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1.  Effect of Purificatory Measures Through Cow's Urine and Milk on Strychnine and Brucine Content of Kupeelu (Strychnos Nuxvomica Linn.) Seeds 
Strychnos nux vomica Linn.(Loganaceae) commonly known as Nux vomica (Kupeelu), is a poisonous plant and its seeds are used widely in Ayurvedic system of medicine since time immemorial. Ayurveda advocates that nux vomica seeds are to be administered in therapeutics only after going through certain purificatory measures (Shodhana). There are more than six media: cow's urine (Go mutra), cow's milk (Go dugdha), cow's ghee (Go ghrita), Kanji (thin gruel), castor oil (Eranda taila) and fresh ginger juice (Ardraka swarasa) etc., which have been reported in different classical texts of Ayurveda for proper processing of nux vomica seeds. In this study, an attempt has been made to purify the seeds by using three different methods as described in ancient treatise by using cow's urine and cow's milk as media alone and together. This study revealed that all the methods studied reduced the toxicity of strychnine and brucine contents in comparison to the raw seeds as determined by HPTLC. Out of these three methods maximum reduction in strychnine and brucine contents was found when the seeds were purified by keeping them in cow's urine for seven days followed by boiling in cow's milk for three hrs.
PMCID: PMC3746529  PMID: 23983327
Kupeelu; Strychnos nuxvomica; Shodhana; strychnine; Ayurveda; brucine; Cow's milk; Cow's urine
2.  Strychnos nux-vomica seeds: Pharmacognostical standardization, extraction, and antidiabetic activity 
Background:
Strychnos nux-vomica, commonly known as kuchla, contains strychnine and brucine as main constituents. Minor alkaloids present in the seeds are protostrychnine, vomicine, n-oxystrychnine, pseudostrychnine, isostrychnine, chlorogenic acid, and a glycoside. Seeds are used traditionally to treat diabetes, asthma, aphrodisiac and to improve appetite.
Objective:
The present study was aimed to evaluate the various pharmacognostical characters and antidiabetic activity of S. nux-vomica seed.
Materials and Methods:
Pharmacognostical characters were performed as per the WHO guideline. Extraction was carried out in petroleum ether, chloroform, alcohol, hydroalcoholic, aqueous, and phytochemical constituents present in extracts were detected by different chemical tests. Among these extracts hydroalcoholic, aqueous extracts were evaluated for antidiabetic activity on the basis of extractive yield and phytoconstituents, in alloxan-induced diabetic rats using gliclazide as standard.
Results:
Various analytical values of S. nux-vomica extract were established. Phytoconstituents present in S. nux-vomica extracts were detected.
Conclusion:
S. nux-vomica extracts show antihyperglycemic activity in experimental animals.
doi:10.4103/0975-9476.96523
PMCID: PMC3371563  PMID: 22707864
Antidiabetic activity; alloxan; extract; kuchla
3.  Simultaneous HPTLC determination of strychnine and brucine in strychnos nux-vomica seed 
Objective:
A simple, sensitive, and specific thin layer chromatography (TLC) densitometry method has been developed for the simultaneous quantification of strychnine and brucine in the seeds of Strychnos nux-vomica.
Materials and Methods:
The method involved simultaneous estimation of strychnine and brucine after resolving it by high performance TLC (HPTLC) on silica gel plate with chloroform–methanol–formic acid (8.5:1.5:0.4 v/v/v) as the mobile phase.
Results:
The method was validated as per the ICH guidelines for precision (interday, intraday, intersystem), robustness, accuracy, limit of detection, and limit of quantitation. The relationship between the concentration of standard solutions and the peak response was linear within the concentration range of 50–1000 ng/spot for strychnine and 100–1000 ng/spot for brucine. The method precision was found to be 0.58–2.47 (% relative standard deviation [RSD]) and 0.36–2.22 (% RSD) for strychnine and brucine, respectively. Accuracy of the method was checked by recovery studies conducted at three different concentration levels and the average percentage recovery was found to be 100.75% for strychnine and 100.52% for brucine, respectively.
Conclusions:
The HPTLC method for the simultaneous quantification of strychnine and brucine was found to be simple, precise, specific, sensitive, and accurate and can be used for routine analysis and quality control of raw material of S. nux-vomica and several unani and ayurvedic formulations containing this as an ingredient.
doi:10.4103/0975-7406.94814
PMCID: PMC3341717  PMID: 22557924
HPTLC; method development; strychnine; brucine; validation
4.  Effect of Shodhana (processing) on Kupeelu (Strychnos nux-vomica Linn.) with special reference to strychnine and brucine content 
Ayu  2011;32(3):402-407.
