he received cobalt radiotherapy. Until now he stayed asymptomatic and free of illness in resonance magnetic images.Conclusions: The best treatment options are obtained with a focus multidisciplinary, in specialized centers and with advanced neurosurgical and radiotherapy techniques(AU)

Uso del lisado plaquetario en artrosis de rodilla / Use of lysed platelets in osteoarthritis of the knee

Introducción: la artrosis es la enfermedad más frecuente de todos los trastornos articulares. Sus tratamientos son variables y van desde los procederes conservadores al quirúrgico pasando por las infiltraciones articulares; últimamente se ha agregado el uso de plasma rico en plaquetas. Objetivo: describir la evolución clínica de 20 pacientes con osteoartritis de rodilla que se les realizó infiltración con lisado plaquetario. Material y métodos: se desarrolló un estudio longitudinal prospectivo, con una muestra de 20 pacientes remitidos a consulta de Ortopedia del Hospital Docente «Dr. Enrique Cabrera¼ con diagnóstico de artrosis de rodilla, de octubre de 2011 a octubre 2012, que cumplieran con los criterios de inclusión establecidos. Se les realizó infiltración articular con lisado plaquetario autólogo y un seguimiento clínico usando variables como dolor, movilidad articular, fuerza muscular y escala funcional. Se conformó una base de datos para el procesamiento estadístico. Se calcularon medidas de resúmenes. Resultados: 14 eran del sexo femenino y 6 masculinos, con una edad que osciló entre los 39 y 81 años, Se infiltraron en total 35 rodillas. Existió una tendencia hacia la mejoría del dolor en 15 pacientes, y ganancia en la fuerza muscular de las rodillas tratadas, en cuanto a la evaluación de actividades físicas, la tendencia fue del paso de las dificultades muy severas y severas a leve o ninguna. Conclusiones: la mejoría en el dolor, la recuperación articular y de la escala funcional en la mayoría de los pacientes es un signo que apoya la mediación de las factores plaquetarios en la acción antiinflamatoria inmediata y su potencial regenerativo.

Introduction: Osteoarthritis is the most common joint ailment. Its treatments vary from conservative procedures to surgery, passing through joint injections and lately the use of plasma enriched in platelets. Objective: describe the clinical evolution of 20 patients with knee osteoarthritis who received joint injection of lysed platelets. Material and Methods: we did a prospective longitudinal study with a 20 patient sample, referred with kneeosteoarthritis to the service of Orthopedics of the «Dr. Enrique Cabrera¼ University Hospital, provided they meet the inclusion requirements. Autologous lysed platelets were injected and a clinical follow up using variables such as pain, joint mobility, muscular strength and functional scale. A database was filled for statistical processing. It were calculated statistical measures. Results: out of the 20 patients 14 were male and 6 female, age 39 to 81. 35 knees were injected. There was a trend to decrease of pain in 15 patients and gain of muscular strength in the injected cases. Regarding evolution of the physical activities, the tendency was improvement from very severe and severe to slight or none. Conclusions: the improvement of pain, joint recovery and functional scale in the vast majority of the treated patients supports the mediation of platelet factors in the immediate anti-inflammatory action and its regenerative potential.

Uso del lisado plaquetario en artrosis de rodilla / Use of lysed platelets in osteoarthritis of the knee

Introducción: la artrosis es la enfermedad más frecuente de todos los trastornos articulares. Sus tratamientos son variables y van desde los procederes conservadores al quirúrgico pasando por las infiltraciones articulares; últimamente se ha agregado el uso de plasma rico en plaquetas.Objetivo: describir la evolución clínica de 20 pacientes con osteoartritis de rodilla que se les realizó infiltración con lisado plaquetario. Material y métodos: se desarrolló un estudio longitudinal prospectivo, con una muestra de 20 pacientes remitidos a consulta de Ortopedia del Hospital Docente «Dr. Enrique Cabrera¼ con diagnóstico de artrosis de rodilla, de octubre de 2011 a octubre 2012, que cumplieran con los criterios de inclusión establecidos. Se les realizó infiltración articular con lisado plaquetario autólogo y un seguimiento clínico usando variables como dolor, movilidad articular, fuerza muscular y escala funcional. Se conformó una base de datos para el procesamiento estadístico. Se calcularon medidas de resúmenes. Resultados: 14 eran del sexo femenino y 6 masculinos, con una edad que osciló entre los 39 y 81 años, Se infiltraron en total 35 rodillas. Existió una tendencia hacia la mejoría del dolor en 15 pacientes, y ganancia en la fuerza muscular de las rodillas tratadas, en cuanto a la evaluación de actividades físicas, la tendencia fue del paso de las dificultades muy severas y severas a leve o ninguna. Conclusiones: la mejoría en el dolor, la recuperación articular y de la escala funcional en la mayoría de los pacientes es un signo que apoya la mediación de las factores plaquetarios en la acción antiinflamatoria inmediata y su potencial regenerativo(AU)

