Repeated hypoxic episodes allow hematological and physiological habituation in rainbow trout.

Introduction: Under climate change, the increase in temperature in aquatic environments may induce oxygen depletion. In extreme cases, low oxygen may become a limiting factor for fish, thus generating stress. In addition, consecutive hypoxic episodes may complicate the recovery of individuals and hinder their ability to modulate physiological and biochemical responses to maintain homeostasis. Thus, the aim of this study was to determine the hematological and physiological responses of rainbow trout under a condition of repeated hypoxic and manipulation stresses at three different time points. Methods: Every hypoxic episode consisted of exposing the fish to low dissolved oxygen concentrations (2 mgO2/L for 1 h). Following the exposure, the fish were allowed to recover for 1 h, after which they were sampled to investigate hematological and physiological parameters. Results and discussion: The results showed a pattern of habituation reflected by values of hematocrit, hemoglobin, and mean corpuscular volume, indicating a certain ability of rainbow trout to resist this type of repeated hypoxic events, provided that the fish can have some recovery time between the exposures.

Programmed Death-Ligand (PD-L1), Epidermal Growth Factor (EGF), Relaxin, and Matrix Metalloproteinase-3 (MMP3): Potential Biomarkers of Malignancy in Canine Mammary Neoplasia.

Gene expression has been suggested as a putative tool for prognosis and diagnosis in canine mammary neoplasia (CMNs). In the present study, 58 formalin-fixed paraffin-embedded (FFPE) paraffined canine mammary neoplasias from 27 different bitches were included. Thirty-seven tumours were classified as benign, whereas thirty-one were classified as different types of canine carcinoma. In addition, mammary samples from three healthy bitches were also included. The gene expression for vascular endothelial growth factor-α (VEGFα), CD20, progesterone receptor (PGR), hyaluronidase-1 (HYAL-1), programmed death-ligand 1 (PD-L1), epidermal growth factor (EGF), relaxin (RLN2), and matrix metalloproteinase-3 (MMP3) was assessed through RT-qPCR. All the assessed genes yielded a higher expression in neoplastic mammary tissue than in healthy tissue. All the evaluated genes were overexpressed in neoplastic mammary tissue, suggesting a role in the process of tumorigenesis. Moreover, PD-L1, EGF, relaxin, and MMP3 were significantly overexpressed in malignant CMNs compared to benign CMNs, suggesting they may be useful as malignancy biomarkers.

Elucidating the Effects of the Lipids Regulators Fibrates and Statins on the Health Status of Finfish Species: A Review.

The most documented fibrates are gemfibrozil, clofibrate and bezafibrate, while for statins, the majority of the published literature focuses on atorvastatin and simvastatin. The present work reviews previously published research concerning the effects of these hypocholesterolaemic pharmaceuticals on fish, with a particular focus on commercially important species, commonly produced by the European aquaculture industry, specifically in recirculated aquaculture systems (RAS). Overall, results suggest that both acute and chronic exposures to lipid-lowering compounds may have adverse effects on fish, disrupting their capacity to excrete exogenous substances, as well as both lipid metabolism and homeostasis, causing severe ontogenetic and endocrinological abnormalities, leading to hampered reproductive success (e.g., gametogenesis, fecundity), and skeletal or muscular malformations, having serious repercussions on fish health and welfare. Nonetheless, the available literature focusing on the effects of statins or fibrates on commonly farmed fish is still limited, and further research is required to understand the implications of this matter on aquaculture production, global food security and, ultimately, human health.

Longitudinal association between subjective and objective memory in older adults: a study with the Virginia Cognitive Aging Project sample.

Using the bivariate dual change score approach, the present study investigated the directionality of the SMC-OMP association in a sample of healthy older adults (N = 2,057) from the Virginia Cognitive Aging Project. The sample was assessed throughout 10 years, five time points, and the impact of education, depressive symptoms, and low-memory functioning was tested. The Memory Functioning Questionnaire was used to assess SMC. There was a lack of longitudinal association with no significant coupling effects found between subjective and objective memory. After including depressive symptoms as a covariate, Frequency of Forgetting significantly predicted subsequent negative changes in OMP . A similar result was found for the low-memory functioning group after the inclusion of depression, with the frequency of memory complaints predicting subsequent memory decline . Our results do not support a predictive value of SMC without accounting for the influence of depressive symptoms and low-memory functioning.

