RESUMO
Like other ciliates, Philasterides dicentrarchi, the scuticociliate parasite of turbot, produces a feeding-only or growing stage called a trophont during its life cycle. Exposure of the trophonts to heat-inactivated serum extracted from the turbot host and containing specific antibodies that induce agglutination/immobilization leads to the production of a mucoid capsule from which the trophonts later emerge. We investigated how these capsules are generated, observing that the mechanism was associated with the process of exocytosis involved in the release of a matrix material from the extrusomes. The extruded material contains mucin-like glycoproteins that were deposited on the surface of the cell and whose expression increased with time of exposure to the heat-inactivated immune serum, at both protein expression and gene expression levels. Stimulation of the trophonts with the immune serum also caused an increase in discharge of the intracellular storage compartments of calcium necessary for the exocytosis processes in the extrusomes. The results obtained suggest that P. dicentrarchi uses the extrusion mechanism to generate a physical barrier protecting the ciliate from attack by soluble factors of the host immune system. Data on the proteins involved and the potential development of molecules that interfere with this exocytic process could contribute to improving the prevention and control of scuticociliatosis in turbot.
Assuntos
Infecções por Cilióforos/veterinária , Doenças dos Peixes/imunologia , Linguados , Imunidade Humoral/imunologia , Oligoimenóforos/fisiologia , Animais , Infecções por Cilióforos/imunologia , Infecções por Cilióforos/parasitologia , Doenças dos Peixes/parasitologiaRESUMO
Scuticociliatosis is a severe disease in farmed flatfish. However, the causative agent is not always accurately identified. In this study, we identified two isolates of scuticociliates from an outbreak in cultured fine flounder Paralichthys adspersus. Scuticociliate identification was based on morphological data, examination of life stages and the use of molecular approaches. The isolates were compared with a strain of Philasterides dicentrachi from turbot Scophthalmus maximus and with a strain deposited in the American Type Culture Collection as Miamiensis avidus ATCC® 50180™. The use of morphological, biological and molecular methods enabled us to identify the isolates from the fine flouder as P. dicentrarchi. Comparison of P. dicentrachi isolates and M. avidus revealed some differences in the buccal apparatus. Unlike P. dicentrarchi, M. avidus has a life cycle with three forms: macrostomes (capable of feeding on P. dicentrarchi), microstomes and tomites. Additionally, we found differences in the 18S rRNA and α- and ß-tubulin gene sequences, indicating that P. dicentrarchi and M. avidus are different species. We therefore reject the synonymy/conspecificity of the two taxa previously suggested. Finally, we suggest that a combination of morphological, biological, molecular (by multigene analysis) and serological techniques could improve the identification of scuticociliates parasites in fish.
RESUMO
The scuticociliatosis is a very serious disease that affects the cultured turbot, and whose causal agent is the anphizoic and marine euryhaline ciliate Philasterides dicentrarchi. Several protozoans possess acidic organelles that contain high concentrations of pyrophosphate (PPi), Ca(2+) and other elements with essential roles in vesicular trafficking, pH homeostasis and osmoregulation. P. dicentrarchi possesses a pyrophosphatase (H(+)-PPase) that pumps H(+) through the membranes of vacuolar and alveolar sacs. These compartments share common features with the acidocalcisomes described in other parasitic protozoa (e.g. acid content and Ca(2+) storage). We evaluated the effects of Ca(2+) and ATP on H (+)-PPase activity in this ciliate and analyzed their role in maintaining intracellular pH homeostasis and osmoregulation, by the addition of PPi and inorganic molecules that affect osmolarity. Addition of PPi led to acidification of the intracellular compartments, while the addition of ATP, CaCl2 and bisphosphonates analogous of PPi and Ca(2+) metabolism regulators led to alkalinization and a decrease in H(+)-PPase expression in trophozoites. Addition of NaCl led to proton release, intracellular Ca(2+) accumulation and downregulation of H(+)-PPase expression. We conclude that the regulation of the acidification of intracellular compartments may be essential for maintaining the intracellular pH homeostasis necessary for survival of ciliates and their adaptation to salt stress, which they will presumably face during the endoparasitic phase, in which the salinity levels are lower than in their natural environment.
