Structural parameters, molecular properties, and biological evaluation of some terpenes targeting Schistosoma mansoni parasite

The use of natural products has a long tradition in medicine, and they have proven to be an important source of lead compounds in the development of new drugs. Among the natural compounds, terpenoids present broad-spectrum activity against infective agents such as viruses, bacteria, fungi, protozoan and helminth parasites. In this study, we report a biological screening of 38 chemically characterized terpenes from different classes, which have a hydroxyl group connected by hydrophobic chain or an acceptor site, against the blood fluke Schistosoma mansoni, the parasite responsible for schistosomiasis mansoni. In vitro bioassays revealed that 3,7-dimethyl-1-octanol (dihydrocitronellol) (10) was the most active terpene (IC50 values of 13-52 mu M) and, thus, we investigated its antischistosomal activity in greater detail. Confocal laser scanning microscopy revealed that compound 10 induced severe tegumental damage in adult schistosomes and a correlation between viability and tegumental changes was observed. Furthermore, we compared all the inactive compounds with dihydrocitronellol structurally by using shape and charge modeling. Lipophilicity (miLogP) and other molecular properties (e.g. molecular polar surface area, molecular electrostatic potential) were also calculated. From the 38 terpenes studied, compound 10 is the one with the greatest flexibility, with a sufficient apolar region by which it may interact in a hydrophobic active site. In conclusion, the integration of biological and chemical analysis indicates the potential of the terpene dihydrocitronellol as an antiparasitic agent. (C) 2015 Elsevier Ireland Ltd. All rights reserved.

Formation of large micellar aggregates before equilibrium in diluted solutions.

We study the formation of premicelles for different values of the concentration of amphiphile molecules in water. Our model consists of a square lattice with water molecules occupying one cell of the lattice while the amphiphilic molecules, represented by chains of five interconnected sites, occupy five cells of the lattice. We perform Monte Carlo simulations in the NVT ensemble, for a fixed temperature and different concentration of amphiphiles, ranging from below to above the critical micelle concentration. We start our simulations from a monomeric state and follow in time all the aggregates sizes until the equilibrium state is reached. We pay particular attention to two aggregate sizes, one related to the minimum and the other to the maximum of the aggregate-size distribution curve obtained at equilibrium. We show that these aggregates evolve in time exhibiting a maximum concentration well before the equilibrium state, revealing the formation of premicelles. The times to reach these maximum concentrations decrease exponentially with the total concentration of the system.

In vitro synergistic interaction between amide piplartine and antimicrobial peptide dermaseptin against Schistosoma mansoni schistosomula and adult worms.

Schistosomiasis is one of the world's major public health problems, and praziquantel is the only available drug to treat this notable neglected disease. Drug combinations have been considered an important strategy for treatment of infectious diseases, which might enhance therapeutic efficacy and delaying resistance. In this study, we have examined the in vitro activities of the amide piplartine and the antimicrobial peptide dermaseptin 01 administered singly or in combination against Schistosoma mansoni of different ages including 3-hour-old and 7-day-old schistosomula and 49-day-old adult schistosomes as well as on egg output by adult worms. We calculated the median lethal concentrations (LC(50)) of 7.87 and 17.99 µM on 49-day-old adults, 11.02 and 71.58 µM on 7-day-old schistosomula, and 70.87 and 98.42 µM on 3-hour-old schistosomula for piplartine and dermaseptin, respectively. Most Piplartine/dermaseptin combinations showed synergistic effect, with combination index (CI) values less than 0.9 when S. mansoni adults or schistosomula were simultaneously incubated with both drugs in vitro; synergy between these two compounds was also indicated using isobolograms. Additionally, we observed alterations on the tegumental surface of schistosomula and adult schistosomes by means of laser scanning confocal microscopy. Furthermore, egg laying of surviving worms was considerably more reduced when exposed to the piplartine/dermaseptin combinations than each drug alone, and this inhibition was irreversible. This is the first report on the synergistic effect between piplartine and dermaseptin against S. mansoni and opens the route to further studies (e.g. in vivo) to characterize this combination in greater detail.

Assessment of the systemic effects of low-level laser therapy (LLLT) on thyroid hormone function in a rabbit model.