Kupeelu (Strychnos nux-vomica Linn.) commonly known as nux vomica is a poisonous plant used extensively in various ayurvedic formulations, with great therapeutic significance. Ayurveda recommended the administration of Kupeelu only after purification in different media like cow's urine (Go mutra), cow's milk (Go dugdha), cow's ghee (Go ghrita), Kanji (sour gruel), and so on. Apart from the classical methods some other methods are also adopted by the traditional practitioners using castor oil (Eranda taila), ginger juice (Ardraka swarasa), in the purification of Kupeelu seeds. In the present study an attempt has been made to purify the seeds by performing two different methods (one classical and another traditional) using Kanji and Ardraka swarasa as Shodhana media. This study reveals that both the methods studied reduce the strychnine and brucine contents in comparison to the raw seeds as determined by high performance thin layer chromatography (HPTLC). After purification in Kanji and Ardraka swarasa, the strychnine content was reduced by 39.25% and 67.82%, respectively, and the brucine content in the purified seeds was also found to have decreased by 17.60% and 40.06%, in comparison to the raw seeds.
doi:10.4103/0974-8520.93923
PMCID: PMC3326892  PMID: 22529660
Ardraka swarasa; brucine; kanji; kupeelu; shodhana; strychnine
5.  Use of Strychnos Nux-Vomica (Azraqi) Seeds in Unani System of Medicine: Role of Detoxification 
Some plants used in Unani system of medicine are toxic, even deadly poisonous. The drugs having such plants as their components are detoxified before they are dispensed to the patients. One such drug, capsule Hudar, has Strychnos nux-vomica L. (Azraqi) seeds as one of its components and is very effectively used to elevate blood pressure. Ancient manuscripts describe many methods of its detoxification. It has been found that the detoxification processes studied reduce the strychnine content, as determined either by using uv-vis spectrophotometer or HPLC, present in Strychnos nux vomica seeds which is responsible for Strychnos nux vomica toxicity. The decrease in strychnine amount was best when the seeds were immersed for detoxification in excess of water for 5 days, in milk for 2 days followed by their boiling in milk. Strychnine in small amounts has been reported to give subjective feeling of stimulation
PMCID: PMC3005396  PMID: 21731158
Strychnos nux vomica; detoxification; Unani medicine
6.  Importance of Media in Shodhana (Purification / Processing) of Poisonous Herbal Drugs 
Ancient Science of Life  2010;30(2):54-57.
In Ayurveda, a series of pharmaceutical procedures which converts a poisonous drug into a therapeutically very effective medicine for various ailments is termed as Shodhana. Various medias are being used for processing the herbal poisonous drugs, are quite interesting to understand with modern scientific technology. The analysis of media before and after Shodhana (purification /processing) will give clear rationale behind the selection of the particular media for the particular drug. The change that takes place during the Shodhana process can be explored by modern analytical methods. Researchers have proved the presence of strychnine and brucine in milk after Shodhana of Nux-vomica highlighting the role media for Shodhana. Importance of Shodhana, the role of media used for Shodhana process of few poisonous drugs is dealt briefly with scientific view.
PMCID: PMC3336272  PMID: 22557427
Shodhana; Purification; Processing; Poisonous herbs; Media
7.  Brucine suppresses ethanol intake and preference in alcohol-preferring Fawn-Hooded rats 
Acta Pharmacologica Sinica  2014;35(7):853-861.
Aim:
Brucine (BRU) extracted from the seeds of Strychnos nux-vomica L is glycine receptor antagonist. We hypothesize that BRU may modify alcohol consumption by acting at glycine receptors, and evaluated the pharmacodynamic profiles and adverse effects of BRU in rat models of alcohol abuse.
Methods:
Alcohol-preferring Fawn-Hooded (FH/Wjd) rats were administered BRU (10, 20 or 30 mg/kg, sc). The effects of BRU on alcohol consumption were examined in ethanol 2-bottle-choice drinking paradigm, ethanol/sucrose operant self-administration paradigm and 5-d ethanol deprivation test. In addition, open field test was used to assess the general locomotor activity of FH/Wjd rats, and conditioned place preference (CPP) was conducted to assess conditioned reinforcing effect.
Results:
In ethanol 2-bottle-choice drinking paradigm, treatment with BRU for 10 consecutive days dose-dependently decreased the ethanol intake associated with a compensatory increase of water intake, but unchanged the daily total fluid intake and body weight. In ethanol/sucrose operant self-administration paradigms, BRU (30 mg/kg) administered before each testing session significantly decreased the number of lever presses for ethanol and the ethanol intake, without affecting the number of sucrose (10%) responses, total sucrose intake, and the number of lever presses for water. Acute treatment with BRU (30 mg/kg) completely suppressed the deprivation-induced elevation of ethanol consumption. Treatment with BRU (10, 20, and 30 mg/kg) did not alter locomotion of FH/Wjd rats, nor did it produce place preference or aversion.
Conclusion:
BRU selectively decreases ethanol consumption with minimal adverse effects. Therefore, BRU may represent a new pharmacotherapy for alcoholism.
doi:10.1038/aps.2014.28
PMCID: PMC4088279  PMID: 24909512
alcoholism; ethanol; brucine; glycine receptor antagonist; Fawn-Hooded (FH/Wjd) rat
8.  TLC – SPECTROPHOTOMETRIC ANALYSIS OF STRYCHNINE AND BRUCINE FROM THE AYURVEDIC PILLS OF NUX VOMICA 
Ancient Science of Life  1986;6(1):47-48.
