Pharmacological and Toxicological Study of Maytenus ilicifolia Leaf Extract Part II-Clinical Study (Phase I).

Maytenus ilicifolia is a plant widely used in South American folk medicine as an effective anti-dyspeptic agent, and the aim of this study was to evaluate their clinical and toxicological effects in healthy volunteers in order to establish its maximum safe dose. We selected 24 volunteers (12 women and 12 men) between 20 and 40 years of age and put them through clinical/laboratory screening and testing to ascertain their psychomotor functions (simple visual reaction, speed and accuracy, finger tapping tests). M. ilicifolia tablets were administered in increasing weekly dosages, from an initial dose of 100 mg to a final dose of 2000 mg. The volunteers' clinical and biochemical profiles and psychomotor functions were evaluated weekly, and they also completed a questionnaire about any adverse reactions. All subjects completed the study without significant changes in the evaluated parameters. The most cited adverse reactions were xerostomia (dry mouth syndrome) (16.7%) and polyuria (20.8%), with reversal of these symptoms without any intervention during the study. The clinical Phase I study showed that the administration of up to 2000 mg of the extract was well tolerated, with few changes in biochemical, hematological or psychomotor function parameters, and no significant adverse reactions. Copyright © 2017 John Wiley & Sons, Ltd.

Evaluation of the antinociceptive activity of Ocimum gratissimum L. (Lamiaceae) essential oil and its isolated active principles in mice.

Ocimum gratissimum is used in popular medicine to treat painful diseases. The antinociceptive properties of O. gratissimum essential oil (OgEO) and two of its active principles (eugenol and myrcene) were tested in classic models of pain (hot plate test and formalin test). Adult male C57BL/6 J mice acutely received corn oil (control group, p.o.), morphine (positive control group, 5 mg/kg, i.p.), OgEO (10, 20, or 40 mg/kg, p.o.), eugenol or myrcene (both at 1, 5, or 10 mg/kg, p.o.). The highest doses of all tested drugs significantly increased the latency to lick the paw(s) in the hot plate test compared with the control group. OgEO at a dose of 40 mg/kg and eugenol and myrcene at a dose of 10 mg/kg were effective in minimizing animal pain in the first and second phases of the formalin test. The antinociceptive effect shown by all drugs tested in hot plate test was reverted by naloxone administration (1 mg/kg), indicating opiod system participation. These results demonstrate the beneficial effects of OgEO and its active principles against neurogenic and inflammatory pain. Our findings demonstrate that OgEO and its isolated active principles exhibited antinociceptive activity in murine pain models.

Empowering knowledge generation through international data network: the IMeCCHI-DATANETWORK.

INTRODUCTION: The International Methodology Consortium for Coded Health Information (IMeCCHI) is a collaboration of health services researchers who promote methodological advances in coded health information. The IMeCCHI-DATANETWORK initiative focuses on developing a multi-purpose distributed data infrastructure and common data model (CDM) to enable cross-border data sharing and international comparisons. METHODS: IMeCCHI consortium partners from six different countries - Canada, Denmark, Italy, New Zealand, South Korea, and Switzerland - used a questionnaire to describe their original databases which differ in size, structure, content and coding systems. To standardize these data, they agreed on a CDM and mapped their population-based databases to meet the CDM specifications. At the end of this process, local data had a more homogenous content and structure, which made them syntactically and semantically interoperable. Data transformation was performed using a common data management software called TheMatrix. RESULTS: The CDM encompasses four tables of structured data (person characteristics, hospitalizations, outpatient prescription medication and death), linked at the individual level through a person identifier. It can be used to answer research questions across countries using locally converted databases, which facilitates study replication in a distributed fashion. As a proof-of-concept study, an initial research question was addressed using an agreed protocol. Local data were transformed in csv files in the CDM structure and TheMatrix was tested to transform the standardized data from each partner into local analytical datasets. This allowed results to be shared between countries, whilst maintaining local control over each region's data. CONCLUSION: The IMeCCHI-DATANETWORK, a model of a distributed data network, demonstrated that it is feasible to analyze international data using standardized analytical methods that enable independent analyses by regions, without relocating datasets thereby protecting local confidentiality obligations. The distributed data infrastructure can produce results that can be generalized to several countries, while facilitating cross-border data sharing and international comparisons. KEYWORDS: Common data model, international comparison, cross-border data sharing, interoperability, observational data.

