Obtainment, selection and characterization of a mutant strain of Kluyveromyces marxianus that displays improved production of 2-phenylethanol and enhanced DAHP synthase activity.

AIMS: Yeasts produce 2-phenylethanol (2-PE) from sugars via de novo synthesis; however, its synthesis is limited due to feedback inhibition on the isofunctional 3-deoxy-d-arabino-heptulosonate-7-phosphate (DAHP) synthases (Aro3p and Aro4p). This work aimed to select Kluyveromyces marxianus mutant strains with improved capacity to produce 2-PE from sugars. METHODS AND RESULTS: Kluyveromyces marxianus CCT 7735 mutant strains were selected from UV irradiation coupled with screening of p-fluoro-dl-phenylalanine (PFP) tolerant strains on culture medium without l-Phe addition. Most of them produced 2-PE titres higher than the parental strain and the Km_PFP41 mutant strain stood out for displaying the highest 2-PE specific production rate. Moreover it showed higher activity of DAHP synthase than the parental strain. We sequenced both ARO3 and ARO4 genes of Km_PFP41 mutant and identified mutations in ARO4 which caused changes in both size and conformation of the Aro4p. These changes seem to be associated with the enhanced activity of DAHP synthase and improved production of 2-PE exhibited by that mutant strain. CONCLUSIONS: The Km_PFP41 mutant strain presented improved 2-PE production via de novo synthesis and enhanced DAHP synthase activity. SIGNIFICANCE AND IMPACT OF THE STUDY: The mutant strain obtained in this work may be exploited as a yeast cell factory for high-level synthesis of 2-PE.

Added sugar and trans fatty acid intake and sedentary behavior were associated with excess total-body and central adiposity in children and adolescents with congenital heart disease.

BACKGROUND: Over the past three decades, the prevalence rate of overweight and obesity has increased in survivors with congenital heart disease, and little is known about the body composition and its association with clinical characteristics and lifestyle factors. OBJECTIVES: To evaluate excess total-body adiposity and central adiposity and, to describe associated factors. METHODS: Cross-sectional study with children and adolescents who underwent procedure to treat congenital heart disease, from January to July 2017. Sociodemographic and clinical characteristics, and lifestyle factors (dietary intake, physical activity, and sedentary behavior) were assessed. Adiposity was assessed using air-displacement plethysmography and waist circumference. Factors associated with excess total-body adiposity and central adiposity were analyzed using logistic regression models. RESULTS: Of 232 patients, 22.4% were identified with excess total-body adiposity and 24.6% with central adiposity. Significant factors positively associated with excess total-body adiposity were intake of added sugar and trans fatty acids, adjusted for confounding factors. Similarly, lifestyle factors were positively associated with central adiposity: intake of added sugar and trans fatty acids, sedentary behavior, and family history of obesity. CONCLUSIONS: Lifestyle factors were associated with excess total-body adiposity and central adiposity. Assessment of body composition and healthy-lifestyle counseling into outpatient care may be the key point to prevent obesity in children and adolescents with congenital heart disease.

Children's perceptions regarding functional limitations due to oral problems.

AIM: To evaluate factors associated with difficulty eating and speaking due to oral problems according to the reports of children. METHODS: A cross-sectional study was conducted with 769 5-year-old children at preschools in a city in northeast Brazil. Parents/caretakers answered a questionnaire addressing socio-demographic data and a history of dental pain. The children answered the scale of oral health outcomes for 5-year-old children, which was used to evaluate the dependent variables. Clinical examinations of the children were performed by examiners who had undergone calibration exercises. Data analysis involved descriptive statistics and Poisson regression analysis (α = 5%). RESULTS: The prevalence of the perception of functional limitations on the part of the children was 35.5% for difficulty eating and 22.9% for difficulty speaking. Difficulty eating was associated with a history of dental pain (PR = 1.68; 95% CI 1.27-2.22) and the occurrence of traumatic dental injury (TDI) (PR = 1.23; 95% CI 1.01-1.52). Difficulty speaking due to oral problems was associated with studying at a public preschool (PR = 1.71; 95% CI 1.19-2.46), a history of dental pain (PR = 1.54; 95% CI 1.07-2.20), and the occurrence of TDI (PR = 1.52; 95% CI 1.12-2.06). CONCLUSIONS: Pain symptoms and the occurrence of TDI can exert an influence on the development of functional limitations. With regard to socioeconomic factors, studying at a public preschool can influence difficulty speaking in children.

