Genetic diversity and differentiation in natural Plasmodium falciparum populations inferred by molecular typing of the merozoite surface proteins 1 and 2

Genetic diversity and differentiation, inferred by typing the polymorphic genes coding for the merozoite surface proteins 1 (Msp-1) and 2 (Msp-2), were compared for 345 isolates belonging to seven Plasmodium falciparum populations from three continents. Both loci yielded similar estimates of genetic diversity for each population, but rather different patterns of between-population differentiation, suggesting that natural selection on these loci, rather than the transmission dynamics of P. falciparum, determines the variation in allele frequencies among populations.

Spectrophotometric and spectrofluorimetric determination of some drugs containing secondary amino group in bulk drug and dosage forms via derivatization with 7-Chloro-4-Nitrobenzofurazon

Sensitive and selective spectrophotometric and spectrofluorimetric methods have been developed for determination of some drugs such as Pramipexole, Nebivolol, Carvedilol, and Eletriptan, which commonly contain secondary amino group. The subject methods were developed via derivatization of the secondary amino groups with 7-Chloro-4-Nitrobenzofurazon in borate buffer where a yellow colored reaction product was obtained and measured spectrophotometrically or spectrofluorimetrically. Concentration ranges were found as 2.0 to 250 μg mL-1 and 0.1 to 3.0 μg mL-1, for spectrophotometric and spectrofluorimetric study, respectively. The described methods can be easily applied by the quality control laboratories in routine analyses of these drugs in pharmaceutical preparations.

Seeds of species from the ‘caatinga’: proteins, oils and fatty acid contents

The seeds of 14 species from the ‘caatinga’, a dry forest ecosystem of the semiarid region of northeast Brazil, were analysed for total protein and total lipid contents, as well as fatty acid distribution. The seeds of Argemone mexicana L., an introduced and naturalized species in Brazil, commonly found in ‘caatingas’ and other vegetation, were also analysed. The protein contents ranged from 123 g.kg-1 to 551 g.kg-1, higher contents being found in species of Leguminosae, but also in Jatropha mollissima (Pohl) Baill. (Euphorbiaceae, 409 g.kg-1). Oil contents ranged from 10 g.kg-1 to 400 g.kg-1. The contents of protein and oil were found to be inversely proportional in the seeds of most species, the figures for proteins being generally higher than those of oils. Most species presented either oleic or linoleic as predominant fatty acids. Cardiospermum cf. corindum L. presented eicosenoic acid as the predominant fatty acid.

The calcium-modulated proteins, S100A1 and S100B, as potential regulators of the dynamics of type III intermediate filaments

The Ca2+-modulated, dimeric proteins of the EF-hand (helix-loop-helix) type, S100A1 and S100B, that have been shown to inhibit microtubule (MT) protein assembly and to promote MT disassembly, interact with the type III intermediate filament (IF) subunits, desmin and glial fibrillary acidic protein (GFAP), with a stoichiometry of 2 mol of IF subunit/mol of S100A1 or S100B dimer and an affinity of 0.5-1.0 µM in the presence of a few micromolar concentrations of Ca2+. Binding of S100A1 and S100B results in inhibition of desmin and GFAP assemblies into IFs and stimulation of the disassembly of preformed desmin and GFAP IFs. S100A1 and S100B interact with a stretch of residues in the N-terminal (head) domain of desmin and GFAP, thereby blocking the head-to-tail process of IF elongation. The C-terminal extension of S100A1 (and, likely, S100B) represents a critical part of the site that recognizes desmin and GFAP. S100B is localized to IFs within cells, suggesting that it might have a role in remodeling IFs upon elevation of cytosolic Ca2+ concentration by avoiding excess IF assembly and/or promoting IF disassembly in vivo. S100A1, that is not localized to IFs, might also play a role in the regulation of IF dynamics by binding to and sequestering unassembled IF subunits. Together, these observations suggest that S100A1 and S100B may be regarded as Ca2+-dependent regulators of the state of assembly of two important elements of the cytoskeleton, IFs and MTs, and, potentially, of MT- and IF-based activities.

