Economic burden of skin and skin structure infections due to Gram-positive bacteria in patients on hospital at home-based outpatient parenteral antibiotic therapy (OPAT).

OBJECTIVE: To describe and quantify resource use and direct health costs associated with skin and skin structure infections (SSSIs) caused by Gram-positive bacteria in adults receiving outpatient parenteral antimicrobial therapy (OPAT), administered by Hospital at Home units (HaH) in Spain. METHODS: Observational, multicenter, retrospective study. We included patients of both sexes included in the HaH-based OPAT Registry during 2011 to 2017 who were hospitalized due to SSSIs caused by Gram-positive bacteria. Resource use included home visits (nurses and physician), emergency room visits, conventional hospitalization stay, HaH stay and antibiotic treatment. Costs were quantified by multiplying the natural units of the resources by the corresponding unit cost. All costs were updated to 2019 euros. RESULTS: We included 194 episodes in 189 patients from 24 Spanish hospitals. The most frequent main diagnoses were cellulitis (26.8%) and surgical wound infection (24.2%), and 94% of episodes resulted in clinical improvement or cure after treatment. The median HaH stay was 13 days (interquartile range [IR]:8-22.7), and the conventional hospitalization stay was 5 days (IR: 1-10.7). The mean total cost attributable to the complete infectious process was €7,326 (95% confidence interval: €6,316-€8,416). CONCLUSIONS: Our results suggest that OPAT administered by HaH is a safe and efficient alternative for the management of these infections and could lead to lower costs compared with hospital admission.

Development of an APC-targeted multivalent E2-based vaccine against Bovine Viral Diarrhea Virus types 1 and 2.

The aim of this study was to develop and test a multivalent subunit vaccine against Bovine Viral Diarrhea Virus (BVDV) based on the E2 virus glycoprotein belonging to genotypes 1a, 1b and 2a, immunopotentiated by targeting these antigens to antigen-presenting cells. The E2 antigens were expressed in insect cells by a baculovirus vector as fusion proteins with a single chain antibody, named APCH I, which recognizes the ß-chain of the MHC Class II antigen. The three chimeric proteins were evaluated for their immunogenicity in a guinea pig model as well as in colostrum-deprived calves. Once the immune response in experimentally vaccinated calves was evaluated, immunized animals were challenged with type 1b or type 2b BVDV in order to study the protection conferred by the experimental vaccine. The recombinant APCH I-tE21a-1b-2a vaccine was immunogenic both in guinea pigs and calves, inducing neutralizing antibodies. After BVDV type 1b and type 2 challenge of vaccinated calves in a proof of concept, the type 1b virus could not be isolated in any animal; meanwhile it was detected in all challenged non-vaccinated control animals. However, the type 2 BVDV was isolated to a lesser extent compared to unvaccinated animals challenged with type 2 BVDV. Clinical signs associated to BVDV, hyperthermia and leukopenia were reduced with respect to controls in all vaccinated calves. Given these results, this multivalent vaccine holds promise for a safe and effective tool to control BVDV in herds.

Polymorphic variants of GSTM1, GSTT1, and GSTP1 genes in childhood acute leukemias: A preliminary study in Argentina.

BACKGROUND AND AIM: Despite recent major advances in leukemia research, the etiopathogenesis of childhood leukemias remains far elusive. Individual predisposing factors, including polymorphisms in detoxification enzymes, have been implicated in the molecular pathogenesis and heterogeneity of the disease. Genetic polymorphisms of glutathione S-transferases (GSTs) that alter enzyme activity could be an additional factor that increases the risk of acute leukemia, but data are lacking in Argentina. We assessed the association of GST polymorphisms and the susceptibility to childhood leukemia in Argentina by conducting an exploratory case-control study and correlated patients' genotype to clinical and biological features. METHODS: Deletion polymorphisms in GSTM1 and GSTT1 genes and the single nucleotide polymorphism in GSTP1 c.313A>G (rs1695; p.105Ile>Val) were genotyped by PCR-RFLP in 36 patients and 133 healthy individuals. RESULTS: GSTM1-null genotype was associated with a lower risk of developing acute leukemia (P = 0.013; OR: 0.31; CI: 0.12-0.80), while GSTP1-GG variants displayed an increased risk (P = 0.01; OR: 3.9; CI: 1.85-8.2). However, no differences were found for GSTT1 gene. Conclusion These preliminary results, to be validated in a larger population from Argentina, suggest that the development of pediatric leukemia may be differentially influenced by polymorphic variants in GST genes.

