Plants utilized as medicines by residents of Quilombo da Fazenda, Núcleo Picinguaba, Ubatuba, São Paulo, Brazil: A participatory survey.

ETHNOPHARMACOLOGICAL RELEVANCE: Participatory research can help to broaden the understanding of medical systems and beliefs of traditional communities. An ethnopharmacological survey in collaboration with local people focused on plants used in quilombos located in Southeast Region in Brazil identified cultural factors that influence plant and recipe choice. AIM OF THE STUDY: To investigate the factors related to the therapeutic efficiency of medicinal plants from the perspective of Quilombo da Fazenda residents. MATERIALS AND METHODS: University researchers collaborated with community residents for both aims and methods of the study. The local partners were trained in the gathering of ethnopharmacological data and then selected and interviewed the residents considered experts on the use of medicinal plants. Data on the use of each species were supported by voucher specimens collected by the local partners and university researchers. Participant observations and field diaries by the university researchers supplemented the data. RESULTS: Eight interviewees mentioned 92 medicinal species with 60 therapeutic uses, applied in 208 recipes or remedies. Asteraceae (13 species), Lamiaceae (5) and Urticaceae (5) contributed most medicinal plant species. Of the 12 etic categories of use, the circulatory system category had the highest number of plants mentioned. Decoction was the most commonly used preparation method (66.8%), and most remedies were administered orally (76.4%). Eighty-six recipes included more than one plant species and/or the addition of other components, such as sugar, salt or animal products. Several cultural factors influence medicinal plant use. Popular beliefs on the quality of blood or the humoral properties of plants and illnesses, characteristics of the plants and other factors determine which plant is used and why. CONCLUSIONS: The participatory method identified a large number of factors that influence medicinal plant use: the patient's blood type; the condition of the plant and the disease (hot-cold system); the route of administration and dosage; the preventive uses of the plants; and the influence of other factors, such as the sun, the moon and dew. The participatory approach is useful for gaining insight on the decision processes of medicinal plant use in traditional societies, and also for those communities wanting to document their knowledge with or without the participation of the academy.

Investigation of urban ethnoveterinary in three veterinary clinics at east zone of São Paulo city, Brazil.

ETHNOPHARMACOLOGICAL RELEVANCE: Urban Ethnoveterinary should be further explored because of its value as it resists conventional medical care. In addition, the comparison between the resources used by Ethnoveterinary and Ethnopharmacology should be investigated in depth, increasing the availability of new bioactive potential in human and veterinary medicines. This project aimed to determine whether plants are used in the health care of dogs and/or cats in urban area and to compare its uses with other ethnoveterinary and ethnopharmacological data. MATERIALS AND METHODS: Three veterinary clinics from east area of São Paulo city were selected, in order to record the offering of plants to pets by their owners. Individual interviews were conducted from May to November 2012 and consisted of application of semi-structured interviews and questionnaires. A literature search was performed to compare our findings with other ethnopharmacological and ethnoveterinarian data from the literature. RESULTS: A total of 273 people were interviewed and 46 (16.84%) of them utilize medicinal plants for their pets. Most plant species are exclusively home grown (57.9%). The plants most cited were Plectranthus barbatus Andrews, Matricaria chamomilla L. and Foeniculum vulgare Mill. The most frequent route of administration, part utilized and mode of preparation was oral (80.35%), leaves (89.47%) and infusion (61.90%), respectively. From 19 cited plants, 14 (73.7%) are mentioned in ethnoveterinary literature, whereas 11 (57.9%) are used for the same purposes. All plants reported in our study have at least one common use with ethnopharmacology. CONCLUSION: The survey provided evidence of ethnoveterinary use of medicinal plants for dogs and/or cats in urban area of São Paulo, complementarily with the official veterinary, and showed that the plants used in pets are also used in humans with the same purposes and routes of administration, as well as in other animals.

Regulation of progestin receptors in medial amygdala: estradiol, phytoestrogens and sex.

Exposure to estrogens during critical developmental periods and in adulthood affects sex differences in the brain. We examined the roles of estradiol (E2) and phytoestrogens, and their interactions, on potential sex differences in brain. We used aromatase knockout (ArKO) mice, which cannot produce endogenous estrogens, along with wild type (WT) littermates. Mice were gestated, raised and maintained on a diet either rich in phytoestrogens or a diet virtually void of soy-derived phytoestrogens. Adult males and females were gonadectomized and received implants filled with 17-beta-estradiol to induce progestin receptors (PR), while controls received empty implants. Mice were sacrificed five days later and brain sections containing the posterodorsal medial amygdala (MePD) were processed for PR immunoreactivity. Activation of sex differences in PR required adult E2 treatment. A diet high in phytoestrogens was required for expression of sex differences in PR after E2 treatment. Our data underscore the important contribution of dietary phytoestrogens for the development of sex differences in PR-ir in the adult mouse medial amygdala. We hypothesize that both aromatization of androgens to estrogens and dietary sources of additional estrogens are part of the normal requirement for sex differences in the rodent brain.