Membrane proteins: functional and structural studies using reconstituted proteoliposomes and 2-D crystals

Reconstitution of membrane proteins into lipid bilayers is a powerful tool to analyze functional as well as structural areas of membrane protein research. First, the proper incorporation of a purified membrane protein into closed lipid vesicles, to produce proteoliposomes, allows the investigation of transport and/or catalytic properties of any membrane protein without interference by other membrane components. Second, the incorporation of a large amount of membrane proteins into lipid bilayers to grow crystals confined to two dimensions has recently opened a new way to solve their structure at high resolution using electron crystallography. However, reconstitution of membrane proteins into functional proteoliposomes or 2-D crystallization has been an empirical domain, which has been viewed for a long time more like "black magic" than science. Nevertheless, in the last ten years, important progress has been made in acquiring knowledge of lipid-protein-detergent interactions and has permitted to build upon a set of basic principles that has limited the empirical approach of reconstitution experiments. Reconstitution strategies have been improved and new strategies have been developed, facilitating the success rate of proteoliposome formation and 2-D crystallization. This review deals with the various strategies available to obtain proteoliposomes and 2-D crystals from detergent-solubilized proteins. It gives an overview of the methods that have been applied, which may be of help for reconstituting more proteins into lipid bilayers in a form suitable for functional studies at the molecular level and for high-resolution structural analysis.

Effect of oral administration of Propionibacterium acnes on growth performance, DTH response and anti-OVA titers in goat kids

Immunostimulants are susbstances that stimuli the response of effector cells to activate the immune response such as antigen uptake, cytokine release or antibody response. These substances can increase resistence to infection by different types of microorganisms, reducing dependence of antibiotics used in livestock animals. Recent reports have demonstrated the positive effect of Propionibacterium acnes (P. acnes) to control animal diseases. In this study, we evaluated the effect of the non-specific immunostimulant P. acnes on immunological functions and growth performance in goat kids. Twenty five goat kids served as control group (A) and another 25 animals received P. acnes being the experimental group (B). Kids were challenged with ovalbumin (OVA) to assess humoral immunity. To assess in vivo cell immunity, delayed type hypersensitivity (DTH) test with phytohemagglutinin (PHA) was used, clinical signs and body weight were recorded each week until 9 weeks of age when the experiment ended. Blood samples were obtained to analyze serum proteins fractions and anti-OVA specific antibodies. No clinical signs of disease and no differences (p>0.05) on body weight between groups were recorded (7.32±0.81 kg in group A, 7.13±0.65 kg in group B). Goat kids from group B had more total protein (59.8±5g/l) and albumin levels (32.8±3.3g/l) than goat kids from group A (56.6±5.7 g/l, 29.6±3.9 g/l respectively) (p<0.05). DTH response in goat kids from group B on day 42 was higher (p<0.05) than group A. At day 63, goat kids from group receiving P. acnes had higher percentage (85.4) of anti-OVA IgM titers (p<0.05) than control group (57.7). In conclusion, the results showed that oral administration of P. acnes to goat kids improved some aspects of the immune system of the animals and it could be used to control goat diseases.

Use of analgesics in adults with pain complaints: prevalence and associated factors, Turkey

OBJECTIVE: Use of analgesics has been increasingly recognized as a major public health issue with important consequences in Turkey. The objective of the study was to determine the prevalence and patterns of analgesics usage and associated factors in adults with pain complaints. METHODS: A cross-sectional study was conducted in 15 cities selected from five demographic regions in Turkey. The study sample population comprised 1.909 adults 18-65 age groups suffering from pain. The sampling method was multi-step stratified weighted quota-adjusted sampling. Data were collected by face-to-face interviews using a semi-structured survey questionnaire consisting of 28 questions. Odds ratios were produced by logistic regression analyses. RESULTS: The prevalence of analgesic use was 73.1%, and it was higher in females (75.7%; p<0.05), in subjects 45-54 years (81.4%; p<0.05), in subjects in rural areas (74.6%; p<0.05), in subjects in northern region (84.3%; p<0.05), in illiterate subjects (79.1%; p>0.05), and in subjects of lower socioeconomic status (74.1%; p>0.05). One in ten of the participants used non-prescription analgesics. Non-prescription analgesics were more prevalent among the 55-65 age groups (18.1%; p<0.05), among female (11.6%; p>0.05), among the urban population (10.7%; p>0.05), and in subjects of lower middle socioeconomic status (13.2%; p<0.05). Logistic regression showed statistically significant ORs only for age groups, duration of education, socioeconomic status, and demographic regions (p<0.05). CONCLUSIONS: The results showed that the prevalence of analgesic use and prescription analgesic use is high in Turkey, and their use is related to sociodemographic characteristics.