Ayurvedic preparations claim on their label only the quantity of crude drugs and not the quantity of active ingredients present therein. So work was taken up to find the percentage of strychnine and brucine from Ayurvedic pills of Nux vomica powder by TLC spectrophotometric analysis, which study has not been reported earlier. However, the literature survey only revealed the following work.
PMCID: PMC3331393  PMID: 22557548
9.  Important Poisonous Plants in Tibetan Ethnomedicine 
Toxins  2015;7(1):138-155.
Tibetan ethnomedicine is famous worldwide, both for its high effectiveness and unique cultural background. Many poisonous plants have been widely used to treat disorders in the Tibetan medicinal system. In the present review article, some representative poisonous plant species are introduced in terms of their significance in traditional Tibetan medicinal practices. They are Aconitum pendulum, Strychnos nux-vomica, Datura stramonium and Anisodus tanguticus, for which the toxic chemical constituents, bioactivities and pharmacological functions are reviewed herein. The most important toxins include aconitine, strychnine, scopolamine, and anisodamine. These toxic plants are still currently in use for pain-reduction and other purposes by Tibetan healers after processing.
doi:10.3390/toxins7010138
PMCID: PMC4303819  PMID: 25594733
poisonous plants; Tibetan ethnomedicine; toxins; aconitine; strychnine; scopolamine; anisodamine
10.  A comparative anti-inflammatory activity of raw and processed Kupeelu (Strychnos nux-vomica Linn.) seeds on albino rats 
Ancient Science of Life  2011;31(2):73-75.
Seeds of Kupeelu (Strychnos nux-vomica Linn.), a known poisonous drug, is used extensively in various Ayurvedic formulations with great therapeutic significance. Ayurveda recommends the administration of Kupeelu only after passing through specific purificatory procedures in different media like cow's urine (Go mutra), cow's milk (Go dugdha), cow's ghee (Go ghrita), Kanji (thin gruel) etc. Strychnos nux vomica seeds are extensively advocated for nervous debility, paralysis, and weakness of limbs, sexual weakness, dyspepsia, and dysentery and in rheumatism where it can be assumed that besides other properties, Kupeelu may have some sort of anti-inflammatory activity too. In the present study, the powder of raw and processed Kupeelu seeds (processed / purified with Kanji i.e sour gruel) as test drugs were assessed for anti-inflammatory activity by employing Carrageenan and Formaldehyde induced hind paw oedema in Wistar strain albino rats at a dose of 22.5 mg/kg body weight orally. This study reveals that both raw and purified Kupeelu showed presence of highly significant anti-inflammatory activity against formaldehyde induced hind paw oedema, but did not have similar activity against Carrageenan induced hind paw oedema.
PMCID: PMC3530271  PMID: 23284209
Kupeelu; Kanji; anti-inflammatory; purification; Shodhana; purificatory procedure
11.  Improved pharmacokinetics and reduced toxicity of brucine after encapsulation into stealth liposomes: role of phosphatidylcholine 
Objective:
Brucine was encapsulated into stealth liposomes using the ammonium sulfate gradient method to improve therapeutic index.
Materials and methods:
Four brucine stealth liposomal formulations were prepared, which were made from different phosphatidylcholines (PCs) with different phase transition temperatures (Tm). The PCs used were soy phosphatidylcholine (SPC), dipalmitoyl phosphatidylcholine (DPPC), hydrogenated soy phosphatidylcholine (HSPC), and distearoyl phosphatidylcholine (DSPC). The stabilities, pharmacokinetics, and toxicities of these liposomal formulations were evaluated and compared.
Results:
Size, zeta potential, and entrapment efficiency of brucine-loaded stealth liposomes (BSL) were not influenced by PC composition. In vitro release studies revealed that drug release rate increased with decreased Tm of PCs, especially with the presence of rat plasma. After intravenous administration, the area under the curve (AUC) values of BSL-SPC, BSL-DPPC, BSL-HSPC, and BSL-DSPC in plasma were 7.71, 9.24, 53.83, and 56.83-fold as large as that of free brucine, respectively. The LD50 values of brucine solution, BSL-SPC, BSL-DPPC, BSL-HSPC, and BSL-DSPC following intravenous injection were 13.17, 37.30, 37.69, 51.18, and 52.86 mg/kg, respectively. It was found in calcein retention experiments that the order of calcein retention in rat plasma was SPC < DPPC << HSPC < DSPC stealth liposomes.