PANCREATECTOMY AND HEPATECTOMY: A COMBINED SURGICAL RESECTION OF PSEUDOPAPILLARY SOLID TUMOR OF PANCREAS ASSOCIATED WITH HEPATIC METASTASIS.

BACKGROUND: Solid pseudopapillary tumor of the pancreas has been frequently reported in the past two decades. Surgery remains the treatment of choice, with the liver being the most frequent site of metastases. AIMS: The study aimed to present an option of surgical treatment for an 18-year-old female patient with a solid lesion in the body and tail of the pancreas associated with metastatic lesions in both hepatic lobes. METHODS: Two surgical procedures were scheduled. In the first procedure, body-caudal pancreatectomy with splenectomy was performed, associated with the resection of three lesions of the liver's left lobe. A right hepatectomy was performed 6 months later, progressing without complications. RESULTS: The patient continues without clinical complaints on the last return, and abdominal magnetic resonance performed 28 months after the second procedure does not show liver or abdominal cavity lesions. CONCLUSIONS: The knowledge on the biological behavior of tumors, evolution, and recurrence risks allows the indication of more rational surgical techniques that best benefit patients.

Bioevaluation of magnetic mesoporous silica rods: cytotoxicity, cell uptake and biodistribution in zebrafish and rodents.

Mesoporous silica nanoparticles (MSN) characterized by large surface area, pore volume, tunable chemistry, and biocompatibility have been widely studied in nanomedicine as imaging and therapeutic carriers. Most of these studies focused on spherical particles. In contrast, mesoporous silica rods (MSR) that are more challenging to prepare have been less investigated in terms of toxicity, cellular uptake, or biodistribution. Interestingly, previous studies showed that silica rods penetrate fibrous tissues or mucus layers more efficiently than their spherical counterparts. Recently, we reported the synthesis of MSR with distinct aspect ratios and validated their use in multiple imaging modalities by loading the pores with maghemite nanocrystals and functionalizing the silica surface with green and red fluorophores. Herein, based on an initial hypothesis of high liver accumulation of the MSR and a future vision that they could be used for early diagnosis or therapy in fibrotic liver diseases; the cytotoxicity and cellular uptake of MSR were assessed in zebrafish liver (ZFL) cells and the in vivo safety and biodistribution was investigated via fluorescence molecular imaging (FMI) and magnetic resonance imaging (MRI) employing zebrafish larvae and rodents. The selection of these animal models was prompted by the well-established fatty diet protocols inducing fibrotic liver in zebrafish or rodents that serve to investigate highly prevalent liver conditions such as non-alcoholic fatty liver disease (NAFLD). Our study demonstrated that magnetic MSR do not cause cytotoxicity in ZFL cells regardless of the rods' length and surface charge (for concentrations up to 50 µg ml-1, 6 h) and that MSR are taken up by the ZFL cells in large amounts despite their length of ∼1 µm. In zebrafish larvae, it was observed that they could be safely exposed to high MSR concentrations (up to 1 mg ml-1 for 96 h) and that the rods pass through the liver without causing toxicity. The high accumulation of MSR in rodents' livers at short post-injection times (20% of the administered dose) was confirmed by both FMI and MRI, highlighting the utility of the MSR for liver imaging by both techniques. Our results could open new avenues for the use of rod-shaped silica particles in the diagnosis of pathological liver conditions.

How modular protein nanoparticles may expand the ability of subunit anti-viral vaccines: The spring viremia carp virus (SVCV) case.