Assuntos
Infecções por Cilióforos/veterinária , Doenças dos Peixes/parasitologia , Linguados/parasitologia , Pirofosfatase Inorgânica/metabolismo , Oligoimenóforos/enzimologia , Trifosfato de Adenosina/fisiologia , Animais , Western Blotting , Cálcio/fisiologia , Infecções por Cilióforos/metabolismo , Infecções por Cilióforos/parasitologia , Doenças dos Peixes/enzimologia , Doenças dos Peixes/metabolismo , Imunofluorescência , Concentração de Íons de Hidrogênio , Pirofosfatase Inorgânica/genética , Camundongos , Camundongos Endogâmicos ICR , Microscopia Eletrônica de Transmissão , Oligoimenóforos/fisiologia , Oligoimenóforos/ultraestrutura , Concentração Osmolar , Reação em Cadeia da Polimerase em Tempo Real , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/enzimologia , Saccharomyces cerevisiae/genética , SalinidadeRESUMO
Blood cultures are the gold standard for detecting bacteremia. We have studied the time to positivity of blood cultures in our neonatal unit to reduce antibiotic doses in patients with a negative blood culture. Empirical antibiotic treatment of neonatal sepsis could be withdrawn 24 hours after obtaining blood cultures.
Assuntos
Bacteriemia , Sepse Neonatal , Recém-Nascido , Humanos , Hemocultura , Bacteriemia/diagnóstico , Bacteriemia/tratamento farmacológico , Sepse Neonatal/diagnóstico , Sepse Neonatal/tratamento farmacológico , Antibacterianos/uso terapêuticoRESUMO
Philasterides dicentrarchi is a free-living microaerophilic scuticociliate that can become a facultative parasite and cause a serious parasitic disease in farmed fish. Both the free-living and parasitic forms of this scuticociliate are exposed to oxidative stress associated with environmental factors and the host immune system. The reactive oxygen species (ROS) generated by the host are neutralized by the ciliate by means of antioxidant defences. In this study we aimed to identify metalloenzymes with superoxide dismutase (SOD) activity capable of inactivating the superoxide anion (â¢O2-) generated during induction of oxidative stress. P. dicentrarchi possesses the three characteristic types of SOD isoenzymes in eukaryotes: copper/zinc-SOD, manganese-SOD and iron-SOD. The Cu/Zn-SOD isoenzymes comprise three types of homodimeric proteins (CSD1-3) of molecular weight (MW) 34-44 kDa and with very different AA sequences. All Cu/Zn-SODs are sensitive to NaCN, located in the cytosol and in the alveolar sacs, and one of them (CSD2) is extracellular. Mn- and Fe-SOD transcripts encode homodimeric proteins (MSD and FSD, respectively) in their native state: a) MSD (MW 50 kDa) is insensitive to H2O2 and NaN3 and is located in the mitochondria; and b) FSD (MW 60 kDa) is sensitive to H2O2, NaN3 and the polyphenol trans-resveratrol and is located extracellularly. Expression of SOD isoenzymes increases when â¢O2- is induced by ultraviolet (UV) irradiation, and the increase is proportional to the dose of energy applied, indicating that these enzymes are actively involved in cellular protection against oxidative stress.
