New synthetic procedure for the antiviral sulfonate carbosilane dendrimer G2-S16 and its fluorescein-labelled derivative for biological studies

The anionic carbosilane (CBS) dendrimer with sulfonate groups G2-S16 is a promising compound for the preparation of a microbicide gel to prevent HIV infection. However, until now its synthesis required aggressive conditions. Hence, a reliable synthetic procedure is very important to face GMP conditions and clinical trials. In this study, G2-S16 has been prepared by a new approach that involves the addition of an amine-terminated dendrimer to ethenesulfonyl fluoride (C₂H₃SO₃F, ESF) and then transformation to the sulfonate dendrimer by treatment with a base. This strategy also makes feasible the synthesis of a labelled sulfonate dendrimer (G2-S16-FITC) to be used as a molecular probe for in vivo experiments. Interestingly, G2-S16-FITC enters into human peripheral blood mononuclear cells (PBMCs).

Ethenesulfonyl fluoride (ESF) is a useful reagent to prepare sulfonate carbosilane dendrimers with antiviral properties and labelled dendrimers for biological studies.     

Accreditation of processes in hepatology

The Spanish Association for the Study of the Liver decided in 2006 to develop a project to assess the quality of the professionals, processes and medical units dealing with the management of patients with liver diseases in Spain. The current article reports the criteria proposed to assess the quality and the accreditation of the processes in hepatology. The processes considered include most patients with liver diseases and the accreditation system designed is highly specific. This document, together with a previous one published in gastroenterología y hepatología concerning the accreditation of the professionals and a third document dealing with the accreditation of liver units that will be published soon, form the basis of the quality assessment of hepatology in our country.     

Bcl-2 and GDNF delivered by HSV-mediated gene transfer act additively to protect dopaminergic neurons from 6-OHDA-induced degeneration

Previous studies have demonstrated that either the neurotrophin glial-derived neurotrophic factor (GDNF) or the antiapoptotic peptide Bcl-2 delivered into striatum by a viral vector protects dopaminergic neurons of the substantia nigra in vivo from degeneration induced by the administration of the neurotoxin 6-hydroxydopamine (6-OHDA). In this study we used recombinant, replication-incompetent, genomic herpes simplex virus-based vectors to deliver the genes coding for Bcl-2 and GDNF into rat substantia nigra (SN) 1 week prior to 6-OHDA injection into the striatum. Vector-mediated expression of either Bcl-2 or GDNF alone each resulted in a doubling in cell survival as measured by retrograde labeling with fluorogold (FG) and a 50% increase in tyrosine hydroxylase-immunoreactive (TH-IR) neurons in the lesioned SN compared to the unlesioned side. Gene transfer of Bcl-2 and GDNF were equivalent in this effect. Coadministration of the Bcl-2-expressing vector with the GDNF-expressing vector improved the survival of lesioned SN neurons as measured by FG labeling by 33% and by the expression of TH-IR by 15%. These results suggest that the two factors delivered together act in an additive fashion to improve DA cell survival in the face of 6-OHDA toxicity.     

Validation of a quality of life questionnaire for critically ill patients

Objective Development and validation of quality of life questionnaire for critical care patients.Design Prospective study.Setting Intensive care unit (ICU) of a general hospital and ICUs of 83 Spanish hospitals.Sample Patients admitted to the ICU>18 years of age; close family members.Method A committee of experts designed a questionnaire with characteristics judged essential for intensive care use: easy, quick administration (5–10 min); capable of completion by patient or close family member, by direct or telephone interview. Fifteen items relevant to critical care patients were grouped in three subscales: basic physiological activities, normal daily activities, and emotional state. Reproducibility of interobserver, intraobserver, patient/family member and telephone/direct interviews was analysed and also internal consistency, responsiveness, and main components.Results Internal consistency (578 patients): Cronbach's alpha coefficient=0.85. Reproducibility: intraobserver reproducibility (n=150): Spearman correlation coefficient=0.92. Interobserver (n=85); correlation=0.92. Patient/family member (n=81): correlation=0.92. Telephone/direct interview (n=54): correlation=0.96. Validity: factorial analysis confirmed that the three subscales were fundamental questionnaire components. There was good concordance between questionnaire/subscale and Glasgow Outcome Scale (GOS) results. Responsiveness: quality of life score changes between preadmission and 6 months' postdischarge correlated with GOS findings (weighted kappa index=0.56).Conclusions Questionnaire meets objectives recommended for critical care use, and fulfills essential requirements of validity and reproducibility when applied to critically ill patients.This study forms part of the PAEEC (Project for the Epidemiological Analysis of Critical Care Patients), and was supported by a grant from the Fondo de Investigaciones Sanitarias (F.I.S.-91/0703), and by the Granada University Research Group (Number 3244)     