Spring viremia of carp (SVC) remains as a vaccine orphan disease mostly affecting juvenile specimens. Young fish are especially difficult to vaccinate and oral administration of vaccine combined with food would be the election system to minimise stress and the vaccination costs associated to injection. However, administration of prophylactics with food pellets faces off several drawbacks mainly related with vaccine degradation and weak protection correlates of oral vaccines. Here we present a platform based on recombinant proteins (subunit vaccines) manufactured as highly resistant nanostructured materials, and providing excellent levels of protection against SVC virus in a preliminary i.p injection challenge. The G3 domain of SVCV glycoprotein G was overexpressed in E. coli together with IFNγ and the modular protein was purified from bacterial aggregates (inclusion bodies) as highly organised nanostructured biomaterial (nanopellets, NP). These SVCV-IFNNP were taken up by zebrafish cells leading to the enhanced expression of different antiviral and IFN markers (e.g vig1, mx, lmp2 or ifngr1 among others) in zebrafish liver cells (ZFL). To monitor if SVCVNP and SVCV-IFNNP can be taken up by intestinal epithelia and can induce antiviral response we performed experiments with SVCVNP and SVCV-IFNNP in 3 days post fertilization (dpf) zebrafish larvae. Both, SVCVNP and SVCV-IFNNP were taken up and accumulated in the intestine without signs of toxicity. The antiviral response in larvae showed a different induction pattern: SVCV-IFNNP did not induce an antiviral response while SVCVNP showed a good antiviral induction. Interestingly ZF4, an embryonic derived cell line, showed an antiviral response like ZFL cells, although the lmp2 and ifngr1 (markers of the IFNγ response) were not overexpressed. Experiments with adult zebrafish indicated an excellent level of protection against a SVCV model infection where SVCV-IFNNP vaccinated fish reached 20% cumulative mortality while control fish reached over 80% cumulative mortality.

Gene expression profiles of beta-adrenergic receptors in canine vascular tumors: a preliminary study.

Beta adrenergic receptors (ß-AR) play a key role in regulating several hallmark pathways of both benign and malignant human and canine tumors. There is scarce information on the expression of ß-AR in canine vascular tumors. Therefore, the purpose of the present research work was to study the mRNA expression levels of the three subtypes of the ß-AR genes (ADRB1, ADRB2, ADRB3) in hemangiosarcoma (HSA) and hemangioma (HA), as well as in vascular hamartomas (VH) from dogs.Fifty samples (n = 50) were obtained from 38 dogs. Twenty-three animals had HSA, eight animals HA and seven animals VH. HSA were auricular (n = 8), splenic (n = 5), cutaneous (n = 6), auricular and splenic (n = 2), cutaneous-muscular (n = 1) and disseminated (n = 1). There were seven cases of HSA that were divided into primary tumor and secondary (metastatic) tumor. Skin and muscle samples with a normal histological study were used as control group. ADRB gene expression was determinate in all samples by real-time quantitative PCR. Results showed that ADRB1, ADRB2 and ADRB3 were overexpressed in HSA when compared to the control group. ADRB2 was overexpressed in HA when compared to the control group. HSA express higher values of ADBR1 (p = 0.0178) compared to VH. There was a high inter-individual variability in the expression of the three subtypes of ADBR. No statistically significant difference in the expression of ADBR genes were observed between HSA primary when compared to metastatic or in different anatomical locations. In conclusion, canine HSA overexpress the three ß-AR subtypes and canine HA ß2-AR. High variability was observed in ß-AR mRNA levels amongst HSA cases.

Magnetic Mesoporous Silica Nanorods Loaded with Ceria and Functionalized with Fluorophores for Multimodal Imaging.

Multifunctional magnetic nanocomposites based on mesoporous silica have a wide range of potential applications in catalysis, biomedicine, or sensing. Such particles combine responsiveness to external magnetic fields with other functionalities endowed by the agents loaded inside the pores or conjugated to the particle surface. Different applications might benefit from specific particle morphologies. In the case of biomedical applications, mesoporous silica nanospheres have been extensively studied while nanorods, with a more challenging preparation, have attracted much less attention despite the positive impact on the therapeutic performance shown by seminal studies. Here, we report on a sol-gel synthesis of mesoporous rodlike silica particles of two distinct lengths (1.4 and 0.9 µm) and aspect ratios (4.7 and 2.2) using Pluronic P123 as a structure-directing template and rendering ∼1 g of rods per batch. Iron oxide nanoparticles have been synthesized within the pores yielding maghemite (γ-Fe2O3) nanocrystals of elongated shape (∼7 nm × 5 nm) with a [110] preferential orientation along the rod axis and a superparamagnetic character. The performance of the rods as T2-weighted MRI contrast agents has also been confirmed. In a subsequent step, the mesoporous silica rods were loaded with a cerium compound and their surface was functionalized with fluorophores (fluorescamine and Cyanine5) emitting at λ = 525 and 730 nm, respectively, thus highlighting the possibility of multiple imaging modalities. The biocompatibility of the rods was evaluated in vitro in a zebrafish (Danio rerio) liver cell line (ZFL), with results showing that neither long nor short rods with magnetic particles caused cytotoxicity in ZFL cells for concentrations up to 50 µg/ml. We advocate that such nanocomposites can find applications in medical imaging and therapy, where the influence of shape on performance can be also assessed.

