Immobilization of a recombinant endo-1,5-arabinanase secreted by Aspergillus nidulans strain A773

An endo-1,5-arabinanase (abnA) encoding gene from Aspergillus niveus was identified, cloned and successfully expressed in Aspergillus nidulans strain A773. Based on amino acid sequence comparison, the 34-kDa enzyme could be assigned to CAZy GH family 43. Characterization of purified recombinant endo-1,5-arabinanase (AbnA) revealed that it is active at a wide pH range (pH 4.0-7.0) and an optimum temperature at 70 degrees C. The immobilization of the AbnA was performed via covalent binding onto agarose-modified supports: glyoxyl iminodiacetic acid-Ni2+, glyoxyl amine, glyoxyl (4% and 10%) and cyanogen bromide activated sepharose. The yield of immobilization was similar on glyoxyl amine and glyoxyl (96%), and higher than glyoxyl iminodiacetic acid-Ni2+ (43%) support. The thermal inactivation of these immobilized preparations showed that the stability of the AbnA immobilized on glyoxyl 4 and 10% was improved by 4.0 and 10.3-fold factor at 70 degrees C. The half-life of glyoxyl 4% derivative at 60 degrees C was >48 h (pH 5), 9 h (pH 7) and 88 min (pH 9). The major hydrolysis product of debranched arabinan or arabinopentaose by glyoxyl agarose-immobilized AbnA was arabinobiose. (C) 2012 Elsevier B.V. All rights reserved.

Dichotomous effects of VEGF-A on adipose tissue dysfunction

Obese fat pads are frequently undervascularized and hypoxic, leading to increased fibrosis, inflammation, and ultimately insulin resistance. We hypothesized that VEGF-A-induced stimulation of angiogenesis enables sustained and sufficient oxygen and nutrient exchange during fat mass expansion, thereby improving adipose tissue function. Using a doxycycline (Dox)-inducible adipocyte-specific VEGF-A overexpression model, we demonstrate that the local up-regulation of VEGF-A in adipocytes improves vascularization and causes a "browning" of white adipose tissue (AT), with massive up-regulation of UCP1 and PGC1 alpha. This is associated with an increase in energy expenditure and resistance to high fat diet-mediated metabolic insults. Similarly, inhibition of VEGF-A-induced activation of VEGFR2 during the early phase of high fat diet-induced weight gain, causes aggravated systemic insulin resistance. However, the same VEGF-A-VEGFR2 blockade in ob/ob mice leads to a reduced body-weight gain, an improvement in insulin sensitivity, a decrease in inflammatory factors, and increased incidence of adipocyte death. The consequences of modulation of angiogenic activity are therefore context dependent. Proangiogenic activity during adipose tissue expansion is beneficial, associated with potent protective effects on metabolism, whereas antiangiogenic action in the context of preexisting adipose tissue dysfunction leads to improvements in metabolism, an effect likely mediated by the ablation of dysfunctional proinflammatory adipocytes.

Impact of the Manaus urban plume on trace gas mixing ratios near the surface in the Amazon Basin: Implications for the NO-NO2-O-3 photostationary state and peroxy radical levels

