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Bacillus subtilisis a valuable industrial production platform for proteins, a bacterial model for cellular differentiation and its endospores have been proposed as a vehicle for vaccine delivery. As suchB. subtilisis a major synthetic biology chassis but, unlikeEscherichia coli, lacks a standardized toolbox for genetic manipulation. EcoFlex is a versatile modular DNA assembly toolkit forE. colisynthetic biology based on Golden Gate cloning. Here we introduce BacilloFlex, an extension of the EcoFlex assembly standard toB. subtilis. Transcription units flanked by sequences homologous to loci in theB. subtilisgenome were rapidly assembled by the EcoFlex standard and subsequently chromosomally integrated. At present, BacilloFlex includes a range of multi-functionalB. subtilisspecific parts with applications including metabolic engineering, biosensors and spore surface display. We hope this work will form the foundation of a widely adopted cloning standard forB. subtilisfacilitating collaboration and the sharing of parts.

The interaction of ochratoxin A (OTA) with heptakis-2,6-di-O-methyl-β-cyclodextrin (DIMEB) in aqueous solutions at two different pHs (3.5 and 9.5) was studied by means of spectroscopic, calorimetric and electrochemical techniques. DIMEB affected the physico-chemical properties of OTA. A stronger interaction was observed at pH = 3.5 where the neutral form of OTA prevails. The spectroscopic information indicated that the OTA/DIMEB inclusion process occurred through the insertion of the phenylalanine group into the cyclodextrin (CD) cavity; moreover, voltammetry experiments showed that the isocoumarinic phenolic group was also involved, probably by non-inclusion interactions. At pH = 3.5, binding constants and thermodynamic parameters of the OTA/DIMEB complex were determined by means of the modified Benesi-Hildebrand equation and van’t Hoff plot. A 1:1 stoichiometry for the OTA/DIMEB complex was observed with a binding constant equal to 530 ± 50 L·mol−1 at 25 °C. The calculated thermodynamic parameters indicate that the interaction is highly endothermic and that the complex formation is driven by entropy. The results provide useful information for potential applications of cyclodextrins in the analysis of mycotoxins and in the field of mycotoxin sequestering agents.

Many factors can interfere with glycemic control in patients with type 2 diabetes mellitus and this patient group has an increased risk of experiencing drug-related negative outcomes (DNO). The aim of this study was to identify the predictive factors of unsatisfactory glycemic control and DNO in this group of patients. A cross-sectional study was conducted collecting data from prescriptions and interviews with 100 patients of the Endocrinology Clinic of the Hospital Complex of Sorocaba, São Paulo, Brazil. Variables associated with unsatisfactory glycemic control were determined and the DNO associated with antidiabetic drugs identified. Age (0.05). Characterization of the profile of patients with uncontrolled diabetes and of aspects associated with drug treatment can contribute to the planning of interventions to improve patient care.

The incidence of abnormal cervical cytology in pregnancy is similar to that reported for non-pregnant women. Furthermore, 1% of pregnant women annually screened for cervical cancer will be diagnosed with cervical intraepithelial neoplasia (CIN) of various degrees. For this reason, Pap smear should be performed in the first trimester of pregnancy. The persistence of HR-HPV infection is related to the development of CIN. However, the relationship between CIN and HR-HPV infection during pregnancy and postpartum can hardly be found. The aim of this work was to assess the proper management of abnormal cytology during and after pregnancy evaluating regression rate, persistence rate and risk of progression and the predictive role of HPV molecular tests.Patients with abnormal cervical cytology were followed-up using colposcopy and colposcopy-directed biopsies every 12 weeks. Molecular tests were performed at the moment of the cytological diagnosis. Patients not treated in pregnancy were re-evaluated with cytology, colposcopy, biopsies, HPV-DNA test and HPV-mRNA test for a final diagnosis 8 weeks postpartum. Women with a persistent CIN 2-3 lesion at this follow-up check, underwent an excisional procedure by LEEP and then re-evaluated every 6 months for a year.HPV-DNA test showed a sensitivity of 90.5% and a negative predictive value of 96.4%. Specificity and positive predictive values were 67.9% and 43.2%, respectively. For HPV-mRNA test, a sensitivity of 76.2% and a NPV of 93.9% were found; specificity and PPV were 98.7% and 94.1% respectively.An observational management based on the use of molecular test and particularly HPV-mRNA test for its higher specificity, is a reasonable possibility in the follow-up of CIN2/3 lesions during pregnancy.