Comparison of strategies for the production of FMDV empty capsids using the baculovirus vector system.

Recombinant FMDV empty capsids have been produced in insect cells and larvae using the baculovirus expression system, although protein yield and efficiency of capsid assembly have been highly variable. In this work, two strategies were compared for the expression of FMDV A/Arg/01 empty capsids: infection with a dual-promoter baculovirus vector coding for the capsid precursor (P12A) and the protease 3C under the control of the polyhedrin and p10 promoters, respectively (BacP12A-3C), or a single-promoter vector coding the P12A3C cassette (BacP12A3C). Expression levels and assembly into empty capsids were analyzed in insect cells and larvae. We observed that the use of the single-promoter vector allowed higher levels of expression both in insect cells and larvae. Recombinant capsid proteins produced by both vectors were recognized by monoclonal antibodies (mAbs) directed against conformational epitopes of FMDV A/Arg/01 and proved to self-assemble into empty capsids (75S) and pentamers (12S) when analyzed by sucrose gradient centrifugation.

Prospective double-blind clinical trial evaluating the effectiveness of Bromelain in the third molar extraction postoperative period.

OBJECTIVES: To evaluate the anti-inflammatory and analgesic effect of Bromelain (pineapple extract) administered orally in the postoperative after extraction of impacted lower molars. STUDY DESIGN: This is a prospective, placebo-controlled, unicentric, double-blind study; the sample size was 34 patients. The pre and postoperative outcomes, evaluated on the third (D3) and eighth day (D8), included inflamtion, pain and oral aperture, as well as the need for analgesics. One group received Bromelain 150mg per day for three days and 100mg on days 4 to 7. The other group received placebo in the same dosage. All outcomes werrecorded quantitatively and analyzed with the Mann-Whitney U test for independent samples. RESULTS: Although there were no statistically significant differences between the treatment groups, a trend towards less inflammation and improved oral aperture was observed in the group that received Bromelain, compared to the group that received placebo. This trend can be attributed completely to random reasons, since there is no statistical difference in the results. CONCLUSIONS: Further studies are necessary to analyze different administration patterns and doses of Bromelain for the use in the postoperative of impacted third molars.

Enantiospecific synthesis of heterocycles linked to purines: different apoptosis modulation of enantiomers in breast cancer cells.

The issue of chiral drug is now a major theme in the design, discovery and development of new drugs. It has been shown for many pharmaceuticals that only one enantiomer contains the desired activity, and the synthesis of such drug molecules in their optically pure form is becoming increasingly important. Mitsunobu reaction was carried out between (R)- and (S)-3,4-dihydro-2H-1,5-benzoxathiepin-3-ol and purines under microwave irradiation. A contraction into a six-membered ring takes place with concomitant inversion at the stereocentre with excellent enatiomeric excesses giving rise to the homochiral 9-(2,3-dihydro-1,4-benzoxathiin-3-ylmethyl)-9H-purines. The anti-tumour activity of all enantiomers is reported against the caspase-3-deficient MCF-7 and the wild type SKBR-3 human breast cancer cells. The most active homochiral compound displays an IC50 of 1.85 µM and induces inhibition of the translation initiation factor eIF2α. All homochiral compounds included in this study show different apoptotic effects between both enantiomers with levels up to 99%. We have analyzed caspase-mediated apoptotic pathways on enantiomers and racemates. We have found a homochiral derivative that activates the canonical intrinsic caspase-8/caspase-3 apoptotic pathway on the MCF-7 cells, and a racemic compound that induces caspase-2 activation. Moreover, we demonstrate the involvement of caspase activation during cell death induced by these compounds in SKBR-3 cells.