We measured the mixing ratios of NO, NO2, O-3, and volatile organic carbon as well as the aerosol light-scattering coefficient on a boat platform cruising on rivers downwind of the city of Manaus (Amazonas State, Brazil) in July 2001 (Large-Scale Biosphere-Atmosphere Experiment in Amazonia-Cooperative LBA Airborne Regional Experiment-2001). The dispersion and impact of the Manaus plume was investigated by a combined analysis of ground-based (boat platform) and airborne trace gas and aerosol measurements as well as by meteorological measurements complemented by dispersion calculations (Hybrid Single-Particle Lagrangian Integrated Trajectory model). For the cases with the least anthropogenic influence (including a location in a so far unexplored region similar to 150 km west of Manaus on the Rio Manacapuru), the aerosol scattering coefficient, sigma(s), was below 11 Mm(-1), NOx mixing ratios remained below 0.6 ppb, daytime O-3 mixing ratios were mostly below 20 ppb and maximal isoprene mixing ratios were about 3 ppb in the afternoon. The photostationary state (PSS) was not established for these cases, as indicated by values of the Leighton ratio, Phi, well above unity. Due to the influence of river breeze systems and other thermally driven mesoscale circulations, a change of the synoptic wind direction from east-northeast to south-southeast in the afternoon often caused a substantial increase of ss and trace gas mixing ratios (about threefold for sigma(s), fivefold for NOx, and twofold for O-3), which was associated with the arrival of the Manaus pollution plume at the boat location. The ratio F reached unity within its uncertainty range at NOx mixing ratios of about 3 ppb, indicating "steady-state" conditions in cases when radiation variations, dry deposition, emissions, and reactions mostly involving peroxy radicals (XO2) played a minor role. The median midday/afternoon XO2 mixing ratios estimated using the PSS method range from 90 to 120 parts per trillion (ppt) for the remote cases (sigma(s) < 11 Mm(-1) and NOx < 0.6 ppb), while for the polluted cases our estimates are 15 to 60 ppt. These values are within the range of XO2 estimated by an atmospheric chemistry box model (Chemistry As A Box model Application-Module Efficiently Calculating the Chemistry of the Atmosphere (CAABA/MECCA)-3.0).

Monsplasty for Women After Massive Weight Loss

Ptosis with excess skin in the pubic area is a very common deformity in patients after massive weight loss. This deformity requires adequate surgical treatment whether combined with abdominoplasty or not. The enlarged pubogenital area may lead to psychosocial distress and impaired quality of life. A series of 23 women with a mean age of 39.5 years who previously underwent bariatric surgeries and later presented with pubogenital ptosis underwent monsplasty. The preoperative surgical markings and the surgical technique presented are easily reproducible. In this prospective study, the surgical outcomes were assessed by questionnaires applied to the patients, who scored the following parameters: movement dynamics, aesthetic appearance, sexual performance, improved hygiene, and use of clothing items. Four of the parameters assessed (movement dynamics, aesthetic appearance, hygiene, and use of clothing items) showed clear improvement, with scores ranging from good to very good. A small percentage of the patients (13%) reported fair improvement in sexual performance. The findings showed monsplasty to be a simple and reproducible technique with favorable outcomes and low morbidity rates.

Post-conventional Political Cultures via Processes of Direct Democracy: Theoretical Considerations Based on Jurgen Habermas and Lawrence Kohlberg

In this article, it is proposed to differentiate political cultures in two dimensions. First, inspired by Habermas' distinction of the contents of discourse, a distinction is suggested between moral, ethical-political and pragmatic elements of political culture as well as of an element of culture of balancing interests. Second, inspired by Kohlberg's stage models for the development of the individual moral consciousness and for moral culture, a distinction is similarly suggested between two pre-conventional, two conventional and two post-conventional collective stages of political culture. It can be shown that from a normative point of view, only deliberations made in a post-conventional political culture can produce reasonable or at least fair results. Conceptual considerations indicate processes of direct democracy as the method for promoting post-conventional political cultures. The more liberty that the citizens have to formulate and trigger processes of direct democracy, the more one can expect from them to generate post-conventional political cultures.

