Interaction of cationic water-soluble meso-tetrakis(4-N-methylpyridiniumyl)porphyrin (TMPyP) with ionic and nonionic micelles: aggregation and binding

The equilibrium of meso-tetrakis(4-N-methylpyridiniumyl)porphyrin (TMPyP) in aqueous solution in the presence of surfactants was studied by optical spectroscopic techniques and SAXS (small angle X-ray scattering). Anionic SDS (sodium dodecyl sulfate), zwitterionic HPS (N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate) and nonionic TRITON X-100 (t-octyl-phenoxypolyethoxyethanol), surfactants were used. TMPyP is characterized by a protonation equilibrium with a pK(a) around 1.0, associated with the diacid-free base transition, and a second pK(a) around 12.0 related with the transition between the free base and the monoanion form. Three independent species were observed for TMPyP at pH 6.0 as a function of SDS concentration: free TMPyP, TMPyP-SDS aggregates and porphyrin monomer bound to micelles. For HPS and TRITON X-100, the equilibrium of TMPyP as a function of pH is quite similar to that obtained in pure aqueous solution: no aggregation was observed, suggesting that electrostatic contribution is the major factor in the interaction between TMPyP and surfactants. SAXS data analysis demonstrated a prolate ellipsoidal shape for SDS micelles; no significant changes in shape and size were observed for SDS-TMPyP co-micelles. Moreover, the ionization coefficient, alpha, decreases with the increase of the porphyrin concentration, suggesting the ""screening"" of the anionic charge of SDS by the cationic porphyrin. These results are consistent with optical absorption, fluorescence and RLS (resonance light scattering) spectroscopies data, allowing to conclude that neutral surfactants present a smaller interaction with the cationic porphyrin as compared with an ionic surfactant. Therefore, the interaction of TMPyP with the ionic and nonionic surfactants is predominantly due to the electrostatic contribution. Copyright (c) 2008 Society of Porphyrins & Phthalocyanines.

Prediction of apparent protein digestibility of ingredients and diets by in vitro pH-stat degree of protein hydrolysis with species-specific enzymes for juvenile Pacific white shrimp Litopenaeus vannamei

Aquafeed production faces global issues related to availability of feed ingredients. Feed manufacturers require greater flexibility in order to develop nutritional and cost-effective formulations that take into account nutrient content and availability of ingredients. The search for appropriate ingredients requires detailed screening of their potential nutritional value and variability at the industrial level. In vitro digestion of feedstuffs by enzymes extracted from the target species has been correlated with apparent protein digestibility (APD) in fish and shrimp species. The present study verified the relationship between APD and in vitro degree of protein hydrolysis (DH) with Litopenaeus vannamei hepatopancreas enzymes in several different ingredients (n = 26): blood meals, casein, corn gluten meal, crab meal, distiller`s dried grains with solubles, feather meal, fish meals, gelatin, krill meals, poultry by-product meal, soybean meals, squid meals and wheat gluten. The relationship between APD and DH was further verified in diets formulated with these ingredients at 30% inclusion into a reference diet. APD was determined in vivo (30.1 +/- 0.5 degrees C, 32.2 +/- 0.4%.) with juvenile L vannamei (9 to 12 g) after placement of test ingredients into a reference diet (35 g kg(-1) CP: 8.03 g kg(-1) lipid; 2.01 kcal g(-1)) with chromic oxide as the inert marker. In vitro DH was assessed in ingredients and diets with standardized hepatopancreas enzymes extracted from pond-reared shrimp. The DH of ingredients was determined under different assay conditions to check for the most suitable in vitro protocol for APD prediction: different batches of enzyme extracts (HPf5 or HPf6), temperatures (25 or 30 degrees C) and enzyme activity (azocasein): crude protein ratios (4 U: 80 mg CP or 4 U: 40 mg CP). DH was not affected by ingredient proximate composition. APD was significantly correlated to DH in regressions considering either ingredients or diets. The relationships between APD and DH of the ingredients could be suitably adjusted to a Rational Function (y = (a + bx)/(1 + cx + dx2), n = 26. Best in vitro APD predictions were obtained at 25 degrees C, 4 U: 80 mg CP both for ingredients (R(2) = 0.86: P = 0.001) and test diets (R(2) = 0.96; P = 0.007). The regression model including all 26 ingredients generated higher prediction residuals (i.e., predicted APD - determined APD) for corn gluten meal, feather meal. poultry by-product meal and krill flour. The remaining test ingredients presented mean prediction residuals of 3.5 points. A model including only ingredients with APD>80% showed higher prediction precision (R(2) = 0.98: P = 0.000004; n = 20) with average residual of 1.8 points. Predictive models including only ingredients from the same origin (e.g., marine-based, R(2) = 0.98; P = 0.033) also displayed low residuals. Since in vitro techniques have been usually validated through regressions against in vivo APD, the DH predictive capacity may depend on the consistency of the in vivo methodology. Regressions between APD and DH suggested a close relationship between peptide bond breakage by hepatopancreas digestive proteases and the apparent nitrogen assimilation in shrimp, and this may be a useful tool to provide rapid nutritional information. (C) 2009 Elsevier B.V. All rights reserved.