Conclusion:
PC composition could exert significant influence on the stabilities, pharmacokinetics, and toxicities of brucine-loaded stealth liposomes. DSPC or HSPC with Tm above 50°C should be used to prepare the stealth liposomal formulation for the intravenous delivery of brucine. However, it was found in the present paper that the pharmacokinetics and toxicity of BSL were not influenced by the PC composition when the Tm of the PC was in the range of −20°C to 41°C.
doi:10.2147/IJN.S32860
PMCID: PMC3418081  PMID: 22904620
brucine; stealth liposomes; phosphatidylcholine; pharmacokinetics; toxicity
12.  Ethnobotanical and Antimicrobial Studies of Some Plants Used in Kibwezi (Kenya) for Management of Lower Respiratory Tract Infections 
Respiratory tract infections have gained worldwide recognition especially due to the increased incidence of HIV/AIDS. The bacteria responsible for these infections have also become increasingly resistant to chemotherapeutic agents in lower respiratory infections in Kibwezi in Kenya. Interviews were conducted using semi-structured questionnaires and detailed discussions with respondents. During the field surveys direct observations were made on how these plants are used. From the ethnobotanical survey the modes of preparation used included chewing and boiling. The plant parts used were mostly bark and root, which implies that the main methods of harvesting these plants are destructive in nature. Water and methanolic extracts of the three most popular plants, Acacia nilotica, Strychnos heninngsii and Microglossa densiflora were tested against three test organism: Staphylococcus aureus, Streptococcus pneumoniae and Escherichia coli for their antimicrobial properties. The efficacy of the extracts was tested by bioassay method using the disk diffusion test. It was interesting to note that none of the tested water extracts showed any activity against the test organisms. This is despite the fact that about 83% of the local people used water for extraction. Methanolic extracts of Acacia nilotica and Strychnos heninningsii showed efficacy against S. aureus, S. pneumoniae and E. coli. The findings of this research indicate that A. nilotica and S. heninngsii have antimicrobial properties and further work especially using mammalian models is recommended.
PMCID: PMC3252686  PMID: 22238495
Medicinal plants; lower respiratory infections; antimicrobial growth inhibitory potential
13.  Alstonia scholaris R. Br. Significantly Inhibits Retinoid-Induced Skin Irritation In Vitro and In Vivo 
Topical retinoids inhibit matrix metalloproteinases and accelerate collagen synthesis, thereby triggering antiaging effects in the skin. However, topical retinoids can cause severe skin reactions, including scaling, erythema, papules, and inflammation. The present study demonstrates that the ethanolic bark extract of Alstonia scholaris R. Br. can significantly inhibit all-trans retinoic acid-induced inflammation in human HaCat keratinocyte cells. Furthermore, two representative retinoid-induced proinflammatory cytokines, monocyte chemoattractant protein-1 and interleukin-8, were significantly suppressed by A. scholaris extract (by 82.1% and 26.3% at 100 ppm, and dose-dependently across the tested concentrations) in vitro. In a cumulative irritation patch test, A. scholaris extract decreased retinol-induced skin irritation, while strengthening the ability of retinoids to inhibit matrix metalloproteinase-1 expression, which is strongly associated with aging effects. These results suggest that A. scholaris is a promising compound that may increase the antiaging function of retinoids while reducing their ability to cause skin irritation.
doi:10.1155/2012/190370
PMCID: PMC3170789  PMID: 21912567
14.  Antimalarial plants of northeast India: An overview 
The need for an alternative drug for malaria initiated intensive efforts for developing new antimalarials from indigenous plants. The information from different tribal communities of northeast India along with research papers, including books, journals and documents of different universities and institutes of northeast India was collected for information on botanical therapies and plant species used for malaria. Sixty-eight plant species belonging to 33 families are used by the people of northeast India for the treatment of malaria. Six plant species, namely, Alstonia scholaris, Coptis teeta, Crotolaria occulta, Ocimum sanctum, Polygala persicariaefolia, Vitex peduncularis, have been reported by more than one worker from different parts of northeast India. The species reported to be used for the treatment of malaria were either found around the vicinity of their habitation or in the forest area of northeast India. The most frequently used plant parts were leaves (33%), roots (31%), and bark and whole plant (12%). The present study has compiled and enlisted the antimalarial plants of northeast India, which would help future workers to find out the suitable antimalarial plants by thorough study.
doi:10.4103/0975-9476.93940
PMCID: PMC3326788  PMID: 22529674
Alkaloids; malaria; medicinal plants; mosquito repellents; northeast India; traditional knowledge of medicine
15.  Formulation and Evaluation of Alstonia boonei Stem Bark Powder Tablets 
The aim of this work was to formulate Alstonia boonei dried stem bark powder into tablets by wet granulation method using acacia, gelatine and sodium carboxymethyl cellulose as binders at concentrations of 1, 2, 4 and 8% w/w. The phytochemistry of the stem bark of Alstonia boonei was evaluated. The micromeritic properties of the granules prepared were studied. The tablets were evaluated using the necessary official and unofficial tests. The results of the phytochemical analysis showed that alkaloids, tannins, steroids, saponins, glycosides, flavonoids and terpenoids were present while anthroquinones and acidcompounds were absent. Micromeritic studies showed that Alstonia boonei granules had good flowability. The formulated tablets complied with British Pharmacopoeial specification for weight uniformity, hardness (≥5 kgf) and tablet friability (<1%). For disintegration test, tablets formulated with gelatine and acacia at concentrations of 1, 2 and 4% w/w complied with Pharmacopoeial specification. However, tablets formulated with SCMC (1-8% w/w) and 8% w/w of acacia and gelatine failed the disintegration tests (Disintegration time more than 15 min).