Evaluation of Aesthetic Abdominal Surgery Using a New Clinical Scale

Evaluation of outcomes after aesthetic surgery still is a challenge in plastic surgery. The evaluation frequently is based on subjective criteria. This study used a new clinical grading scale to evaluate aesthetic results for plastic surgeries to the abdomen. The method scores each of the following five parameters: volume of subcutaneous tissue, contour, excess of skin, aspect of the navel, and quality of the scar on the abdominal wall. The scale options are 0 (poor), 1 (fair), and 2 (good), and the total rate can range from 0 to 10. The study included 40 women ages 18-53 years. Of these 40 women, 20 underwent traditional abdominoplasty, and 20 had liposuction alone. Preoperatively and at least 1 year later, photographic results were analyzed and scored by three independent plastic surgeons. In the abdominoplasty group, the average grade rose from 2.9 +/- A 0.4 to 6.8 +/- A 0.4 postoperatively. In the liposuction group, the average grade was 5.3 +/- A 0.5 preoperatively and 7.7 +/- A 0.4 postoperatively. In both groups, the average postoperative grade was significantly higher than the preoperative grade. The mean scores for groups A and L were significantly different, demonstrating that the scale was sensitive in identifying different anatomic abnormalities in the abdomen. The rating scale used for the aesthetic evaluation of the abdomen was effective in the analysis of two different procedures: conventional abdominoplasty and liposuction. Abdominoplasty provided the greater gain according to a comparison of the pre- and postoperative scores.

The antioxidative effect of de novo generated vitamin B-6 in Plasmodium falciparum validated by protein interference

The malaria parasite Plasmodium falciparum is able to synthesize de novo PLP (pyridoxal 5'-phosphate), the active form of vitamin B-6. In the present study, we have shown that the de novo synthesized PLP is used by the parasite to detoxify O-1(2) (singlet molecular oxygen), a highly destructive reactive oxygen species arising from haemoglobin digestion. The formation of O-1(2) and the response of the parasite were monitored by live-cell fluorescence microscopy, by transcription analysis and by determination of PLP levels in the parasite. Pull-down experiments of transgenic parasites overexpressing the vitamin B-6-biosynthetic enzymes PfPdx1 and PfPdx2 clearly demonstrated an interaction of the two proteins in vivo which results in an elevated PLP level from 12.5 mu M in wild-type parasites to 36.6 mu M in the PfPdx1/PfPdx2-overexpressing cells and thus to a higher tolerance towards O-1(2). In contrast, by applying the dominant-negative effect on the cellular level using inactive mutants of PfPdx1 and PfPdx2, P. falciparum becomes susceptible to O-1(2). Our results demonstrate clearly the crucial role of vitamin B-6 biosynthesis in the detoxification of O-1(2) in P falciparum. Besides the known role of PLP as a cofactor of many essential enzymes, this second important task of the vitamin B-6 de novo synthesis as antioxidant emphasizes the high potential of this pathway as a target of new anti-malarial drugs.

Duplications of BHLHA9 are associated with ectrodactyly and tibia hemimelia inherited in non-Mendelian fashion

Background Split-hand/foot malformation (SHFM)-also known as ectrodactyly-is a congenital disorder characterised by severe malformations of the distal limbs affecting the central rays of hands and/or feet. A distinct entity termed SHFLD presents with SHFM and long bone deficiency. Mouse models suggest that a defect of the central apical ectodermal ridge leads to the phenotype. Although six different loci/mutations (SHFM1-6) have been associated with SHFM, the underlying cause in a large number of cases is still unresolved. Methods High resolution array comparative genomic hybridisation (CGH) was performed in patients with SHFLD to detect copy number changes. Candidate genes were further evaluated for expression and function during limb development by whole mount in situ hybridisation and morpholino knock-down experiments. Results Array CGH showed microduplications on chromosome 17p13.3, a locus previously associated with SHFLD. Detailed analysis of 17 families revealed that this copy number variation serves as a susceptibility factor for a highly variable phenotype with reduced penetrance, particularly in females. Compared to other known causes for SHFLD 17p duplications appear to be the most frequent cause of SHFLD. A similar to 11.8 kb minimal critical region was identified encompassing a single gene, BHLHA9, a putative basic loop helix transcription factor. Whole mount in situ hybridisation showed expression restricted to the limb bud mesenchyme underlying the apical ectodermal ridge in mouse and zebrafish embryos. Knock down of bhlha9 in zebrafish resulted in shortening of the pectoral fins. Conclusions Genomic duplications encompassing BHLHA9 are associated with SHFLD and non-Mendelian inheritance characterised by a high degree of non-penetrance with sex bias. Knock-down of bhlha9 in zebrafish causes severe reduction defects of the pectoral fin, indicating a role for this gene in limb development.

