Primer registro de anisakis sp. (nematoda, anisakidae) l3 en la cavidad corporal deatlantoraja platana (chondrichthyes, rajidae)

En la presente nota se registra por primera vez el tercer estadio larval de Anisakis sp. infectando a Atlantoraja platana (Günther, 1880). Los hospederos fueron obtenidos en desembarques pesqueros de plantas de procesado de los puertos de San Antonio Oeste (40° 44' S 64° 57' O) y San Antonio Este (40° 49' S 64° 57' O), Provincia de Río Negro, Argentina. Las larvas fueron colectadas en la cavidad visceral de los peces, cerca del órgano epigonal, un tejido linfomieloide estrechamente asociado a las gónadas y exclusivo de los peces cartilaginosos. Hay evidencias documentadas que las altas concentraciones de urea en tejidos y fluídos corporales tornan inhabitable el medioambiente celómico para ser colonizado por helmintos. Los resultados expuestos en este trabajo constituyen el primer reporte de L de Anisakis sp. en la cavidad corporal 3 de un elasmobranquio, en particular A. platana, y demuestran la capacidad de este anisákido para sobrevivir en la masa visceral de estos hospederos.This communication is the first record of the presence of a third stage larva of Anisakis sp. infecting Atlantoraja platana. The hosts were collected from fishery landings at processing plants of San Antonio Oeste (40° 44' S 64° 57' O) and San Antonio Este ports (40° 49' S 64° 57' O), Rio Negro province, Argentina. They were found in the visceral cavity near the epigonal organ, a lymphomyeloid tissue closely associated with gonads and only in cartilaginous fish. The high concentrations of urea in the body fluid and tissues of elasmobranch hosts made an inhospitable environment to the colonization of helminthes. The results produced by this work constitute the first report of L of Anisakis sp. in the body cavity of an elasmobranch, in particular 3 A. platana, and show the capability of this anisakid to survive in the visceral mass of these hosts.Fil: Moya, Ana Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaFil: Galíndez, Elena Juana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaFil: Di Giacomo, Edgardo Ernesto. Universidad Nacional del Comahue; ArgentinaFil: Tanzola, Rubén Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; Argentin

Oscillating Magnet Array-Based Nanomagnetic Gene Transfection: A Valuable Tool for Molecular Neurobiology Studies

To develop treatments for neurodegenerative disorders, it is critical to understand the biology and function of neurons in both normal and diseased states. Molecular studies of neurons involve the delivery of small biomolecules into cultured neurons via transfection to study genetic variants. However, as cultured primary neurons are sensitive to temperature change, stress, and shifts in pH, these factors make biomolecule delivery difficult, particularly non-viral delivery. Herein we used oscillating nanomagnetic gene transfection to successfully transfect SH-SY5Y cells as well as primary hippocampal and cortical neurons on different days in vitro. This novel technique has been used to effectively deliver genetic material into various cell types, resulting in high transfection efficiency and viability. From these observations and other related studies, we suggest that oscillating nanomagnetic gene transfection is an effective method for gene delivery into hard-to-transfect neuronal cell types

Urinary tract infections due to multiresistant microorganisms in hospitalized renal transplant recipients

Introduction: There is currently an increase in urinary tract infections in kidney transplant recipients due to multidrug-resistant organisms (MRO), which have become a medical challenge.Objective: To describe the prevalence of urinary tract infection (UTI) due to RMO in hospitalized renal transplant patients (PTxR), their risk factors, treatment and evolution at 1 year.Material and methods: medical records and cultures of hospitalized PTxR infectious with OMR in the period between 1/1/2016 and 31/12/2017 were reviewed. Risk factors such as: gender, advanced age, prolonged presence of double J catheter, surgical complications and prolonged hospitalization and renal function at hospitalization, at discharge and at one year and the occurrence of rejections at one year were evaluated. Results: The presence of multidrug-resistant germs was found in 58 PTxR (31.18%) who presented 105 episodes of UTI, 36 had a single infection and 22 P had more than one. 55.17% (32) were male and the mean age was 50.52 ±14.24 years. Of the total number of patients, 43 (74.15%) had risk factors such as: late removal of the double J catheter in 8 (13.8%), surgical complications in 11 (18.9%), prolonged inter- nation in 12 (20.7%) and 18 (31.03%) were older than 60 years. Nine patients required dialysis, 4 of whom recovered renal function. Creatinine at hospitalization in patients who did not require dialysis was 1.8 (1.39 - 3.01) mg/dl; at discharge 1.5 (1.1 - 2.1) mg/dl (p=0.025) and at one year it was 1.5 (1.18 - 2.1) mg/dl with no significant difference with respect to that at discharge (p=0.089). In the annual follow-up 5 patients died and 5 lost the graft. The incidence of rejection was 15.51%. The germs rescued were 13 A. baumanii cpx. (ABA) (11.92%), E. coli (ECO) 24 (22.01%), Enterobacter spp. 4 (3.66%), Enterococ- cus spp. 3 (2.75%), Klebsiella spp. 58 (53.21%), Serratia spp. 5 (4.58%), Proteus spp. 1 (0.91%) and Pseudomonas aeruginosa (PAE)1 (0.91). Of the 105 episodes of UTI, 79 were treated with monotherapy: 57 with carbapenem (54.28%), 10 with Colistin (9.51%), 4 with Linezolid (3.8%), 4 with Piperacillin+Tazobactam (3.8%), 3 with Ciprofloxacin (2.85%) and 1 with Nitrofurantoin (0.95%). In 26 episodes combined therapies of Carbapenem were used in 21 cases, colis- tin in 14, amikacin in 13, fosfomycin in 2 and tigecycline in 1 and ciprofloxacin in another. Conclusion: ORM UTIs were frequent and similar to those described in other series. No differences were found in the evolution of renal function, in rejections, in mortality in ORM UTIs with or without associated risk factors, nor was there any influence of recurrent or recurrent UTIs. Further studies with a larger number of patients are needed to evaluate the prognosis and evolution of patients with these infections