Pancreatectomy and hepatectomy: a combined surgical resection of pseudopapillary solid tumor of pancreas associated with hepatic metastasis / Pancreatectomy and hepatectomy: ressecção cirúrgica combinada de tumor pseudopapilar sólido do pâncreas com metástases hepáticas

ABSTRACT BACKGROUND: Solid pseudopapillary tumor of the pancreas has been frequently reported in the past two decades. Surgery remains the treatment of choice, with the liver being the most frequent site of metastases. AIMS: The study aimed to present an option of surgical treatment for an 18-year-old female patient with a solid lesion in the body and tail of the pancreas associated with metastatic lesions in both hepatic lobes. METHODS: Two surgical procedures were scheduled. In the first procedure, body-caudal pancreatectomy with splenectomy was performed, associated with the resection of three lesions of the liver's left lobe. A right hepatectomy was performed 6 months later, progressing without complications. RESULTS: The patient continues without clinical complaints on the last return, and abdominal magnetic resonance performed 28 months after the second procedure does not show liver or abdominal cavity lesions. CONCLUSIONS: The knowledge on the biological behavior of tumors, evolution, and recurrence risks allows the indication of more rational surgical techniques that best benefit patients.

RESUMO RACIONAL: O tumor sólido pseudopapilar do pâncreas tem sido relatado na literatura com mais frequência nas últimas duas décadas. A cirurgia continua sendo o tratamento indicado, sendo o fígado o local mais frequente de metástases. OBJETIVOS: Apresentar a opção de tratamento cirúrgico de paciente feminino, 18 anos, com lesão sólida no corpo e cauda do pâncreas associada a lesões metastáticas em ambos os lobos hepáticos. MÉTODOS: Dois procedimentos cirúrgicos foram agendados. No primeiro procedimento foi realizada pancreatectomia corpo-caudal com esplenectomia, associada à ressecção de 3 lesões do lobo esquerdo do fígado. A hepatectomia direita foi realizada seis meses após, evoluindo sem complicações. RESULTADOS: O paciente continua sem queixas clínicas no último retorno, e a ressonância magnética do abdomem realizada 28 meses após o segundo procedimento não mostra lesões hepáticas ou na cavidade abdominal. CONCLUSÕES: O conhecimento do comportamento biológico dos tumores, a evolução e os riscos de recidiva, permitem a indicação de técnicas cirúrgicas mais racionais e que melhor beneficiam os pacientes.

Depressive symptoms as a predictor of memory decline in older adults: A longitudinal study using the dual change score model.

The directionality of the longitudinal association between depressive symptoms and memory remains a topic of intense debate. A unidirectional association where depression impacts the change in memory (or vice-versa) and a bidirectional association where the trajectories of both dimensions affect each other lead to different clinical implications. METHOD: This study investigated the depression-memory longitudinal association in a sample of 2057 older adults aged between 60 and 99 years old from the Virginia Cognitive Aging Project. The bivariate dual change score model was used to investigate the directionality of the association between episodic memory and three dimensions of depression (somatic symptoms, depressed affect, and positive affect) throughout ten years (five measurement points), controlling for education and sex. RESULTS: the bidirectional model showed the best fit between somatic symptoms and memory, with a significant coupling effect observed from initial somatic symptoms to subsequent changes in memory. For depressed and positive affect, the unidirectional model with initial levels of depression predicting following changes in memory showed the best fit to the data, with significant coupling effects observed. Higher initial levels of somatic symptoms and depressed affect predicted a subsequent decline in memory, and higher initial levels of positive affect predicted subsequent better memory performance. Statistical adjustments for covariates (education and sex) had no significant influence on these associations. CONCLUSIONS: Our findings support a unidirectional association with higher depressive symptoms preceding a steeper decline in memory in older adults. We discuss the clinical implications for depressive symptoms as a predictor of subsequent memory decline.