PMCID: PMC3757864  PMID: 24019574
Alstonia boonei tablets; antimalarial; micromeritic; phyllotaxy; phytochemical analysis
16.  Qualitative Behavior of Spontaneous Potentials from Explants of 15 Day Chick Embryo Telencephalon in Vitro 
The Journal of General Physiology  1962;45(6):1065-1076.
The apparatus and technique used in the preparation and observation of explants of brain tissue capable of producing spontaneous potentials in vitro are described. The magnitude and pattern of spontaneous potentials from explants of telencephalon of 15 day chick embryos (measured using external bare platinum electrodes) and some aspects of their "normal" behavior during 12 days in vitro are also described. No change was noted in these potentials with change of amplifiers, recorders, or electrodes. The response of the potentials to change in temperature and proportionate composition of the atmosphere around the explant was such as to suggest that the potentials arise as a result of a living process. The changes brought about by the administration of anesthetics, strychnine, brucine, and barbiturates were those that might be anticipated in a normal functional activity of the central nervous system. It is concluded that these potentials are a true physiological phenomenon and arise from living cells of the central nervous system.
PMCID: PMC2195244  PMID: 13882640
17.  Study on Phytochemical Composition, Antibacterial and Antioxidant Properties of Different Parts of Alstonia scholaris Linn. 
Advanced Pharmaceutical Bulletin  2013;3(2):379-384.
Purpose: To evaluate phytochemical composition, antibacterial and antioxidant properties of methanolic extracts of different parts viz., leaves, follicles and latex of Indian devil tree (Alstonia scholaris Linn.) R. Br. Methods: Antibacterial activities of the methanol extracts against Gram +ve (Bacillus subtilis and Staphylococcus aureus) and Gram -ve (Escherichia coli, Pseudomonas aeruginosa) bacteria were determined by well diffusion techniques. Aantioxidant profiles of methanol extracts were determined by 1,1-diphenyl-2-picryl-hydrazil (DPPH) free radical scavenging, superoxide anion radial scavenging and ferric thiocyanate reducing assays. Results: Phytochemical composition revealed abundance of flavonoids (97.3 mg QE/g DW), proanthocynidins (99.3 mg CE/g DW) and phenolics (49.7 mgGAE/g DW) in the leaf extract. Extracts of follicles and latex had comparatively very content of phenolics, flavonoids and proanthocyanidins. However, in follicle extract level of proanthocyanidins was significantly higher (46.8 mg CE/gDW). Latex extract among others exhibited most potent antibacterial activity. All the extracts displayed strong DPPH free radical and superoxide anion scavenging activities, only leaf extract displayed powerful reducing and ferrous ion chelating activities. Conclusion: Study revealed significant antioxidant activities of A. scholaris leaf, follicles and latex extracts and potential antibacterial activity of latex extract.
doi:10.5681/apb.2013.061
PMCID: PMC3848240  PMID: 24312864
Alstonia scholaris; Antibacterial; Antioxidant; Follicles; Latex; Phenolics
18.  Nonlinear Dynamical Systems Effects of Homeopathic Remedies on Multiscale Entropy and Correlation Dimension of Slow Wave Sleep EEG in Young Adults with Histories of Coffee-Induced Insomnia 
Homeopathy  2012;101(3):182-192.
Background
Investigators of homeopathy have proposed that nonlinear dynamical systems (NDS) and complex systems science offer conceptual and analytic tools for evaluating homeopathic remedy effects. Previous animal studies demonstrate that homeopathic medicines alter delta electroencephalographic (EEG) slow wave sleep. The present study extended findings of remedy-related sleep stage alterations in human subjects by testing the feasibility of using two different NDS analytic approaches to assess remedy effects on human slow wave sleep EEG.
Methods
Subjects (N=54) were young adult male and female college students with a history of coffee-related insomnia who participated in a larger 4-week study of the polysomnographic effects of homeopathic medicines on home-based all-night sleep recordings. Subjects took one bedtime dose of a homeopathic remedy (Coffea cruda or Nux vomica 30c). We computed multiscale entropy (MSE) and the correlation dimension (Mekler-D2) for stage 3 and 4 slow wave sleep EEG sampled in artifact-free 2-minute segments during the first two rapid-eye-movement (REM) cycles for remedy and post-remedy nights, controlling for placebo and post-placebo night effects.
Results
MSE results indicate significant, remedy-specific directional effects, especially later in the night (REM cycle 2) (CC: remedy night increases and post-remedy night decreases in MSE at multiple sites for both stages 3 and 4 in both REM cycles; NV: remedy night decreases and post-remedy night increases, mainly in stage 3 REM cycle 2 MSE). D2 analyses yielded more sporadic and inconsistent findings.