Assuntos
Mitocôndrias/metabolismo , Oligoimenóforos/enzimologia , Oligoimenóforos/metabolismo , Superóxido Dismutase/metabolismo , Superóxidos/química , Sequência de Aminoácidos/genética , Animais , Doenças dos Peixes/parasitologia , Linguados/parasitologia , Peróxido de Hidrogênio/metabolismo , Peróxido de Hidrogênio/toxicidade , Camundongos , Camundongos Endogâmicos ICR , Oligoimenóforos/genética , Estresse Oxidativo/fisiologia , Resveratrol/toxicidade , Azida Sódica/toxicidade , Superóxido Dismutase/genética , Superóxido Dismutase/isolamento & purificação , Raios UltravioletaRESUMO
BACKGROUND: We aim to delineate the progression of cerebellar syndrome in children with phosphomannomutase-deficiency (PMM2-CDG) using the International Cooperative Ataxia Rating Scale (ICARS). We sought correlation between cerebellar volumetry and clinical situation. We prospectively evaluated PMM2-CDG patients aged from 5 to 18 years through ICARS at two different time points set apart by at least 20 months. We reviewed available MRIs and performed volumetric analysis when it was possible. RESULTS: From the eligible 24, four patients were excluded due to severe mental disability (n = 2) and supratentorial lesions (n = 2). Two different ICARS evaluations separated by more than 20 months were available for 14 patients showing an improvement in the cerebellar syndrome: ICARS1: 35.71 versus ICARS2: 30.07 (p < 0.001). When we considered time, we saw an improvement of 2.64 points in the ICARS per year with an SD of 1.97 points (p < 0.001). The ICARS subscales results improved with time, reaching statistical significance in "Posture and gait" (p < 0.001), "Kinetic functions" (p = 0.04) and "Speech abnormalities" (p = 0.045). We found a negative correlation between the ICARS results and total cerebellar volume (r = -0.9, p = 0.037) in a group of five patients with available volumetric study, meaning that the higher the ICARS score, the more severe was the cerebellar atrophy. CONCLUSIONS: Our study shows a stabilization or mild improvement in the cerebellar functions of paediatric PMM2-CDG patients despite cerebellar volume loss. ICARS is a valid scale to quantify the evolution of cerebellar syndrome in PMM2-CDG patients. The availability of ICARS and other reliable and sensitive follow-up tools may prove essential for the evaluation of potential therapies.
Assuntos
Doenças Cerebelares/diagnóstico , Doenças Cerebelares/enzimologia , Fosfotransferases (Fosfomutases)/deficiência , Adolescente , Doenças Cerebelares/patologia , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Neuroimagem , Fosfotransferases (Fosfomutases)/genéticaRESUMO
OBJECTIVE: To report the clinical features, comorbidities, receptor subunit targets, and outcome in patients with anti-GABAA receptor (GABAAR) encephalitis. METHODS: Clinical study of 26 patients, including 17 new (April 2013-January 2016) and 9 previously reported patients. Antibodies to α1, ß3, and γ2 subunits of the GABAAR were determined using reported techniques. RESULTS: Patients' median age was 40.5 years (interquartile range 48.5 [13.75-62.35] years; the youngest 2.5 months old; 13 female). Symptoms included seizures (88%), alteration of cognition (67%), behavior (46%), consciousness (42%), or abnormal movements (35%). Comorbidities were identified in 11 (42%) patients, including 7 tumors (mostly thymomas), 2 herpesvirus encephalitis (herpes simplex virus 1, human herpesvirus 6; coexisting with NMDAR antibodies), and 2 myasthenia without thymoma. Brain MRI was abnormal in 23 (88%) patients, showing in 20 (77%) multifocal, asynchronous, cortical-subcortical T2/fluid-attenuated inversion recovery abnormalities predominantly involving temporal (95%) and frontal (65%) lobes, but also basal ganglia and other regions. Immunologic or tumor therapy resulted in substantial improvement in 18/21 (86%) assessable patients; the other 3 (14%) died (2 status epilepticus, 1 sepsis). Compared with adults, children were more likely to have generalized seizures (p = 0.007) and movement disorders (p = 0.01) and less likely to have a tumor (p = 0.01). The main epitope targets were in the α1/ß3 subunits of the GABAAR. CONCLUSIONS: Anti-GABAAR encephalitis is characterized by frequent seizures and distinctive multifocal cortical-subcortical MRI abnormalities that provide an important clue to the diagnosis. The frequency of symptoms and comorbidities differ between children (more viral-related) and adults (more tumor-related). The disorder is severe but most patients respond to treatment.
