Presence of cholinomimetic and acetylcholinesterase inhibitory constituents in betel nut

In this investigation, we report the presence of cholinomimetic and acetylcholinesterase (AChE) inhibitory constituents in betel nut, the most commonly used drug in the world after tobacco, ethanol and caffeine. The crude extract of betel nuts or Areca catechu (Ac.Cr) caused a dose-dependent (0.3-300 microg/mL) spasmogenic effect in the isolated rabbit jejunum. The spasmogenic effect was blocked by atropine, similar to that of acetylcholine (ACh), suggestive of muscarinic receptor mediated effect. Both the extract (0.3-10 microg/mL) and physostigmine (0.1-3.0 microM) potentiated the effect of a fixed dose of ACh (10 microM) in a dose-dependent fashion, suggesting acetylcholinesterase (AChE) inhibitory effect. This effect was confirmed in the in vitro assay where both the crude extract (1-100 microg/mL) and physostigmine inhibited the enzyme. In the in vivo model of gastrointestinal transit, Ac.Cr (10-30 mg/kg) enhanced the travel of charcoal meal and also exhibited a laxative effect in mice. The plant extract was subjected to activity-directed fractionation and all resultant fractions showed atropine-sensitive spasmogenicity in rabbit jejunum and also AChE inhibitory effect at doses similar to that for the parent crude extract, the ethyl acetate fraction being slightly less potent. Some of the known constituents of betel nut, including arecoline, were tested for the possible inhibitory effect on AChE, none were found active. The study provides first evidence for the presence of AChE inhibitory constituents in betel nut, though additional direct muscarinic stimulatory effect cannot be ruled out and this study provides sound scientific basis for some of the folkloric uses associated with betel nut chewing.     

Transplacental micronucleus inducing ability of arecoline, a betel nut alkaloid, in mice

Arecoline, a major betel nut alkaloid, was tested for its effectiveness in inducing micronuclei in fetal mouse blood after transplacental exposure late in the gestation period. Positive results were obtained and a linear dose-response relationship was expressed when pregnant mice were treated with arecoline at dose levels of 20, 40 and 80 mg/kg and micronucleated polychromatic erythrocytes from fetal blood were subsequently analysed.     

Effect of chewing betel nut on measurements of salivary progesterone and estradiol

The measurement of steroids in saliva is both simple and non-invasive and has been widely used in field and clinical-based research. The observance of particular cultural practices by some populations, however, may hamper accurate hormonal analyses. The present study evaluated the effects of one such practice-the chewing of betel nut-on the accurate measurement of salivary progesterone and estradiol. A time series experiment was conducted among Bangladeshi women who are regular users of betel nut. Salivary steroids were analyzed by radioimmunoassay in samples collected prior to and then 30, 60, 120, and 240 min following betel quid use. Results show no significant difference between basal steroid levels and those obtained 60, 120, and 240 min after chewing betel nut. We conclude that with specific collection protocols that take into account time since chewing, salivary steroid analyses can be undertaken in populations among whom the practice of chewing betel nut is endemic.     

A monograph of the betel nut palms (Areca: Arecaceae) of East Malesia

The palm genus Areca is widespread in tropical Asia and includes the economically important betel nut palm, A. catechu. The genus has three centres of high species diversity: the Sunda Region, the Philippines and East Malesia (to the east of Wallace's line). The taxonomy of the genus in East Malesia has been neglected. Prior to this study, 19 species were accepted for this area, all but one endemic, but their limits and differences were not understood. Here, we provide a taxonomic monograph of East Malesian Areca spp., based on an extensive study of the genus in herbaria and in the field. We recognize six species of Areca in East Malesia, including the widespread cultivated A. catechu. Five wild species are accepted, namely A. macrocalyx, A. mandacanii, A. novohibernica, A. oxycarpa and A. vestiaria. We place 12 of the previously accepted species into synonymy, and provide additional new synonymy in A. catechu. © 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 168 , 147–173.     

Vitamin D: a clinical perspective

Vitamin D has been the focus of considerable recent scientific study and media attention because of a growing appreciation of its potential importance in extra‐skeletal health. This ‘Perspective’ presents data supporting the viewpoint that physicians should not routinely test vitamin D levels in the general population but should routinely endorse vitamin D supplementation at levels recommended by the Institute of Medicine. It further reasons that it is unadvisable to recommend intentional limited sun exposure as a source of vitamin D.     