Complex networks analysis of language complexity

Methods from statistical physics, such as those involving complex networks, have been increasingly used in the quantitative analysis of linguistic phenomena. In this paper, we represented pieces of text with different levels of simplification in co-occurrence networks and found that topological regularity correlated negatively with textual complexity. Furthermore, in less complex texts the distance between concepts, represented as nodes, tended to decrease. The complex networks metrics were treated with multivariate pattern recognition techniques, which allowed us to distinguish between original texts and their simplified versions. For each original text, two simplified versions were generated manually with increasing number of simplification operations. As expected, distinction was easier for the strongly simplified versions, where the most relevant metrics were node strength, shortest paths and diversity. Also, the discrimination of complex texts was improved with higher hierarchical network metrics, thus pointing to the usefulness of considering wider contexts around the concepts. Though the accuracy rate in the distinction was not as high as in methods using deep linguistic knowledge, the complex network approach is still useful for a rapid screening of texts whenever assessing complexity is essential to guarantee accessibility to readers with limited reading ability. Copyright (c) EPLA, 2012

Linker for Activation of T-cell Family Member2 (LAT2) a Lipid Raft Adaptor Protein for AKT Signaling, Is an Early Mediator of Alkylphospholipid Anti-leukemic Activity

Lipid rafts are highly ordered membrane domains rich in cholesterol and sphingolipids that provide a scaffold for signal transduction proteins; altered raft structure has also been implicated in cancer progression. We have shown that 25 mu M 10-(octyloxy) decyl-2-(trimethylammonium) ethyl phosphate (ODPC), an alkylphospholipid, targets high cholesterol domains in model membranes and induces apoptosis in leukemia cells but spares normal hematopoietic and epithelial cells under the same conditions. We performed a quantitative (SILAC) proteomic screening of ODPC targets in a lipid-raft-enriched fraction of leukemic cells to identify early events prior to the initiation of apoptosis. Six proteins, three with demonstrated palmitoylation sites, were reduced in abundance. One, the linker for activation of T-cell family member 2 (LAT2), is an adaptor protein associated with lipid rafts in its palmitoylated form and is specifically expressed in B lymphocytes and myeloid cells. Interestingly, LAT2 is not expressed in K562, a cell line more resistant to ODPC-induced apoptosis. There was an early loss of LAT2 in the lipid-raft-enriched fraction of NB4 cells within 3 h following treatment with 25 mu M ODPC. Subsequent degradation of LAT2 by proteasomes was observed. Twenty-five mu M ODPC inhibited AKT activation via myeloid growth factors, and LAT2 knockdown in NB4 cells by shRNA reproduced this effect. LAT2 knockdown in NB4 cells also decreased cell proliferation and increased cell sensitivity to ODPC (7.5X), perifosine (3X), and arsenic trioxide (8.5X). Taken together, these data indicate that LAT2 is an early mediator of the anti-leukemic activity of alkylphospholipids and arsenic trioxide. Thus, LAT2 may be used as a target for the design of drugs for cancer therapy. Molecular & Cellular Proteomics 11: 10.1074/mcp.M112.019661, 1898-1912, 2012.

Limitations in activities of people affected by leprosy after completing multidrug therapy: application of the SALSA scale

Objectives: To identify people affected by leprosy with impairments after completing multidrug therapy for leprosy, and to assess their limitations in conducting daily activities by applying the Screening of Activity Limitation and Safety Awareness (SALSA) scale. Methods: A cross-sectional study was performed of all residents of a medium-sized city who were treated for leprosy from 1998 to 2006. A specific questionnaire was applied to obtain general and clinical data and the SALSA scale was used to assess limitations in activities. Impairments were assessed using the 'World Health Organization leprosy disability grading system' (WHO-DG). Findings: Of the 335 people affected by leprosy treated in the period, 223 (62.1%) were located and interviewed. A total of 51.6% were female with a mean age of 54 years (SD +/- 15.72) and 67.9% had up to 6 years formal education. The borderline form predominated among interviewees (39.9%) and 54.3% suffer from associated diseases with hypertension (29.1%) and diabetes (10.3%) being the most common. Pain was reported by 54.7% of interviewees. By multiple logistic regression analysis, associations were found between limitations in activities and being female (P < 0.025), family income <= 3 minimum wages (P-value < 0.003), reports of major lesions (P-value < 0.004), pain (P-value < 0.001), associated diseases (P-value < 0.023) and the WHO-DG (P-value < 0.001). Disabilities, as identified using the WHO-DG, were less common (32%) than limitations in activities as evaluated by the SALSA scale (57.8%). Conclusion: Limitations in activities proved to be common in people affected by leprosy and were. associated with low income, being female, reported major lesions, disability, disease and pain.