Conclusions
Homeopathic medicines Coffea cruda and Nux vomica in 30c potencies alter short-term nonlinear dynamic parameters of slow wave sleep EEG in healthy young adults. MSE may provide a more sensitive NDS analytic method than D2 for evaluating homeopathic remedy effects on human sleep EEG patterns.
doi:10.1016/j.homp.2012.05.007
PMCID: PMC3402916  PMID: 22818237
homeopathy; polysomnography; electroencephalography; Coffea cruda; Nux vomica; nonlinear dynamics, multiscale entropy; correlation dimension; slow wave sleep; complex systems; time-dependent sensitization
19.  The Natural History of the Arboreal Ant, Crematogaster ashmeadi 
The arboreal ant, Crematogaster ashmeadi Emery (Hymenoptera: Formicidae), is the most dominant arboreal ant in the pine forests of the coastal plain of northern Florida. The majority of pine trees harbor a colony of these ants. The colonies inhabit multiple chambers abandoned by bark-mining caterpillars, especially those of the family Cossidae, in the outer bark of living pines. They also inhabit ground level termite galleries in the bark, often locating the queen in galleries. The density of chambers and ants is highest in the base of the tree and drops sharply with height on the trunk. Because chambers are formed in the inner layer of bark, they gradually move outward as more bark layers are laid down, eventually sloughing off the tree's outer surface. Chambers have a mean lifetime of about 25 yr. The abundant chambers in pine bark are excavated by a small population of caterpillars and accumulate over decades. Ant colonies also inhabit abandoned galleries of woodboring beetles in dead branches in the crowns of pines. Because newly mated queens found colonies in abandoned woodboring beetle galleries in the first dead branches that form on pine saplings, C. ashmeadi is dependent on cavities made by other insects throughout its life cycle, and does little if any excavation of its own. Mature colonies nest preferentially in chambers greater than 10 cm2 in area, a relatively rare chamber size. In natural pine forests, this does not seem to limit the ant's populations.
Founding queens containabout 50% fat and lose about half of their dry weight during the claustral period, converting approximately half of this lost weight into progeny. The claustral period is about 40 to 50 days at 27°C. Mature colonies contain several tens of thousands of workers (est. up to 80,000), and have a life expectancy of 10 to 15 years. Each colony occupies an entire tree, and sometimes two trees if they are close together. Within a colony, there is a single queen capable of laying up to 450 eggs/day during the warm season. Such queens weigh 12 to 18 mg, have 50 to 60 active ovarioles and 120 to 600 vitellogenic oocytes in their ovaries.
Mature colonies begin producing sexual brood in late April or early May. Sexual adults are present from late May through June. Mating flights commence in June and most sexuals have left their natal nests by late July. Female sexuals are an especially large investment; the energetic content of a single, flight-ready female sexual is almost 20 times that of a worker. The newly mated queen sequesters a mean of 2.64 × 106 sperm in her spermatheca, a supply that should last her for 16 years at the observed reproductive rates.
PMCID: PMC355912  PMID: 15455046
Formicidae; nest chambers; arboreal ants; colony founding; seasonality; colony longevity; Cossidae; Givira francesca (Dyar); Buprestidae; Cerambicidae; woodboring beetles; longleaf pine; slash pine; Pinus palustris; Pinus elliotti; red-cockaded woodpecker
20.  In vitro and in vivo antidermatophytic activity of the dichloromethane-methanol (1:1 v/v) extract from the stem bark of Polyscias fulva Hiern (Araliaceae) 
Background
During the last decades, the number of people suffering from dermatophytoses has seriously increased, mainly due to the development of resistant strains of microorganisms to a range of formally efficient antibiotics. Polyscias fulva, a medium size tree which grows in the West Region of Cameroon is traditionally used for local application against dermatoses and orally against venereal infections. The dichloromethane-methanol (1:1 v/v) extract from the stem bark of Polyscias fulva was evaluated for its in vitro and in vivo antifungal activities.
Methods
The plant extract was prepared by maceration of its stem bark powder in CH2Cl2-MeOH (1:1 v/v). The extract obtained was successively partitioned in hexane, ethyl acetate and n-butanol. Phytochemical screening was performed using standard methods. In vitro antidermatophytic activity was assayed by the well diffusion and broth microdilution methods. The degree of dermal irritation of the crude extract was determined in guinea pigs using the occluded dermal irritation test method. The in vivo antidermatophytic activity of the extract-oil formulation (1.25, 2.5 and 5% w/w concentrations) was evaluated using Trichophyton mentagrophytes-induced dermatophytosis in a guinea pigs model.
Results
Phytochemical screening indicated that, the crude extract, ethyl acetate, n-butanol and residue fractions contain in general saponins, tannins, alkaloids, anthraquinones and phenols while the hexane fraction contains only alkaloids. The ethyl-acetate, n-butanol and residue fractions displayed higher antifungal activities (MIC = 0.125-0.5 mg.mL-1) against eight dermatophytes as compared to the crude extract (MIC = 0.5-1 mg.mL-1). This latter appeared to have slight perceptible erythema effects on guinea pigs as the primary irritation index (PII) was calculated to be 0.54. In vivo, the antidermatophytic activities of the extract-oil formulations were dose-dependent. Griseofulvin-oil 5% at 0.01 g/kg and formulated extract-oil (5%) at 0.1 g/kg eradicated the microbial infection after thirteen and fourteen days of daily treatment respectively.