The aim of this study was to assess the effects of low-level laser therapy (LLLT) applied to a dental extraction socket on thyroid gland function in a rabbit model, based on serum triiodothyronine and thyroxine levels. Sixteen male New Zealand rabbits were randomly distributed into two groups: a control group (non-irradiated animals) and an experimental group (irradiated animals: one irradiation point in the extraction socket of the lower incisor). Animals in the experimental group were irradiated with an aluminium gallium arsenide diode laser (AlGaAs; wavelength 830 nm, 40 mW, CW laser), for 13 days, every 48 h, at a dose of 6 J/cm(2) per session, resulting in a total dose of 42 J/cm(2). Serum triiodothyronine and thyroxine levels were measured in both groups before extraction and on the last day of observation (day 15). There were no statistically significant differences between the groups in pre- and post-irradiation triiodothyronine and thyroxine values. With the irradiation protocol used in this study, LLLT did not affect thyroid function in rabbits as assessed by circulating serum triiodothyronine and thyroxine levels.

Influence of the simultaneous addition of bentonite and cellulose fibers on the mechanical and barrier properties of starch composite-films.

The addition of nanoclay or cellulose fibers has been presented in the literature as a suitable alternative for reinforcing starch films. The aim of the present work was to evaluate the effect of the simultaneous incorporation of nanoclay (bentonite) and cellulose fibers on the mechanical and water barrier properties of the resultant composite-films. Films were prepared by casting with 3% in weight of cassava starch, using glycerol as plasticizer (0.30 g per g of starch), cellulose fibers at a concentration of 0.30 g of fibers per g of starch and nanoclay (0.05 g clay per g starch and 0.10 g clay per g starch). The addition of cellulose fibers and nanoclay increased the tensile strength of the films 8.5 times and the Young modulus 24 times but reduced the elongation capacity 14 times. The water barrier properties of the composite-films to which bentonite and cellulose fibers were added were approximately 60% inferior to those of starch films. Diffractograms showed that the nanoclay was intercalated in the polymeric matrix. These results indicate that the simultaneous addition of bentonite and cellulose fibers is a suitable alternative to increase the tensile strength of the films and decrease their water vapor permeabilities.

Cell proliferation and death in the gastric epithelium of developing rats after glucocorticoid treatments.

Glucocorticoids take part in the intense morphofunctional modifications that occur in the gastric mucosa during fetal and postnatal development. Two studies were designed to evaluate corticoids role in gastric cell proliferation and apoptosis in developing rats: in vivo, using suckling animals; in vitro, using gastric explants obtained from 20-day fetuses. These explants were cultured in DMEM/F12 medium treated or not with 50 ng/ml of corticosterone; after 22 hr, vincristine was added to the medium for 2 hr to block mitosis. The metaphasic index decreased significantly after the 24-hr treatment (controls: 1.52 +/- 0.53; treated: 0.40 +/- 0.21) and apoptotic cells were visualized under light and electron microscopy. Fifteen-day-old rats were treated with hydrocortisone (25 mg/Kg) for 3 days, and injected with BrDU (100 mg/Kg) 1 hr before sacrifice on the 18th day. BrDu-labeled and non-labeled cells were counted to determine the labeling index of epithelial cells. As apoptotic cells are rapidly eliminated, other animals were treated for only 2-3 hr. Sections were investigated for the presence of apoptotic cells, using morphological criteria and TUNEL labeling. Hydrocortisone significantly reduced the labeling index (controls: 15.6 +/- 1.6 vs. treated: 11.7 +/- 1.1), besides altering the body weight gain. Hydrocortisone treatment doubled the number of apoptotic cells after 2 hr, and quadruplicated it after 3 hr. The results demonstrated that glucocorticoids inhibit cell proliferation in the gastric epithelium of fetuses and suckling rats and increase apoptotic rates, suggesting the exit from cell cycle.

Predictors related to the occurrence of a measles epidemic in the city of São Paulo in 1997.