Assuntos
Anticorpos/sangue , Anticorpos/líquido cefalorraquidiano , Encefalite/imunologia , Receptores de GABA-A/imunologia , Adolescente , Adulto , Animais , Encéfalo/metabolismo , Transtornos Cognitivos/etiologia , Estado de Consciência , Encefalite/metabolismo , Encefalite/terapia , Feminino , Células HEK293 , Humanos , Imunoterapia , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Transtornos dos Movimentos/etiologia , Ratos , Receptores de GABA-A/genética , Receptores de GABA-A/metabolismo , Estudos Retrospectivos , Convulsões/etiologia , Transfecção , Adulto JovemRESUMO
BACKGROUND: Phosphomannomutase deficiency (PMM2-CDG) is the most frequent congenital disorder of glycosylation. The cerebellum is nearly always affected in PMM2-CDG patients, a cerebellar atrophy progression is observed, and cerebellar dysfunction is their main daily functional limitation. Different therapeutic agents are under development, and clinical evaluation of drug candidates will require a standardized score of cerebellar dysfunction. We aim to assess the validity of the International Cooperative Ataxia Rating Scale (ICARS) in children and adolescents with genetically confirmed PMM2-CDG deficiency. We compare ICARS results with the Nijmegen Pediatric CDG Rating Scale (NPCRS), neuroimaging, intelligence quotient (IQ) and molecular data. METHODS: Our observational study included 13 PMM2-CDG patients and 21 control subjects. Ethical permissions and informed consents were obtained. Three independent child neurologists rated PMM2-CDG patients and control subjects using the ICARS. A single clinician administered the NPCRS. All patients underwent brain MRI, and the relative diameter of the midsagittal vermis was measured. Psychometric evaluations were available in six patients. The Mann-Whitney U test was used to compare ICARS between patients and controls. To evaluate inter-observer agreement in patients' ICARS ratings, intraclass correlation coefficients (ICC) were calculated. ICARS internal consistency was evaluated using Cronbach's alpha. Spearman's rank correlation coefficient test was used to correlate ICARS with NPCRS, midsagittal vermis relative diameter and IQ. RESULTS: ICARS and ICARS subscores differed between patients and controls (p < 0.001). Interobserver agreement of ICARS was "almost perfect" (ICC = 0.99), with a "good" internal reliability (Cronbach's alpha = 0.72). ICARS was significantly correlated with the total NPCRS score (rs 0.90, p < 0.001). However, there was no agreement regarding categories of severity. Regarding neuroimaging, inverse correlations between ICARS and midsagittal vermis relative diameter (rs -0.85, p = 0.003) and IQ (rs -0.94, p = 0.005) were found. Patients bearing p.E93A, p.C241S or p.R162W mutations presented a milder phenotype. CONCLUSIONS: ICARS is a reliable instrument for assessment of PMM2-CDG patients, without significant inter-rater variability. Despite our limited sample size, the results show a good correlation between functional cerebellar assessment, IQ and neuroimaging. For the first a correlation between ICARS, neuroimaging and IQ in PMM2-CDG patients has been demonstrated.