Conclusions
The results of preclinical in vitro and in vivo evaluations indicate that the extract-oil formulation at 5% may constitute an alternative means to alleviate fungal infections caused by dermatophytes.
doi:10.1186/1472-6882-13-95
PMCID: PMC3658906  PMID: 23641972
Antidermatophytic activity; Polyscias fulva; Extract-oil formulation; Irritation test; Primary irritation index; Guinea pigs
21.  An Ethnobotanical study of Medicinal Plants in high mountainous region of Chail valley (District Swat- Pakistan) 
Background
This paper represents the first ethnobotanical study in Chail valley of district Swat-Pakistan and provides significant information on medicinal plants use among the tribal people of the area. The aim of this study was to document the medicinal uses of local plants and to develop an ethnobotanical inventory of the species diversity.
Methods
In present study, semi-structured interviews with 142 inhabitants (age range between 31–75 years) were conducted. Ethnobotanical data was analyzed using relative frequency of citation (RFC) to determine the well-known and most useful species in the area.
Results
Current research work reports total of 50 plant species belonging to 48 genera of 35 families from Chail valley. Origanum vulgare, Geranium wallichianum and Skimmia laureola have the highest values of relative frequency of citation (RFC) and are widely known by the inhabitants of the valley. The majority of the documented plants were herbs (58%) followed by shrubs (28%), trees (12%) and then climbers (2%). The part of the plant most frequently used was the leaves (33%) followed by roots (17%), fruits (14%), whole plant (12%), rhizomes (9%), stems (6%), barks (5%) and seeds (4%). Decoction was the most common preparation method use in herbal recipes. The most frequently treated diseases in the valley were urinary disorders, skin infections, digestive disorders, asthma, jaundice, angina, chronic dysentery and diarrhea.
Conclusion
This study contributes an ethnobotanical inventory of medicinal plants with their frequency of citations together with the part used, disease treated and methods of application among the tribal communities of Chail valley. The present survey has documented from this valley considerable indigenous knowledge about the local medicinal plants for treating number of common diseases that is ready to be further investigated for biological, pharmacological and toxicological screening. This study also provides some socio-economic aspects which are associated to the local tribal communities.
doi:10.1186/1746-4269-10-36
PMCID: PMC4022037  PMID: 24739524
Geo-ethnographical; Ethnoflora; Khyber Pakhtunkhwa; Pharmacological studies
22.  Phenotypic differentiation between wild and domesticated varieties of Crescentia cujete L. and culturally relevant uses of their fruits as bowls in the Yucatan Peninsula, Mexico 
Background
Selection criteria are important for analyzing domestication of perennial plant species, which experience a selection pressure throughout several human generations. We analyze the preferred morphological characteristics of Crescentia cujete fruits, which are used as bowls by the Maya of Yucatan, according to the uses they are given and the phenotypic consequences of artificial selection between one wild and three domesticated varieties.
Methods
We performed 40 semi-structured interviews in seven communities. We calculated Sutrop’s salience index (S) of five classes of ceremonial and daily life uses, and of each item from the two most salient classes. We sampled 238 bowls at homes of people interviewed and compared their shape, volume and thickness with 139 fruits collected in homegardens and 179 from the wild. Morphology of varieties was assessed in fruit (n = 114 trees) and vegetative characters (n = 136 trees). Differences between varieties were evaluated through linear discriminant analysis (LDA).
Results
Use of bowls as containers for the Day of the Dead offerings was the most salient class (S = 0.489) with chocolate as its most salient beverage (S = 0.491), followed by consumption of daily beverages (S = 0.423), especially maize-based pozol (S = 0.412). The sacred saka’ and balche' are offered in different sized bowls during agricultural and domestic rituals. Roundness was the most relevant character for these uses, as bowls from households showed a strong selection towards round shapes compared with wild and homegarden fruits. Larger fruits from domesticated varieties were also preferred over small wild fruits, although in the household different sizes of the domesticated varieties are useful. LDA separated wild from domesticated trees (p < 0.001) according to both fruit and vegetative variables, but domesticated varieties were not different among themselves.
Conclusions
The association between C. cujete bowls and traditional beverages in ritual and daily life situations has driven for centuries the selection of preferred fruit morphology in this tree. Selection of fruit roundness and volume has allowed for the differentiation between the wild variety and the three domesticated ones, counteracting gene flow among them. By choosing the best fruits from domesticated varieties propagated in homegardens, the Maya people model the domestication process of this important tree in their culture.
doi:10.1186/1746-4269-9-76
PMCID: PMC3842825  PMID: 24229087
Calabash; Crescentia; Domestication; Gourd tree; Maya; Mesoamerica; Morphology; Phenotypic variation
23.  Eurasian Tree Sparrows, Risk for H5N1 Virus Spread and Human Contamination through Buddhist Ritual: An Experimental Approach 
PLoS ONE  2011;6(12):e28609.
Background
The Highly Pathogenic Avian Influenza H5N1 virus has dramatically spread throughout Southeast Asia since its first detection in 1997. Merit Release Birds, such as the Eurasian Tree Sparrow, are believed to increase one's positive karma when kissed and released during Buddhist rituals. Since these birds are often in close contact with both poultry and humans, we investigated their potential role in the spread of H5N1 virus.
