Magnolia officinalis reduces the long-term effects of the status epilepticus induced by kainic acid in immature rats.

During critical periods of neurodevelopment, the immature brain is susceptible to neuronal hyperexcitability, alterations such as hyperthermia, hypoxia, brain trauma or a preexisting neuroinflammatory condition can trigger, promote and prolong epileptiform activity and facilitate the development of epilepsy. The goal of the present study was to evaluate the long-term neuroprotective effects Magnolia officinalis extract, on a model of recurrent status epilepticus (SE) in immature rats. Sprague-Dawley rats were treated with kainic acid (KA) (3 mg/kg, dissolved in saline solution) beginning at day 10 P N every 24 h for five days (10 P N-14PN). Two experimental groups (KA) received two treatments for 10 days (14-24 P N): one group was treated with 300 mg/kg Magnolia Officinalis (MO) (KA-MO), and another was treated with 20 mg/kg of celecoxib (Clbx) (KA-Clbx) as a control drug. A SHAM control group at day 90 P N was established. Seizure susceptibility was analyzed through an after-discharge threshold (ADT) evaluation, and electroencephalographic activity was recorded. The results obtained from the ADT evaluation and the analysis of the electroencephalographic activity under basal conditions showed that the MO and Clbx treatments protected against epileptiform activity, and decreases long-term excitability. All rats in the KA-MO and KA-Clbx groups presented a phase I seizure on the Racine scale, corresponding to the shaking of a wet dog. In contrast, the KA group showed phase V convulsive activity on the Racine scale. Similarly, MO and Clbx exerted neuroprotective effects on hippocampal neurons and reduced gliosis in the same areas. Based on these results, early intervention with MO and Clbx treatments to prevent the inflammatory activity derived from SE in early phases of neurodevelopment exerts neuroprotective effects on epileptogenesis in adult stages.

[Impact of Lean methodology to improve care processes and levels of satisfaction in patient care in a clinical laboratory]. / Impacto de la metodología Lean en la mejora de procesos asistenciales y niveles de satisfacción en la atención de pacientes en un laboratorio clínico.

INTRODUCTION: The application of the Lean methodology in health institutions is an effective tool to improve the capacity and workflow, as well as to increase the level of satisfaction of patients and employees. OBJECTIVE: To optimise the time of outpatient care in a clinical laboratory, by implementing a methodology based on the organisation of operational procedures to improve user satisfaction and reduce the number of complaints for delays in care. MATERIAL AND METHODS: A quasi-experimental before and after study was conducted between October 2011 to September 2012. XBar and S charts were used to observe the mean service times and standard deviation. The user satisfaction was assessed using service questionnaires. RESULTS: A reduction of 17 minutes was observed in the time of patient care from arrival to leaving the laboratory, and a decrease of 60% in complaints of delay in care. Despite the high staff turnover and 38% increase in the number of patients seen, a culture of empowerment and continuous improvement was acquired, as well as greater efficiency and productivity in the care process, which was reflected by maintaining standards 12 months after implementation. CONCLUSION: Lean is a viable methodology for clinical laboratory procedures, improving their efficiency and effectiveness.

Enhanced CD8+ T cell immune response against a V3 loop multi-epitope polypeptide (TAB13) of HIV-1 Env after priming with purified fusion protein and booster with modified vaccinia virus Ankara (MVA-TAB) recombinant: a comparison of humoral and cellular immune responses with the vaccinia virus Western Reserve (WR) vector.

The humoral and cytotoxic T-lymphocyte (CTL) responses have been shown to be determinant in the clearance of many viral infections and because of those characteristics, vaccine candidates against AIDS are designed to enhance both arms of the immune system. While a protocol of immunization able to confer protection in humans against HIV will have to await the results of current clinical trials, it remains important to identify protocols of immunization in animals that achieve significant levels of humoral and cellular immune responses to HIV. In this study we have carried out a comparative analysis of the immune responses elicited in mice immunized with recombinants based on the modified vaccinia virus Ankara strain (rMVA) versus the Western Reserve strain (WR) of vaccinia virus (rVV), both expressing a V3 loop multi-epitopic protein from eight different HIV isolates (TAB13). We found that during priming, rMVA elicited a two- to three-fold higher specific CD8+ T cell response than rVV. Similar enhancement was observed during priming with purified protein TAB13 followed by a booster with rMVA. The epitopes LR150, MN and IIIB, located at the ends and in the middle of the chimeric protein, were able to induce a specific CD8+ T cell response, both after priming or prime/booster with the recombinant viruses but not after prime/booster with TAB13. By examining the cytokine pattern, the immune response triggered by these vectors was of Th-1 type. Humoral immune responses were higher in animals immunized with TAB13/TAB13 or TAB13/rVV than in animals immunized with TAB13/rMVA. These findings demonstrate that during priming or in a prime/booster immunizations, rMVA is superior to rVV in the ability to enhance specific cellular responses to an HIV-1 protein, and that both humoral and cellular immune responses to theV3 loop epitope of HIV-1 Env can be obtained by priming with TAB13 followed by a booster with viral vectors.