Hemodynamics changes in children and adolescents with mitral valve replacement]

The authors analyze the pre and post-operative hemodynamic picture of 30 children and adolescents with mitral valve replacement, which corresponds to a group of 106 patients operated on from 1964 to 1974. At variable time periods, between 9 days and 48 months (average 28 months) from the valve replacement, a new catheterization was done. The majority of the studies were done in the period between 13 and 36 months. Of the 30 cases, 21 were carriers of the Starr-Edwards valve, and the rest of different types. The Starr-Edwards valves produced a lawering of the CVP from 23.0 to 11.0 mm. Hg (average values) whereas with the other types the average pre-operative figure was 21.0 and in the post-operative period it fell to 11.0 mm. Hg. In the group with SE valves, pre-operative SPP was an average of 64.3 and in the post-operative period it was 32.0 mm. Hg. In the cases of SE valves the pre-operative total pulmonary resistances were 12.3 units and post-operatively they fell to 3.7 units. With the other types of valves the pre-opeative average was 10.9 units and post-operatively it was 5.3 units. The hemodynamic results obtained over a period of time with valvular substitution were similar in the distinct types of valves. However in the SE the benefits resulted more constant. The value of radiological and electrocardiographic study in order to predict the early post-operative hemodynamic changes is discussed. These studies serve essentially for a long range evaluation, whereas the hemodynamic, and clinical improvement maintained a strict relationship. This work shows that, in children and adolescents, the presence of severe venous-capillary and arterial pulmonary hypertension, and the elevated pulmonary resistences, are not contraindications for surgical treatment. It equally proves the eficacy of the valvular replacement, when the surgical indication and the control of the patients are handled in the proper way.     

Neurocognitive and clinical predictors of functional outcome in patients with schizophrenia and bipolar I disorder at one-year follow-up

OBJECTIVE: Many studies have reported that cognitive ability may be predictive of the functional outcome for patients with schizophrenia. However, no study has prospectively examined these aspects in schizophrenia and bipolar disorders simultaneously. The present study attempted to analyze if neurocognition and clinical status predicts the real-life functioning for patients with schizophrenia or bipolar I disorder, using a longitudinal design. METHOD: Forty-seven schizophrenic and 43 bipolar I outpatients were assessed twice with a neurocognitive battery (Executive Functions, Working Memory, Verbal Memory, Visual Memory, Visual-Motor Processing, Vigilance, Vocabulary and Motor Speed tasks), clinical scales (the Positive and Negative Symptom Scale, the Hamilton Rating Scale for Depression and the Clinician Administered Rating Scale for Mania) and functional outcome measures (the Global Assessment of Functioning Scale, the WHO's Disability Assessment Scale and occupational adaptation level) over a one-year follow-up period. The cognitive performance of the patients was compared, at baseline and one year later, with that of 25 healthy subjects. RESULTS: In schizophrenia patients, global functioning one year later was predicted by a composite neurocognitive score and three specific domain (verbal memory, motor speed, vocabulary). Symptoms appeared to explain less of the variance in functioning. In bipolar I patients, changes in the composite neurocognitive score over one year, deficits in the visual/motor processing domain, severity of symptoms (psychotic, excitatory and affective symptoms) and premorbid adjustment at the first assessment were the variables that better predicted functioning or disability changes over follow-up period. CONCLUSIONS: Although the relationships between cognition, symptoms and functional capacity differ for schizophrenia or bipolar I patients, neuropsychological performance seems to be a principal longitudinal predictor of functioning in both disorders. Baseline neurocognition and cognitive changes over 12 months predicted changes in functioning over the same period, but only in bipolar I patients. These cognitive domains could be potential neurocognitive endophenotypes (endophenocognitypes) with regard to bipolar I disorder.     