Structure, properties and enhanced expression of galactose-binding C-type lectins in mucous cells of gills from freshwater Japanese eels (Anguilla japonica).

Using a Japanese-eel (Anguilla japonica) gill cDNA subtraction library, two novel beta-d-galactose-binding lectins were identified that belong to group VII of the animal C-type lectin family. The eel C-type lectins, termed eCL-1 and eCL-2, are simple lectins composed of 163 amino acid residues, including a 22-residue signal peptide for secretion and a single carbohydrate-recognition domain (CRD) of approximately 130 residues typical of C-type lectins. The galactose specificity of the CRD was suggested by the presence of a QPD motif and confirmed by a competitive binding assay. Using Ruthenium Red staining, the lectins were shown to bind Ca(2+) ions. SDS/PAGE showed that native eCL-1 and eCL-2 have an SDS-resistant octameric structure (a tetramer of disulphide-linked dimers). Northern and Western blot analyses demonstrated high-level expression of eCL-1 and eCL-2 mRNAs and their protein products in gills from freshwater eels, which decreased markedly when the eels were transferred from freshwater to seawater. Immunohistochemistry showed that the eel lectins are localized in the exocrine mucous cells of the gill.

Metastin suppresses the motility and growth of CHO cells transfected with its receptor.

We recently reported having identified of the ligand for an orphan G-protein-coupled receptor, hOT7T175, as the gene product (68-121)-amide of the metastasis suppressor gene KiSS-1. We further showed that the ligand, which we named "metastin," inhibits chemotaxis and invasion of Chinese hamster ovary (CHO) cells transfected with hOT7T175 cDNA (CHO/h175) in vitro, and pulmonary metastasis of hOT7T175-transfected B16-BL6 melanomas in vivo. In the present study, we investigated the activity of metastin in CHO/h175 cells in greater detail. Metastin significantly suppressed motility in a chemotaxis assay and wound healing assay at 10-100 nM order concentrations. Two N-terminally truncated peptides, metastin(40-54) and metastin(45-54) inhibited the migration of CHO/h175 cells as potently as metastin itself. Metastin also inhibited the spreading, monolayer growth and colony formation in agar (0.8%) of CHO/h175 cells at 10-100 nM concentrations. These results indicate that metastin is a potent inhibitor of cell motility, leading to suppression of cell growth and antimetastatic activity, and suggest that low molecular chemical compounds could replace its activity as a novel antimetastatic agent.

The first clinical trial of topical application of procyanidin B-2 to investigate its potential as a hair growing agent.

Procyanidin B-2 is a compound we have identified in apple which acts as a growth-promoting factor on murine hair epithelial cells. This report describes our investigation of the hair-growing effects of 1% procyanidin B-2 tonic after sequential use for 4 months. A double-blind clinical trial was performed, involving a total of 29 subjects (procyanidin B-2, 19 men; placebo, 10 men). No adverse side effects were observed in either group. In the procyanidin B-2 group, 78.9% showed an increased mean value of hair diameter, whereas only 30.0% in the placebo group showed any increase (p < 0.02, Fisher's exact probability test). The increased ratio of hairs measuring more than 40 microm in diameter after 4 months of procyanidin B-2 treatment was significantly higher than that of the placebo controls (p < 0.05, two-sample-t-test). The increase in number of total hairs in the designated scalp area (0.25 cm(2)) of procyanidin B-2 subjects after a 4 month trial was significantly greater than that of the placebo controls (procyanidin-B-2, 3.67 +/- 4.09 (mean +/- SD)/0.25 cm(2); placebo, -2.54 +/- 4.00/0.25 cm(2); p < 0.001, two-sample t-test). Procyanidin B-2 therapy shows potential as a promising cure for male pattern baldness.

[Regulation of microglial cell function by ATP].

Accumulated evidence suggests that extracellular ATP functions occur in neurons and glial cells in the nervous systems. Besides well-documented roles as a neurotransmitter or neuromodulator, ATP has also been demonstrated to have effects on glial cells. Reports have shown that ATP stimulates microglia to release various biologically active substances, such as interleukin-1 beta, tumor necrosis factor-alpha, and plasminogen. Microglial cell death was also demonstrated after stimulation with high-dose ATP. Although these responses were known to occur, via P2X7, we have recently found that ATP and ADP induced the formation of membrane ruffles and chemotaxis through Gi/o-coupled P2Y receptors. Taken together, it is suggested that two distinct P2X and P2Y receptor subtypes are involved in the diverse function of microglia in both physiological and pathological states.