High-Performance Liquid Chromatography Under Partially Denaturing Conditions (dHPLC) is a Fast and Cost-Effective Method for Screening Molecular Defects: Four Novel Mutations Found in X-Linked Chronic Granulomatous Disease

Implementing precise techniques in routine diagnosis of chronic granulomatous disease (CGD), which expedite the screening of molecular defects, may be critical for a quick assumption of patient prognosis. This study compared the efficacy of single-strand conformation polymorphism analysis (SSCP) and high-performance liquid chromatography under partially denaturing conditions (dHPLC) for screening mutations in CGD patients. We selected 10 male CGD patients with a clinical history of severe recurrent infections and abnormal respiratory burst function. gDNA, mRNA and cDNA samples were prepared by standard methods. CYBB exons were amplified by PCR and screened by SSCP or dHPLC. Abnormal DNA fragments were sequenced to reveal the nature of the mutations. The SSCP and dHPLC methods showed DNA abnormalities, respectively, in 55% and 100% of the cases. Sequencing of the abnormal DNA samples confirmed mutations in all cases. Four novel mutations in CYBB were identified which were picked up only by the dHPLC screening (c.904 insC, c.141+5 g>t, c.553 T>C, and c.665 A>T). This work highlights the relevance of dHPLC, a sensitive, fast, reliable and cost-effective method for screening mutations in CGD, which in combination with functional assays assessing the phagocyte respiratory burst will contribute to expedite the definitive diagnosis of X-linked CGD, direct treatment, genetic counselling and to have a clear assumption of the prognosis. This strategy is especially suitable for developing countries.

Artificial and natural radioactivity in edible mushrooms from Sao Paulo, Brazil

Environmental biomonitoring has demonstrated that organisms such as crustaceans, fish and mushrooms are useful to evaluate and monitor both ecosystem contamination and quality. Particularly, some mushroom species have a high capacity to retain radionuclides and some toxic elements from the soil and the air. The potential of mushrooms to accumulate radionuclides in their fruit-bodies has been well documented. However, there are no studies that determine natural and artificial radionuclide composition in edible mushrooms, in Brazil. Artificial (Cs-137) and natural radioactivity (K-40. Ra-226. Ra-228) were determined in 17 mushroom samples from 3 commercialized edible mushroom species. The edible mushrooms collected were Agaricus sp., Pleurotus sp. and Lentinula sp. species. The activity measurements were carried out by gamma spectrometry. The levels of Cs-137 varied from 1.45 +/- 0.04 to 10.6 +/- 0.3 Bq kg(-1), K-40 levels varied from 461 +/- 2 to 1535 +/- 10 Bq kg(-1), Ra-226 levels varied from 14 +/- 3 to 66 +/- 12 Bq kg(-1) and Ra-228 levels varied from 6.2 +/- 0.2 to 54.2 +/- 1.7 Bq kg(-1). Cs-137 levels in Brazilian mushrooms are in accordance with the radioactive fallout in the Southern Hemisphere. The artificial and natural activities determined in this study were found to be below the maximum permissible levels as established by national legislation. Thus, these mushroom species can be normally consumed by the population without any apparent risks to human health.

Selection of Top 10 Quality Indicators for Nutrition Therapy

Background: The identification of useful quality indicators for nutrition therapy (QINTs) is of great interest and a challenge. This study attempted to identify the 10 QINTs that best suit the practice of quality control in nutrition therapy (NT) by evaluating the opinion of experts in NT with the use of psychometric techniques and statistical tools. Methods: Thirty-six QINTs available for clinical application in Brazil were assessed in 2 distinct phases. In phase 1, 26 nutrition experts ranked QINTs by scoring 4 attributes (utility, simplicity, objectivity, low cost) to assess each QINT on a 5-point Likert scale. The top 10 QINTs were identified from the 10 best scores obtained, and the reliability of expert opinion for each indicator was assessed by Cronbach's alpha. In phase 2, experts provided feedback regarding the selected top 10 QINTs by answering 2 closed questions. Results: The top 10 QINTs, in descending order, are the frequency of nutrition screening of hospitalized patients, diarrhea, involuntary withdrawal of enteral feeding tubes, feeding tube obstruction, fasting longer than 24 hours, glycemic dysfunction, estimated energy expenditure and protein needs, central venous catheter infection, compliance of NT indication, and frequency of application of subjective global assessment. Opinions were consistent among the interviewed experts. During feedback, 96% of experts were satisfied with the top 10 QINTs, and 100% had considered them in accordance with their previous opinion. Conclusion: The top 10 QINTs were identified according to their usefulness in clinical practice by obtaining adequate agreement and representativeness of opinion of nutrition experts. (Nutr Clin Pract. 2012;27:261-267)