Longitudinal impact and effects of booster sessions in a cognitive training program for healthy older adults.

This paper reports the results from a 3-year follow-up study to measure the long-term efficacy of a cognitive training for healthy older adults and investigates the effects of booster sessions using an entropy-based metric. DESIGN: semi-randomized quasi-experimental controlled design. PARTICIPANTS: 50 older adults, (M = 73.3, SD = 7.77) assigned into experimental (N = 25; Mean age = 73.9; SD = 8.62) and control groups (N = 25; mean age = 72.9; SD = 6.97). INSTRUMENTS: six subtests of WAIS and two episodic memory tasks. PROCEDURES: the participants were assessed on four occasions: after the end of the original intervention, pre-booster sessions (three years after the original intervention), immediately after the booster sessions and three months after the booster sessions. RESULTS: the repeated measures ANOVA showed that two of the cognitive gains reported in the original intervention were also identified in the follow-up: Coding (F(1, 44) = 11.79, MSE = 0.77, p = .001, eta squared = 0.084) and Picture Completion (F(1, 47) = 10.01, MSE = 0.73, p = .003, eta squared = 0.060). After the booster sessions, all variables presented a significant interaction between group and time favorable to the experimental group (moderate to high effect sizes). To compare the level of cohesion of the cognitive variables between the groups, an entropy-based metric was used. The experimental group presented a lower level of cohesion on three of the four measurement occasions, suggesting a differential impact of the intervention with immediate and short-term effects, but without long-term effects.

Educação problematizadora nas ações de integração ensino - serviço e promoção à saúde nos territórios / Problem-solving education in the teaching-service integration and health promotion actions in the territories / Educación problematizadoraen la integración del servicio docente y acciones de promoción de la salud en los territorios

Introdução:A disciplina de Saúde e Cidadania da Universidade Federal do Rio Grande do Norte preconiza, pelo Arco de Maguerez, a aprendizagem vivencial para oportunizar intervenções nos territóriosonde a disciplina ocorre.Objetivo:Relatar experiências deações de promoção à saúde realizadas em equipamentos sociais de um território adscrito a uma Unidade de Saúde da Família do Distrito Sanitário Oeste do Município de Natal, Rio Grande do NorteentreMaio a Junho de 2018.Metodologia:Estudo descritivo sobreintervenções realizadas e planejadas por dinâmicas da disciplina voltadas às demandas de sensibilizar crianças em idade pré-escolar sobre a importância da higiene; viabilizar recursos financeiros e alimentícios para uma associação beneficente; estimular o autoconhecimento e a autovalorização dos idosos para reduzir abusos intrafamiliares e depressão,e apreender o contexto familiar de crianças de 2 a 5 anos. Contemplou-se essas demandas, respectivamente, com a realizaçãode uma peça teatral e um bazar, ambos em Associação beneficente; exposição de filme e dinâmica "para quem você tira o chapéu?" na Associação de Idosos e realização de um Sandplay(jogo de areia) em um Centro Municipal de Educação Infantil.Resultados:O bazar arrecadoualimentos e produtos doados eo teatro de fantoches sensibilizou as crianças sobre o cuidado comhigiene bucal e corporal. A atividade com idosos enalteceu a importância da escuta, do incentivo às relações inter geracionais saudáveis para o crescimento mútuo dos envolvidos e o Sandplayapresentoudensa participação das crianças. Conclusões:As intervenções obtiveram resultado positivo, possibilitando sensibilização dosdiscentes para o cuidado à comunidade e estimulando habilidades de comunicação oral, corporal e empatia (AU).