Code of Ethics and Conduct of the Brazilian Association of Embryologists - PRONÚCLEO.

For more than three decades, Brazilian Clinical Embryologists have been working without specific regulations and following the standards adopted by other healthcare professionals. This document aims to guide behavior and decision-making, while providing directions to embryologist with the purpose of aiding professionals involved with assisted reproduction procedures and their patients. The Code of Ethics and Conduct is an important breakthrough and the first step toward regulating Clinical Embryology as a profession.

Accuracy of octa-polar bioelectrical impedance analysis for the assessment of total and appendicular body composition in children and adolescents with HIV: comparison with dual energy X-ray absorptiometry and air displacement plethysmography.

BACKGROUND: Body composition analysis has been used to investigate fat mass (FM) and bone mineral content (BMC) in children and adolescents diagnosed with HIV. Investigating the validity of bioelectrical impedance analysis (BIA) is interesting with respect to testing useful techniques for monitoring body composition in children and adolescents in clinical practice. The present study aimed to determine the validity of body composition analysis by BIA compared to dual-energy X-ray absorptiometry (DXA) and air displacement plethysmography (ADP) in children and adolescents an HIV diagnosis. METHODS: Sixty-four children and adolescents (35 females and 29 males) with a mean (SD) age of 12.22 (2.13) years and with an HIV diagnosis participated in the study. Fat-free mass (FFM), FM and body fat percentage (%BF) were obtained by BIA for comparison with DXA and ADP. Segmented FM (trunk, legs and arms), lean soft tissue mass (LSTM) (total and segmented) and BMC were obtained by BIA for comparison with DXA. RESULTS: BIA presented a clinically acceptable correlation with DXA and ADP for FFM. Values found by BIA were underestimated compared to ADP, and overestimated compared to DXA. BIA presented a clinically acceptable correlation with DXA for LSTM estimates (total and segmented parameters) in both sexes (underestimating FM and overestimating LSTM). For other components (%BF, FM and BMC), BIA had a clinically unacceptable correlation with the reference methods in both sexes. CONCLUSIONS: BIA was suitable for evaluating FFM and LSTM in children and adolescents with an HIV diagnosis. For FM, %BF and BMC, BIA was not suitable for performing an evaluation in both sexes.

Degradation of organic contaminants in effluents-synthetic and from the textile industry-by Fenton, photocatalysis, and H2O2 photolysis.

In this study, the oxidation of the dye rhodamine B (RhB), present in a synthetic effluent, and the degradation of organic matter present in a textile effluent, were assessed by photolysis (H2O2, UV), homogeneous Fenton (Fe2+, H2O2), and photocatalysis (TiO2, UV). The results showed that photolysis and Fenton had an efficiency of 100 % and photocatalysis, 96 %, to discoloration 10 mg L-1 RhB, present in the synthetic effluent. The best experimental conditions determined for these reactions showed that the one performed with 51 mg L-1 H2O2 and UV light had the best results, where 100 % of RhB was discolored in only 6 min of reaction. The optimum conditions determined in the first part of this study for the RhB oxidation did not show satisfactory results for the degradation of organic matter present in the textile effluent sample, and it was necessary to increase the amount of reagents in the three processes. After resizing the concentration of the reagents for the reactions with the textile effluent, the following reductions of color, total organic carbon (TOC), and total soluble solids (SS) were obtained: photocatalysis 29, 25, and 32 %; photolysis 85, 69, and 35 %; Fenton 98, 90, and 23 %; and biological (followed by physicochemical) treatment carried out by the textile industry 96, 48, and 9 %. It is observed that the Fenton reaction showed the best result, followed by photolysis reaction, a treatment carried out by industry and, at last, photocatalysis.