T-2 and HT-2 toxins are mycotoxins produced by several Fusarium species that are commonly found in various cereal grains, including oats, barley, wheat and maize. Intake estimates indicate that the presence of these mycotoxins in the diet can be of concern for public health. In this work, the inclusion processes occurring between fluorescent anthracene-derivatives of T-2 and HT-2 toxins and different cyclodextrin (CD) molecules were investigated in aqueous solutions by means of UV-Vis absorption, fluorescence emission and dynamic light scattering. Binding constant values and chemico-physical parameters were calculated. It was found that b-CDs give stronger inclusion reactions with both T-2 and HT-2 derivatives, as stated by important emission intensity increments. Such interactions were found to be fundamentally enthalpy-driven. Among b-CDs, the effect of the methylation at hydroxyl groups was tested: as a result, the di-methyl form of b-CD was found to induce the best fluorescence intensity enhancements.

Hypothyroidism is associated with the risk of development of the metabolic syndrome (MS) and hypercholesterolemia. Direct evidence that hypothyroidism might be associated with advanced chronic liver disease via nonalcoholic steatohepatitis (NASH) is limited. We studied the relationship between thyroid hormones, thyroid stimulating hormone (TSH), cholesterol, and NASH. In consecutive euthyroid patients with biopsy-proven nonalcoholic fatty liver disease, TSH and thyroid hormone (FT3 and FT4) concentrations were compared in 25 patients with steatosis and 44 non-cirrhotic NASH patients featuring concurrent ballooning, lobular inflammation and steatosis. The MS was diagnosed according to ATP III criteria. A meta-analysis of previously published studies was performed to evaluate whether NASH, compared to simple steatosis, is associated with lower cholesterol levels. At univariate analysis, compared to those with steatosis, patients with NASH have a wider waist, elevated levels of BMI, ALT, AST, fasting insulin, HOMA-IR, ferritin, TSH and a lower serum cholesterol. At stepwise multivariable logistic regression analysis, the independent predictors of NASH are high HOMA and TSH and lower total cholesterol (Model 1); MS and high TSH (Model 2). At meta-analysis, serum total cholesterol levels are significantly lower in predominantly non-cirrhotic NASH than in simple steatosis. This study provides cross-sectional and meta-analytic evidence that, in euthyroid patients, high-though-normal TSH values are independently associated with NASH. Further work is needed to ascertain the role, if any, of lower cholesterol serum levels in assisting in the diagnosis of NASH.

Ochratoxin A (OTA) is a nephrotoxic mycotoxin produced by several fungal species, mainly Aspergillus ochraceus, A. carbonarius and Penicillium verrucosum. It contaminates many foodstuffs, particularly cereals and their derivatives, coffee, beer, wine and cocoa, and represents a serious health threat both to humans and animals. Spectroscopic properties of OTA solutions depend on the pH, solvent polarity and can be influenced by the presence of cyclodextrins (CDs). In this work, the effect of beta-CD on spectroscopic properties of OTA in aqueous solutions has been investigated by means of absorption and steady-state fluorescence at different pHs (range 3.5-9.5). Binding constants of OTA/beta-CD inclusion complexes have been determined by applying modified Benesi-Hildebrand equation. A 1:1 stoichiometry of OTA/beta-CD complexes has been observed at all tested pHs.