Preclinical toxicological assessment of a phytotherapeutic product--CPV (based on dry extracts of Crataegus oxyacantha L., Passiflora incarnata L., and Valeriana officinalis L.).

Associations of plants have been widely used, for centuries, in Ayurveda and in Chinese medicine and have been increasingly acknowledged in Western medicine. The objective of this study is to assess the level of toxicity of an association of three plants: Crataegus oxyacantha, Passiflora incarnata, and Valeriana officinalis (CPV extract). This association was administered to rats, mice, and dogs, both acute and chronically for 180 days. The tests used in the acute experiments were: observational pharmacological screening, LD(50), motor coordination and motor activity. Chronic tests carried out were: weight gain/loss and behavioral parameters in rats and in mice; estrus cycle, effects on fertility, and teratogenic studies in rats and of mutagenic features in mice, in addition to the Ames test. The following parameters were assessed in dogs: weight gain/loss, general physical conditions, water/food consumption and anatomopathological examination of the organs subsequent to the 180 days of treatment. All of the results were negative, showing that CPV administered in high doses and over a long period of time presents no toxicity, suggestive of the fact that this is an association devoid of risk for human beings.

Use of alcohol among the inhabitants of the 107 largest cities in Brazil--2001.

Alcohol is part of the history of humanity, seemingly as a result of countless factors including the easy production of alcoholic beverages in practically all regions of the world. The authors studied aspects of the use of and the dependence on alcohol in Brazil, through a household survey conducted by Centro Brasileiro de Informações sobre Drogas Psicotrópicas (CEBRID). A total of 8,589 interviews were held in 107 of the largest cities in Brazil, all of them with more than 200 thousand inhabitants. The study was planned to gather information within the household environment about a stratified probabilistic sample obtained in three selection phases: 1) the censitaire sectors for each municipality, 2) a systematic randomized sampling, and 3) drafting a respondent by lot in each household to provide information. Approximately 11.2% of the subjects were concerned with their own consumption of alcohol. The signs/symptoms of the syndrome of dependence evident in a greater percentage were the desire to stop or reduce the use of alcohol and to stop or reduce resorting to alcoholic beverages more often than desired, as reported by 14.5 and 9.4% of the respondents, respectively. The regions in Brazil with the highest percentage of dependents were the North (16.3%) and the Northeast (19.9%). According to the estimates obtained in the survey, 5.2% of the teenagers were concerned about the use of alcohol. The estimates obtained in this survey reveal a need to implant specific preventive programs for the problem of alcohol, especially for the very young.

Decrease in tobacco use among Brazilian students: a possible consequence of the ban on cigarette advertising?

In 2000, cigarette advertising was banned from the Brazilian media [LEI N degrees 10.167, de 27 de dezembro de dezembro. (2000). http://www.planalto.gov.br/ccivil_03/Leis/L10167.htm]. Nevertheless, not enough surveys have been carried out to measure the impact of the prohibition. The objective of the present survey was to compare the use of tobacco by primary and secondary school students in 1997 and 2004, that is, before and after the ban on tobacco advertising in Brazil. Two surveys were conducted (in 1997 and 2004) using the same methodology, with a target population of primary (from the fifth grade on) and secondary public schools in ten different Brazilian capitals. The sampling was done by conglomerates, stratified, and obtained in two stages. In total, 15,501 students were surveyed in 1997, and 21,712 in 2004. The questionnaire was adapted from an instrument developed by WHO, anonymous, self-administered, and applied collectively in the classroom. In a comparison of the two surveys (1997 and 2004) tobacco lifetime use (used any psychotropic drug at least once in their life) decreased significantly in seven out of ten capitals surveyed. The 11-12 year old age group experienced the largest decrease in tobacco lifetime use. There was a decrease in lifetime use for males in nine capitals, and in eight capitals for females. A decrease was also observed in heavy tobacco use also in eight capitals. In conclusion there was a significant decrease in tobacco consumption among the surveyed students, suggesting that the decrease is related to changes in public policy in Brazil over the surveyed period.