Introduction: Osteoarthritis is the most common joint ailment. Its treatments vary from conservative procedures to surgery, passing through joint injections and lately the use of plasma enriched in platelets. Objective: describe the clinical evolution of 20 patients with knee osteoarthritis who received joint injection of lysed platelets. Material and Methods: we did a prospective longitudinal study with a 20 patient sample, referred with kneeosteoarthritis to the service of Orthopedics of the Dr. Enrique Cabrera University Hospital, provided they meet the inclusion requirements. Autologous lysed platelets were injected and a clinical follow up using variables such as pain, joint mobility, muscular strength and functional scale. A database was filled for statistical processing. It were calculated statistical measures. Results: out of the 20 patients 14 were male and 6 female, age 39 to 81. 35 knees were injected. There was a trend to decrease of pain in 15 patients and gain of muscular strength in the injected cases. Regarding evolution of the physical activities, the tendency was improvement from very severe and severe to slight or none. Conclusions: the improvement of pain, joint recovery and functional scale in the vast majority of the treated patients supports the mediation of platelet factors in the immediate anti-inflammatory action and its regenerative potential(AU)

Ontogenetic variation in parasite infracommunities of the clingfish Sicyases sanguineus (Pisces: Gobiesocidae).

This study analyzed the variation of the parasite infracommunities and their relationship with the diet and spatial distribution of the clingfish Sicyases sanguineus during its ontogeny. In total, 154 clingfish were collected from the intertidal and the upper subtidal zone of Pacific Ocean along the central Chilean coast. A wide range in body length (1.4-34.5 cm) was observed, including juvenile and adult specimens. Eleven parasite species were found in the clingfish, i.e., 2 Copepoda, 2 Hirudinea, 1 Monogenea, 5 Digenea, and 1 Cestoda. The prevalence, total abundance, Brillouin's diversity index, and the infracommunity species richness increased with fish body length. The parasite communities, the diet, and the spatial distribution changed during clingfish ontogeny, specifically between juveniles (<20 cm body length [BL]) and adults (>20 cm BL). When fish reached a 20-cm BL, a considerable increment in abundance and species richness of parasites was observed; this coincided with an increase in the number of prey items in the diet and with a change of the fish from the intertidal into the subtidal zone. Therefore, the ontogeny of S. sanguineus is important for the variation of parasite infracommunities, which also was associated with dietary and spatial distribution shifts of this fish.

Epidemiología del suicidio en la segunda región de Chile / Epidemiology of suicide at the second region of Chile

Como problema de salud pública el suicidio ha adquirido especial relevancia estas últimas décadas. El los países occidentales ocupa las primeras cinco a diez causas de muerte. Desde un punto de vista epidemiológico el estudio de los patrones de muerte por suicidio podría arrojar luz sobre acciones que pudieran ser preventivas y por lo tanto salvadores de vidas. El presente es un estudio descriptivo destinado a conocer las características epidemiológicas del suicidio en la II Región, en el período comprendido entre los años 1989-1995. Se utilizó como metodología la revisión de la totalidad de los certificados de defunción que resume la mortalidad general de la Región en el período antes citado. Para recolectar la información se confeccionó un instrumento especialmente diseñado para este efecto. Se encontró un total de 194 casos: 161 hombres y 33 mujeres (relación H/M de 4,8-1). El promedio de edad de los hombres fue 39,05 y de las mujeres 43,12 años. A diferencia de otros estudios no se observó el efecto edad, ya que la población afectada es esencialmente joven en edad productiva. Las estaciones de mayor frecuencia de suicidio en los hombres son el invierno y el verano. No se encontró tendencia alguna respecto de los días de la semana. A partir de 1991 se observó un alza importante de suicidios en la Región (aumentó 14,6 por ciento) a expensas de los hombres, lo cual se reveló en lastasas que muestran valores altos comparados con el resto del país y Santiago. En las mujeres, a pesar de que no se observó alza en el suicidio, las tasas son igualmente elevadas hecha la comparación anterior. Se concluye la necesidad de efectuar otros estudios destinados a clarificar este problema

An automated system for detection and analysis of locomotor behavior in crustaceans

An efficient and simple system in presented for the analysis of crustacean locomotor behavior. The system is composed by six dual-compartment actographic chambers with photocoupling circuit for movement detection, and a device for acquisition and analysis of cata. Such device is made by a digital interface which feeds in to a microcomputer with disc unit and printer. Information is processed in real time during the experiment, with a simultaneous printout and storage in a floppy disc