A study of pent-up demand in ophthalmology: Divinolandia Hospital/Unicamp

Objective: To assess the waiting time for eye care identifying the number of patients with each complaint; to investigate how the waiting time may worsen the patient's condition; to check the screening of urgent cases for effectiveness; and to devise means of increasing the medical-surgical care capacity. Methods: A retrospective descriptive survey was conducted using data obtained on 12 occasions during collaborative team visits to provide eyecare services. These initiatives were designed to decrease the waiting time and to treat urgent cases that occurred on each occasion; eyecare services were provided every Saturday, in the period from June to August 2006, in 16 cities of the region covered by Conderg (Consortium for the Development of the Sao Joao da Boa Vista Administrative Region). Results: Referrals used 1,743 (87.1%) of the 2,000 places available. The most frequent diagnoses were refractive errors, with 683 cases, corresponding to 39.1% of the total, followed by cataracts, with 296 cases, corresponding to 20.9%. Of the 238 surgeries indicated, 54.6% were phakectomies. Thirty-five (2.0%) cases were considered urgent. Conclusion: The most common diagnoses made during the team visits to manage the excess demand for eyecare were refractive errors and cataracts, which, together, accounted for the majority of the cases. The Divinolandia Hospital has the necessary human and material resources to meet the demand left unattended by the local SUS network. Immediate referral of urgent cases by the primary units' screeners proved effective.

Seroprevalence and isolation of Toxoplasma gondii from free-range chickens from Espirito Santo state, southeastern Brazil

Prevalence of Toxoplasma gondii infection in 510 free-range (FR) chickens (380 from 33 small farms, and 130 from a slaughter house for FR chickens) from Espirito Santo state, southeastern Brazil, was investigated. Antibodies to T. gondii were sought using commercial indirect haemagglutination (IHAT, Imuno-HAI Toxo (R), Wama Diagnostica, Sao Paulo, Brazil, cut-off 1:16) and the modified agglutination test (MAT, cut-off 1:25) tests. Attempts were made to isolate viable T. gondii from seropositive chickens by bioassay in mice. Pooled samples of brain, heart and quadriceps muscle of one thigh (total 40 g) from 64 chickens with IHAT titers of >= 1:16 were minced, digested in pepsin and bioassayed in mice. Antibodies to T. gondii were found in 40.4% (206/510) FR chickens by IHAT (titer >= 1:16) and 38.8% (198/510) by MAT (titer >= 1:25); concordance between IHAT and MAT was 81.6% (kappa index = 0.614). Viable T. gondii was isolated (designated TgCkBr234-281) from 48 of 64(75%) seropositive (IHAT titers >= 1:32) FR chickens. Most isolates of T. gondii were virulent for mice; 100% of mice inoculated with 44 of 48 isolates died of toxoplasmosis within 30 days post inoculation (p.i). An epidemiological investigation revealed that people living in rural areas have little knowledge about the parasite and about the risk of acquiring it from raw meat. Results indicated that the locally available IHAT was useful for screening of chicken sera for T. gondii antibodies. (C) 2012 Elsevier B.V. All rights reserved.

Recipient of kidney from donor with asymptomatic infection by Paracoccidioides brasiliensis

The increase in solid organ transplantations may soon create a rise in the occurrence of endemic fungal diseases, such as paracoccidioidomycosis, due to the lack of rigorous screening of donors from endemic areas. Here we present the first case of an immunocompetent and asymptomatic kidney donor who had Paracoccidioides brasiliensis infected-adrenal tissue but no glandular dysfunction.

