Pharmacokinetics of articaine hydrochloride and its metabolite articainic acid after subcutaneous administration in red deer (Cervus elaphus)

AIM: To develop and validate a simple and sensitive method using liquid chromatography-mass spectrometry (LC-MS) for quantification of articaine, and its major metabolite articainic acid, in plasma of red deer (Cervus elaphus), and to investigate the pharmacokinetics of articaine hydrochloride and articainic acid in red deer following S/C administration of articaine hydrochloride as a complete ring block around the antler pedicle.

METHODS: The LC-MS method was validated by determining linearity, sensitivity, recovery, carry-over and repeatability. Articaine hydrochloride (40?mg/mL) was administered S/C to six healthy male red deer, at a dose of 1?mL/cm of pedicle circumference, as a complete ring block around the base of each antler. Blood samples were collected at various times over the following 12 hours. Concentrations in plasma of articaine and articainic acid were quantified using the validated LC-MS method. Pharmacokinetic parameters of articaine and articainic acid were estimated using non-compartmental analysis.

RESULTS: Calibration curves were linear for both articaine and articainic acid. The limits of quantifications for articaine and articainic acid were 5 and 10?ng/mL, respectively. Extraction recoveries were >72% for articaine and >68% for articainic acid. After S/C administration as a ring block around the base of each antler, mean maximum concentrations in plasma (C_max) of articaine were 1,013.9 (SD 510.1) ng/mL, detected at 0.17 (SD 0.00) hours, and the C_max for articainic acid was 762.6 (SD 95.4) ng/mL at 0.50 (SD 0.00) hours. The elimination half-lives of articaine hydrochloride and articainic acid were 1.12 (SD 0.17) and 0.90 (SD 0.07) hours, respectively.

CONCLUSIONS AND CLINICAL RELEVANCE: The LC-MS method used for the quantification of articaine and its metabolite articainic acid in the plasma of red deer was simple, accurate and sensitive. Articaine hydrochloride was rapidly absorbed, hydrolysed to its inactive metabolite articainic acid, and eliminated following S/C administration as a ring block in red deer. These favourable pharmacokinetic properties suggest that articaine hydrochloride should be tested for efficacy as a local anaesthetic in red deer for removal of velvet antlers. Further studies to evaluate the safety and residues of articaine hydrochloride and articainic acid are required before articaine can be recommended for use as a local anaesthetic for this purpose.     

Isolation,purification, and biochemical characterization of a novel water soluble protein from Inca peanut (Plukenetia volubilis L.)

A water soluble storage albumin from Inca peanut (IPA) accounted for approximately 25% (w/w) of defatted seed flour weight, representing 31% of the total seed protein. IPA is a 3S storage protein composed of two glycosylated polypeptides, with estimated molecular weights (MW) of 32800 and 34800 Da, respectively. IPA has an estimated sugar content of 4.8% +/- 0.92% (n = 6). IPA is a basic protein (pI of approximately 9.4) and contains all of the essential amino acids in adequate amounts when compared to the FAO/WHO recommended pattern for a human adult. The tryptophan content of IPA is unusually high (44 mg/g of protein), whereas the phenylalanine content is low (9 mg/g of protein). IPA is a highly digestible protein in vitro.     

Development of an efficient protocol for plant regeneration from nodal explants of recalcitrant bamboo (Bambusa arundinacea Retz. Willd) and assessment of genetic fidelity by DNA markers

