Protein recovery, separation and purification: selection of optimal techniques using an expert system

The paper discusses the utilization of new techniques ot select processes for protein recovery, separation and purification. It describesa rational approach that uses fundamental databases of proteins molecules to simplify the complex problem of choosing high resolution separation methods for multi component mixtures. It examines the role of modern computer techniques to help solving these questions.

The Influence of Self-fertilization performance and Copulation Behaviour in Reproduction by Cross-fertilization in Groups of Biomphalaria tenagophila (Mollusca, Planorbidae)

The following hypotheses were tested for groups of simultaneous hermaphrodites Biomphalaria tenagophila: (a) snails that have low reproductive success during the process of self-fertilization do not increase their reproductive success after the end of grouping; (b) the copulation behaviour and the presence of one snail whose eggs have a low viability rate influence the partner's reproductive success by cross-fertilization. Groups were constituted by a homozygous pigmented snail and two albinos: one with a viability rate higher than 70% ("good reproducers") and the other less than 10% ("bad reproducers"). All pigmented snails had viability rates higher than 70%. The "good" and "bad" reproducer albino snails had similar copulation behaviour. However, after the end of grouping, the "bad reproducers" continued to have viability rates less than 10% over 30 days. In 100% of the cases that pigmented snails copulated (performing either a male role or simultaneously male and female roles) exclusively with "good" reproducer albinos, they presented high reproductive success (producing, on average of 8.4 pigmented embryos/egg-mass). However, in 100% of the cases that pigmented snails copulated with both partners, the "good" reproducer albino snails produced none or very few embryos (the highest average was 2.2 pigmented embryos/egg-mass). Therefore, the production of viable embryos by cross-fertilization was more influenced by self-fertilization performance than by copulation behaviour. The presence of a snail whose eggs have a low viability rate could decrease their partners reproductive success

Purification and Partial Characterization of Trypanosoma cruzi Triosephosphate Isomerase

The enzyme triosephosphate isomerase (TPI, EC 5.3.1.1) was purified from extracts of epimastigote forms of Trypanosoma cruzi. The purification steps included: hydrophobic interaction chromatography on phenyl-Sepharose, CM-Sepharose, and high performance liquid gel filtration chromatography. The CM-Sepharose material contained two bands (27 and 25 kDa) with similar isoelectric points (pI 9.3-9.5) which could be separated by gel filtration in high performance liquid chromatography. Polyclonal antibodies raised against the porcine TPI detected one single polypeptide on western blot with a molecular weight (27 kDa) identical to that purified from T. cruzi. These antibodies also recognized only one band of identical molecular weight in western blots of several other trypanosomatids (Blastocrithidia culicis, Crithidia desouzai, Phytomonas serpens, Herpertomonas samuelpessoai). The presence of only one enzymatic form of TPI in T. cruzi epimastigotes was confirmed by agarose gel activity assay and its localization was established by immunocytochemical analysis. The T. cruzi purified TPI (as well as other trypanosomatid' TPIs) is a dimeric protein, composed of two identical subunits with an approximate mw of 27,000 and it is resolved on two dimensional gel electrophoresis with a pI of 9.3. Sequence analysis of the N-terminal portion of the 27 kDa protein revealed a high homology to Leishmania mexicana and T. brucei proteins

Purification and biochemical characterization of four iron superoxide dismutases in Trypanosoma cruzi

Four superoxide dismutase (SOD) activities (SOD I, II, III, and IV) have been characterized in the epimastigote form of Trypanosoma cruzi. The total extract was subjected to two successive ammonium sulphate additions between 35 and 85%, and the resulting fraction was purified using two continuous chromatography processes (ion exchange and filtration). Enzymes were insensitive to cyanide but sensitive to hydrogen peroxide, properties characteristic of iron-containing SODs. The molecular masses of the different SODs were 20 kDa (SOD I), 60 kDa (SOD II), 50 kDa (SOD III) and 25 kDa (SOD IV), whereas the isoelectric points were 6.9, 6.8, 5.2 and 3.8, respectively. Subcellular location and digitonin experiments have shown that these SODs are mainly cytosolic, with small amounts in the low-mass organelles (SOD II and SOD I) and the mitochondrion (SOD III), where these enzymes play an important role in minimizing oxidative damage.