Ceramide-enhanced urokinase-type plasminogen activator (uPA) release is mediated by protein kinase C in cultured microglia.

As described previously, a relatively high dose of neurotrophins increased the release of urokinase-type plasminogen activator (uPA) from cultured microglia. This biological response is suggested to be caused by ceramide, which is a metabolite of nerve growth factor low-affinity receptor (NGFRp75)-associated sphingomyelin turnover. Therefore, in the present study, we examined the effect of ceramide on the release of uPA from cultured microglia. Treatment of the cells with permeable C8-ceramide (D-erythro-Sphingosine, N-octanoyl-) enhanced uPA release in a dose-dependent manner. This effect of C8-ceramide was mimicked by treatment with bacterial sphingomyelinase. A pharmacological study using a specific PKC activator, phorbol-12-myristate-13-acetate, and a protein kinase C (PKC) inhibitor, bisindolylmaleimide, showed that PKC activation is required in order to release uPA from ceramide-stimulated microglia as well as from nonstimulated microglia. Further study using a specific conventional PKC (cPKC) activator, 1-oleoyl-2-acetyl-sn-glycerol (OAG), and a specific cPKC inhibitor, Gö 6976, suggested that PKC-delta and/or -epsilon is involved in uPA release. As opposed to the apoptotic pathway, however, no activation of c-Jun N-terminal kinase and nuclear factor kappa B was observed in C8-ceramide-stimulated microglia. The findings suggest that uPA release from microglia is regulated by a mechanism in which PKC-delta and/or -epsilon are activated and further signals are transduced subsequently.

Preoperative diagnosis of ileal lipoma by endoscopic ultrasonography probe.

The occurrence of tumor in the small intestine is relatively rare. It has been demonstrated that lipoma of the ileum is a cause of intussusception. We report a 59-year-old man admitted to our hospital for lower abdominal pain. Diagnosis of intussusception was made by abdominal x-ray and ultrasonography. Enema contrast studies revealed ileocolic intussusception. Colonoscopy revealed a tumor with an submucosal tumor (SMT)-like head and coil-spring appearance in the ascending colon. Endoscopic ultrasonography (EUS) revealed a hyperechoic submucosal lesion with features compatible with lipoma. Subsequently, this was confirmed histopathologically after resection. To our knowledge, this is the first report of preoperative diagnosis of ileal lipoma by EUS.

Pharmacokinetic and pharmacodynamic evaluation for tissue-selective inhibition of cholesterol synthesis by pravastatin.

The tissue-selective inhibition of cholesterol synthesis by pravastatin was evaluated pharmacokinetically and pharmacodynamically. Plasma, tissue, urine, and bile concentrations were measured after i.v. bolus injection of pravastatin to rats at various doses. The total body clearance and steady state volume of distribution decreased with increasing dose. A saturable biliary excretion was also observed. The time course of plasma and liver concentrations was described by a three-compartment model, consisting of a central compartment, a deep compartment with an nonsaturable uptake process, and a shallow compartment with saturable uptake and nonsaturable elimination processes. It suggests that a mechanism for the decrease in the total body clearance and distribution volume might be explained by a saturation of pravastatin uptake into the liver. Plasma concentration data after oral administration was also fitted to the same model by connecting an absorption compartment to the shallow compartment. The inhibitory activity of pravastatin against cholesterol synthesis in liver could be related to the concentration in the shallow compartment via a sigmoidal Emax model and the obtained pharmacodynamic parameters were comparable to those in vitro. Results suggest that the carrier-mediated hepatic uptake of pravastatin is actually responsible for the hepatoselective inhibition of cholesterol synthesis under physiological conditions.

[The feasibility of CEF (cyclophosphamide, epirubicin, 5-FU) regimen in the adjuvant setting of primary breast cancer].

In Japan, 5-FU/5-FU derivatives or the combination therapy of CAF (cyclophosphamide, CPA; adriamycin, ADM; 5-fluorouracil; 5-FU) have been commonly used for the adjuvant treatment of breast cancer. Recently, a combination of CEF (CPA; Epirubicin, EPI; 5-FU) has come to the stage of adjuvant setting, because the cardiotoxicity was reduced in EPI. In this study, we investigated the feasibility of 6 cycles of CEF (CPA 700 mg/m2, EPI 70 mg/m2, 5-FU 700 mg/m2; day 1 iv every 3-4 weeks) in the adjuvant treatment of primary breast cancer patients with nodal involvements. All 12 patients completed 6 cycles of CEF within 8 months. The median treatment duration was 6.2 months. More than Grade III side effects of neutropenia, nausea/vomiting and alopecia were observed in 7/12 (58.3%), 5/12 (41.7%) and 12/12 (100%), respectively. No serious side effects, including cardiotoxicity, were shown. CEF seems to be feasible regimen as an adjuvant treatment for breast cancer.