Diet and the risk of head and neck cancer: a pooled analysis in the INHANCE consortium

We investigated the association between diet and head and neck cancer (HNC) risk using data from the International Head and Neck Cancer Epidemiology (INHANCE) consortium. The INHANCE pooled data included 22 case-control studies with 14,520 cases and 22,737 controls. Center-specific quartiles among the controls were used for food groups, and frequencies per week were used for single food items. A dietary pattern score combining high fruit and vegetable intake and low red meat intake was created. Odds ratios (OR) and 95% confidence intervals (CI) for the dietary items on the risk of HNC were estimated with a two-stage random-effects logistic regression model. An inverse association was observed for higher-frequency intake of fruit (4th vs. 1st quartile OR = 0.52, 95% CI = 0.43-0.62, p (trend) < 0.01) and vegetables (OR = 0.66, 95% CI = 0.49-0.90, p (trend) = 0.01). Intake of red meat (OR = 1.40, 95% CI = 1.13-1.74, p (trend) = 0.13) and processed meat (OR = 1.37, 95% CI = 1.14-1.65, p (trend) < 0.01) was positively associated with HNC risk. Higher dietary pattern scores, reflecting high fruit/vegetable and low red meat intake, were associated with reduced HNC risk (per score increment OR = 0.90, 95% CI = 0.84-0.97).

Measurement of prompt J/psi and beauty hadron production cross sections at mid-rapidity in pp collisions at root s=7 TeV

The ALICE experiment at the LHC has studied J/psi production at mid-rapidity in pp collisions at root s = 7 TeV through its electron pair decay on a data sample corresponding to an integrated luminosity L-int = 5.6 nb(-1). The fraction of J/psi from the decay of long-lived beauty hadrons was determined for J/psi candidates with transverse momentum p(t) > 1,3 GeV/c and rapidity vertical bar y vertical bar < 0.9. The cross section for prompt J/psi mesons, i.e. directly produced J/psi and prompt decays of heavier charmonium states such as the psi(2S) and chi(c) resonances, is sigma(prompt J/psi) (p(t) > 1.3 GeV/c, vertical bar y vertical bar < 0.9) = 8.3 +/- 0.8(stat.) +/- 1.1 (syst.)(-1.4)(+1.5) (syst. pol.) mu b. The cross section for the production of b-hadrons decaying to J/psi with p(t) > 1.3 GeV/c and vertical bar y vertical bar < 0.9 is a sigma(J/psi <- hB) (p(t) > 1.3 GeV/c, vertical bar y vertical bar < 0.9) = 1.46 +/- 0.38 (stat.)(-0.32)(+0.26) (syst.) mu b. The results are compared to QCD model predictions. The shape of the p(t) and y distributions of b-quarks predicted by perturbative QCD model calculations are used to extrapolate the measured cross section to derive the b (b) over bar pair total cross section and d sigma/dy at mid-rapidity.

A clinical experience of the supraclavicular flap used to reconstruct head and neck defects in late-stage cancer patients

The supraclavicular island flap has been widely used in head and neck reconstruction, providing an alternative to the traditional techniques like regional or free flaps, mainly because of its thin skin island tissue and reliable vascularity. Head and neck patients who require large reconstructions usually present poor clinical and healing conditions. An early experience using this flap for late-stage head and neck tumour treatment is reported. Forty-seven supraclavicular artery flaps were used to treat head and neck oncologic defects after cutaneous, intraoral and pharyngeal tumour resections. Dissection time, complications, donor and reconstructed area outcomes were assessed. The mean time for harvesting the flaps was 50 min by the senior author. All donor sites were closed primarily. Three cases of laryngopharyngectomy reconstruction developed a small controlled (salivary) leak that was resolved with conservative measures. Small or no strictures were detected on radiologic swallowing examinations and all patients regained normal swallowing function. Five patients developed donor site dehiscence. These wounds were treated with regular dressing until healing was complete. There were four distal flap necroses in this series. These necroses were debrided and closed primarily. The supraclavicular flap is pliable for head and neck oncologic reconstruction in late-stage patients. High-risk patients and modified radical neck dissection are not contraindications for its use. The absence of the need to isolate the pedicle offers quick and reliable harvesting. The arc of rotation on the base of the neck provides adequate length for pharyngeal, oral lining and to reconstruct the middle and superior third of the face. (C) 2012 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.