Development of a level A in vitro-in vivo correlation for extended release dosage forms of quetiapine fumarate.

Quetiapine is an atypical antipsychotic recommended as first-line treatment for acute bipolar depression. The extended-release quetiapine formulation is intended to be administered as an once-daily dosing. The development of an in vitro-in vivo correlation (IVIVC) and the use of in vitro data to predict in vivo bioavailability parameters has been of great interest for the rational development and evaluation process for extended release dosage forms. The aim of this study was to develop an IVIVC for quetiapine extended release formulation. In vitro dissolution rate data were obtained using USP apparatus 2 at 50 rpm, in 3 bio-relevant dissolution media with different pH values (1.2, 4.5 and 6.8). The drug release profiles of the 2 extended release dosage forms were compared using the similarity factor (f 2). The relative bioavailability of quetiapine was evaluated by a single-dose, randomized-sequence, open-label, 2 period cross over study with 16 healthy volunteers. A linear level A IVIVC model was established using percentage of absorbed and dissolved data obtained at pH 1.2. The developed IVIVC model was employed to predict quetiapine concentration-time profiles, as well as the bioequivalence parameters for test formulation. Percent prediction errors were estimated for Cmax and AUC to evaluate the validity of the correlation. The values did not exceed 15%, proving the predictability of the correlation model. In conclusion, the established level A IVIVC model proved to be an excellent tool for predicting the rate and extent of quetiapine absorption as characterized by Cmax and AUC for test formulation.

The trans-sialidase, the major Trypanosoma cruzi virulence factor: Three decades of studies.

Chagas' disease is a potentially life-threatening disease caused by the protozoan parasite Trypanosoma cruzi. Since the description of Chagas'disease in 1909 extensive research has identified important events in the disease in order to understand the biochemical mechanism that modulates T. cruzi-host cell interactions and the ability of the parasite to ensure its survival in the infected host. Exactly 30 years ago, we presented evidence for the first time of a trans-sialidase activity in T. cruzi (T. cruzi-TS). This enzyme transfers sialic acid from the host glycoconjugates to the terminal ß-galactopyranosyl residues of mucin-like molecules on the parasite's cell surface. Thenceforth, many articles have provided convincing data showing that T. cruzi-TS is able to govern relevant mechanisms involved in the parasite's survival in the mammalian host, such as invasion, escape from the phagolysosomal vacuole, differentiation, down-modulation of host immune responses, among others. The aim of this review is to cover the history of the discovery of T. cruzi-TS, as well as some well-documented biological effects encompassed by this parasite's virulence factor, an enzyme with potential attributes to become a drug target against Chagas disease.

Formulary availability and regulatory barriers to accessibility of opioids for cancer pain in Latin America and the Caribbean: a report from the Global Opioid Policy Initiative (GOPI).

The nations of the Caribbean, Central America and South America form a heterogeneous region with substantial variability in economic, social and palliative care development. Palliative care provision is at varied stages of development throughout the region. The consumption of opioids in Latin America and the Caribbean is variable with moderate levels of consumption by international standards (1-10 mg morphine equivalents/capita/year) observed in Argentine, Brazil, Chile, Colombia, Cuba, Mexico, Costa Rica, Uruguay and most of the Caribbean but relatively low levels of consumption in other countries particularly Guatemala, Honduras and Bolivia. Data for Latin American and Caribbean is reported on the availability and accessibility of opioids for the management of cancer pain in 24 of the 33 countries surveyed. The results of this survey are relevant to 560 million of the region's 595 million people (94%). Opioid availability continues to be low throughout most of Latin America and the Caribbean. While formularies in this region generally include all recommended morphine formulations, access is significantly impaired by widespread over-regulation that continues to be pervasive across the region.

Note: An ion source for alkali metal implantation beneath graphene and hexagonal boron nitride monolayers on transition metals.

The construction of an alkali-metal ion source is presented. It allows the acceleration of rubidium ions to an energy that enables the penetration through monolayers of graphene and hexagonal boron nitride. Rb atoms are sublimated from an alkali-metal dispenser. The ionization is obtained by surface ionization and desorption from a hot high work function surface. The ion current is easily controlled by the temperature of ionizer. Scanning Tunneling Microscopy measurements confirm ion implantation.