Introduction:The discipline of Health and Citizenship of the Federal University of Rio Grande do Norte advocates, through the Arco de Maguerez, experiential learning to provide interventions in the territories where the discipline occurs. Objective:To report experiences of health promotion actions carried out in social facilities in a territory assigned to a Family Health Unit in the Western Sanitary District of the city of Natal, Rio Grande do Norte between May to June 2018. Methodology:Descriptive study on interventionscarried out and planned by discipline dynamics aimed at the demands of sensitizing children of preschool age about the importance of hygiene; make financial and food resources available to a charitableorganization; encourage self-knowledge and self-worth of the elderly to reduce intrafamily abuse and depression, and learn the family context of children aged 2 to 5 years. These demands were met, respectively, with the realization of a play and a bazaar, both in a charityorganization; film anddynamic exhibition "Who do you take your hat off to?" at the Elderly Association and performing a Sandplay (sand game) at a Municipal Center for Early Childhood Education. Results:The bazaar collected donated food and products and the puppet theater sensitized the children about the care with oral and body hygiene. The activity with the elderly praised the importance of listening, encouraging healthy intergenerational relationships for the mutualgrowth of those involved, and Sandplayshowed a dense participation by children. Conclusions:The interventions obtained a positive result, making students aware of the care for the community and stimulating oral, body and empathy communication skills (AU).

Introducción: La disciplina de Salud y Ciudadanía de la Universidad Federal de Río Grande del Norte aboga, a través del Arco de Maguerez, por el aprendizaje experimental para proporcionar intervenciones en los territorios donde ocurre la disciplina. Objetivo: Informar sobre experiencias de acciones de promoción de la salud realizadas en establecimientos sociales en un territorio asignado a una Unidad de Salud Familiar en el DistritoSanitarioOeste de Natal, Rio Grande do Norteentre mayo y junio de 2018. Metodología: Estudio descriptivo sobre las intervenciones realizadas y planificadas por la dinámica dirigidas a las demandas de sensibilizar a los niños en edad preescolar sobre la importancia de la higiene; poner recursos financieros y alimentarios a disposición de una asociación benéfica; Fomentar el autoconocimiento y la autoestima de las personas mayores para reducir el abuso y la depresión intrafamiliar, y aprender el contexto familiar de los niños de 2 a 5 años. Estas demandas se cumplieron, respectivamente, con la realización de una obra de teatro y un bazar, ambos en una asociación de caridad; película y exhibición dinámica "¿A quién le quitas el sombrero?" en la Asociación de Ancianos y realizando un Juego de arena en un Centro Municipal para la Educación de la Primera Infancia. Resultados: El bazar recolectó alimentos y productos donados y el teatro de marionetas sensibilizó a los niños sobre el cuidado con la higiene bucal y corporal. La actividad con los ancianos alabó la importancia de escuchar, alentando las relaciones intergeneracionales saludables para el crecimiento mutuo de los involucrados, y el Juego de arena mostró una densa participación de los niños. Conclusiones: Las intervenciones obtuvieron un resultado positivo, sensibilizando a los estudiantes sobre el cuidado de la comunidad y estimulando las habilidades de comunicación oral, corporal y de empatía (AU).

Effect of nanoplastics on fish health and performance: A review.

Small plastic particles are considered emerging pollutants, and this has motivated a considerable number of studies to establish their environmental consequences. At present, the study of the effects of nanoplastics (NPs) on aquatic organisms is still scarce, especially in organisms from higher trophic levels such as fish. This review describes the effects reported in different fish species after exposure to plastic particles smaller than 100 nm. Studies show that NPs can adversely affect fish at different stages of development, with reported accumulation in tissues, decreased locomotor and foraging activities, effects on growth and the immune system and alterations on lipid metabolism and neurotoxicity. However, mortality, effects on hatching success or malformations related to NPs have not been reported to this date.

Transport and Recovery of Gilthead Sea Bream (Sparus aurata L.) Sedated With Clove Oil and MS222: Effects on Oxidative Stress Status.