[ES] El presente trabajo final de grado aborda el cálculo y diseño de una nave industrial para la producción de queso fresco, ubicada en el término municipal de Oliva (Valencia). La estructura se corresponde con una nave a dos aguas de 2240 metros cuadrados de superficie y posee, además, un altillo para oficinas. Para abarcar dicha superficie, se han dispuesto 11 pórticos de 28 metros de luz, separados 8 metros entre sí, llegando a los 80 metros de longitud. El desarrollo de este TFG se ha realizado mediante el programa de cálculo de estructuras CYPE, gran parte en las versiones de CYPECAD y CYPE 3D. Por otro lado, el programa de dibujo Autocad ha sido utilizado en la realización de todos los planos de la estructura. Para finalizar, el presupuesto del proyecto ha sido realizado con el software Arquímedes, complementado con el “Generador de precios” de CYPE., [CA] El present treball final de grau aborda el càlcul i disseny d'una nau industrial per a la producció de formatge fresc, situada en el terme municipal d'Oliva (València). L'estructura es correspon amb una nau a dues aigües de 2240 metres quadrats de superfície i posseeix, a més, un altell per a oficines. Per abastar aquesta superfície, s'han disposat 11 pòrtics de 28 metres de llum, separats 8 metres entre si, arribant als 80 metres de longitud. El desenvolupament d'aquest TFG s'ha realitzat mitjançant el programa de càlcul d'estructures CYPE, gran part en les versions de CYPECAD i CYPE 3D. D'altra banda, el programa de dibuix Autocad ha estat utilitzat en la realització de tots els plànols de l'estructura. Per finalitzar, el pressupost del projecte ha estat realitzat amb el programa Arquímedes, complementat amb el "Generador de preus" de CYPE., [EN] This project addresses the calculation and design of an industrial building for the production of fresh cheese, located in the town of Oliva (Valencia). The structure corresponds to a gable roof 2240 square meters industrial plant and also has a mezzanine floor for offices. To cover this surface, eleven frames are arranged with a 28 meters width, spaced 8 meters apart, reaching 80 meters in length. The development of this project was performed using the program CYPE, mostly with the versions CYPECAD and 3D CYPE. On the other hand, the program Autocad was used for drawing the plans of the structure. Finally, the budget of the project was made with the Archimedes software, complemented with "Builder Price" CYPE.

Made available in DSpace on 2018-12-11T17:23:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-01 Background: Studies on diabetic foot and its complications involving a significant and representative sample of patients in South American countries are scarce. The main objective of this study was to acquire clinical and epidemiological data on a large cohort of diabetic patients from 19 centers from Brazil and focus on factors that could be associated with the risk of ulcer and amputation. Methods: This study presents cross sectional, baseline results of the BRAZUPA Study. A total of 1455 patients were included. Parameters recorded included age, gender, ethnicity, diabetes and comorbidity-related records, previous ulcer or amputation, clinical symptomatic score, foot classification and microvascular complications. Results: Patients with ulcer had longer disease duration (17.2 ± 9.9 vs. 13.2 ± 9.4 years; p < 0.001), and poorer glycemic control (HbA1c 9.23 ± 2.03 vs. 8.35 ± 1.99; p < 0.001). Independent risk factors for ulcer were male gender (OR 1.71; 95 % CI 1.2-3.7), smoking (OR 1.78; 95 % CI 1.09-2.89), neuroischemic foot (OR 20.34; 95 % CI 9.31-44.38), region of origin (higher risk for those from developed regions, OR 2.39; 95 % CI 1.47-3.87), presence of retinopathy (OR 1.68; 95 % CI 1.08-2.62) and absence of vibratory sensation (OR 7.95; 95 % CI 4.65-13.59). Risk factors for amputation were male gender (OR 2.12; 95 % CI 1.2-3.73), type 2 diabetes (OR 3.33; 95 % CI 1.01-11.1), foot at risk classification (higher risk for ischemic foot, OR 19.63; 95 % CI 3.43-112.5), hypertension (lower risk, OR 0.3; 95 % CI 0.14-0.63), region of origin (South/Southeast, OR 2.2; 95 % CI 1.1-4.42), previous history of ulcer (OR 9.66; 95 % CI 4.67-19.98) and altered vibratory sensation (OR 3.46; 95 % CI 1.64-7.33). There was no association between either outcome and ethnicity. Conclusions: Ulcer and amputation rates were high. Age at presentation was low and patients with ulcer presented a higher prevalence of neuropathy compared to ischemic foot at risk. Ischemic disease was more associated with amputations. Ethnical differences were not of great importance in a miscegenated population. Faculty of Medical Sciences State University of Campinas Unity of Diabetes State University of Rio de Janeiro Department of Nutrology Federal University of Rio de Janeiro Centre for Diabetes and Endocrinology in the State of Bahia (CEDEBA) Centro Integrado de Diabetes e Hipertensão Do Ceará State University of Feira de Santana Federal University of Campina Grande Hospital das Clínicas de Porto Alegre Federal University of Rio Grande Do sul Santa Casa de Porto Alegre Hospital das Clínicas da Faculdade de Medicina Universidade de São Paulo Hospital Regional de Taguatinga Faculdade de Medicina Federal University of Santa Catarina Federal University of Sergipe Pontifícia Universidade Católica de Sorocaba Division of Endocrinology Hospital Agamenon Magalhães Faculdade de Medicina de Catanduva Faculdade de Medicina de Botucatu UNESP Universidade de São Paulo Faculdade de Medicina Santa Casa de Belo Horizonte Faculdade de Medicina de Botucatu UNESP