Modulation of platelet aggregation by areca nut and betel leaf ingredients: roles of reactive oxygen species and cyclooxygenase

There are 2 to 6 billion betel quid (BQ) chewers in the world. Areca nut (AN), a BQ component, modulates arachidonic acid (AA) metabolism, which is crucial for platelet function. AN extract (1 and 2 mg/ml) stimulated rabbit platelet aggregation, with induction of thromboxane B2 (TXB2) production. Contrastingly, Piper betle leaf (PBL) extract inhibited AA-, collagen-, and U46619-induced platelet aggregation, and TXB2 and prostaglandin-D2 (PGD2) production. PBL extract also inhibited platelet TXB2 and PGD2 production triggered by thrombin, platelet activating factor (PAF), and adenosine diphosphate (ADP), whereas little effect on platelet aggregation was noted. Moreover, PBL is a scavenger of O2(*-) and *OH, and inhibits xanthine oxidase activity and the (*)OH-induced PUC18 DNA breaks. Deferoxamine, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) and neomycin prevented AN-induced platelet aggregation and TXB2 production. Indomethacin, genistein, and PBL extract inhibited only TXB2 production, but not platelet aggregation. Catalase, superoxide dismutase, and dimethylthiourea (DMT) showed little effect on AN-induced platelet aggregation, whereas catalase and DMT inhibited the AN-induced TXB2 production. These results suggest that AN-induced platelet aggregation is associated with iron-mediated reactive oxygen species production, calcium mobilization, phospholipase C activation, and TXB2 production. PBL inhibited platelet aggregation via both its antioxidative effects and effects on TXB2 and PGD2 production. Effects of AN and PBL on platelet aggregation and AA metabolism is crucial for platelet activation in the oral mucosa and cardiovascular system in BQ chewers.     

A preliminary assessment of possible mutagenicity of betel nut and ingredients of the betel quid when administered alone or in combinations to larvae of Drosophila melanogaster

The “carcinogenic” betel nut and constituents of betel quid were tested for possible mutagenicity in Drosophila. The test compounds were administered either alone or in combinations by larval feeding. The data on sex-chromosome loss, sex-linked recessive lethals and autosomal translocations suggest lack of mutagenicity.     

Towards an understanding of the role of forces in carcinogenesis: a perspective with therapeutic implications.

Carcinogenesis may aberrantly have as its source an underlying dynamic which likely operates in accord with or is expressive of a universal principle of force. This drive can be seen as a type of necessary completion of force configurations giving forth a force-based, completing guidance or dyneotaxis. This completing guidance is a completing communication. Aberrations of this drive can manifest as incomplete communication arising from incomplete or non-uniform force-configurations throughout different levels of organization within the organism. Such non-uniformities of force-configuration manifest themselves through intra- and intercellular distortion. This is represented by changes or distortions within cellular and tissue architecture, or shape, occurring through and enabling a non-uniform loosening. Cycles of non-uniform loosening and distortion would generate a progressive disruption or aberration in an otherwise further adhesively completing, thereby further stabilizing, dyneotaxis. Carcinogenesis ensues as an aberrant means to re-establish a uniformity or symmetry of adhesive and mechanical forces in such tissues, and within the genome. However, it rather results in a further loosening-based reorganization or reshaping. This would be a manifestation of a still further non-uniformity of forces. Uncontrolled mutagenesis, itself stemming from genomic distortion, is also seen as a process that could lead to loosening and further distortion throughout the cell, and thereby, a contributor to carcinogenesis if a transcellular, globally completive or uniform adhesiveness cannot be maintained. A therapeutic approach could be employed to reverse such a cycle of loosening and distortion within cellular and tissue organization. The developmental reversal of carcinogenesis would be the consequence. Such an approach could be through the application of a coherent, uniform and united array of guiding mechanical and electromagnetic forces. The deeper consequence would be the re-establishment of a global, cancer-countering, uniform or symmetrical, guiding adhesiveness via and dependent on a complete and non-distorted network of cellular and intercellular structures, further completing, and thus further stabilizing, such structures in the process.     

A preliminary assessment of possible mutagenicity of betel nut and ingredients of the betel quid when administered alone or in combinations to larvae of Drosophila melanogaster.

The "carcinogenic" betel nut and constituents of betel quid were tested for possible mutagenicity in Drosophila. The test compounds were administered either alone or in combinations by larval feeding. The data on sex-chromosome loss, sex-linked recessive lethals and autosomal translocations suggest lack of mutagenicity.     

Companion animal parasitology: a clinical perspective

In recent years there have been many changes to the ways that clinical veterinary science is conducted and nowhere is this more evident than in companion animal practice. Veterinarians working with pet dogs and cats are facing new challenges associated with the emergence and re-emergence of parasitic diseases. Some, such as Neospora caninum, have been recently recognised; others like Giardia and Cryptosporidium have been reported with increasing frequency, in part as a result of laboratory tests with improved sensitivity and specificity. In many regions, the emergence of parasitic diseases has been a consequence of pet travel and exotic diseases pose a unique diagnostic challenge for the veterinarian, as the index of suspicion for these conditions may be absent. The ranges of certain vector-borne diseases such as babesiosis, hepatozoonosis, ehrlichiosis, leishmaniasis and dirofilariasis are extending due to ecological and climatic changes and enhanced by animals with subclinical infection returning home from endemic areas. In companion animal practice, veterinarians have the additional responsibility of providing accurate information about the zoonotic transmission of parasite infections from pets, especially to those most vulnerable such as children, the elderly and the immunocompromised. Effective education is vital to allay public concerns and promote responsible pet ownership.     