Características de actividad, iniciativa personal y pasividad en la construcción de proyectos laborales: un estudio en jóvenes escolarizados de la Ciudad Autónoma de Buenos Aires / Activity, personal initiative and passivity characteristics en work projects construction: a study in school attending young people in Ciudad Autónoma de Buenos Aires

La inserción laboral de los jóvenes escolarizados se ha modificado sustancialmente en los últimos años. Uno de los requerimientos que se les exige para ingresar y mantenerse en el trabajo es que tengan características personales de actividad. Se trata de un estudio cuantitativo en el que se trabajó con una muestra de quinto y sexto (n=399) de jóvenes de seis escuelas medias públicas de la Ciudad Autónoma de Buenos Aires. Se evaluaron las características de actividad e iniciativa con la escala Self-reported initiative y la característica de pasividad con la escala Passivity (Frese et al., 1997). Se han realizado análisis descriptivos y análisis de varianzas (ANOVA). Asimismo se ha estudiado mediante una regresión lineal múltiple qué variables explican mejor la iniciativa personal.Se han encontrado diferencias significativas en las características personales de iniciativa en la variable sexo a favor de las mujeres. Además, los alumnos de la especialidad técnica son más activos en relación a los alumnos de la comercial y los que tienen experiencia laboral respecto de los que no la poseen.

Características de actividad, iniciativa personal y pasividad en la construcción de proyectos laborales: un estudio en jóvenes escolarizados de la Ciudad Autónoma de Buenos Aires / Activity, personal initiative and passivity characteristics en work projects construction: a study in school attending young people in Ciudad Autónoma de Buenos Aires

La inserción laboral de los jóvenes escolarizados se ha modificado sustancialmente en los últimos años. Uno de los requerimientos que se les exige para ingresar y mantenerse en el trabajo es que tengan características personales de actividad. Se trata de un estudio cuantitativo en el que se trabajó con una muestra de quinto y sexto (n=399) de jóvenes de seis escuelas medias públicas de la Ciudad Autónoma de Buenos Aires. Se evaluaron las características de actividad e iniciativa con la escala Self-reported initiative y la característica de pasividad con la escala Passivity (Frese et al., 1997). Se han realizado análisis descriptivos y análisis de varianzas (ANOVA). Asimismo se ha estudiado mediante una regresión lineal múltiple qué variables explican mejor la iniciativa personal.Se han encontrado diferencias significativas en las características personales de iniciativa en la variable sexo a favor de las mujeres. Además, los alumnos de la especialidad técnica son más activos en relación a los alumnos de la comercial y los que tienen experiencia laboral respecto de los que no la poseen.(AU)

Two tautomeric polymorphs of 2,6-dichloropurine.

Two polymorphs of 2,6-dichloropurine, C(5)H(2)Cl(2)N(4), have been crystallized and identified as the 9H- and 7H-tautomers. Despite differences in the space group and number of symmetry-independent molecules, they exhibit similar hydrogen-bonding motifs. Both crystal structures are stabilized by intermolecular N-H···N interactions that link adjacent molecules into linear chains, and by some nonbonding contacts of the C-Cl···π type and by π-π stacking interactions, giving rise to a crossed two-dimensional herringbone packing motif. The main structural difference between the two polymorphs is the different role of the molecules in the π-π stacking interactions.