Assessment of using digital manipulation tools for diagnosing mandibular radiolucent lesions.

OBJECTIVE: The purpose of this study was to analyse the use of digital tools for image enhancement of mandibular radiolucent lesions and the effects of this manipulation on the percentage of correct radiographic diagnoses. METHODS: 24 panoramic radiographs exhibiting radiolucent lesions were selected, digitized and evaluated by non-experts (undergraduate and newly graduated practitioners) and by professional experts in oral diagnosis. The percentages of correct and incorrect diagnoses, according to the use of brightness/contrast, sharpness, inversion, highlight and zoom tools, were compared. All dental professionals made their evaluations without (T1) and with (T2) a list of radiographic diagnostic parameters. RESULTS: Digital tools were used with low frequency mainly in T2. The most preferred tool was sharpness (45.2%). In the expert group, the percentage of correct diagnoses did not change when any of the digital tools were used. For the non-expert group, there was an increase in the frequency of correct diagnoses when brightness/contrast was used in T2 (p=0.008) and when brightness/contrast and sharpness were not used in T1 (p=0.027). The use or non-use of brightness/contrast, zoom and sharpness showed moderate agreement in the group of experts [kappa agreement coefficient (κ) = 0.514, 0.425 and 0.335, respectively]. For the non-expert group there was slight agreement for all the tools used (κ ≤ 0.237). CONCLUSIONS: Consulting the list of radiographic parameters before image manipulation reduced the frequency of tool use in both groups of examiners. Consulting the radiographic parameters with the use of some digital tools was important for improving correct diagnosis only in the group of non-expert examiners.

Leishmanicidal effects of piperine, its derivatives, and analogues on Leishmania amazonensis.

Leishmaniasis is a tropical disease caused by protozoan parasites of the genus Leishmania which affects 12 million people worldwide. The discovery of drugs for the treatment of leishmaniasis is a pressing concern in global health programs. The aim of this study aim was to evaluate the leishmanicidal effect of piperine and its derivatives/analogues on Leishmania amazonensis. Our results showed that piperine and phenylamide are active against promastigotes and amastigotes in infected macrophages. Both drugs induced mitochondrial swelling, loose kinetoplast DNA, and led to loss of mitochondrial membrane potential. The promastigote cell cycle was also affected with an increase in the G1 phase cells and a decrease in the S-phase cells, respectively, after piperine and phenylamide treatment. Lipid analysis of promastigotes showed that piperine reduced triglyceride, diacylglycerol, and monoacylglycerol contents, whereas phenylamide only reduced diacylglycerol levels. Both drugs were deemed non toxic to macrophages at 50 µM as assessed by XTT (sodium 2,3,-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)-carbonyl]-2H-tetrazolium inner salt), Trypan blue exclusion, and phagocytosis assays, whereas low toxicity was noted at concentrations higher than 150 µM. None of the drugs induced nitric oxide (NO) production. By contrast, piperine reduced NO production in activated macrophages. The isobologram analysis showed that piperine and phenylamide acted synergistically on the parasites suggesting that they affect different target mechanisms. These results indicate that piperine and its phenylamide analogue are candidates for development of drugs for cutaneous leishmaniasis treatment.

Determination of pH turning point with pH mapping of the gastroesophageal junction: an alternative technique to orientate esophageal pH monitoring.