The effect of betel nut extract on cell growth and prostaglandin endoperoxide synthase in human epidermoid carcinoma cells

The objective of this study was to find out whether prostaglandin endoperoxide synthase (PHS) involves the action of betel nut extract (BNE) on the growth of oral cancers. Therefore, growth and PHS activity were examined in two human oral carcinoma cell lines (OEC-M1 and KB) and one normal fibroblast cell line (NF) in the presence of increasing BNE concentration. BNE at concentrations above 50 microg/ml significantly inhibited the cell growth of OEC-M1 after 72 h in culture, of KB and NF after 48 h in culture. The IC50 of BNE in OEC-M1, KB and NF at 24 h in culture was about 406, 37.5 and 140 microg/ml respectively. PHS activity in OEC-M1 was significantly increased by low BNE concentrations (50 microg/ml, 114%; 100 microg/ml, 33%; 150 microg/ml, 30%) but significantly reduced at higher BNE concentrations (300 microg/ml, 33%; 500 microg/ml, 61%). The PHS activity in KB was significantly inhibited by BNE and this effect was intensified as concentrations increased (50 microg/ml, 31%; 100 microg/ml, 24%; 150 microg/ml, 43%; 300 microg/ml, 60%; 500 microg/ml, 92%). Similar to that in OEC-M1, the PHS activity in NF was significantly increased at low BNE concentrations (50 microg/ml, 139%; 100 microg/ml, 87%;150 microg/ml, 77%) but reduced at higher concentrations (300 microg/ml, 55%; 500 microg/ml, 72%). The PHS activity in all cell lines was almost completely blocked by indomethacin (5 x 10(-6) M). We conclude that these findings suggest that PHS may be an important biochemical mediator of the effect of BNE on the growth of two human oral carcinoma cell lines.     

Study of unscheduled DNA synthesis following exposure of human cells to arecoline and extracts of betel nut in vitro.

Aqueous, acetic acid, hydrochloric acid and ethanol extracts of betel nut (Areca catechu L.) have been found to induce unscheduled DNA synthesis in Hep 2 cells obtained from human larynx carcinoma, in vitro. Different concentrations of extracts of betel nut induced dose-dependent unscheduled DNA synthesis in Hep 2 cells. Together with the viability of the Hep 2 cells, our results indicate that the aqueous and acetic acid extracts of betel nut induce relatively more unscheduled DNA synthesis than the hydrochloric acid and ethanol extracts and arecoline. The carcinogenic potency of raw and unprocessed betel nut of North-East India used in this study is discussed.     

Phylogeographic analysis of the Bufo gargarizans species complex: a revisit

Using mtDNA sequencing and allozyme electrophoresis data, we tested the "vicariance followed by dispersal" hypothesis of the Bufo gargarizans species group and re-evaluated the species status in the general lineages species concept. A phylogenetic analysis suggested that dispersal, instead of vicariance, dominated the history of the species group. There was a general trend of west to east dispersal, while some lineages from the east subsequently returned to the west. The secondary admixture of those previously allopatric lineages produced substantial levels of sympatric genetic diversity, often as high as 7.0% pairwise difference within populations. The phylogenetic hypothesis does not support the current two species designation. Neither B. andrewsi nor B. gargarizans represents an independent evolutionary lineage, and monophyletic groups did not correspond to geographically discrete groups. Allozyme data also failed to reveal any fixed allelic difference among the populations. Therefore, we recommend regarding the complex as a single species, Bufo gargarizans, without subspecies division.     

Is fever beneficial to the host: a clinical perspective

Fever is a phylogenetically ancient host response to infection, being found in fish and lizards, and conserved, with all its metabolic costs, in the higher mammals, including man. The conservation of the fever response in evolution is used as an argument for its survival value and, indeed, in experiments with cold-blooded animals "behavioral fever" has been demonstrated to reduce mortality associated with infection. Recent advances in the biology of interleukin-1 and other cytokines have allowed the testing, in vitro, of components of mammalian host defense (such as immune cell function) at temperatures typical of fever, and marked effects have been found. It remains to be demonstrated, however, that the hyperthermia of fever has survival value in man, and though it might be predicted that fever would be beneficial in infections, it is quite possible that in some circumstances even mild fever could be construed as harmful. In autoimmunity, for example, increased T-cell activation at febrile temperatures may well accelerate disease progress.     

Pathogenesis of lymphatic disease in bancroftian filariasis: a clinical perspective

The pathogenesis of lymphatic filariasis has been a matter of debate for many decades. Here, Gerusa Dreyer and colleagues propose a dynamic model of bancroftian filariasis, integrating clinical, parasitological, surgical, therapeutic, ultrasonographic and histopathological data. This model has profound implications for filariasis control programs and the management of the individual patient.     

The value of juvenile animal studies: a pediatric clinical perspective

With the emphasis of US American and European legislators on consideration of children in the drug development process regulatory authorities ask increasingly for additional non-clinical data to elucidate the safety of a given drug in development in future pediatric use. Juvenile animal studies are increasingly requested. These requests should never be tick box requests. Companies, academic toxicologists, clinicians, and regulatory authorities need a dialogue to differentiate between the perceived need to do "something" and the request for studies that have clinically meaningful results.