Lesión de nervios periféricos en anestesia obstétrica: descripción de cuatro casos / No disponible

El abordaje neuroaxial es de elección para el control del dolor en obstetricia, pero no está exento de riesgos. Las neuropatías periféricas posparto pueden producirse por diferentes condiciones médicas, quirúrgicas y anestésicas, por lo que debemos tenerlas presentes para diagnosticarlas y tratarlas precozmente y así evitar que se atribuyan, por defecto, a la técnica anestésica. Presentamos cuatro casos que se diagnosticaron en el periodo periparto, 2 meralgias parestésicas, 1 neuropatía del femoral y 1 lumbociatalgia, de las cuales solo la lumbociatalgia pudo ser atribuida parcialmente a la técnica anestésica. Tras confirmar su diagnóstico se instauró tratamiento conservador que resolvió ad integrum, dentro de los primeros siete días, todas las neuropatías salvo la lumbociatalgia. La baja incidencia de neuropatías periféricas tras técnicas neuroaxiales en obstetricia puede ser superior a la que se diagnostica. Resulta imprescindible una buena anamnesis preanestésica y conocer los diferentes mecanismos fisiopatológicos que pueden desencadenar neuropatías periféricas (AU)

A neuroaxial approach is of choice for the management of pain in obstetrics but is not exempt of risks. Postpartum peripheral neuropathy may occur because of various medical, surgical, and anesthetic conditions, hence we should have them in mind in order to recognize them and treat them early enough to prevent their deffault attribution to the anesthetic technique. We report four cases diagnosed during the peripartum – 2 paresthetic meralgias, 1 femoral neuropathy, and 1 lumbosciatalgia, of which only the lumbosciatalgia could be partly attributed to the anesthetic technique. Once the diagnosis was confirmed a conservative therapy was initiated that solved ad integrum all neuropathies, except for the lumbosciatalgia episode, within seven days. The low incidence of peripheral neuropathy following neuroaxial techniques in obstetrics may be higher than diagnosed. Adequate history taking before anesthesia is crucial, as is an understanding of the various pathophysiological mechanisms that may trigger peripheral neuropathy (AU)

Insect larvae biofactories as a platform for influenza vaccine production.

Increased production capacity is one of the most important priorities for seasonal and pandemic influenza vaccines. In the present study, we used a baculovirus-insect larvae system (considered small, living biofactories) to improve the production of recombinant influenza virus H1N1 hemagglutinin (HA). Insect larvae produced four-fold more HA protein than insect cells per biomass unit (1 g of fresh larvae weight). A single infected Trichoplusia ni larva produced up to 113 µg of soluble and easily purified recombinant HA, an amount similar to that produced by 1.2×10(8) Sf21 insect cells infected by the same baculovirus. The use of the KDEL endoplasmic reticulum retention signal fused to the HA protein further increased recombinant protein production. Larvae-derived HA was immunogenically functional in vaccinated mice, inducing the generation of hemagglutination inhibition antibodies and a protective immune response against a lethal challenge with a highly virulent virus. The productivity, scalability and cost efficiency of small, living biofactories based on insect larvae suggest a broad-based strategy for the production of recombinant subunit vaccines against seasonal or pandemic influenza as an alternative to fermentation technologies.

Elemental carbon exposure and lung function in school children from Mexico City.

Though exposure to air pollution has a detrimental effect on respiratory health, few studies have examined the association between elemental carbon exposure and lung function among schoolchildren. The aim of the present study was to present the association between short-term elemental carbon exposure and lung function in schoolchildren from Mexico City. 55 asthmatic and 40 non-asthmatic children were followed for an average of 22 weeks. A spirometry test was performed every 15 days during follow-up. Portable air samplers collected particulate matter onto Teflon filters. Gravimetric analysis was conducted and elemental carbon was quantified using transmission densitometry. The association between the main variables was analysed using linear mixed effects models. The mean ± sd of elemental carbon light absorption was 92.7 ± 54.7 Mm(-1). An increase of one interquartile range in the 24-h average of elemental carbon (100.93 Mm(-1)) was associated with a significant negative impact on forced expiratory volume in 1 s (FEV(1)) (-62.0 (95% CI -123.3- -1.2) mL) and forced expiratory flow at 25-75% of forced vital capacity (FVC) (FEF(25-75%)) (-111 (95% CI -228.3- -4.1) mL) among asthmatic children, equal to 3.3% and 5.5%, respectively; and on FEV(1) (-95.0 (95% CI -182.3- -8.5) mL) and FVC (-105.0 (95% CI -197.0- -13.7) mL) among non-asthmatic children. Exposure to elemental carbon resulted in an important negative effect on lung function in atopic schoolchildren, regardless of asthma status.