Factors affecting biodegradation of 2-chlorophenol by Alcaligenes sp. in aerobic reactors.

The influence of variations in carbon source concentration, cell inocula, pH, presence of other substrates, and other organisms on the biodegradation of 2-chlorophenol (2-CP) was studied for Alcaligenes sp. isolated from natural sources. Assays of biodegradation were performed in batch and continuous-flow fluidized-bed aerobic reactors. Evaluation of biodegradation was performed by determining total phenols, chemical oxygen demand (COD), and 2-CP by ultraviolet (UV) spectrophotometry. Measurement of microbial growth was carried out by the plate count method. Bioassays of acute toxicity were performed to evaluate detoxification by using Daphnia magna. Results obtained show that under batch conditions with initial inocula of 10(6) cells/mL the strain grew exponentially with 100, 200, and 300 mg/L of 2-CP within 48 hr. A lag period was observed with low cell density inocula (10(5) cells/mL). The strain showed marked delay in the biodegradation of 2-CP at pH 5. Removal of target substrate from mixtures containing other carbon sources demonstrated the possibility of concurrent growth. Mineralization of 2-CP was assessed by gas chromatography carried out at the end of the batch assays and at the exit of the continuous-flow reactor. The presence of other organisms (bacteria, rotifers, ciliate, and algae) that developed in the fluidized-bed reactor did not affect the efficacy of the biodegradation of 2-CP. The removal of 2-CP in the two assayed systems was over 97% in all cases. Toxicity was not detected at the exit of the continuous reactor.

Recombinant proteins produced by vaccinia virus vectors can be incorporated within the virion (IMV form) into different compartments.

Vaccinia virus (VV) is one of the largest and most complex of animal viruses, with a virion that contains about 100 different polypeptides. Assembly of the viral proteins occurs in discrete cytoplasmic sites leading to formation of two infectious forms, an abundant (>90%) intracellular mature virus (IMV) with an envelope, and a minor extracellular enveloped virus (EEV) with an extra membrane acquired from the trans-Golgi network. It has been shown that while EEV contains in the outer membrane cellular proteins probably acquired during virus release from cells, however, IMV exclude host proteins during assembly. Since VV recombinants (VVr) expressing genes of interest are candidates as potential vaccines against pathogens and cancer, it becomes important to know if VVr can acquire foreign proteins during morphogenesis. In this investigation we show that purified virions (IMVs) from VVr can incorporate foreign proteins into different sites in the virus particle. By sequential fractionation of virion compartments with detergents, we found foreign proteins in the lipid envelope (cytokine IL-12 and CS antigen of Plasmodium yoelii), as part of a protein matrix-like membrane (HIV-1 gp41 of env), or more closely associated with the core containing the DNA complexes (HIV-1 gp160; a multiepitopic protein with the V3 loop of HIV-1 Env from different isolates, and beta-galactosidase). Similar findings were observed with purified virions derived from the WR strain as well as from the highly attenuated modified vaccinia virus Ankara (MVA) strain. These observations should be taken into consideration when VVr are used in clinical trials or in other vaccination approaches.

Toxicity tests to assess pollutants removal during wastewater treatment and the quality of receiving waters in Argentina.