A comparison of plants utilized in ritual healing by two Brazilian cultures: Quilombolas and Kraho Indians.

The present study deals with two ethnobotanical surveys carried out in two different segments of the Brazilian population: the first among the Krahô Indians living in Tocantins State, a Cerrado region, and the second one among the descendants of former black slaves, the Quilombolas, living in Mato Grosso State. Both populations use plants which may have effects on the central nervous system (CNS) in their ritual healing ceremonies. Field work was performed during two years by one of the authors (E. Rodrigues) utilizing methods from botany and anthropology. Information was obtained on a total of 169 plants which were utilized in the preparation of 345 prescriptions for 68 ailments seemingly of the CNS, classified as tonics, analgesics, anorectics, hallucinogens, and anxiolytics. The taxonomic families of plants used, the more common therapeutic indications and types of healing rituals are discussed.

Plants with possible psychoactive effects used by the Krahô Indians, Brazil.

OBJECTIVE: In spite of the richness of the Brazilian biodiversity, no phytomedicines have been developed from this flora with the purpose of being used in psychiatric treatments. The objective of the present study was to document the use of plants with possible psychoactive effects in rituals performed by the Krahô Indians, who live in the cerrado savannahs biome in the central region of Brazil. Also, the present data were compared with the data obtained during a review of the literature on the use of psychoactive plants by 25 Brazilian indigenous groups. METHOD: The study was carried out during two years of fieldwork during which anthropological and botanical methods were employed. RESULTS: Seven local shamans were interviewed and they indicated 98 formulas, consisting of 45 plant species that appear to have psychoactive properties and were used in 25 different treatments. Some of the psychoactive properties were "prevention of madness", "stimulant effect", "tranquilizing effect", "prevention of tremors", "longer sleeping period", "open mind" and "induction of sleep". This article also describes the review of literature, which recorded 58 plants that may have psychoactive effects used by 25 Brazilian Indian cultures. CONCLUSION: The treatment of psychological/psychiatric disorders based on the plants used by the Krahô Indians is very rich. It is also observed among other Brazilian indigenous groups. Future phytochemical and pharmacological studies on these plants may develop new medicines to treat psychiatric disorders.

Prostasome to sperm transfer of CD13/aminopeptidase N (EC 3.4.11.2).

Prostasomes are membranous vesicles (150-200 nm in diameter) that are present in human semen. They are secreted by the prostate gland and contain large amounts of cholesterol, sphingomyelin and Ca2+. In addition, some of their proteins are enzymes. Prostasomes enhance the motility of ejaculated spermatozoa and are involved in a number of additional biological functions. In previous papers, we demonstrated that lipid can be transferred from prostasomes to sperm by a fusion process occurring at slightly acidic pH. CD (cluster antigens) are ubiquitous proteins; in this paper, we demonstrate that CD13/aminopeptidase N is present is semen, where it is bound to prostasomes. Upon mixing prostasomes and sperm at slightly acidic pH (7 or less), aminopeptidase is transferred from prostasomes to sperm. This evidence comes from enzymatic activity determinations and from the use of the monoclonal antibody, anti-human CD13. The transfer was about 8% of total prostasomal activity at pH 5 and with a prostasome to sperm ratio of 2 (on a protein basis). The transfer did not occur at pH 8.0, but was measurable at pH 7. Therefore, this mechanism may represent a means of modifying the composition and the biological properties of ejaculated sperm.

Transfer of aminopeptidase activity from prostasomes to sperm.

Prostasomes are membranous vesicles (150-200 nm diameter) present in human semen. They are secreted by the prostate and contain large amounts of cholesterol, sphingomyelin and Ca2+. In addition, some of their proteins are enzymes. Prostasomes enhance the motility of ejaculated spermatozoa and are involved in a number of additional biological functions. It has been demonstrated that lipid can be transferred from prostasomes to sperm by a fusion process occurring at slightly acidic pH. In this paper, we show that an aminopeptidase activity is transferred from prostasome to sperm. This may be of particular interest since it indicates the involvement of protein in the process of fusion and because sperm may acquire new membrane-bound proteins by this procedure.