Purification and biochemica characterisation of endoplasmic reticulum α 1,2-mannosidase from Sporothrix schenckiil

Alpha 1,2-mannosidases from glycosyl hydrolase family 47 participate in N-glycan biosynthesis. In filamentous fungi and mammalian cells, α1,2-mannosidases are present in the endoplasmic reticulum (ER) and Golgi complex and are required to generate complex N-glycans. However, lower eukaryotes such Saccharomyces cerevisiae contain only one α1,2-mannosidase in the lumen of the ER and synthesise high-mannose N-glycans. Little is known about the N-glycan structure and the enzyme machinery involved in the synthesis of these oligosaccharides in the dimorphic fungus Sporothrix schenckii. Here, a membrane-bound α-mannosidase from S. schenckii was solubilised using a high-temperature procedure and purified by conventional methods of protein isolation. Analytical zymograms revealed a polypeptide of 75 kDa to be responsible for enzyme activity and this purified protein was recognised by anti-α1,2-mannosidase antibodies. The enzyme hydrolysed Man9GlcNAc2 into Man8GlcNAc2 isomer B and was inhibited preferentially by 1-deoxymannojirimycin. This α1,2-mannosidase was localised in the ER, with the catalytic domain within the lumen of this compartment. These properties are consistent with an ER-localised α1,2-mannosidase of glycosyl hydrolase family 47. Our results also suggested that in contrast to other filamentous fungi, S. schenckii lacks Golgi α1,2-mannosidases and therefore, the processing of N-glycans by α1,2-mannosidases is similar to that present in lower eukaryotes.

Good agreements between self and clinician-collected specimens for the detection of human papillomavirus in Brazilian patients

Women infected with human papillomavirus (HPV) are at a higher risk of developing cervical lesions. In the current study, self and clinician-collected vaginal and cervical samples from women were processed to detect HPV DNA using polymerase chain reaction (PCR) with PGMY09/11 primers. HPV genotypes were determined using type-specific PCR. HPV DNA detection showed good concordance between self and clinician-collected samples (84.6%; kappa = 0.72). HPV infection was found in 30% women and genotyping was more concordant among high-risk HPV (HR-HPV) than low-risk HPV (HR-HPV). HPV16 was the most frequently detected among the HR-HPV types. LR-HPV was detected at a higher frequency in self-collected; however, HR-HPV types were more frequently identified in clinician-collected samples than in self-collected samples. HPV infections of multiple types were detected in 20.5% of clinician-collected samples and 15.5% of self-collected samples. In this study, we demonstrated that the HPV DNA detection rate in self-collected samples has good agreement with that of clinician-collected samples. Self-collected sampling, as a primary prevention strategy in countries with few resources, could be effective for identifying cases of HR-HPV, being more acceptable. The use of this method would enhance the coverage of screening programs for cervical cancer.

Critical thinking and accuracy of nurses' diagnoses. Part II: application of cognitive skills and guidelines for self-development

Part I of this article, the author explained the difficulties of achieving accuracy of nurses' diagnoses, the relevance of critical thinking to the achievement of accuracy, and newer views of critical thinking. In Part II, the critical thinking dimensions identified as important for nursing practice are applied in the diagnostic process using a case study of a 16 year old girl with type 1 diabetes. Application of seven cognitive skills and ten habits of mind illustrate the importance of using critical thinking for accuracy of nurses' diagnoses. Ten strategies are proposed for self-development of critical thinking abilities.

Ability for self-care and its association with sociodemographic factors of people with spinal cord injury

This study aimed to verify the association between self-care ability and sociodemographic factors of people with spinal cord injury (SCI). It was a cross-sectional study, conducted in 2012, in all 58 Basic Health Units of Natal/RN, Brazil. Seventy-three subjects completed a sociodemographic form andSelf-Care Agency Scale. Statistical analyses were performed using SPSS,including Cronbach’s Alpha, Chi-square, Fisher’s and contingency coefficient tests. The Cronbach's alpha was 0.788. The result verified that sex (p = 0.028), religion (p <0.001), education (p = 0.046), current age (p = 0.027), SCI time (p = 0.020) and the SCI type (p = 0.012) were variables associated with self-care ability of the subjects. It was concluded that sociodemographic factors may interfere with the self-care ability of persons with SCI, and nurses should consider this aspect during the execution of the nursing process.