Sofalcone for treatment of Helicobacter pylori infection.

It is difficult to regard sofalcone as a single drug for the treatment of Helicobacter pylori infection. However, sofalcone exerts multiple effects against H. pylori: it has antibacterial activity, induces morphological changes, inhibits adhesion to gastric mucin and inhibits lipolytic activity. The safety profile of sofalcone even on long-term administration is well established. Therefore, it may be possible to establish a new triple therapy for H. pylori infection using sofalcone combined with antibacterial drugs and proton pump inhibitors.

The alternative exons 1 of the mouse aromatase cytochrome P-450 gene.

Aromatase cDNA clones were isolated from cDNA libraries of mouse hypothalamus, amygdala and ovary. Analysis of the nucleotide sequences of the 5' regions of the obtained cDNAs suggested that the mouse aromatase gene is tissue-specifically regulated by alternative exons 1. There were obvious differences between the 5' regions of the brain and ovary aromatase cDNAs, but no difference was found between the sequences of the hypothalamus and amygdala ones. We further isolated a mouse genomic DNA clone containing brain- and ovary-specific exons 1. The brain specific exons 1 and their promoters were highly homologous in the human and mouse aromatase genes. In contrast there were several differences in the sequences among the promoter regions of the ovary-specific exons 1 of the mouse, human and rat aromatase genes, significant homology between their sequences was also observed. The present results demonstrate that expression of the mouse aromatase gene is also tissue-specifically regulated through the use of alternative exons 1 and promoters, as reported for man.

[Studies on Rehmanniae radix. III. The relation between changes of constituents and improvable effects on hemorheology with the processing of roots of Rehmannia glutinosa].

The relation between constituents and effects on hemorheology with processing of root of Rehmannia glutinosa was investigated. With the processing to dried or steamed root from crude root, the content of stachyose, which was the highest component in crude root, was decreased and the content of manninotriose was increased. Iridoid glycoside, catalpol, was gradually decreased with the processing. From these results, the content of carbohydrates and catalpol made it possible to estimate the quality of Rehmanniae Radix by processings. On the other hand, the change of pharmacological activities with the processing was investigated. The 50% ethanolic extract (BJ-ext) from steamed root of R. glutinosa increased erythrocyte deformability and erythrocyte ATP contents, inhibited polybrene-induced erythrocyte aggregation, and promoted an activity of fibrinolytic system. The extracts from crude or dried root had weak or no effect. The crude or dried root and steamed root of R. glutinosa showed the different pharmacological activities, and the quality of Rehamanniae Radix by processings may be estimated by investigation of correlation between the changes of constituents and improvable effects of hemorheology.

The distribution fields of sensory neurons in the human thalamus.

Thalamic neurons were identified by activity related to passive joint movement, active joint movement, tapping stimulation, and light touch stimulation during surgery to treat movement disorders. The neurons were classified into three types: movement-related neurons, tapping-related neurons, or superficial sensory neurons. Tapping-related neurons had characteristics of lemniscal sensory neurons and occupied the border area of movement-related neurons and superficial sensory neurons. Tapping-related neurons showed a laminar distribution 1.0-1.5 mm in width in the anterodorsal region of the nucleus ventrocaudalis. Results suggest that the distribution pattern of tapping-related neurons in the human thalamus is consistent with the functional distribution pattern found in the monkey thalamus.

Molecular cloning and in situ localization in the brain of rat sky receptor tyrosine kinase.

We previously identified a human cDNA encoding a novel receptor tyrosine kinase, termed Sky, which is predominantly expressed in the brain and has a unique extracellular domain consisting of two immunoglobulin (Ig)-like and two fibronectin type III (FN III) motifs. In attempts to define the functional role of the Sky receptor, we cloned a rat sky cDNA, and localized the sites of expression of sky transcript in the adult rat brain by Northern blot and in situ hybridization analyses using the cloned rat cDNA as a probe. The deduced amino acid sequence of rat Sky has an overall sequence and a domain topology highly conserved with human Sky (90% overall identity and 98% identity within the tyrosine kinase domain). Northern blot analysis revealed that a single 3.8-kb sky mRNA is expressed in PC12 pheochromocytoma and Neuro-2a neuroblastoma cell lines and in various regions of the adult rat brain. In situ hybridization analysis revealed widespread but confined neuronal populations in adult rat brain that express sky transcript; prominent hybridization signals were detected in the inner granular layer of the olfactory bulb, CA-1 area of the hippocampus, granule cell layer of the cerebellum, tenia tectum and cingulate gyrus neurons, and wide regions of cortex layers II-VI. The high level of expression of sky mRNA in neurons in restricted brain regions suggests that the Sky receptor may play an important role in development, function, and maintenance of specific neuronal populations in the central nervous system.