Characterization of Polydimethylsiloxane (PDMS) Properties for Biomedical Micro/Nanosystems

Polydimethylsiloxane (PDMS Sylgard 184, Dow Corning Corporation) pre-polymer was combined with increasing amounts of cross-linker (5.7, 10.0, 14.3, 21.4, and 42.9 wt.%) and designated PDMS1, PDMS2, PDMS3, PDMS4, and PDMS5, respectively. These materials were processed by spin coating and subjected to common micro-fabrication, micro-machining, and biomedical processes: chemical immersion, oxygen plasma treatment, sterilization, and exposure to tissue culture media. The PDMS formulations were analyzed by gravimetry, goniometry, tensile testing, nano-indentation, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and X-ray photoelectron spectroscopy (XPS). Spin coating of PDMS was formulation dependent with film thickness ranging from 308 microm on PDMS1 to 171 microm on PDMS5 at 200 revolutions per minute (rpm). Ultimate tensile stress (UTS) increased from 3.9 MPa (PDMS1) to 10.8 MPa (PDMS3), and then decreased down to 4.0 MPa (PDMS5). Autoclave sterilization (AS) increased the storage modulus (sigma) and UTS in all formulations, with the highest increase in UTS exhibited by PDMS5 (218%). PDMS surface hydrophilicity and micro-textures were generally unaffected when exposed to the different chemicals, except for micro-texture changes after immersion in potassium hydroxide and buffered hydrofluoric, nitric, sulfuric, and hydrofluoric acids; and minimal changes in contact angle after immersion in hexane, hydrochloric acid, photoresist developer, and toluene. Oxygen plasma treatment decreased the contact angle of PDMS2 from 109 degrees to 60 degrees. Exposure to tissue culture media resulted in increased PDMS surface element concentrations of nitrogen and oxygen.     

Effect of poly(lactic-co-glycolic acid) contact on maturation of murine bone marrow-derived dendritic cells

Understanding of biomaterial adjuvant effect and its mechanisms is essential for the effective design and selection of appropriate materials for specific applications. We have previously shown that poly(lactic-co-glycolic acid) (PLGA), one of the most commonly studied polymers in tissue engineering, supports an adjuvant effect as measured by enhanced immune response against a co-delivered model antigen, which was dependent on the form of the biomaterial. Furthermore, we have shown that PLGA induces the maturation of human peripheral blood mononuclear cell-derived dendritic cells (DCs) in vitro. In this study, the effect of PLGA contact on the maturation of murine bone marrow-derived DCs was investigated in part to explain the biomaterial adjuvant effect observed. Treatment of bone marrow-derived DCs from C57BL6 mice with PLGA microparticles or films lead to maturation of these cells as exemplified by increased expression of co-stimulatory molecules CD80 and CD86 and production of proinflammatory cytokines TNF-alpha and IL-6. These results suggest that PLGA contact induces maturation of murine DCs, supporting our observations with human DCs. With the techniques developed in this study and given the results, our future goal is to utilize transgenic murine models to delineate the mechanisms of biomaterial-induced DC maturation.     

Developmental distribution of plasma membrane Ca2+-ATPase isoforms in chick cerebellum.

The plasma membrane Ca(2+)-ATPase (PMCA) is highly expressed in the nervous system, but little information is available about its implication in neuronal development. We have analyzed the expression and localization of different isoforms of PMCA in membrane vesicles and sections of chick cerebellum from embryonic day 10 to hatching. We found that the relative amount of each PMCA isoform and their spatiotemporal distribution in the cerebellum are directly linked to precise cellular types during the cerebellar maturation, even in a non-neural tissue as choroid plexus. Purkinje cells contain the highest diversity of PMCA isoforms of the cerebellar cortex since the moment of its morphogenesis. From embryonic day 15, the PMCA2 was highly expressed in the whole Purkinje cell, while PMCAs 1 and 3 had a more restricted distribution in the soma and dendritic branches, and these distributions were evolving according with cell maturation. Other cellular types seem to contain a specific combination of isoforms, but with a well-defined distribution pattern at late moments of development. Thus, PMCAs 1 and 3 were located in the soma of molecular layer interneurons, and only the PMCA2 was observed in granule cells at hatching. Furthermore, PMCA isoforms are also expressed in cellular compartments characterized by a high amount of synapses, suggesting a key role of these proteins in synaptogenesis and in the maturation of neuronal electrophysiological properties.