The use of anesthesia is a common practice in aquaculture to sedate fish and mitigate handling stress. Although the employ of anesthesia is considered beneficial for fish, as it reduces stress and improves welfare, at the same time it may induce hazardous side-effects. The aim of the present study was to investigate the effects of clove oil (CO) and tricaine methanesulfonate (MS222), two of the most used anesthetics, on several oxidative stress related parameters in gilthead sea bream (Sparus aurata), as these types of effects of anesthetics have been seldom investigated. To assess these effects, S. aurata juveniles were placed in a setup of mobile water tanks and were transported during 6 h with either 2.5 mg/L CO or 5 mg/L MS222. After transport, half of the fish were sampled, whereas the remaining fish were transferred to tanks without anesthetics where they were allowed to recover for 18 h before sampling. Changes in the expression levels of several target genes related with the antioxidant response and cell-tissue repair were evaluated in the gills, liver and brain. Those transcripts included glutathione peroxidase 1 (gpx1), catalase (cat), glutathione S-transferase 3 (gst3), glutathione reductase (gr), superoxide dismutase [Zn] (sod2), heat shock protein-70 (hsp70), and metallothionein (mt). Antioxidant enzymatic activities glutathione S-transferase, GST; catalase, CAT; and glutathione reductase, GR, levels of non-enzymatic antioxidants (non-protein thiols - NPT), and pro-oxidative damage, assessed as lipid peroxidation (LPO), were determined in gills, liver and brain. Acetylcholinesterase activity (AChE) was determined in plasma, gills, brain, muscle and heart as an indicator of neuro-muscular alterations. In plasma, the total antioxidant capacity (TAC) and total oxidative status (TOS) were also measured. Results showed that the use of both anesthetic agents, CO and MS222, interferes with fish antioxidant status. All tested biological matrices displayed alterations in antioxidant endpoints, confirming that these substances, although minimizing the effects of transport stress, may have long term effects on fish defenses. This result is of high relevance to aquaculture considering that the oxidative stress, may increase the susceptibility to different environmental or biotic stress and different types of pathologies.

Comparative study of stress and immune-related transcript outcomes triggered by Vibrio anguillarum bacterin and air exposure stress in liver and spleen of gilthead seabream (Sparus aurata), zebrafish (Danio rerio) and rainbow trout (Oncorhynchus mykiss).

The stress and immune-related effects of short-term (1, 6 and 24 h) air exposure stress (1 min), bath vaccination with Vibrio anguillarum bacterin, and both stressors combined were evaluated in liver and spleen of Sparus aurata, Danio rerio and Onchorhynchus mykiss. Expression profiles of immune (interleukin 1 beta: il1ß; tumor necrosis factor alpha: tnfα; interleukin 10: il10; tumor growth factor beta: tgfß1; immunoglobulin M: igm; lysozyme: lys; complement protein c3: c3) and stress-related genes (glucocorticoid receptor: gr; heat shock protein 70: hsp70; and enolase) were analysed by RT-qPCR. Cortisol level was assessed by radioimmunoassay. The gene expression patterns in liver and spleen were found to be differentially regulated in a time- and organ-dependent manner among species. In seabream, a higher il1ß-driven inflammatory response was recorded. In zebrafish, air exposure stress but not bath vaccination alone modulated most of the changes in liver and spleen immune transcripts. Stressed and vaccinated trout showed an intermediate pattern of gene expression, with a lower upregulation of immune-related genes in liver and the absence of changes in the expression of hsp70 and enolase in spleen (as it was observed in seabream but not in zebrafish). Following air exposure, cortisol levels increased in plasma 1 h post-stress (hps) and then decreased at 6 hps in O. mykiss and D. rerio. By contrast, in S.aurata the cortisol level remained higher at 6 hps suggesting a greater degree of responsiveness to this stressor. When fish were exposed to combined air exposure plus bath vaccination cortisol levels were also augmented at 1 and 6 hps in O. mykiss and S.aurata and restored to basal level at 24 hps, whereas in D. rerio the response was higher in response to the combination of both stressors. In addition, V. anguillarum bacterin vaccination triggered cortisol secretion only in D. rerio, suggesting a greater responsiveness of D. rerio hypothalamic-pituitary-interrenal axis. Overall, comparing the tissue transcription responsiveness, liver was found to be more implicated in the response to handling stress compared to spleen. These results also indicate that a species-specific response accounts for the deviations of stress and immune onset in the liver and spleen in these fish species.

Gold nanoparticles exposure modulates antioxidant and innate immune gene expression in the gills of Sparus aurata.