The present study describes an efficient method for in vitro plant regeneration in B. arundinacea through axillary shoot bud proliferation. Nodal explants were excised, cultured on MS medium containing different concentrations of 6-benzylaminopurine (BAP), kinetin (KIN) (0.5–5.0 mg l^?1) alone and/or in combinations with KIN/BAP (0.5 mg l^?1). The highest frequency (91.5 %) of multiple shoot bud induction with maximum number of shoots (85 shoots/explant) was noticed on MS medium + 3.0 mg l^?1 BAP + 0.5 mg l^?1 KIN. The regenerated multiple shoots were elongated on MS medium + 4.0 mg l^?1 KIN + 2.0 mg l^?1 gibberellic acid (GA₃) with maximum shoot length (4.9 cm). The elongated shoots were transferred to MS medium containing indole-3 butyric acid (IBA; 0.5–5.0 mg l^?1) alone and/or in combination with 0.5 mg l^?1 KIN and BAP. Highest frequency of rooting (75 %) was obtained on half-strength MS medium + 2.0 mg l^?1 IBA + 0.5 mg l^?1 KIN. After hardening, the plantlets were shifted to the green house and subsequently established in the field conditions with 90 % survival rate. random amplified polymorphic DNA (RAPD) markers were used to evaluate the genetic stability of the regenerants. RAPD profiles generated from the regenerated plants were found to be monomorphic, similar to the control. Results confirmed that the regenerated plants were true-to-type in nature and the developed micropropagation protocol could be used for large scale plant production of B. arundinacea.     

Biochemical characterization of amandin,the major storage protein in almond (Prunus dulcis L.)

The almond major storage protein, amandin, was prepared by column chromatography (amandin-1), cryoprecipitation (amandin-2), and isoelectric precipitation (amandin-3) methods. Amandin is a legumin type protein characterized by a sedimentation value of 14S. Amandin is composed of two major types of polypeptides with estimated molecular weights of 42-46 and 20-22 kDa linked via disulfide bonds. Several additional minor polypeptides were also present in amandin. Amandin is a storage protein with an estimated molecular weight of 427,300 +/- 47,600 Da (n = 7) and a Stokes radius of 65.88 +/- 3.21 A (n = 7). Amandin is not a glycoprotein. Amandin-1, amandin-2, and amandin-3 are antigenically related and have similar biochemical properties. Amandin-3 is more negatively charged than either amandin-1 or amandin-2. Methionine is the first essential limiting amino acid in amandin followed by lysine and threonine.     

Large scale in vitro propagation of Stevia rebaudiana (bert) for commercial application: Pharmaceutically important and antidiabetic medicinal herb

Stevia rebaudiana is a valuable medicinal plant species and it is being used for the treatment of diabetes. Currently, there is a high demand for raw material of this medicinal herb due to ever increasing diabetes disorder among the population. In order to meet the increased demand an efficient in vitro propagation of S. rebaudiana was established. Nodal explants collected from the field were cultured on MS basal medium fortified with different concentrations of BAP (0.5-3.0 mg/l) and KIN (0.5-3.0 mg/l) individually for shoot bud induction. In vitro derived nodal buds were cultured on MS medium supplemented with different concentrations (0.5-3.0 mg/l) of BAP and KIN for multiple shoot bud regeneration. In the second experiment, in vitro derived buds were placed on MS medium supplemented with different concentrations of BAP (0.5-3.0 mg/l) in combination with 0.5 mg/l IAA or IBA or NAA for shoot bud multiplication. The highest frequency (94.50%) of multiple shoot regeneration with maximum number of shoots (15.69 shoots/explant) was noticed on MS medium supplemented with 1.0 mg/l BAP. For large scale plant production, in vitro derived nodal bud explants were cultured on MS medium fortified with 1.0 mg/l BAP, in which about 123 shoots/explant were obtained after three subcultures on the same media composition. Elongated shoots (>2 cm) dissected out from the in vitro proliferated shoot clumps were cultured on half-strength MS medium containing different concentrations of NAA (0.1-0.5 mg/l) and/or MS medium fortified with various concentrations (0.5-2.0 mg/l) of auxins (NAA, IAA and IBA) for root induction. Highest frequency of rooting (96%) was noticed on half-strength MS medium augmented with 0.4 mg/l NAA. The rooted plantlets were successfully transferred into plastic cups containing sand and soil in the ratio of 1:2 and subsequently established in the greenhouse. The present in vitro propagation protocol would facilitate an alternative method for rapid and large-scale production of this important antidiabetic medicinal plant.     