PAM-2 decreases neuropathic pain in mice and modulates chemokine/cytokine production in human microglial cells

Background: People worldwide suffer from neuropathic pain, and indicated medications are often either not effective or induce tolerance and abuse. Therefore, there is an urgent need to identify additional therapeutic options to treat this form of pain. Nicotinic acetylcholine receptors (nAChRs), particularly a7-nAChRs, have been implicated in pain signaling. Therefore, this study was designed to investigate the extent to which the selective positive allosteric modulator (PAM) of a7 AChRs, PAM-2, modulates neuropathic pain. The working hypothesis, that PAM-2 inhibits inflammatory signaling and neuropathic pain, was tested using animal and cellular models.Methods: The anti-neuropathic pain activity of PAM-2 was assessed in two independent murine models of neuropathic pain. Briefly, neuropathic pain was induced in adult, male CD-1 mice (n=10/condition) via i.p. administration of either streptozotocin (STZ) or oxaliplatin (OXA). After 14 days, when neuropathic pain was present, mice were administrated with PAM-2 (1.0 or 3.0 mg/kg, p.o.) or vehicle. The pain threshold was subsequently determined by the cold plate test before and 15, 30, 45, and 60 min after treatment. In addition, C20 human microglial cells were exposed to interleukin (IL)-1B (20 ng/ml) or vehicle alone, and in combination with nicotine (3 uM), PAM-2 (1-100 uM), or nicotine + PAM-2 for 24 h. After 24 h, cytokine/chemokine levels in the culture media were measured by ELISA.Results: A single dose of PAM-2 (3.0 mg/kg) decreased both STZ- and OXA-induced neuropathic pain in mice. Repeated treatment with an inactive dose (1.0 mg/kg) of PAM-2 showed anti-pain activity in OXA-treated mice after 14, but not 7, days of treatment. Additionally, methyllycaconitine blocked the anti-pain effects elicited by PAM-2, supporting the view that a7 AChRs are instrumental in the anti-pain actions of PAM-2. Cellular experiments revealed that nicotine minimally inhibited IL-1B-induced IL-6 and interferon-gamma-induced chemotactic protein 10 expression in C20 human microglial cells, and that this inhibition was potentiated by PAM-2 (100 uM). However, we cannot rule out the possibility that PAM- 2 was cytotoxic in this cell culture model.Conclusions: These findings indicate that a7 AChRs are involved in neuropathic pain signaling and that a7-PAMs may potentially be used therapeutically. The extent to which these protective effects involve reduced neuroinflammation remains to be determined

Drv Concentrations And Viral Load In Csf In Patients On Drv/r 600/100 Or 800/100mg Once Daily Plus Two Nrti