Traditional surgery presents some disadvantages, such as the necessity for general anesthesia, hemorrhage, recurrence of pathology, and the possible onset of dyspareunia due to an excessive scarring. CO2 laser surgery might resolve these problems and might be employed in a wider range of clinical indications than usual. We examined the results of CO2 laser surgery in patients affected by benign pathologies and congenital malformations of the female lower genital tract.In this observational study, we enrolled 49 women who underwent CO2 laser surgery for the following indications: Bartholin's gland cyst, imperforate hymen, vaginal septum, Nabothian cyst, and vaginal polyps. Feasibility, cost-effectiveness, complication rate, recurrence rate, short- and long-term outcomes were assessed.All procedures were carried out in a short operative time, without any intraoperative complications. Only 1 (2.0%) out of 49 patients required a hemostatic suture for bleeding. Postoperative period was uneventful in all patients, except 6 (12.2%) out of 49 patients who reported pain one day after surgery, successfully treated with paracetamol. Healing was rapid and excellent in all cases; no wound infection, scarring or stenosis were noticed. Preoperative symptoms reduced or disappeared in all cases. No recurrence was observed and no re-intervention was needed.CO2 laser surgery provides several advantages over traditional surgery, as its systematic use in treating pre-invasive, benign, and congenital pathologies of the female lower genital tract reduces patient discomfort, improves short- and long-term outcomes, and optimizes cost-effectiveness.

Background: Insulin resistance is a risk factors for non-alcoholic fatty liver disease (NAFLD) and for gallstone disease (GD). Aims of the present study were to assess the prevalence of and factors associated with GD in unselected patients with NAFLD. Methods: A total of 161 consecutive patients with NAFLD diagnosed through compatible ultrasonography in the absence of known etiologies of liver disease (in all patients) and/or confirmed histologically (in 61 patients), was studied. Gallstone disease was diagnosed through ultrasound scanning or on the basis of previous cholecystectomy. Anthropometric and biochemical variables and concurrent diseases were compared in 32 NAFLD-GD patients and in 129 NAFLD patients without GD (controls) according to gender. Results: The overall prevalence of GD was 19.88%, higher in female patients (P

The `charge state breeder' BRIC (BReeding Ion Charge) is based on an EBIS source and it is designed to accept Radioactive Ion Beam (RIB) with charge state +1, in a slow injection mode, to increase their charge state up to +n. BRIC has been developed at the INFN section of Bari (Italy) during these last 3 years with very limited funds. Now, it has been assembled at the LNL (Italy) where are in progress the first tests as stand alone source and where, in the future, with some implementation, it will be tested as charge breeder at ISOL/TS facility of that laboratory. BRIC could be considered as a solution for the charge state breeder of the SPES project under study also at the LNL. The new feature of BRIC, with respect to the classical EBIS, is given by the insertion, in the ion drift chamber, of a radio frequency (RF) - quadrupole aiming to filter the unwanted masses and then making a more efficient containment of the wanted ions. In this paper, the first ion charge state measurements and analysis and the effect of the RF field applied on the ion chamber will be reported and discussed. The first RF test measurements seem confirm, as foreseen by simulation results carried out previously, that a selective containment can be obtained. However, most accurate measurements needed to study with more details the effect. For this reason, few implementations of the system are in order to improve the accuracy of the measurements. The proposed modifications of the BRIC device, then, will be also presented and shortly discussed.