Manometric location of the lower esophageal sphincter (LES) has been mandatory before esophageal pH monitoring, despite costs and discomfort related with esophageal manometry. The aims of the study were: (i) to map the pH of the gastroesophageal junction (GEJ) to determine a pH turning point (PTP) and its relation with LES; and (ii) to test the feasibility of this technique to orientate esophageal pH monitoring. We studied 310 adult patients who underwent esophageal manometry and pH monitoring off acid-suppressive therapy. GEJ pH mapping was carried out by step-pulling the pH sensor from 5 cm below to 5 cm above LES, and a PTP was determined when pH changed from below to above 4, in centimeters from the nostril. Thirty-six patients referred only for pH monitoring were studied with pH sensor placed at 5 cm above the PTP. Out of 310 patients, a PTP was found in 293 (94.5%): inside LES in 86.3%, into the stomach in 8.2% and in the esophageal body in 5.5% of patients. The median distance between PTP and place where pH sensor monitored reflux was 8 cm. Among 36 patients who performed pH monitoring without LES manometry, there was no gastric monitoring during reflux testing. In adult patients investigated off acid suppressive therapy, GEJ pH mapping with reflux monitoring 5 cm above the PTP can be an alternative technique to perform esophageal pH monitoring when LES manometry is not available. Additional studies are needed before the widespread use of GEJ pH mapping in the clinical practice.

Barriers to the development of palliative care in Western Europe.

UNLABELLED: The Eurobarometer Survey of the EAPC Task Force on the Development of Palliative Care in Europe is part of a programme of work to produce comprehensive information on the provision of palliative care across Europe. AIM: To identify barriers to the development of palliative care in Western Europe. METHOD: A qualitative survey was undertaken amongst boards of national associations, eliciting opinions on opportunities for, and barriers to, palliative care development. By July 2006, 44/52 (85%) European countries had responded to the survey; we report here on the results from 22/25 (88%) countries in Western Europe. ANALYSIS: Data from the Eurobarometer Survey were analysed thematically by geographical region and by the degree of development of palliative care in each country. RESULTS: From the data contained within the Eurobarometer, we identified six significant barriers to the development of palliative care in Western Europe: (i) lack of palliative care education and training programmes; (ii) lack of awareness and recognition of palliative care; (iii) limited availability of/knowledge about opioid analgesics; (iv) limited funding; (v) lack of coordination amongst services; and (vi) uneven palliative care coverage. CONCLUSION: Findings from the EAPC Eurobarometer Survey suggest that barriers to the development of palliative care in Western Europe may differ substantially from each other in both their scope and context and that some may be considered to be of greater significance than others. A number of common barriers to the development of the discipline do exist and much work still remains to be done in the identified areas. This paper provides a road map of which barriers need to be addressed.

Performance of indirect immunofluorescence assay, immunochromatography assay and reverse transcription-polymerase chain reaction for detecting human respiratory syncytial virus in nasopharyngeal aspirate samples.

Comparison of the use of indirect immunofluorescence assay (IFA), immunochromatography assay (ICA-BD) and reverse transcription-polymerase chain reaction (RT-PCR) for detecting human respiratory syncytial virus (HRSV) in 306 nasopharyngeal aspirates samples (NPA) was performed in order to assess their analytical performance. By comparing the results obtained using ICA-BD with those using IFA, we found relative indices of 85.0% for sensitivity and 91.2% for specificity, and the positive (PPV) and negative (NPV) predictive values were 85.0% and 91.2%, respectively. The relative indices for sensitivity and specificity as well as the PPV and NPV for RT-PCR were 98.0%, 89.0%, 84.0% and 99.0%, respectively, when compared to the results of IFA. In addition, comparison of the results of ICA-BD and those of RT-PCR yielded relative indices of 79.5% for sensitivity and 95.4% for specificity, as well as PPV and NPV of 92.9% and 86.0%, respectively. Although RT-PCR has shown the best performance, the substantial agreement between the ICA-BD and IFA results suggests that ICA-BD, also in addition to being a rapid and facile assay, could be suitable as an alternative diagnostic screening for HRSV infection in children.

Performance of indirect immunofluorescence assay, immunochromatography assay and reverse transcription-polymerase chain reaction for detecting human respiratory syncytial virus in nasopharyngeal aspirate samples