A matched case-control study was performed to identify risk factors for measles during an epidemic that occurred in 1997 in the city of São Paulo, in the Brazilian state of the same name. Measles cases from the city of São Paulo from 1 January 1997 to 15 August 1997 were included in the study. The criteria for case definition were age below 30 years, having received no measles vaccine 5-21 days before the onset of rash, and laboratory confirmation by IgM antibodies detection. From a bank of confirmed measles cases, 130 cases for each of five age ranges (under 1 year, 1-5 years, 6-20 years, 21-24 years, and 25-29 years) were picked at random according to a systematic criterion proportional to the number of cases in seven areas of the city. Data were collected through a home survey, and for each measles case studied two controls matched by age and place of residence were selected. The matched conditional logistic regression analysis for the potential risk factors from the univariate analysis showed that the best predictors for acquiring measles during the epidemic were: lack of measles vaccination, previous contact with a measles-like disease at home or on the job, having been born either outside the state of São Paulo or in a rural area, being employed, and spending time in a semiclosed institution, such as a nursery, day care center, or school. The risk factors were not homogeneous for the different age groups. The data in the present survey suggest that, in addition to lack of vaccination, other risk factors should be considered when planning a measles vaccination strategy for a developing country.

Marginal fit of nickel-chromium copings before and after internal adjustments with duplicated stone dies and disclosing agent.

STATEMENT OF PROBLEM: Despite careful procedures, master stone dies may be damaged during laboratory procedures. The dentist routinely adjusts castings because the marginal fit of casting is not as accurate as on the dies. PURPOSE: This study evaluated the technique of internal adjustment of castings with use of duplicated stone dies and a disclosing agent to improve marginal fit discrepancy. MATERIAL AND METHODS: Thirty-two nickel-chromium copings were fabricated and simulated standard clinical and laboratory procedures with 2 variables: tooth preparation convergence angles of 6 and 18 degrees, with or without internal relief. Master stone dies and their duplicates were selected for coping construction and internal adjustment, respectively. A specimen positioning device was coupled with a Toolmakers microscope to allow reproducibility of measurements. Each coping was evaluated at 8 locations of its marginal perimeter, before and after internal adjustment. RESULTS: Marginal fit discrepancy of copings were significantly reduced with an internal adjustment technique (mean > 52%) for all experimental groups. Tooth preparations with greater convergence and internally relieved castings recorded a better marginal fit. CONCLUSION: The casting internal adjustment technique with use of duplicated stone dies and a disclosing agent substantially reduced marginal fit discrepancy.

Method for marginal measurements of restorations: accessory device for toolmakers microscope.

Standardization of measurements for marginal fit of castings is critical. This study describes the fabrication of a device that allowed fixation of specimens on a Toolmakers microscope with identical conditions according to tri-dimensional positioning of specimens, measuring location, and seating force. The device also allows mapping of the marginal discrepancies on the entire marginal perimeter of the tooth preparation.

[Vaccination coverage in the first year of life in 4 cities of the state of São Paulo, Brazil]. / Cobertura vacinal no primeiro ano de vida em quatro cidades do Estado de São Paulo, Brasil.

Immunization is an important disease prevention measure, and evaluating the effectiveness of immunization programs is crucial to ensuring their success. This study describes the results of a household survey in four cities in the state of São Paulo, Brazil: Francisco Morato, Guarulhos, Osasco, and São Paulo. The survey was done in order to estimate immunization coverage for the cohort of children born in 1996. The city of São Paulo was divided into five strata, according to socioeconomic and living conditions. The survey followed the methodology that the Pan American Health Organization recommends for immunization coverage surveys. The proportion of children who had received a complete set of the recommended vaccinations at the time of the interview, taking into account both oral reports and information recorded on the children's immunization cards, was above 90% for all the cities except Francisco Morato, which had the worst living conditions. In the city of São Paulo, the worst coverage was found in the lowest and highest strata. When only the doses received during the first year of life were considered, the coverage was not adequate to produce herd immunity. The use of private vaccination services was higher in the areas with better living conditions. The difference between the coverage calculated based on data from health services and the coverage calculated based on the survey was inversely proportional to living conditions. Our results suggest that surveys similar to the one described here should be carried out in other cities. Employees who provide vaccination services should be trained to correctly record vaccination data. In addition, it is important to make health professionals aware of the official immunization calendar, and to facilitate the public's access to health services.