Virtual Guide On Ocular Self-Examination To Support The Self-Care Practice For People With HIV/AIDS

Objetivou-se descrever processo de desenvolvimento da cartilha virtual sobre autoexame ocular para pessoas com HIV/aids. A proposta metodológica seguiu as cinco etapas preconizadas por Falkembach: análise e planejamento, modelagem, implementação, avaliação e distribuição. A adequação da versão impressa para virtual requereu a construção de um vídeo tutorial, agregação de fotos ilustrativas para visualização de possíveis alterações oculares e ferramenta de interatividade com demonstração do resultado do exame ao usuário. Na avaliação inicial do material, foram diagnosticadas falhas no layout. Assim, comandos foram recolocados, unificados, dispostos em local de fácil visualização e foi feita a adequação da linguagem. Considera-se possível promover aproximação do usuário com métodos de prevenção na área da saúde ocular por meio de cartilha virtual, contribuindo para desenvolvimento de habilidades e divulgação do autoexame.


Aromatherapy with ylang ylang for anxiety and self-esteem: a pilot study

Objective: To verify if the use of ylang ylang essential oil by cutaneous application or inhalation alters the anxiety and self-esteem perception and physiological parameters as blood pressure and temperature. Method : A pilot study with 34 professionals from a nursing group randomized in three groups: one received the ylang ylang essential oil by cutaneous application, the second received through inhalation and the third (placebo) received the ylang ylang essence through cutaneous application. The assessment was done by an Anxiety Inventory (IDATE) and the Dela Coleta self-esteem scale, applied on baseline, after 30, 60 and 90 days and after 15 days post-intervention (follow up). Results : In the pre and post-intervention intergroup analysis, there was a significant difference in self-esteem for the three groups (p values: G1=0.014; G2=0.016; G3=0.038). There were no differences in the analysis between groups for anxiety or for physiological parameters. Conclusion : It was found significant alterations only to the intergroup perception of self-esteem for the three groups.


Perceived self-image in adolescent idiopathic scoliosis: an integrative review of the literature

Objective: To learn about the experiences of adolescents diagnosed with idiopathic scoliosis. Method: Integrative review of the literature published within a specified time frame. Results: For both sexes, the predominant clinical symptom of this condition appears to be the negative effect that the deformity exerts on perceived self-image. Quantitative studies used numerical scores to assess perceptions of body image but did not analyse emotional aspects. Patients treated surgically were found to have a better self-image than patients treated with a brace. Quality of life was improved by a reduction in the magnitude of the curve. Conclusion: Spinal deformity exerts a psychological effect on adolescent girls.


Self-perceived health and clinical characteristics in young adult students from the brazilian northeast

Objective To analyze the association between socioeconomic situation, clinical characteristics referred and the family history of cardiovascular disease, with the Self-perceived health of young adults education and their implications for clinical characteristics observed. Method Analytical study conducted with 501 young adults who are students in countryside city in the Brazilian Northeast. We used binary logistic regression. Results The final model explained 83.3% of the self-perceived positive health, confirming the association of Self-perceived health with male, residence in the community, have excellent/very good lifestyle and does not have or do not know that there are cases of stroke in the family. Conclusion Health perception was often optimistic, being important to identify devices to be worked closer to their perception of their actual health condition, increasing the effectiveness of health promotion activities undertaken by professionals.

Association between self-reported health and sociodemographic characteristics with cardiovascular diseases in adults

OBJECTIVE To assess the association of sociodemographic and self-rated health in the presence of cardiovascular diseases and the association of this perception with the type of disease. METHODS A cross-sectional population survey study carried out with 1,232 individuals aged between 20 and 59 years of both genders living in the metropolitan region of Maringá-PR. Data were analyzed using multiple and simple logistic regression. RESULTS In multivariate analysis, the age range and self-rated health were associated with cardiovascular disease, and in the univariate analysis self-rated regular health was associated with arterial hypertension, while self-rated poor health was associated to heart failure, stroke, and to acute myocardial infarction (heart attack). CONCLUSION The differences in association of self-rated health with these diseases can indicate how individuals with certain characteristics cope with the disease, allowing for more individualized and specific health care.