This study aimed to assess antioxidant and immune gene transcription alterations in the gills of Sparus aurata exposed during 96 h to 4, 80, and 1600 µg/L of gold nanoparticles (AuNPs) coated with citrate or polyvinyl pyrrolidone (PVP). After 96 h of exposure, gr and cat mRNA levels decreased for all tested concentrations of AuNPs, for both coatings. Instead, gst3 mRNA increased after exposure to 1600 µg/L AuNPs (both coatings) and prdx6 increased after exposure to 1600 µg/L AuNPs-citrate. Concerning immune genes, il1ß mRNA levels increased after exposure to 80 µg/L AuNPs-citrate and 1600 µg/L AuNPs-PVP and cox2 mRNA showed increased levels in fish exposed to 1600 µg/L AuNPs-citrate. Results indicate that AuNPs with distinct coatings induced different gene expression profiles in gills, though most of the studied genes remained unaltered for the tested conditions.

Comparison of traditional statistical quality control using commercially available control materials and two patient-based quality control procedures for the ADVIA 120 Hematology System.

BACKGROUND: Quality control procedures are an important part of the overall quality assurance for production of accurate and reliable hematologic results. OBJECTIVES: This study aimed to validate a quality control material-based procedure and assess two patient-based quality control procedures (repeat patient testing [RPT] and average of normals [AoN]) with the ADVIA 120 Hematology System. METHODS: Requirements for quality control procedures were obtained with the computerized statistical and quality program, EZRules3. The procedures were evaluated comparing the probability of error detection (Ped), probability of false rejection (Pfr), and sigma metrics. RESULTS: All three of the quality control procedures could be applied with 1-3s control rules, achieving the desired quality requirements. Validation of the quality control materials achieved values for Ped and Pfr of ≥90% and 0%, respectively. Patient-based procedures obtained a ≥85% Ped and a 0% Pfr, except for platelets in the AoN procedure, which achieved a 77% Ped. The RPT achievable total errors were similar to those of the traditional quality control materials and the AoN procedures, except for platelets, which had an achievable total error of 75%. CONCLUSIONS: Patient-based procedures are suitable for veterinary laboratories. The RPT approach may benefit laboratories with limited budgets and low hematology caseloads. The AoN procedure may benefit laboratories with higher hematology caseloads.

Cyclooxygenase-2 immunoexpression in intestinal epithelium and lamina propria of cats with inflammatory bowel disease and low grade alimentary lymphoma.

BACKGROUND: Cyclooxygenase 2 (COX-2) is an inducible isoform by cellular activation, proinflammatory cytokines and growth factors. The aims of the current study were to evaluate COX-2 immunoexpression in epithelial and lamina propria (LP) of cats with inflammatory bowel disease (IBD) and low grade alimentary lymphoma (LGAL), as well as to correlate them with clinical signs and histopathological scoring. Cats diagnosed with IBD and LGAL (2007-2013) were included in the current study. Feline chronic enteropathy activity index (FCEAI) was calculated for all cases. Control group was composed by 3 healthy indoor cats and 5 sick cats died or were euthanized (non-gastrointestinal illness). Diagnosis and classification of IBD and LGAL was established according to the WSAVA gastrointestinal standardization group template and the National Cancer Institute formulation, respectively. Furthermore, a modified WSAVA template was applied for LGAL evaluation. Immunolabelling for COX-2 (polyclonal rabbit anti-murine antibody) was performed on biopsy samples. Epithelial and LP (inflammatory or neoplastic cells) COX-2 immunolabelling was calculated according to the grade and intensity. The most representative segment scored by the WSAVA and the modified WSAVA were used for statistical analysis. RESULTS: Significant difference was found regarding COX-2 intensity overexpression in the epithelial cells of IBD and LGAL groups when compared to control cats, but not between the groups of sick cats, whereas no differences were found regarding the grade of immunoreactivity between groups. No difference was found for COX-2 immunoexpression at the LP between all groups. However, 3 cats from LGAL group showed COX-2 expression in neoplastic cells at the LP. There were no correlations between epithelial or LP COX-2 expression and FCEAI and histological alterations. CONCLUSIONS: Increased COX-2 intensity at the epithelial cells observed in cats with IBD and LGAL may be secondary to the inflammatory response or a protective function in the intestinal reparation. COX-2 expression at the LP was presented in 33% of LGAL. This result provides a reason for further investigation concerning the role of COX-2 expression in feline alimentary lymphoma.