Interactions between prostasomes and leukocytes.

Prostasomes are membranous vesicles (150-200 nm diameter) present in human semen. They are secreted by the prostate gland and contain large amounts of cholesterol, sphingomyelin and Ca2+. In addition, some of their proteins are enzymes. Prostasomes enhance the motility of ejaculated sperm and are involved in a number of biological functions. In a previous work, we found that prostasome can fuse to spermatozoa at slightly acidic pH values, as demonstrated by the transfer of the lipophilic octadecylrhodamine probe. In this paper, we study the interactions of two leukocyte populations (polymorphonuclear and mononuclear) with prostasomes and find a pH-dependent adhesion (revealed by microscopic observation), but no fusion. These phenomena may be relevant for the functions of leukocytes in human reproduction.

Household survey on drug abuse in Brazil: study involving the 107 major cities of the country--2001.

The prevalence of the use in Brazil of illicit drugs, as well as of alcohol and tobacco, was determined. Further, illicit use of psychotropic medicines, and anabolic steroids were also surveyed. This study was carried out in 107 Brazilian cities with more than 200,000 inhabitants aged 12-65 years. The sampling design adopted was that of sampling per aggregates in three stages: first, the census sectors were selected; second, homes were selected among the sectors; finally, in each home, a respondent was selected in a manner independent of the interviewer. There were 8589 persons interviewed. The questionnaire utilized was that of the SAMHSA (Substance Abuse and Mental Health Services Administration) which was translated and adapted for Brazilian conditions. The lifetime use of alcohol in the 107 major cities of the country was 68.7%, which was close to numbers observed for Chile at 70.8% and the USA at 81.0%. Yet, tobacco lifetime use was 41.1% of the total, which is lower than the prevalence observed in the USA (70.5%). The data on the lifetime use of marijuana in Brazil (6.9%) approximated the findings for Colombia (5.4%), however, being much lower than that observed in the USA (34.2%) and the United Kingdom (25.0%). The prevalence of lifetime use of cocaine was 2.3%, well below the levels for the USA with 11.2% of the total population. The abuse of inhalants was 5.8% of the total, greater than that found in Colombia (1.4%) and about four times less than that in the United Kingdom with 20.0%. Among the medicaments, stimulants had a 1.5% prevalence of lifetime use, and that of benzodiazepines had similar percentages in Brazil (3.3%) and in the USA (5.8%). In this survey, only four individuals reported lifetime use of heroin, which was equivalent to about 0.04% of the sample and much lower than that in the USA with 1.2% and in Colombia reaching 1.5%. These findings will allow the implementation of public policies fitted to the situation with psychotropic drugs in Brazil.

[Use of psychotropic drugs in Brazil: household survey in the 107 biggest Brazilian cities--2001]. / Uso de drogas psicotrópicas no Brasil: pesquisa domiciliar envolvendo as 107 maiores cidades do país--2001.

The objective was to estimate the prevalence of the use of drugs, alcohol, tobacco and the use of non-medical psychotropics. This study enclosed the 107 biggest cities in Brazil; sample: ages between 12 and 65 years. Sampling in three periods: tax sectors; household and the respondent. Were interviewed 8,589 people. The lifetime use of the alcohol was 68.7%, closer to 70.8% in Chile. The lifetime use of the tobacco was of 41.1%, lower than USA (70.5%). The lifetime use of the marijuana was of 6.9% closer to Colombia (5.4%) and lower than USA (34.2%). The lifetime use of the cocaine was 2.3%, lower than USA (11.2%). The lifetime use of solvent was of 5.8%, much lower than the United Kingdom (20.0%). The stimulants have had 1.5% of lifetime use and the anxiolytics with 3.3%. These findings will allow the implantation of adjusted public politics to our reality in the field of the psychotropics drugs.

Increase of human spermatozoa intracellular Ca2+ concentration after fusion with prostasomes.