Assuntos
Ataxia Cerebelar/diagnóstico , Cerebelo/patologia , Defeitos Congênitos da Glicosilação/diagnóstico , Fosfotransferases (Fosfomutases)/deficiência , Índice de Gravidade de Doença , Adolescente , Ataxia Cerebelar/enzimologia , Ataxia Cerebelar/genética , Criança , Pré-Escolar , Defeitos Congênitos da Glicosilação/enzimologia , Defeitos Congênitos da Glicosilação/genética , Feminino , Humanos , Masculino , Fosfotransferases (Fosfomutases)/genéticaRESUMO
OBJECTIVE: In this study, we assessed the efficacy of 2 pharmacodynamically different antidepressants, citalopram (a selective serotonin reuptake inhibitor) and reboxetine (a norepinephrine reuptake inhibitor), as adjunctive therapy to risperidone and olanzapine for the treatment of negative symptoms in schizophrenia. METHOD: We performed a 6-month, multicenter, double-blind, randomized, placebo-controlled clinical trial. The recruitment period was from November 2008 to December 2011.The sample comprised 90 patients with a diagnosis of schizophrenia (DSM-IV criteria) who exhibited negative symptoms. The patients were recruited from 10 centers in different cities of the Spanish State. The primary efficacy measure was change in score on the negative subscale of the Positive and Negative Syndrome Scale (PANSS) between baseline and 6-month assessment. Other efficacy measures were changes in the PANSS subscales and total score, as well as the Scale for the Assessment of Negative Symptoms (SANS) subscales and total score. RESULTS: For statistical analysis, we employed mixed-effects models. We did not find statistically significant differences between the placebo group and the 2 treatment groups at 6-month assessments for the PANSS total (P=.6511), any PANSS subscale (negative [P=.5533], positive [P=.1723], or general psychopathology [P=.2083]), or the SANS (P= .5884). Cohen d measure showed a small effect size below the 0.5 threshold for all comparisons. CONCLUSIONS: In conclusion, our results do not support adjunctive use of citalopram or reboxetine with risperidone or olanzapine for the treatment of negative symptoms in schizophrenia. TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT01300364.
Assuntos
Antidepressivos/uso terapêutico , Antipsicóticos/uso terapêutico , Benzodiazepinas/uso terapêutico , Citalopram/uso terapêutico , Depressão/tratamento farmacológico , Morfolinas/uso terapêutico , Risperidona/uso terapêutico , Esquizofrenia/tratamento farmacológico , Psicologia do Esquizofrênico , Adulto , Antipsicóticos/efeitos adversos , Benzodiazepinas/efeitos adversos , Citalopram/efeitos adversos , Depressão/diagnóstico , Método Duplo-Cego , Quimioterapia Combinada , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Morfolinas/efeitos adversos , Olanzapina , Escalas de Graduação Psiquiátrica , Reboxetina , Risperidona/efeitos adversos , Esquizofrenia/diagnósticoRESUMO
Se analiza el comportamiento de los costos de un legrado uterino y la actividad asistencial de la sala, en el cuatrienio 1983-1986. El estudio se realizó en el Servicio de Ginecología del Hospital Ginecoobstétrico Docente Provincial "Mariana Grajales" de Santa Clara. En él se proponen una serie de medidas administrativas con el objetivo de reducir los costos de un legrado. Se registró un costo de $24,33 en 1983 y una reducción en los dos últimos años analizados de $18,39 y $17,27
Assuntos
Aborto Induzido , Custos e Análise de CustoRESUMO
Se proponen indicadores de gastos por pacientes ginecológicos de algunos productos relacionados como materiales de curaciones, termómetros, jeringuillas y antibióticos. El estudio se realizó en el servicio de ginecología del hospital ginecoobstétrico provincial docente "Mariana Grajales" de Santa Clara, en 1982. Se plantean medidas organizativas y económicas en el ahorro mediante un plan de los productos señalados, se hace un análisis de los costos hospitalarios que contemplan estos epígrafes y la utilidad de las medidas y normas de gastos propuestos. Los objetivos fundamentales de este trabajo son contribuir al conocimiento de las demandas que se operan en el orden económico y perfeccionar el sistema de dirección y planificación de la economía. Se formulan conclusiones
Assuntos
Economia Hospitalar , Unidade Hospitalar de Ginecologia e Obstetrícia/organização & administração , Custos e Análise de Custo , Administração de Materiais no HospitalRESUMO
Se propone una serie de medidas administrativas para la reducción de los costos hospitalarios en el epigrafe de materiales de curaciones y medicamentos. El estudio realizado en el Servicio de Ginecología del Hospital Ginecoobstétrico de Santa Clara contempla la introducción de normas unitarias por pacientes, de productos seleccionados de importación cuyo control sistemático logre disminuir los gastos por estos conceptos