In Argentina, legislation to control adverse impacts of effluent discharges and the quality of receiving waters is scant and relies mainly on the physicochemical characteristics of the effluents and receiving waters. Objectives of this study were to use standardized acute toxicity tests to assess treatment of petrochemical industry effluents and the toxicity of various treated industrial effluents in the Buenos Aires metropolitan area and their receiving waters. Tests for the first objective used Daphnia magna and Ceriodaphnia dubia; those for the second used D. magna, Spirillum volutans, and Scenedesmus spinosus. Chemical analyses demonstrated that the removal of aromatic hydrocarbon compounds (benzene, toluene, ethylbenzene, xylene, styrene, and naphthalene) from the petrochemical effluents ranged between 77 and 93%, but toxicity removal was significantly lower: untreated effluents were very toxic and treated effluents were very toxic to toxic [acute toxicity units (TUa) > 3]. Physicochemical parameters measured according to current Argentinian regulations indicated that industrial effluents (e.g., from textile and paper industries) were within established guidelines, but 25% of the samples were moderately to highly toxic (TUa > 1.33). However, for the receiving waters, toxicity tests were moderate to very toxic. The results show the need of including tests for toxicity of discharged effluents, and their effects on receiving waters of Argentina, especially for regulatory purposes.

Isolates from four different HIV type 1 clades circulating in Cuba identified by DNA sequence of the C2-V3 region.

According to the epidemiology of human immunodeficiency virus infection in Cuba, the main sources of infection have been persons coming from foreign countries, mainly from Africa, and individuals who have had sexual contacts with foreigners in Cuba. However, the first Cuban HIV-1 isolates sequenced have been all classified as subtype B. In this note we report the sequence of the gp120 C2/V3 region from 11 HIV-1 isolates from Cuban patients. DNA was isolated either directly from blood PBMC or from primary isolates, PCR amplified and sequenced. Six isolates were classified as subtype B and three of them had the atypical sequences GRGR, GWGR, and TPGR on the tip of the V3 loop. Besides, two other sequences were classified as subtype A, two as subtype H, and one as subtype C. These results confirm that although subtype B seems to be predominant, HIV-1 isolates from various subtypes do circulate in Cuba.

A prime-boost regime that combines Montanide ISA720 and Alhydrogel to induce antibodies against the HIV-1 derived multiepitope polypeptide TAB9.

A phase I clinical trial with the HIV-1-derived multi-epitope polypeptide (MEP) TAB9 in the oil adjuvant Montanide ISA720 (M-ISA720) was recently performed. Although severe local reactions were reported after the second and third injections of this vaccine candidate, the first inoculation was well tolerated. In this article we evaluated a prime-boost regime consisting of one inoculation of TAB9 in M-ISA720 followed by a booster with the same antigen in aluminum hydroxide. This combination of adjuvants elicited similar antibody levels in rabbits than the traditional two-dose schedule with M-ISA720. A control group injected three times with TAB9 in aluminum hydroxide developed markedly lower antibody titers. These results showed that although oil adjuvants are better than alum for priming the immune system for antibody production against TAB9, both kinds of adjuvants can be equally effective in booster immunizations. Therefore, by using the more reactogenic oil adjuvant only for priming, we should be able to eliminate the undesirable reactions characteristic of these compounds while achieving equivalent levels of specific antibodies.

The V3 loop based multi-epitope polypeptide TAB9 adjuvated with montanide ISA720 is highly immunogenic in nonhuman primates and induces neutralizing antibodies against five HIV-1 isolates.

In a previous work we selected montanide ISA720 (M-ISA720) among different adjuvants for the vaccination with a V3 loop based multi-epitope polypeptide TAB9. In this paper we present the evaluation of the toxicity and immunogenicity of this formulation in non-human primates. TAB9 in M-ISA720 was highly immunogenic in macaques (Macaca fascicularis) inducing antibodies against TAB9 in all animals after one inoculation and a strong anamnestic response after booster injections. Furthermore 97% of the V3 peptides included were recognized by TAB9 sera. No differences between doses of 200 microg and 1 mg of TAB9 in M-ISA720 were observed after four immunizations. Neutralizing antibodies against five HIV-1 isolates were detected in most animals. Animals remain healthy throughout the study and did not show lesions at the inoculation site.

An immunoassay with bovine serum albumin coupled peptides for the improved detection of anti V3 antibodies in HIV-1 positive human sera.