Prostasomes are membranous vesicles (150-200 nm diameter) present in human semen. They are secreted by the prostate gland and contain large amounts of cholesterol, sphingomyelin and calcium, and some of their proteins are enzymes. Prostasomes are involved in a number of biological functions. In previous work, we discovered that prostasomes may fuse to sperm at neutral or at slightly acidic pH values. This mechanism may deliver calcium to sperm, thereby influencing sperm functions. We measured sperm [Ca2+]i with the fura-2 AM method and found that it increased after mixing prostasomes and sperm at pH values allowing fusion (pH 5-7). The increase of [Ca2+]i was proportional to the extent of fusion as measured through the relief of R18 self-quenching. We also examined the increase of sperm [Ca2+]i and the extent of fusion as a function of sperm to prostasome ratio and, also in this case, there was proportionality between the extent of fusion and the increase of [Ca2+]i that reached its maximal values in about 10-20 min. However, a detectable increase of [Ca2+]i was attained after 2 min of fusion. This would represent a new mechanism to influence sperm [Ca2+]i besides ion-exchange systems and ATP-dependent pumps. The value of [Ca2+]i remained elevated, unless Na+ was also present in the external medium. Therefore, the mechanism of fusion might influence deeply the physiology of sperm by producing a transient increase of [Ca2+]i.

Progesterone-induced increase of sperm cytosolic calcium is enhanced by previous fusion of spermatozoa to prostasomes.

Ejaculated spermatozoa must undergo a number of modifications before becoming able to fertilize the oocyte. The interaction of sperm with other semen components may influence these phenomena; human semen contains vesicles of prostatic origin, called prostasomes that may fuse to sperm at slightly acidic to neutral pH values. Prostasomes contain calcium and it has been demonstrated that their fusion with spermatozoa produces a transient increase (wave) of [Ca(2+)](i) in these cells. The fusion process also transfers protein and lipid to spermatozoa. These phenomena may induce long-lasting changes of sperm properties. We test the hypothesis that spermatozoa, as modified by fusion, change their ability to undergo the progesterone-induced increase of [Ca(2+)](i) and we find that the increase of [Ca(2+)](i) produced by the fusion with prostasomes and by the stimulation with progesterone are independent and additive phenomena. We also find that spermatozoa present a stronger response to the progesterone-induced increase of [Ca(2+)](i) if they are previously made to fuse with prostasomes. This effect does not depend directly on the [Ca(2+)](i) increase due to fusion, since it is still present after the [Ca(2+)](i) has returned to resting values.

Sulfation pharmacogenetics: SULT1A1 and SULT1A2 allele frequencies in Caucasian, Chinese and African-American subjects.

Sulfotransferase (SULT) enzymes catalyze the sulfate conjugation of drugs, other xenobiotics, neurotransmitters and hormones. The genes for SULT1A1 and SULT1A2 contain common genetic polymorphisms that are associated with individual variations in levels of enzyme activity as well as variations in biochemical and physical properties. We set out to compare the frequencies of common SULT1A1 and SULT1A2 alleles in Caucasian, Chinese and African-American subjects. Allele frequencies for SULT1A1*1, *2 and *3 in 242 Caucasian subjects were 0.656, 0.332 and 0.012, respectively. Frequencies of those same alleles were significantly different in 290 Chinese subjects: 0.914, 0.080 and 0.006, respectively, as were frequencies in 70 African-American subjects: 0.477, 0.294 and 0.229, respectively. Ethnic variation in allele frequencies was also observed for SULT1A2, with frequencies in Caucasian subjects for SULT1A2*1, *2 and *3 of 0.507, 0.389 and 0.104; frequencies in Chinese of 0.924 and 0.076 with no *3 alleles observed; and, finally, in African-Americans frequencies of 0.637, 0.249 and 0.114, respectively. We also found that SULT1A1*2 and SULT1A2*2, the most common variant alleles for these two genes, were in positive linkage disequilibrium in all three populations studied, with D' values of 0.776 in Caucasian (P < 0.001), 0.915 in Chinese (P < 0.001) and 0.864 in African-American subjects (P < 0.001). These observations represent a step towards determining the possible functional implications for individual variations in sulfate conjugation of common genetic polymorphisms for SULT1A1 and SULT1A2.

Transfer of CD26/dipeptidyl peptidase IV (E.C. 3.5.4.4) from prostasomes to sperm.