Background: Molecular engineering of the intermediary physiology of cyanobacteria has become important for the sustainable production of biofuels and commodity compounds from CO2 and sunlight by "designer microbes." The chemical commodity product L-lactic acid can be synthesized in one step from a key intermediary metabolite of these organisms, pyruvate, catalyzed by a lactate dehydrogenase. Synthetic biology engineering to make "designer microbes" includes the introduction and overexpression of the product-forming biochemical pathway. For further optimization of product formation, modifications in the surrounding biochemical network of intermediary metabolism have to be made.Results: To improve light-driven L-lactic acid production from CO2, we explored several metabolic engineering design principles, using a previously engineered L-lactic acid producing mutant strain of Synechocystis sp. PCC6803 as the benchmark. These strategies included: (i) increasing the expression level of the relevant product-forming enzyme, lactate dehydrogenase (LDH), for example, via expression from a replicative plasmid; (ii) co-expression of a heterologous pyruvate kinase to increase the flux towards pyruvate; and (iii) knockdown of phosphoenolpyruvate carboxylase to decrease the flux through a competing pathway (from phosphoenolpyruvate to oxaloacetate). In addition, we tested selected lactate dehydrogenases, some of which were further optimized through site-directed mutagenesis to improve the enzyme’s affinity for the co-factor nicotinamide adenine dinucleotide phosphate (NADPH). The carbon partitioning between biomass and lactic acid was increased from about 5% to over 50% by strain optimization.Conclusion: An efficient photosynthetic microbial cell factory will display a high rate and extent of conversion of substrate (CO2) into product (here: L-lactic acid). In the existing CO2-based cyanobacterial cell factories that have been described in the literature, by far most of the control over product formation resides in the genetically introduced fermentative pathway. Here we show that a strong promoter, in combination with increased gene expression, can take away a significant part of the control of this step in lactic acid production from CO2. Under these premises, modulation of the intracellular precursor, pyruvate, can significantly increase productivity. Additionally, production enhancement is achieved by protein engineering to increase co-factor specificity of the heterologously expressed LDH.

Objective To determine whether an intraoral stent may decrease radiation dose to health tissues during intensity-modulated radiotherapy (IMRT) and to evaluate the effect on mucositis. Study Design A total of 33 patients with tongue or floor of the mouth cancer were retrospectively evaluated and divided into 2 groups: group 1 (with stent, n = 19) and group 2 (without stent, n = 14). Data were collected on dosimetric and mucositis outcomes. Results The mean dose to the maxilla was significantly lower in group 1 (20.9 Gy) than in group 2 (35.8 Gy) ( P = .05). The mean dose to the ipsilateral parotid was 35.0 Gy in group 1 vs 41.8 Gy in group 2 ( P = .05). No difference was seen in the severity of mucositis between groups ( P = .82). However, grade III mucositis was present in group 1 at 4 weeks after IMRT, 1 week after its occurrence in group 2. Conclusions A stent was effective in decreasing doses to healthy structures and delaying the emergence of mucositis.