Gill infection of Leporinus macrocephalus Garavello & Britski, 1988 (Osteichthyes: Anostomidae) by Henneguya leporinicola n. sp. (Myxozoa: Myxobolidae). Description, histopathology and treatment.

Piauçus (Leporinus macrocephalus), were raised in 300 m2 ponds (density of 10 fish/m2) presenting asphyxia signals and daily mortality of 27 fishes. Specimens with 8-cm total body length, were collected for necropsy. Mucus of body surface and pieces of organs were collected and examined microscopically, in wet mounts, stained or in histological sections. The smears examination showed the presence of several spores in the secondary lamellae of the gill filaments, identified as Henneguya leporinicola n.sp (Myxozoa: Myxobolidae). Histopathological study showed epithelial hyperplasia and fulfilling of the spaces between the secondary lamellae, congestion and telangiectasia sinusoidal. It was also observed hyperplasia of the goblet cells and several cysts of parasite with 70.3 microns diameter. Such cysts were situated among the secondary lamellae, covered or not by the hyperplasic epithelium. With this diagnostic, three applications of formalin solution 10 ml/m3 were carried out. Fifteen days after that, fish were examined again to ascertain whether the treatment was efficient on disease caused by the protozoa. The tissue alterations present in the gills after the treatment were just a moderate sinusoidal congestion and a slight epithelial hyperplasia on the base of the secondary lamellae.

Pattern of immune response to GP43 from Paracoccidioides brasiliensis in susceptible and resistant mice is influenced by antigen-presenting cells.

Paracoccidioidomycosis (PCM), endemic in Latin America, is a progressive systemic mycosis caused by Paracoccidioides brasiliensis. The infection can evolve to different clinical forms that are associated with various degrees of suppressed cell-mediated immunity. In the murine model, A/Sn and B10.A isogenic strains of mice are known to be resistant and susceptible, respectively, to this fungal infection. Assuming that the effector immune response is a consequence of the preferential activation of either Th1 or Th2 subsets, in the present work we evaluated the importance of two antigen-presenting cells (APCs), macrophages and B cells, in the development of the immune response to P. brasiliensis. In resistant mice, purified gp43, the main antigenic component of P. brasiliensis, seems to have been preferentially presented by macrophages and stimulated Th1 lymphokine production. On the other hand, in susceptible animals gp43 was distinguishably presented by B cells, which led to stronger activation of Th2 subsets. Moreover, T cells from resistant mice responded as those from susceptible animals when stimulated by gp43 presented by APCs from susceptible mice and vice versa, indicating that there are no significant differences in the T cell repertoires from A/Sn and B10.A mice. When T cells from F1 (A/Sn x B10.A) mice were stimulated by gp43 presented by APCs from A/Sn or B10.A, impaired behavior of B10.A macrophages in activating Th1 cells and a B10.A B cell tendency to stimulate T cells that secrete higher levels of IL-10 were observed. Taken together, our results suggest that APCs may be implicated in the outcome of P. brasiliensis infection.

Schistosomiasis mansoni in the "Baixada Ocidental Maranhense", state of Maranhão, Brazil: cross-sectional studies performed in 1987 and 1993.

A cross-sectional study on the prevalence of schistosomiasis mansoni in three sites of the "Baixada Ocidental Maranhense" was carried out in 1993 in: Alegre (in the municipality of São Bento), Aliança (in Cururupu) and Coroatá II (in the municipality of São João Batista). Results were compared to those of another study performed at the same sites and in similar conditions, in 1987. The entire population of the three sites, with few exceptions, was submitted to fecal tests using the Kato-Katz method and immediate intradermal tests for schistosomiasis in both studies. Subjects with positive results in one of these tests were clinically evaluated by a physical examination. In 1993, the total of 827 subjects were submitted to fecal examination and 826 to intradermal test. Schistosoma mansoni eggs were found in the feces of 154 (18.6%) subjects, while 478 (57.9%) subjects presented a positive intradermal test. Stool examination was carried out in 367 subjects in Alegre with a positivity rate of 14.9%; the intradermal test, performed in 366 subjects, was positive in 47.5% of the cases. In Aliança, 277 subjects had their feces examined and were submitted to an intradermal test, with a positivity rate of 34.4% and 70.7%, respectively. Finally in Coroatá II, 183 inhabitants submitted to fecal and intradermal tests had positivity rates of 2.2% and 59.0%, respectively. When the present data were compared to those obtained in the survey performed in 1987, a significant decrease in the prevalence of infection by S. mansoni was observed in Alegre and Coroatá II, and a prevalence increase in Aliança.