Functional characterization and oligomerization of a recombinant xyloglucan-specific endo-beta-1,4-glucanase (GH12) from Aspergillus niveus

Xyloglucan is a major structural polysaccharide of the primary (growing) cell wall of higher plants. It consists of a cellulosic backbone (beta-1,4-linked glucosyl residues) that is frequently substituted with side chains. This report describes Aspergillus nidulans strain A773 recombinant secretion of a dimeric xyloglucan-specific endo-beta-1,4-glucanohydrolase (XegA) cloned from Aspergillus niveus. The ORF of the A. niveus xegA gene is comprised of 714 nucleotides, and encodes a 238 amino acid protein with a calculated molecular weight of 23.5 kDa and isoelectric point of 4.38. The optimal pH and temperature were 6.0 and 60 degrees C, respectively. XegA generated a xyloglucan-oligosaccharides (XGOs) pattern similar to that observed for cellulases from family GH12, i.e., demonstrating that its mode of action includes hydrolysis of the glycosidic linkages between glucosyl residues that are not branched with xylose. In contrast to commercial lichenase, mixed linkage beta-glucan (lichenan) was not digested by XegA, indicating that the enzyme did not cleave glucan beta-1,3 or beta-1,6 bonds. The far-UV CD spectrum of the purified enzyme indicated a protein rich in beta-sheet structures as expected for GH12 xyloglucanases. Thermal unfolding studies displayed two transitions with mid-point temperatures of 51.3 degrees C and 81.3 degrees C respectively, and dynamic light scattering studies indicated that the first transition involves a change in oligomeric state from a dimeric to a monomeric form. Since the enzyme is a predominantly a monomer at 60 degrees C. the enzymatic assays demonstrated that XegA is more active in its monomeric state. (c) 2012 Elsevier B.V. All rights reserved.

Mastopexia após perda ponderal maciça: suspensão dérmica, remodelação do parênquima e aumento com tecido autógeno

INTRODUÇÃO: Pacientes com perda ponderal significativa podem apresentar mamas com ptose acentuada, perda de projeção do polo superior e excesso de tecido na porção toracolateral. Rubin & Khachi descreveram técnica de mastopexia com suspensão dérmica e remodelação do parênquima associada a aumento com tecido autógeno, tratando a deformidade mamária e o excesso toracolateral em um só estágio. Neste trabalho, é ilustrada essa técnica cirúrgica e demonstradas sua reprodutibilidade e suas complicações. MÉTODO: Foram operadas 14 pacientes com deformidade graus 2 e 3 pela Escala de Pittsburgh, no Hospital Estadual de Sapopemba (São Paulo, SP, Brasil), no período de dezembro de 2008 a dezembro de 2009, utilizando a técnica referida. Foram analisados os seguintes dados: tipos de deformidade das mamas, translocação do complexo areolopapilar (CAP), dimensões dos retalhos, tempo cirúrgico, tempo de permanência do dreno e incidência de complicações. RESULTADOS: A média de idade das pacientes foi de 41,21 + 7,67 anos e o índice de massa corporal médio foi de 29,30 + 2,77. O tempo de seguimento das pacientes variou de 3 meses a 18 meses, com média de 8 meses. Dentre as 14 pacientes operadas, 4 (28,6%) apresentavam deformidade grau 3 e 10 (71,4%), grau 2. A média de translocação do CAP foi de 6,38 cm. As dimensões médias do retalho foram de 25,21 cm x 6,92 cm. O tempo cirúrgico médio foi de 188,57 minutos. Os drenos permaneceram, em média, por 6,21 dias. Foram observadas as seguintes complicações: epiteliose de CAP, deiscência na junção do T, hematoma pequeno e linfedema toracolateral. CONCLUSÕES: A mastopexia com suspensão dérmica, remodelação do parênquima e aumento com tecido autólogo é uma técnica reprodutível, rápida e com baixo índice de complicações.