Introduction Darunavir/r (DRV/r) is currently used at a dose of 800/100 mg once daily (OD) in a high proportion of patients. Pharmacokinetic data suggest that 600/100 OD may be effective, reducing toxicity and cost. However, drug concentrations in reservoirs such as cerebrospinal fluid (CSF) might not be adequate to inhibit viral replication. We aimed to evaluate concentrations of DRV and HIV‐1 viral load (VL) in CSF patients receiving DRV 600/100 mg OD. Methods DRV600 is an ongoing randomized open study comparing DRV/r 800/100 mg (DRV800) vs 600/100 mg (DRV600) OD plus TDF/FTC or ABC/3TC in 100 virologically suppressed patients (eudraCT 2011‐006272‐39). Here we present the results of a CSF sub‐study. A lumbar puncture (LP) was performed in participating patients after at least six months of inclusion in the study, 20–28 hours after a dose of DRV/r. VL (PCR, LOD 40 copies/mL) was determined in CSF and in plasma. DRV concentrations were quantified in CSF by liquid chromatography mass spectrometry (LC/MS/MS) and in plasma using high‐performance liquid chromatography (HPLC). Results Sixteen patients were included (eight in each arm). All DRV600 patients and four out of eight DRV800 patients received TDF/FTC, and the other four ABC/3TC. 75% were males, median (range) age was 48 (17–71) years, CD4 cell count 532 cells/mL (190–1,394). Median total time on DRV/r was 30 (11–57) months, and since the beginning of the study 8 (6–12) months in DRV800 and 10 (7–12) months in DRV600 patients. LP was performed a median of 26 (24–28) hours after the last DRV/r+TVD or KVX dose. In DRV600 patients the median DRV plasma levels were 1,674 (326–3,742) ng/mL, CSF levels 17.08 (5.79–30.19) ng/mL and DRV CSF:plasma ratio 0.0084 (0.0028–0.0388), while in the DRV800 arm, median DRV plasma levels were 1,707 (958–3,910) ng/mL, CSF levels 13.23 (3.47–32.98) ng/mL and DRV CSF:plasma ratio 0.0104 (0.0018–0.0262). All patients had VL<40 copies/mL in plasma and 14 patients VL<40 copies/mL in CSF. Two patients (1 in each arm, and taking TDF/FTC) had detectable VL in CSF (280 and 159 c/mL). These patients had the lowest CSF DRV concentrations (5.47 and 3.47 ng/mL), with plasma DRV concentrations of 802 and 958 ng/mL respectively. Conclusions CSF DRV concentrations and CSF VL were similar between patients receiving DRV/r 800/100 mg or 600/100 mg OD. Low CSF DRV concentrations might be associated with viral escape in CNS. This may be taken into account in patients receiving OD DRV/r. Larger studies should confirm these findings

In vivo role of different domains and of phosphorylation in the transcription factor Nkx2-1

Background: The transcription factor Nkx2-1 (also known as TTF-1, Titf1 or T/EBP) contains two apparently redundant activation domains and is post-translationally modified by phosphorylation. We have generated mouse mutant strains to assess the roles of the two activation domains and of phosphorylation in mouse development and differentiation. Results: Mouse strains expressing variants of the transcription factor Nkx2-1 deleted of either activation domain have been constructed. Phenotypic analysis shows for each mutant a distinct set of defects demonstrating that distinct portions of the protein endow diverse developmental functions of Nkx2-1. Furthermore, a mouse strain expressing a Nkx2-1 protein mutated in the phosphorylation sites shows a thyroid gland with deranged follicular organization and gene expression profile demonstrating the functional role of phosphorylation in Nkx2-1. Conclusions: The pleiotropic functions of Nkx2-1 are not all due to the protein as a whole since some of them can be assigned to separate domains of the protein or to specific post-translational modifications. These results have implication for the evolutionary role of mutations in transcription factors. © 2011 Silberschmidt et al; licensee BioMed Central Ltd

Treatment of Chronic Myelogenous Leukemia by Blocking Cytokine Alterations Found in Normal Stem and Progenitor Cells

SummaryLeukemic cells disrupt normal patterns of blood cell formation, but little is understood about the mechanism. We investigated whether leukemic cells alter functions of normal hematopoietic stem and progenitor cells. Exposure to chronic myelogenous leukemia (CML) caused normal mouse hematopoietic progenitor cells to divide more readily, altered their differentiation, and reduced their reconstitution and self-renewal potential. Interestingly, the normal bystander cells acquired gene expression patterns resembling their malignant counterparts. Therefore, much of the leukemia signature is mediated by extrinsic factors. Indeed, IL-6 was responsible for most of these changes. Compatible results were obtained when human CML were cultured with normal human hematopoietic progenitor cells. Furthermore, neutralization of IL-6 prevented these changes and treated the disease

In vitro quantitative imaging assay for phagocytosis of dad neuroblastoma cells by iPSC-macrophages