Pertussis booster vaccine for adolescents and young adults in São Paulo, Brazil

OBJECTIVE: To develop a model to assess different strategies of pertussis booster vaccination in the city of São Paulo. METHODS: A dynamic stationary age-dependent compartmental model with waning immunity was developed. The "Who Acquires Infection from Whom" matrix was used to modeling age-dependent transmission rates. There were tested different strategies including vaccine boosters to the current vaccination schedule and three of them were reported: (i) 35% coverage at age 12, or (ii) 70% coverage at age 12, and (iii) 35% coverage at age 12 and 70% coverage at age 20 at the same time. RESULTS: The strategy (i) achieved a 59% reduction of pertussis occurrence and a 53% reduction in infants while strategy (ii) produced 76% and 63% reduction and strategy (iii) 62% and 54%, respectively. CONCLUSION: Pertussis booster vaccination at age 12 proved to be the best strategy among those tested in this study as it achieves the highest overall reduction and the greatest impact among infants who are more susceptible to pertussis complications.

Prescription, dispensation and marketing patterns of methylphenidate

OBJECTIVE To analyze the patterns and legal requirements of methylphenidate consumption. METHODS We conducted a cross-sectional study of the data from prescription notification forms and balance lists of drugs sales – psychoactive and others – subject to special control in the fifth largest city of Brazil, in 2006. We determined the defined and prescribed daily doses, the average prescription and dispensation periods, and the regional sales distribution in the municipality. In addition, we estimated the costs of drug acquisition and analyzed the individual drug consumption profile using the Lorenz curve. RESULTS The balance lists data covered all notified sales of the drug while data from prescription notification forms covered 50.6% of the pharmacies that sold it, including those with the highest sales volumes. Total methylphenidate consumption was 0.37 DDD/1,000 inhabitants/day. Sales were concentrated in more developed areas, and regular-release tablets were the most commonly prescribed pharmaceutical formulation. In some regions of the city, approximately 20.0% of the prescriptions and dispensation exceeded 30 mg/day and 30 days of treatment. CONCLUSIONS Methylphenidate was widely consumed in the municipality and mainly in the most developed areas. Of note, the consumption of formulations with the higher abuse risk was the most predominant. Both its prescription and dispensation contrasted with current pharmacotherapeutic recommendations and legal requirements. Therefore, the commercialization of methylphenidate should be monitored more closely, and its use in the treatment of behavioral changes of psychological disorders needs to be discussed in detail, in line with the concepts of the quality use of medicines.

Identification and characterization of sex-linked proteins of Schistosoma mansoni

The proteins of adults worms (male and female) of two isolates (BH and RJ) of Shistosoma mansoni were extracted using Triton X-114 phase separation. The SDS-polyacrilamide gel electrophoresis profiles of the three phases (detergent, aqueous and insoluble proteins) obtained were compared after Coomassie blue and silver staining, surface radioiodination and Western blotting. No major differences were detected between the 2 isolates. Of the 25 or more proteins which partitioned into the detergent phase, only about 8 proteins could be surface radiodinated on live adult worms. A comparison was also made between the profiles of mael and females worms, isolated from bisexually infected mice. Two major female-specific and one male-specific band were detected by silver and/or Coomassie staining. The female bands, 32 KDa and 18 KDa, partitioned into the detergent and aqueous phase, respectively. The male-specific band of 42 KDa remained in the insoluble phase. Antigenic differences between male and females protins were detected by Western vlotting using a sera from infected Nectomys squamipes.