Novel exon 1 of the aromatase gene specific for aromatase transcripts in human brain.

Aromatase in the brain is supposed to participate in sexual differentiation of the brain. The presence of a brain-specific exon 1 and promoter in the human aromatase gene was examined in the 5' regions of aromatase cDNAs isolated from the amygdala and hypothalamus areas. The DNA sequences of all the isolated clones were different in the 5' regions encoded by exon 1 from those encoded by any alternative exons 1 of human aromatase gene. This unique sequence was also found in the clones isolated from a human genomic library and indicated to be that of a new exon 1. It involved putative TATA and CAAT boxes, Ad4 sequence, and the androgen responsive element in the promoter region. Reverse transcriptase-polymerase chain reaction analysis of mRNAs in various tissues revealed that expression of this unique species of aromatase mRNA is specific for the brain, in particular in the fetal brain. These show that the aromatase gene in the human brain is expressed by means of the brain-specific exon 1 and promoter, and may be responsible for sexual differentiation of the brain in the fetal period.

Dietary fish oil alters rat milk composition and liver and brain fatty acid composition of fetal and neonatal rats.

This study was performed to determine the effects of dietary fish oil rich in docosahexaenoic acid [22:6(n-3)] on the milk composition of maternal rats and tissue fatty acid composition of fetal and neonatal rats. The fatty acid composition of dam's milk was affected by the dietary fat. The level of 22: 6(n-3) in the milk of dams fed the fish oil-containing diet was severalfold higher than that of dams fed the diet that did not contain fish oil, and the levels of 22: 6(n-3) in the phospholipids of the livers and brains of fetuses and pups of the fish oil-fed group were higher than those in fetuses and pups of the group that did not receive fish oil. These findings suggest that a high level of 22:6(n-3) in the livers and brains of fetal and neonatal rats can be achieved by intake of fish oil by the dams.

Ad4BP regulating steroidogenic P-450 gene is a member of steroid hormone receptor superfamily.

Bovine cytochrome P-450(11 beta) gene (CYP11B) has six different cis-acting elements, Ad1, Ad2, Ad3, Ad4, Ad5, and Ad6, in the promoter region. The Ad4 site also exists in the promoter regions of other steroidogenic P-450 genes as well as in CYP11B1. An Ad4-binding protein (Ad4BP) which specifically binds to the Ad4 site was purified from bovine adrenal cortex nuclear extract, and its molecular mass was 53 kDa. A complementary DNA encoding Ad4BP was isolated from a bovine adrenal cortex cDNA library. The cDNA clone contained an open reading frame of 1383 base pairs encoding 461 amino acids, whose calculated molecular weight was 51,020. The predicted amino acid sequence of Ad4BP revealed that the protein has a zinc finger domain and a ligand binding/dimerization domain. Ad4BP is a novel member of the steroid hormone receptor superfamily. Comparison of the primary structures of the hormone receptor superfamily showed that Ad4BP was highly homologous to FTZ-F1, which regulates the fushi tarazu gene, and ELP, which is expressed in the murine embryonal carcinoma cells. Transfection of a Ad4BP expression plasmid into CV-1 cells activated the transcription of the CAT reporter gene carrying the Ad4 sequence in the promoter region.

High-dose intravenous gammaglobulin therapy for neonatal immune haemolytic jaundice due to blood group incompatibility.

Three newborn infants who developed hyperbilirubinemia due to blood group incompatibility were treated with high-dose gammaglobulin. Hyperbilirubinemia was caused by Rhesus (Rh) incompatibility (anti-E + anti-c) in Infant 1 and ABO incompatibility (anti-B) in Infants 2 and 3. Hyperbilirubinemia was refractory to conventional phototherapy but responded well to intravenous gammaglobulin (IVGG) at a dose of 1 g/kg in all infants. No adverse effects were observed. These findings suggest that high-dose IVGG may be useful in the treatment of hyperbilirubinemia due to isoimmune haemolytic disease resistant to phototherapy.