Prostasomes are vesicles present in human semen. They are secreted by the prostate and contain large amounts of cholesterol and sphingomyelin. Some of their proteins are enzymes. Prostasomes are involved in a number of biological functions. In previous papers we demonstrated that lipid can be transferred from prostasomes to sperm by a fusion process occurring at neutral or slightly acidic pH. In this paper we demonstrate that CD26/dipeptidyl peptidase IV, an enzymatic activity absent in sperm, is transferred to sperm from prostasomes. This may be of particular interest since, by this procedure, sperm may acquire new membrane-bound enzymes and modify the catalytic activity of their surface.

Factors influencing the aggressiveness elicited by marihuana in food-deprived rats.

1. Aggressive behaviour was elicited in rats that had been deprived of food for 20 h daily (starved), by chronic administration of Cannabis sativa extract or (-)-Delta(9)-trans-tetrahydrocannabinol.2. The influence of intraperitoneal (i.p.) or oral glucose administration, cold environment, acidosis, and corn, and protein-free diets on this aggressiveness was studied.3. Intraperitoneal injections of glucose (100-1,600 mg/kg) did not alter the aggressiveness induced by marihuana in starved rats; glucose given orally, however, blocked this behaviour.4. Low temperature (14 degrees C) strongly potentiated the aggressive behaviour induced by marihuana in the starved rats.5. Lactic acid in doses capable of potentiating thiopental anaesthesia, failed to alter the marihuana-aggressiveness of starved rats or to facilitate this effect of marihuana in rats fed ad libitum. The same negative results were obtained with ammonium chloride.6. In rats fed ad libitum with protein-free or corn diets, marihuana administered chronically did not elicit aggressive behaviour. However, aggressiveness appeared when rats were fed for only 2 h daily on those diets.7. The results suggest that the stress of hunger (and not hypoglycaemia, acidosis or lack of specific nutrients due to starvation) is the factor that facilitates the development of aggressive behaviour by chronic administration of marihuana.

Cannabis, catecholamines, rapid eye movement sleep and aggressive behaviour.

1. Previous work from our laboratory has shown that cannabis induces aggressive behaviour in rats that have been deprived of rapid eye movement (REM) sleep. It was suggested that this effect was related to brain catecholamines, with dopamine playing an agonist role and noradrenaline an inhibitory one. The present paper describes new experiments dealing with this subject. 2. Previous REM sleep-deprivation enhanced both delta9-tetrahydrocannabinol (THC)-induced hypothermia and nomifensine effects on aggressive behaviour. 3. A marihuana extract decreased brain dopamine turnover in REM sleep-deprived rats, an effect not observed in non-deprived rats. Noradrenaline metabolism was not altered. 4. Fighting behaviour was elicited in REM sleep-deprived rats treated with 4 different dopamine-beta-hydroxylase inhibitors. 5. Apomorphine, nomifensine and delta9-THC administered to non-deprived rats pretreated with bis(4-methyl-1-homopiperanzinyl-thiocarbonyl) disulphide (Fla-63), induced fighting behaviour. 6. Nomifensine and apomorphine induced fighting in non-deprived rats pretreated with delta9-THC. 7. Clonidine inhibited the fighting elicited in REM sleep-deprived rats by either delta9-THC or Fla-63 pretreatment. 8. The data are discussed in terms of the influence of REM sleep-deprivation (or the stress associated with deprivation) on the response to dopaminergic drugs and cannabis. Taken together they emphasize the participation of brain dopamine and noradrenaline systems in the aggressive behaviour studied.

Effects of chronic administration of (+)-amphetamine on maze performance of the rat.

1. The influence of (+)-amphetamine, given 1 min after each training session, on the performance of 124 rats in a Lashley III maze was measured every 48 hr.2. The first three injections of the drug significantly improved the learning ability of naive rats.3. With prolonged treatment, (+)-amphetamine strongly impaired the maze performance of these rats.4. The chronic administration of (+)-amphetamine to previously trained rats produced the same adverse effect.5. Amylobarbitone sodium given to previously trained rats 30 min before the training sessions completely blocked the adverse effect of (+)-amphetamine.6. (+)-Amphetamine did not produce impairment of performance when given chronically 30 min before training sessions, to previously trained rats.