Infecções de corrente sanguínea por bactérias multirresistentes em UTI de tratamento de queimados: experiência de 4 anos

INTRODUÇÃO: O conhecimento do perfil de resistência aos antibióticos das bactérias de um nosocômio é essencial para orientar tratamento adequado dos pacientes. Isso é especialmente importante para os pacientes mais graves, já que o tratamento deve ser instituído antes do resultado das culturas. O objetivo deste estudo foi analisar o perfil das bactérias multirresistentes encontradas nas hemoculturas de pacientes admitidos na Unidade de Tratamento Intensivo (UTI) da Unidade de Queimados do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo. MÉTODO: Foram analisados 178 pacientes internados na UTI para tratamento de queimados, no período de 2009 a 2011, sendo 131 do sexo masculino, com média de idade de 29,2 anos. RESULTADOS: Entre os pacientes analisados, 80 (44,9%) apresentaram hemocultura periférica positiva, sendo 66 (82,5%) casos com bactérias multirresistentes. Em 48 pacientes, foram isoladas Staphylococcus sp., que se apresentaram resistentes à oxacilina em 33 deles. Em 11 pacientes, foram isoladas Acinetobacter baumanii, que se apresentaram resistentes a imipenem em 8 casos. Em 19 pacientes, foram isoladas Pseudomonas sp., resistentes a imipenem em 16 casos. Em 10 pacientes foram isoladas Enterobacter sp., resistentes a amicacina e ciprofloxacina em 2 casos. A presença de bactérias multirresistentes não foi associada a maior ocorrência de óbitos, porém foi verificado maior tempo de internação (52,6 dias vs. 36,3 dias para os grupos com e sem bactérias multirresistentes, respectivamente; P = 0,0306). Não foi encontrada interação significante entre superfície corpórea queimada e presença de bactérias MR. CONCLUSÕES: A presença de bactérias multirresistentes é um problema grave, tanto pela prevalência como pela morbidade e mortalidade associadas.

Análise comparativa da evolução e das complicações pós-operatórias nas cirurgias plásticas do contorno corporal em pacientes idosos e jovens com perda ponderal maciça

INTRODUÇÃO: Os procedimentos aplicados a pacientes ex-obesos idosos após terapêuticas bariátricas estão em ascensão. Dados fidedignos quanto à evolução e às complicações nesse grupo populacional ainda são escassos na literatura. O objetivo do presente estudo é analisar a evolução e as complicações em abdominoplastias realizadas em pacientes com idade mais avançada após perda ponderal maciça, e compará-las às de pacientes mais jovens. MÉTODO: Foram analisados, retrospectivamente, pacientes com perda ponderal maciça submetidos a cirurgia para contorno da região abdominal, entre julho de 2005 e julho de 2011, no Hospital Estadual Sapopemba (HESAP). Como critério para divisão dos grupos, a fim de analisar o período pós-operatório e as complicações das abdominoplastias realizadas após perda ponderal maciça, foi estabelecida idade > 60 anos. RESULTADOS: Foram analisados 264 pacientes, 19 deles com idade entre 60 anos e 75 anos (grupo I) e 245 entre 22 anos e 59 anos (grupo J). O grupo I apresentou 10,5% de complicações maiores (P > 0,999) e 41,1% de complicações menores (P = 0,280), enquanto o grupo J obteve 10,6% de complicações maiores (P > 0,999) e 30,2% de complicações menores (P = 0,280). CONCLUSÕES: Os pacientes com > 60 anos de idade não apresentaram maior número de complicações que o grupo mais jovem.