Comparison of the use of indirect immunofluorescence assay (IFA), immunochromatography assay (ICA-BD) and reverse transcription-polymerase chain reaction (RT-PCR) for detecting human respiratory syncytial virus (HRSV) in 306 nasopharyngeal aspirates samples (NPA) was performed in order to assess their analytical performance. By comparing the results obtained using ICA-BD with those using IFA, we found relative indices of 85.0 percent for sensitivity and 91.2 percent for specificity, and the positive (PPV) and negative (NPV) predictive values were 85.0 percent and 91.2 percent, respectively. The relative indices for sensitivity and specificity as well as the PPV and NPV for RT-PCR were 98.0 percent, 89.0 percent, 84.0 percent and 99.0 percent, respectively, when compared to the results of IFA. In addition, comparison of the results of ICA-BD and those of RT-PCR yielded relative indices of 79.5 percent for sensitivity and 95.4 percent for specificity, as well as PPV and NPV of 92.9 percent and 86.0 percent, respectively. Although RT-PCR has shown the best performance, the substantial agreement between the ICA-BD and IFA results suggests that ICA-BD, also in addition to being a rapid and facile assay, could be suitable as an alternative diagnostic screening for HRSV infection in children.

Mechanical evaluation of a respiratory device.

This article aims to characterize the mechanical behaviour of the Flutter VRP1, a respiratory physiotherapy device designed to aid sputum clearance of the airways of patients. The device resembles a smoking pipe with a conical cavity containing a stainless steel sphere which floats up and down while the patient comes with a forced expiration through it. The sphere's oscillatory movement is function of the air flow rate and angular orientation of the device. When the sphere's oscillatory frequency matches the natural frequency of the patient's chestwall+abdomen system, it will produce resonance which, in turn, will enhance sputum clearance. A dynamical model of the Flutter was formulated and an experimental setup was assembled in order to study the oscillatory frequency of the sphere under different conditions of air flow rate, fluid pressure, angular orientation and sphere's material and weight. Interesting results presented by this article point to eventual mechanical optimization of the device and show information that could be beneficial to the professional of the respiratory physiotherapy.

[Optimization of implant surfaces: titanium plasma spray and acid-etched sandblasting -- current status]. / Otimização das superfícies dos implantes: plasma de titânio e jateamento com areia condicionado por ácido--estado atual.

The aim of this review was to update the concepts regarding the preparation of the surfaces of titanium implants, focusing on TPS (titanium plasma-sprayed implants) and SLA (sandblasted and acid etched implants). Texture was the most remarkable isolated feature, regarded as an osseointegration promoter. In a comprehensive review of the effects of implant surface topography on cell behavior, one can verify that there is bone apposition onto the implant surface regardless of its characteristics: polished or rough, made of titanium or ceramic. Roughness is not mandatory for bone apposition. However, it has been shown that roughness may play an important role in the percentage of bone apposition as well as in the velocity of apposition. In this review, a quite promising type of surface called SLA is pointed out, showing that either roughness or acid conditioning of the surfaces can significantly improve shear strength. Besides optimizing the procedure, these surface characteristics may, eventually, allow for an earlier loading of the implant and extend the indications for implants in low-density alveolar bone and in regenerated bone.

Influence of the pineal gland on the physiology, morphometry and morphology of pancreatic islets in rats.

To investigate the influence of the pineal gland through melatonin secretion on the physiological and morphological parameters of pancreatic islets, we studied the plasma biochemistry and morphological and morphometric characteristics of the endocrine pancreas of male Wistar rats. The animals were distributed into five groups of ten rats each: NC - normal control group; NS -- sham-operated group; Px (25) -- pinealectomised group, studied 15--25 days after surgery; Px (70) -- pinealectomised group, studied 60-70 days after surgery; ALX - alloxan monohydrate-treated group. Data are analyzed statistically by ANOVA and by the Kruskal-Wallis test. Although there was no significant difference in plasma glucose or insulin levels between the Px (25), Px (70) and NC groups, Px (25) animals showed a tendency to increased glucose and reduced insulin levels. The ALX group showed a clear elevation of plasma glucose and a reduction of plasma insulin compared to the other groups. Morphometric analysis showed a larger pancreatic islet area and a lower pancreatic islet density in the pancreas of Px (70) animals and an increase in degenerative pathological processes in the pancreatic islets of the Px (25) and ALX groups. The present results suggest that melatonin, in addition to acting on tissue sensitivity to insulin (as reported in other studies), affects the secretory action of beta cells, as demonstrated by the morphological and morphometric changes observed in pinealectomised animals.