The V3 loop of the gp120 envelope glycoprotein contains the principal neutralizing determinant of HIV-1. Many serological studies have been carried out to assess the reactivity of HIV-1 infected individuals against V3 loop synthetic peptides from different HIV-1 subtypes. V3 directed serology has also been used to demonstrate the association between ELISA reactivity and progression to AIDS in HIV patients, and to study the reactivity against the V3 region in sera from vaccinated animals and human volunteers. The advantage of the use of bovine serum albumin (BSA) conjugated V3 peptides over free V3 peptides for ELISA is described. 15 meric V3 peptides representing several HIV-1 isolates were synthesized, chemically coupled to BSA, and used to coat ELISA microplates. Conjugated peptides were compared with free peptides for the detection of anti V3 antibodies in the sera from rabbits immunized with V3 containing chimeric proteins and from HIV-1 infected individuals. No differences in reactivity against free or BSA-peptide were found for most rabbit sera, however human plasma recognized preferentially the BSA conjugated peptides. Although technically more complex, ELISA with BSA coupled V3 peptides is more sensitive and appropriate for serological studies of HIV-1 infected persons.

Biodegradación de 2-clorofenol en reactores aeróbicos empleando microorganismos autóctonos

Se describen los resultados obtenidos en ensayos de biodegradación de 2-CP, empleando reactores "batch" y de lecho fluidizado (LF), operados aerobiamente y con microorganismos autóctonos especificamente seleccionados y optimizados

Biodegradación de 2-clorofenol en reactores aeróbicos empleando microorganismos autóctonos

Se describen los resultados obtenidos en ensayos de biodegradación de 2-CP, empleando reactores "batch" y de lecho fluidizado (LF), operados aerobiamente y con microorganismos autóctonos especificamente seleccionados y optimizados

Biodegradación de 2-clorofenol en reactores aeróbicos empleando microorganismos autóctonos

Los objetivos del presente trabajo fueron: Seleccionar e identificar microorganismos capaces de biodegradar 2-clorofenol en distintos procesos aeróbios; Determinar la cinética y la velocidad de degradación a escala laboratorio empleando reactores batch; Establecer la efectividad de un reactor de lecho fluidizado aeróbico relleno con carbón activado granular para remover 2-CP como única fuente de carbono, suministrando como nutrientes un fertilizante foliar agrícola de uso comercial; Evaluar la remoción de toxicidad a través de bioensayos

Non-extraction therapy in maxillary deficiency cases.

Combined orthodontic and surgical management of maxillary deficiency with a Class III profile has accomplished efficient treatment and predictable results. Also, orthodontic/orthopedic therapy can give good results; but, in any case, if one of either or a combined therapy is selected, the orthodontist has to be sure of the correct diagnosis. The cephalometric analyses have to be done with care as it is possible to find contradictions among different analyses, especially in the A/P aspect. The Wits appraisal shows good capacity for determining the antero-posterior maxillomandibular relationship because it is sensitive to the osseous dimensional variants and lower anterior facial height. Nevertheless, it is a supplementary evaluation requiring additional data for the localization of the deformity.

[Mothers' attitudes and behaviors regarding vaccination in the outpatient clinic of Hospital Robert Reid Cabral]. / Actitudes, practicas de las madres sobre la vacunacion en la consulta externa del Hospital Robert Reid Cabral.

PIP: Concern over relatively low rates of vaccination in the Dominican Republic prompted a study of the attitudes and practices of mothers attending outpatient clinics at a children's hospital in Santo Domingo. 200 mothers were prospectively interviewed in late 1991. 57% of the children were incompletely vaccinated and 7.5% were completely unvaccinated. 98.5% of the mothers stated their belief that vaccination protects children. 125 of the mothers reported they had access to orientation about vaccination. 80% of the children whose mothers had received orientation were completely vaccinated, compared to only 18.7% of children whose mothers had not received orientation. 9% of the study mothers had children who died of diseases preventable by vaccination, primarily measles. Only 59% of the children of sample mothers had been vaccinated against measles.^ieng

Stimulus-response latencies of two instruments delivering transcutaneous electrical neuromuscular stimulation (TENS).

Stimulus-response latencies of low-frequency transcutaneous electrical neuromuscular stimulation (TENS) were studied in 15 healthy subjects, applying the two different pulse configurations delivered by the Biotens and the Myomonitor instruments. Latencies, in milliseconds, were determined on bipolar raw surface electromyograms (EMG) of the suprahyoid muscles, using the skin surface over the sigmoid notches of the mandible as the site of stimulation. Stimulus-response times were measured from the onset of the stimulus artefact to the first response peak on EMG, and their mean values showed ranges of 3.79-4.49 ms for Biotens and 5.10-5.34 ms for Myomonitor. It was concluded that low-frequency TENS caused direct stimulation of motor nerves, and that the timing of the contraction response was not affected by altered electrode placement, lead-wire reversal or unbalanced (right/left) stimulation.