Novel substituted quinazolines for potent EGFR tyrosine kinase inhibitors.

The type I receptor tyrosine kinases (RTKs) are involved in various aspects of cell growth, survival, and differentiation. Among the known RTKs, the epidermal growth factor receptor (EGFR) and ErbB-2 (HER-2) are two widely studied proteins that are prototypic members of the ErbB family which also includes ErbB-3 (Her-3) and ErbB-4 (Her-4). Overexpression of ErbB-2 and EGFR has been associated with aggressive disease and poor patient prognosis in a range of human tumour types (e.g. breast, lung, ovarian, prostate, and squamous carcinoma of head and neck). Disruption of signal transduction of these kinases has been shown to have an antiproliferative effect. Various approaches have been developed to target the ErbB signalling pathways including monoclonal antibodies (trastuzumab/Herceptin™ and cetuximab/Erbitux™) directed against the receptor, and synthetic tyrosine kinase inhibitors (gefitinib/Iressa™ and erlotinib/Tarceva™). Since many tumours overexpress ErbB receptors, simultaneous targeting of multiple ErbB receptors therefore becomes a promising approach to cancer treatment. Lapatinib (Tykerb™), a potent dual EGFR/ErbB-2 inhibitor, was approved for the treatment of ErbB-2-positive breast cancer. Despite years of intensive research on EGFR inhibitors, there is a surprising dearth of chemically distinct small inhibitors with a high degree of selectivity. There is also a need for new scaffolds due to the recent finding of EGFR mutations which render the kinase resistant to gefinitib and erlotinib. The structures under study will be quinazolines with different substituents. The structure-activity relationships and biological evaluation of compounds published during the last four years will be reviewed herein.

DIVA diagnostic of Aujeszky's disease using an insect-derived virus glycoprotein E.

Commercial vaccines against Aujeszky's disease are mainly formulated using deleted versions of attenuated or inactivated Pseudorabies virus (PRV) particles lacking of the structural glycoprotein E (gE). Complementary diagnostic assays used to differentiate infected from vaccinated animals (DIVAs), are based on the detection of serum antibodies against gE. A recombinant version of the PRV gE protein was expressed in a baculovirus vector system in Trichoplusia ni insect larvae in order to obtain this diagnostic reagent for large scale diagnosis at reduced costs. A recombinant gE gene (gEr), lacking of signal peptide and transmembrane domains, was cloned into a modified baculovirus vector to allow glycosylation of the protein and its subsequent exportation to the extracellular space. Analysis by SDS-PAGE, Western-blotting and glycoprotein staining revealed that a glycosylated protein of the expected electrophoretic mobility was obtained in infected larvae. Time course experiments revealed that maximum expression levels were reached 72h post-infection using 10(4)pfu of the recombinant baculovirus (BACgEr) per inoculated larva. An indirect PRV gE-ELISA was developed using gEr as a coating antigen. A comparison between larvae-derived PRV gE-ELISA and two commercially available PRV diagnostic kits showed good correlation between assays and better sensitivity when testing certain sera pig samples using the gEr ELISA. More than 30,000 ELISA determinations could be performed from crude extracts obtained from a single larva infected with the recombinant baculovirus, indicating the feasibility of this strategy for inexpensive production of glycosylated antigens for PRV diagnosis.