Methodology/Principal Findings
Seven series of experiments were conducted in order to investigate the possible interactions between inoculated and exposed birds, including sparrow/sparrow, sparrow/chicken, duck/sparrow. Daily and post-mortem samples collected were tested for H5N1 virus by real-time RT-PCR and egg inoculation. When directly inoculated, Eurasian Tree Sparrows were highly susceptible to the H5N1 virus, with a fatality rate approaching 100% within 5 days post-inoculation. Although transmission of fatal infection between sparrows did not occur, seroconversion of the exposed birds was observed. Up to 100% chickens exposed to inoculated sparrows died of H5N1 infection, depending on the caging conditions of the birds, while a fatality rate of 50% was observed on sparrows exposed to infected ducks. Large quantities of H5N1 virus were detected in the sparrows, particularly in their feathers, from which infectious particles were recovered.
Conclusions/Significance
Our study indicates that under experimental conditions, Eurasian Tree Sparrows are susceptible to H5N1 infection, either by direct inoculation or by contact with infected poultry. Their ability to transmit H5N1 infection to other birds is also demonstrated, suggesting that the sparrows may play a role in the dissemination of the virus. Finally, the presence of significant quantities of H5N1 virus on sparrows' feathers, including infectious particles, would suggest that Merit Release Birds represent a risk for human contamination in countries where avian influenza virus is circulating and where this religious ritual is practiced.
doi:10.1371/journal.pone.0028609
PMCID: PMC3229601  PMID: 22164310
24.  Traditional management of ear, nose and throat (ENT) diseases in Central Kenya 
Diseases of ear, nose and throat (ENT) often have serious consequences including hearing impairment, and emotional strain that lower the quality of life of patients. In Kenya, upper respiratory infections are among the most common infections encountered in outpatient facilities. Some of these infections are becoming difficult to control because some of the causing microorganisms have acquired antibiotic resistance and hence the need to develop new drugs with higher efficacy. Ethnobotanical studies have now been found to be instrumental in improving chances of discovering plants with antimicrobial activity in new drug development. In Kenya the majority of local people are turning to herbal remedies for primary health care needs. In most cases the sources of these remedies are undocumented and the knowledge about them passed orally form generation to generation, hence under threat of disappearing with current rates of modernisation.
This study explored the traditional remedies used in managing various ENT diseases in seven districts of the Central Province of Kenya. The most common ENT conditions managed using traditional therapies include: common cold, cough, tonsillitis, otitis-media, chest pains and asthma. The results indicate that 67 species belonging to 36 plant families were utilized in this region. These plants were of varying habits; herbs (37.3%), shrubs (34.4%), trees (25.4%) as well as some grasses and sedges (3%). The traditional preparations were found to be made mainly from leaves (49%), roots (20.5%) and barks (12.5%). For each of the ENT conditions multiple species are utilized mainly as individual preparations but occasionally as polyherbal concoctions. In the case of common cold for example, 30 different species are used. Plants reported in this survey are important candidates for antimicrobial tests against ENT disease causing micro-organisms, especially those with antibiotic resistance.
doi:10.1186/1746-4269-2-54
PMCID: PMC1770906  PMID: 17192184
25.  Leaf Life Span Plasticity in Tropical Seedlings Grown under Contrasting Light Regimes 
Annals of Botany  2006;97(2):245-255.
• Background and Aims The phenotypic plasticity of leaf life span in response to low resource conditions has a potentially large impact on the plant carbon budget, notably in evergreen species not subject to seasonal leaf shedding, but has rarely been well documented. This study evaluates the plasticity of leaf longevity, in terms of its quantitative importance to the plant carbon balance under limiting light.
• Methods Seedlings of four tropical tree species with contrasting light requirements (Alstonia scholaris, Hevea brasiliensis, Durio zibethinus and Lansium domesticum) were grown under three light regimes (full sunlight, 45 % sunlight and 12 % sunlight). Their leaf dynamics were monitored over 18 months.
• Results All species showed a considerable level of plasticity with regard to leaf life span: over the range of light levels explored, the ratio of the range to the mean value of life span varied from 29 %, for the least plastic species, to 84 %, for the most. The common trend was for leaf life span to increase with decreasing light intensity. The plasticity apparent in leaf life span was similar in magnitude to the plasticity observed in specific leaf area and photosynthetic rate, implying that it has a significant impact on carbon gain efficiency when plants acclimate to different light regimes. In all species, median survival time was negatively correlated with leaf photosynthetic capacity (or its proxy, the nitrogen content per unit area) and leaf emergence rate.
• Conclusions Longer leaf life spans under low light are likely to be a consequence of slower ageing as a result of a slower photosynthetic metabolism.
doi:10.1093/aob/mcj023
PMCID: PMC2803358  PMID: 16299004
Alstonia scholaris; carbon balance; Durio zibethinus; Hevea brasiliensis; Lansium domesticum; leaf life span; light; plasticity

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