Frequency of fear of needles and impact of a multidisciplinary educational approach towards pregnant women with diabetes

PURPOSE: To evaluate the frequency of fear of needles and the impact of a multidisciplinary educational program in women with pre-gestational and gestational diabetes taking insulin during pregnancy. METHODS: The short Diabetes Fear of Injecting and Self-testing Questionnaire (D-FISQ), composed by two subscales that access fear of self injection (FSI) and fear of self testing (FST), was administered twice during pregnancy to 65 pregnant women with pre-gestational and gestational diabetes: at the first endocrine consult and within the last two weeks of pregnancy or postpartum. An organized multidisciplinary program provided diabetes education during pregnancy. Statistical analysis was carried out by Wilcoxon and McNemar tests and Spearman correlation. A p<0.05 was considered to be significant. RESULTS: Data from the short D-FISQ questionnaire shows that 43.1% of pregnant women were afraid of needles in the first evaluation. There was a significant reduction in scores for FSI and FST subscales between the first and second assessments (first FSI 38.5% compared with second 12.7%, p=0.001; first FST 27.7% compared with second FST 14.3%, p=0.012). CONCLUSIONS: The fear of needles is common in pregnant women on insulin therapy and an organized multidisciplinary educational diabetes program applied during pregnancy reduces scores of such fear.

Insecticidal and antifungic proteins of the latex from Manihot glaziovii Muell. Arg.

Analysis of the latex from Manihot glaziovii showed the presence of various enzymatic and inhibitory activities. The latex also presented an inhibitory effect on the development of cowpea weevil (Callosobruchus maculatus) in an artificial seed system and on the development, in an in vitro assay, of phytopathogenic fungi Colletotrichum gloesporioides, Fusarium solani and Macrophomina phaseolina. These results suggest the presence of substances, some of them of protein nature, involved in plant defense mechanisms in this exudation product.

Dietary fiber, energy intake and nutritional status during the treatment of children with chronic constipation

The present prospective study was carried out to determine dietary fiber and energy intake and nutritional status of children during the treatment of chronic constipation. Twenty-five patients aged 2 to 12 years with chronic constipation were submitted to clinical evaluation, assessment of dietary patterns, and anthropometry before and after 45 and 90 days of treatment. The treatment of chronic constipation included rectal disimpaction, ingestion of mineral oil and diet therapy. The standardized diet prescribed consisted of regular food without a fiber supplement and met the nutrient requirements according to the recommended daily allowance. The fiber content was 9.0 to 11.9 g for patients aged less than 6 years and 12.0 to 18.0 g for patients older than 6 years. Sixteen patients completed the 90-day follow-up and all presented clinical improvement. The anthropometric variables did not change, except midarm circumference and triceps skinfold thickness which were significantly increased. Statistically significant increases were also found in percent calorie intake adequacy in terms of recommended daily allowance (55.5 to 76.5% on day 45 and to 68.5% on day 90; P = 0.047). Percent adequacy of minimum recommended daily intake of dietary fiber (age + 5 g) increased during treatment (from 46.8 to 52.8% on day 45 and to 56.3% on day 90; P = 0.009). Food and dietary fiber intake and triceps skinfold thickness increased during follow-up. We conclude that the therapeutic program provided a good clinical outcome.

Different responses of the GlnB and GlnZ proteins upon in vitro uridylylation by the Azospirillum brasilense GlnD protein

Azospirillum brasilense is a diazotroph found in association with important agricultural crops. In this organism, the regulation of nitrogen fixation by ammonium ions involves several proteins including the uridylyltransferase/uridylyl-removing enzyme, GlnD, which reversibly uridylylates the two PII proteins, GlnB and GlnZ, in response to the concentration of ammonium ions. In the present study, the uridylylation/deuridylylation cycle of A. brasilense GlnB and GlnZ proteins by GlnD was reconstituted in vitro using the purified proteins. The uridylylation assay was analyzed using non-denaturing polyacrylamide gel electrophoresis and fluorescent protein detection. Our results show that the purified A. brasilense GlnB and GlnZ proteins were uridylylated by the purified A. brasilense GlnD protein in a process dependent on ATP and 2-oxoglutarate. The dependence on ATP for uridylylation was similar for both proteins. On the other hand, at micromolar concentration of 2-oxoglutarate (up to 100 µM), GlnB uridylylation was almost twice that of GlnZ, an effect that was not observed at higher concentrations of 2-oxoglutarate (up to 10 mM). Glutamine inhibited uridylylation and stimulated deuridylylation of both GlnB and GlnZ. However, glutamine seemed to inhibit GlnZ uridylylation more efficiently. Our results suggest that the differences in the uridylylation pattern of GlnB and GlnZ might be important for fine-tuning of the signaling pathway of cellular nitrogen status in A. brasilense.