Enterovirus 71 infection and acute neurological disease among children in Brazil (1988-1990).

Surveillance for Enterovirus 71 (EV-71) infection in children up to 15 years of age was carried out in Brazil, from 1988 to 1990. Patients with acute neurological diseases (AND) such as flaccid paralysis, Bell's palsy, acute cerebellar ataxia and Guillain-Barré syndrome were included in the study. EV-71 infection was detected in 24 of 426 children (5.6%) with AND. EV-71 infection was confirmed only by virus isolation in 13 children, by virus isolation and seroconversion in 4, and by seroconversion alone in 7. EV-71 was also isolated from 15 of the 427 household contacts (3.5%) of 165 AND patients. There was some evidence of high infectivity of EV-71: household clusters were detected in the case of 7 of 24 children (29.1%) infected with EV-71 and manifesting AND; EV-71 was isolated from 11/40 household contacts (27.5%) of the infected patients but from only 4/387 household contacts (1.0%) of children in whom it was not possible to demonstrate EV-71 infection. Seven of the 24 children infected with EV-71 exhibited residual motor deficiency when examined 6 months after the disease onset. The relevance of these results for the Plan for Global Eradication of Wild Poliovirus is discussed, as well as the need to increase knowledge about the behaviour of this virus and its possible association with AND.

Intra-urban differentials in death rates from homicide in the city of São Paulo, Brazil, 1988-1994.

The objective of this research was to analyze the relationship between socioeconomic statistics and homicide mortality rates in the city of São Paulo between 1988 and 1994. City districts were grouped into five geographic areas. A socioeconomic indicator (ISE) was constructed with census information combining average income of the family head, illiteracy rate for the population over 5 yr of age, average number of rooms and number of persons per household. The higher the score, the better the socioeconomic situation (possible values: 4 to 384). Deaths from homicide were grouped by residential areas, and the rates for these areas were calculated. The association between homicide rates and the socioeconomic situation was analyzed with Spearman correlation coefficients. Average ISE scores varied from 291 in the Central area (homicide rate = 27.96 deaths per 100000) to 119.9 in the East area (homicide rate = 40.38). The Spearman coefficient between ISEs and homicide rates was -0.98 (p < 0.05). Due to the heterogeneity inside the areas, the median ISE is a better indicator of the socioeconomic conditions yielding an rs = -1.0. Almost half the population resides in areas with the highest risk of homicide mortality (East and South). Taking the Central area as a reference, we found risks of 1.36 in the Western, 1.37 in the Northern, 1.44 in the Eastern and 2.67 in the Southern areas.

Prevalence of serotypes and antimicrobial resistance of streptococcus pneumoniae strains isolated from Brazilian children with invasive infections. Pneumococcal Study Group in Brazil for the SIREVA Project. Regional System for Vaccines in Latin America.

A laboratory surveillance study was developed in Brazil in 1993 to determine capsular types and antimicrobial susceptibility of Streptococcus pneumoniae strains. By studying 360 strains isolated from children with invasive infections in three different cities, 8 out of 34 types were identified as being the most prevalent and considered as the reference group for further analyses. This group comprised 77.7% of all strains studied, and includes the types 1, 5, 6A/B, 9V, 14, 19F, 19A, and 23F. The prevalence of this reference group was significantly higher among strains isolated from children with pneumonia than meningitis. Similarly, this group was more prevalent among strains isolated from children 3 to 6 years of age than from children under 2 years of age. Most strains (78.6%) were found to be susceptible to penicillin and only 1.4% showed high resistance to this antibiotic. However, intermediate resistance to penicillin was detected in 20% of the strains. This laboratory surveillance will be maintained and extended to other cities of Brazil to better define and monitor the trends of pneumococcal infections for proper control and prevention.