Within-group repeated measures.To determine the effect of a semi-rigid prophylactic ankle stabilizer (PAS) on performance of subjects with post-acute, unilateral ankle sprains who have mechanically stable ankles, but are functionally impaired.Most studies on PAS performance to date are limited to subjects with noninjured, nonimpaired ankles. No research has been reported to determine the effect PASs have on performance in subjects who have a mechanically stable, nonacute ankle sprain with functional impairment.Twenty-five subjects (8 men and 17 women; average height = 168.91 +/- 33.02 cm, average weight = 61.10 +/- 29.5 kg, and average age = 16.2 +/- 6 years) met the qualification criteria of the study. Subjects had unilateral grade I or II lateral ankle sprains of 3 to 4 weeks duration and were cleared medically to return to activities of daily living. Each subject's injured ankle was mechanically stable as determined by physical exam but was functionally impaired as determined by instability during the modified Rhomberg test. Separate 2 x 2 ANOVAs with repeated measures on brace condition (Aircast SportStirrup and nonbraced control) and test session (test session 1 and test session 2) were completed to assess the effect of ankle stabilization on 2 dependent variables (36.58-m shuttle-run and vertical-jump).Shuttle-run time was significantly faster for the braced condition (mean = 9.43 +/- .72 seconds) than for the nonbraced control (mean = 9.57 +/- .75 seconds), regardless of test session. Shuttle-run time was significantly faster for test session 2 (mean = 9.43 +/- .79 seconds) than for test session 1 (mean = 9.57 +/- .68 seconds), regardless of brace condition. No significant main or interaction effects occurred for vertical-jump.Use of a semi-rigid PAS significantly increased shuttle-run but not vertical-jump performance. This effect was immediate and did not require PAS acclimation.

Background: Mycosis fungoides (MF) is rare in young patients. Its clinical behavior is still uncertain, as some reports have suggested that it has a more aggressive course than does the adult-onset type. Aim: To ascertain if early-onset MF represents a heterogeneous group of cutaneous T cell lymphomas. Materials and Methods: Clinical, immunohistopathological and follow-up data of early-onset (Results: The majority of the 49 early-onset MF patients had patch-plaque stage disease at diagnosis. Ten had hypopigmented lesions. The predominant phenotype was CD3+ CD4+CD7–CD8–. Seven patients had a stage progression, 6 with extracutaneous involvement. Five- and 10-year survival rates were 93 and 74%, respectively. Conclusions: No statistically significant differences were found in the disease course between early- and adult-onset MF.

Development of multiple primary melanomas is a rare but well recognized disease, with an estimated incidence ranging from 1.75% to 8.5% in several series. The clinical, histological and epidemiological characteristics of 49 patients, identified from 2470 with histologically confirmed melanoma, are described in this study. Thirty-five of these patients had two primary melanomas, 11 had three melanomas and three had four, five and six melanomas, respectively. Diagnosis was concurrent in 22 patients (45%); in the remaining cases the median time interval between the first and second melanoma was 22.6 months and the longest interval was 21.5 years. The mean Breslow's thickness decreased significantly (P0.001) from the first melanoma to the second and third lesion. The multiple melanoma patients had a higher percentage of subjects over 70 years of age or with lentigo maligna melanoma than single melanoma patients. The mean follow-up time was 12 years (range 4 23 years). The 5-year survival rate from first melanoma excision (83%) does not differ from that of patients with a single melanoma. In conclusion, the presence of multiple primary melanomas does not appear to be a negative prognostic factor; our data show the importance of close follow-up in melanoma patients in order to detect not only metastases, but also subsequent primaries in their earliest phases.

Summary CD56-positive (CD56+) lymphomas, characterized by the expression of the neural cell adhesion molecule on pathological lymphocytes, share a frequent extranodal involvement and a generally aggressive course. Five CD3- CD56+ lymphoma patients presenting with nodular lesions were identified among 180 immunophenotyped cutaneous lymphomas. All the patients were men, with ages ranging from 55 to 78 years. After staging, two patients were diagnosed as having primary cutaneous lymphomas: the remaining three had the secondary cutaneous type. The clinical course was aggressive and four patients died within 8 months from diagnosis. The remaining patient is still alive after a 17-month follow-up. The histological diagnosis was immunoblastic lymphoma in two patients, and medium and large cell pleomorphic lymphoma in three. The angiocentric infiltrate was located mainly in the dermis: azurophilic granules were present in three of the five patients. Immunogenotypic analyses suggested the natural killer cell origin of these neoplasias: all cases exhibited a CD56+ CD3- CD5- T-cell receptor (TCR) silent phenotype, and Southern blot analysis showed a germline configuration of the TCR β-chain gene.