Clinical and sociodemographic factors in a sample of older subjects experiencing depressive symptoms

Objectives This study aims to determine the frequency of clinically significant depressive symptoms (CSDS) in a community sample of older Brazilians and to examine their relationship with sociodemographic factors, cognitive and functional impairment (CFI), and medical illness. Methods A total of 1145 subjects aged 60?years or older living in the City of Ribeirao Preto, State of Sao Paulo, Brazil, were interviewed. The following instruments were used: a 10-item scale for screening of depressive symptoms in older people, the mini mental state examination, the Fuld Object Memory Evaluation, the Informant Questionnaire on Cognitive Decline in the Elderly, the Bayer Activities of Daily Living Scale, and a sociodemographic and clinical questionnaire. Results The frequency of CSDS was 15.7%. Logistic regression analysis indicated that being previously depressed, having CFI, having lower level of education, using psychotropics, and not engaging in physical exercise were related to CSDS. On the other hand, being a woman, older, medically ill, employed, or married was not associated with CSDS. Conclusions Consistent with previous reports, lower education, lack of physical activity, and CFI were significantly associated with higher frequencies of CSDS. Further investigations are necessary to clarify the occurrence of depression and possible modifiable factors in developing countries such as Brazil. Copyright (C) 2011 John Wiley & Sons, Ltd.

Pseudopolymorphism in hydroxybenzophenones: the dihydrate of 2,2 ',4,4 '-tetrahydroxybenzophenone

A dihydrate pseudopolymorph of bis(2,4-dihydroxyphenyl)methanone, C13H10O5 center dot 2H2O, (I), was obtained during polymorphism screening of hydroxybenzophenone derivatives. This structure, in which the molecule sits on a twofold axis, was compared with the known anhydrous form of (I) [Schlemper (1982). Acta Cryst. B38, 554559]. The role of water in the crystal assembly was established on the basis of the known monohydrate pseudopolymorph of 3,4-dihydroxybenzophenone [Landre, Souza, Correa, Martins & Doriguetto (2010). Acta Cryst. C66, o463o465].

A study of pent-up demand in ophthalmology: Divinolândia Hospital/Unicamp

OBJECTIVE: To assess the waiting time for eye care identifying the number of patients with each complaint; to investigate how the waiting time may worsen the patient's condition; to check the screening of urgent cases for effectiveness; and to devise means of increasing the medical-surgical care capacity. METHODS: A retrospective descriptive survey was conducted using data obtained on 12 occasions during collaborative team visits to provide eyecare services. These initiatives were designed to decrease the waiting time and to treat urgent cases that occurred on each occasion; eyecare services were provided every Saturday, in the period from June to August 2006, in 16 cities of the region covered by Conderg (Consortium for the Development of the São João da Boa Vista Administrative Region). RESULTS: Referrals used 1,743 (87.1%) of the 2,000 places available. The most frequent diagnoses were refractive errors, with 683 cases, corresponding to 39.1% of the total, followed by cataracts, with 296 cases, corresponding to 20.9%. Of the 238 surgeries indicated, 54.6% were phakectomies. Thirty-five (2.0%) cases were considered urgent. CONCLUSION: The most common diagnoses made during the team visits to manage the excess demand for eyecare were refractive errors and cataracts, which, together, accounted for the majority of the cases. The Divinolândia Hospital has the necessary human and material resources to meet the demand left unattended by the local SUS network. Immediate referral of urgent cases by the primary units' screeners proved effective.

Trypanocidal activity of Brazilian plants against epimastigote forms from Y and Bolivia strains of Trypanosoma cruzi

Chagas disease is one of the main public health problems in Latin America. Since the available treatments for this disease are not effective in providing cure, the screening of potential antiprotozoal agents is essential, mainly of those obtained from natural sources. This study aimed to provide an evaluation of the trypanocidal activity of 92 ethanol extracts from species belonging to the families Annonaceae, Apiaceae, Cucurbitaceae, Lamiaceae, Lauraceae, Moraceae, Nyctaginaceae, and Verbenaceae against the Y and Bolivia strains of Trypanosoma cruzi. Additionally, cytotoxic activity on LLCMK2 fibroblasts was evaluated. Both the trypanocidal activity and cytotoxicity were evaluated using the MTT method, in the following concentrations: 500, 350, 250, and 100 µg/mL. Benznidazole was used for positive control. The best results among the 92 samples evaluated were obtained with ethanol extracts of Ocotea paranapiacabensis (Am93) and Aegiphila lhotzkiana (Am160). Am93 showed trypanocidal activity against epimastigote forms of the Bolivia strain and was moderately toxic to LLCMK2 cells, its Selectivity Index (SI) being 14.56, while Am160 showed moderate trypanocidal activity against the Bolivia strain and moderate toxicicity, its SI being equal to 1.15. The screening of Brazilian plants has indicated the potential effect of ethanol extracts obtained from Ocotea paranapiacabensis and Aegiphila lhotzkiana against Chagas disease.