Enhancement of the recollision rate in diffusion-influenced reactions in an inhomogeneous medium.

Brownian dynamics simulations were performed to determine the first collision and recollision rates of spherical reagent particles in a reaction medium made heterogeneous by the presence of randomly located inert spherical obstacles in a continuum solvent. The recollision rate vp (and hence the overall reactive collision rate when activation energy is high) was always enhanced by the presence of obstacles, the degree of enhancement increasing with the volume fraction occupied by obstacles (phi) and with decreasing reagent concentration phiR. Enhancement increased with obstacle size at high phiR, and fell with increasing obstacle size at low phiR. The vp-phiR data follow a power law, where the scaling factor betap decreased with decreasing obstacle size and increasing phi, and the prefactor kp initially increased with phi and then fell (except for large obstacles). The behavior of betap appears to be largely due to the obstacles reducing the probability that reagent particles escape from each other after collision, while the dominant factors responsible for the behavior of kp appear to be initially the effect of obstacles in enhancing effective local reagent concentration, and then (for small obstacles), their reduction of the reagent-particle coordination number. As the energy of activation falls, the reactive collision rate becomes less influenced by the reagent recollision rate and more influenced by the rate of first collision. For low-activation-energy reactions, the presence of obstacles depresses the reactive collision rate if reagent concentration is low or if the obstacles are small and their concentration high. The fall in the reactive collision rate with decreasing activation energy is steeper, the lower the reagent concentration and the smaller the obstacles.

QSAR-derived choline kinase inhibitors: how rational can antiproliferative drug design be?

This review presents an overview of Choline Kinase (ChoK) inhibitors with antiproliferative activity. The consideration of ChoK as a novel target for the development of new anticancer drugs is justified. The synthesis of several derivatives based on structural modifications of hemicholinium-3 (HC-3) is not accompanied by potentiation of the neurological toxicity of HC-3. The increment of both ChoK inhibitory and antiproliferative activities was successfully obtained by the two following changes: a) substitution of the oxazonium moiety of HC-3 by several aromatic heterocycles, and b) using the 1,2-ethylene(bisbenzyl) moiety instead of the 4,4'-biphenyl fragment. In an attempt to understand the ChoK inhibitory activity, a quantitative structure-activity relationship was developed. The QSAR equations have described the forces involved in quantitative terms. The electron characteristic of the substituent at position 4 of the heterocycle and the lipophilic character of the whole molecule were found to significantly affect the antitumour activity in compounds 17-95. Trispyridinium compounds 91-95 are more potent than the bispyridinium ones 87-89 as ChoK inhibitors. Nevertheless, 91-95 are less active than 87-89 as antiproliferative agents because the latter show better lipophilicities to cross the cytosolic membranes. Inhibition of the growth of human tumours in nude mice has been demonstrated: Antitumour activity of compound 64 against human HT-29 produced a decrease of up to 70% in the size of the tumour in nude mice. These results indicate that ChoK can be used as a general target for anticancer drug design against Ras-dependent tumourigenesis.

Characterization of a Mycobacterium intracellulare variant strain by molecular techniques.

This paper describes a Mycobacterium intracellulare variant strain causing an unusual infection. Several isolates obtained from an immunocompromised patient were identified as members of the Mycobacterium avium complex (MAC) by the commercial AccuProbe system and biochemical standard identification. Further molecular approaches were undertaken for a more accurate characterization of the bacteria. Up to seven different genomic sequences were analyzed, ranging from conserved mycobacterial genes such as 16S ribosomal DNA to MAC-specific genes such as mig (macrophage-induced gene). The results obtained identify the isolates as a variant of M. intracellulare, an example of the internal variability described for members of the MAC, particularly within that species. The application of other molecular approaches is recommended for more accurate identification of bacteria described as MAC members.