Microglia orchestrate neuroimmune responses in several neurodegenerative diseases, including Parkinson's disease and Alzheimer's disease. Microglia clear up dead and dying neurons through the process of efferocytosis, a specialized form of phagocytosis. The phagocytosis function can be disrupted by environmental or genetic risk factors that affect microglia. This paper presents a rapid and simple in vitro microscopy protocol for studying microglial efferocytosis in an induced pluripotent stem cell (iPSC) model of microglia, using a human neuroblastoma cell line (SH-SY5Y) labeled with a pH-sensitive dye for the phagocytic cargo. The procedure results in a high yield of dead neuroblastoma cells, which display surface phosphatidylserine, recognized as an "eat-me" signal by phagocytes. The 96-well plate assay is suitable for live-cell time-lapse imaging, or the plate can be successfully fixed prior to further processing and quantified by high-content microscopy. Fixed-cell high-content microscopy enables the assay to be scaled up for screening of small molecule inhibitors or assessing the phagocytic function of genetic variant iPSC lines. While this assay was developed to study phagocytosis of whole dead neuroblastoma cells by iPSC-macrophages, the assay can be easily adapted for other cargoes relevant to neurodegenerative diseases, such as synaptosomes and myelin, and other phagocytic cell types

Evaluation of expression and function of the H+/myo-inositol transporter HMIT;

BACKGROUND: The phosphoinositide (PIns) signalling pathway regulates a series of neuronal processes, such as neurotransmitter release, that are thought to be altered in mood disorders. Furthermore, mood-stabilising drugs have been shown to inhibit key enzymes that regulate PIns production and alter neuronal growth cone morphology in an inositol-reversible manner. Here, we describe analyses of expression and function of the recently identified H+/myo-inositol transporter (HMIT) investigated as a potential regulator of PIns signalling. RESULTS: We show that HMIT is primarily a neuronal transporter widely expressed in the rat and human brain, with particularly high levels in the hippocampus and cortex, as shown by immunohistochemistry. The transporter is localised at the Golgi apparatus in primary cultured neurones. No HMIT-mediated electrophysiological responses were detected in rat brain neurones or slices; in addition, inositol transport and homeostasis were unaffected in HMIT targeted null-mutant mice. CONCLUSION: Together, these data do not support a role for HMIT as a neuronal plasma membrane inositol transporter, as previously proposed. However, we observed that HMIT can transport inositol triphosphate, indicating unanticipated intracellular functions for this transporter that may be relevant to mood control

Caracterización del ovario de Jenynsia Lineata mediante técnicas histológicas

El género Jenynsia sp. (madrecitas) pertenece al orden Cyprinodontiformes, familia Anablepidae, se encuentra ampliamente distribuido en el territorio argentino. En las provincias de Córdoba y Buenos Aires se encuentran las poblaciones más numerosas. Son peces dulceacuícolas, vivíparos, que actualmente se utilizan como modelo experimental para estudios toxicológicos, de impacto ambiental y para el control de larvas de insectos vectores de enfermedades. El objetivo del presente trabajo fue describir la estructura histológica del ovario y el gonoducto de Jenynsia lineata. Las muestras fueron fijadas en formol bufferado al 10% y procesadas mediante la técnica histológica tradicional de inclusión en parafina. Se efectuaron cortes de 5-6 μm, que posteriormente se colorearon con H-E y tricrómico de Masson. En base a la observación microscópica y a las técnicas histológicas utilizadas se ha podido realizar una descripción detallada del ovario. En J. lineata, el ovario se caracterizó por ser un órgano hueco cuya luz central disminuyó cuando las hembras se encontraban en período de gestación, además durante este período, se observó una gran cantidad de células del sistema inmunológico. En la época reproductiva se observaron folículos ováricos en distintos estadios de maduración gonadal. Cada uno de ellos rodeados por un epitelio plano simple, que en el caso de los folículos embrionados se observó altamente vascularizado. La pared del gonoducto, en las regiones craneal, media y caudal, presentó un epitelio cilíndrico simple, conectivo laxo muy vascularizado, una capa longitudinal de musculatura lisa y peritoneo en la periferia. El análisis de la estructura ovárica y gonoductal y de las características embrionarias de J. lineata ofrece valiosos elementos morfológicos y fisiológicos.Fil: Di Cesare, Luca Sebastian Guido. Universidad Nacional de la Plata. Facultad de Ciencias Veterinarias. Laboratorio de Histología y Embriología Descriptiva, Experimental y Comparada (LHYEDEC); . Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Montes, Martin Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; ArgentinaFil: Garcia, Ignacio Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Limnología "Dr. Raúl A. Ringuelet". Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Limnología; ArgentinaFil: Barbeito, Claudio Gustavo. Universidad Nacional de la Plata. Facultad de Ciencias Veterinarias. Laboratorio de Histología y Embriología Descriptiva, Experimental y Comparada (LHYEDEC); . Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Plaul, Silvia Elena. Universidad Nacional de la Plata. Facultad de Ciencias Veterinarias. Laboratorio de Histología y Embriología Descriptiva, Experimental y Comparada (LHYEDEC); . Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaVI Simposio Argentino de IctiologíaBarilocheArgentinaUniversidad Nacional del ComahueConsejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Nort