Risk factors in relation to human deaths and other tsunami (2004) impacts in Sri Lanka: the fishers'‐eye view

• 1. This study examines the perceptions of 500 Sri Lankan fishers about influences on the outcome of the 2004 Asian tsunami. It is based upon analysis of questionnaire data on 13 natural environmental and development risk factors, in relation to human deaths and house damage (impact indicators).
• 2. Mangroves, coral reefs and sand dunes afforded protection against tsunami damage (67–94% of fisher responses), as did housing and roads.
• 3. Fishers overall believed rivers/estuaries, concave coastlines and hotels exacerbated impacts. However, a significantly greater proportion of fishers living within 100 m of the coast reported that rivers/estuaries had a protective role than those living further inland. Rivers seemingly diverted ‘tsunami water’ far inland, where it overflowed and caused damage.
• 4. Risk and damage are multi‐faceted concepts and measurable in different ways. Findings are considered in the light of ecological studies and modelling, with special reference to mangroves, whose alleged protective role has become equivocal during post‐tsunami research.
• 5. Insights of fishers and other communities with intuitive knowledge add a valuable perspective to the understanding of natural disasters and environmental change. This approach is seen as complementary rather than an alternative approach to purely ‘scientific’ research.

Copyright © 2008 John Wiley & Sons, Ltd.     

Use of ground based hyperspectral remote sensing for detection of stress in cotton caused by leafhopper (Hemiptera: Cicadellidae)

Detection of crop stress is one of the major applications of hyperspectral remote sensing in agriculture. Many studies have demonstrated the capability of remote sensing techniques for detection of nutrient stress on cotton with only few on pest damage but none so far on leafhopper (LH) severity. Subsequent to introduction of Bt cotton, leafhopper is emerging as a key pest in several countries. In view of its wide host range, geographical distribution and damage potential, a study was initiated to characterise leafhopper stress on cotton, identify sensitive bands, and derive hyperspectral vegetation indices specific to this pest. Cotton plants with varying levels of LH severity were selected from three locations across major cotton growing regions of India. About 57-58 cotton plants from each location exhibiting different levels of LH damage symptoms were selected. Reflectance measurements in the spectral range of 350-2500 nm were made using hyperspectral radiometer. Simultaneously chlorophyll (Chl) and relative water content (RWC) were also estimated from the selected plants. Reflectance from healthy and leafhopper infested plants showed a significant difference in VIS and NIR regions. Decrease in Chl a pigment was more significant than Chl b in the infested plants and the ratio of Chl a/b showed a decreasing trend with increase in LH severity. Regression analysis revealed a significant linear relation between LH severity and Chl (R² = 0.505^∗∗), and a similar fit was also observed for RWC (R² = 0.402^∗∗). Plotting linear intensity curves between reflectance at each waveband with infestation grade resulted in six sensitive bands that exhibited maximum correlation at different regions of the electromagnetic spectrum (376, 496, 691, 761, 1124 and 1457 nm). Regression analysis of several ratio indices formulated with two or more of these sensitive bands led to the identification of new leaf hopper indices (LHI) with a potential to detect leafhopper severity. These new indices along with 20 other stress related hyperspectral indices compiled from literature were further tested for their ability to detect LH severity. Two novel indices LHI 2 and LHI 4 proposed in this study showed significantly high coefficients of determination across locations (R² range 0.521 to 0.825^∗∗) and hence have the potential use for detection of leafhopper severity in cotton.     

Changes in frigate tuna populations on the south coast of Sri Lanka: evidence of the shifting baseline syndrome from analysis of fisher observations

• 1. This study examines changes in frigate tuna populations in southern Sri Lanka, based on reports from fishers in three age classes. Significantly higher values for best day's catch and largest specimen ever caught were obtained by older fishers than younger ones. Values were also significantly higher during early years, providing clear evidence of a decrease in the resource over time (1951–2007).
• 2. Older fishers reported best catches further inshore and in shallower waters which, on becoming depleted, forced younger generations to fish in less exploited areas further offshore. Heavy harvesting is also evident from the significantly greater number of sites reported by older fishers as being depleted, compared with observations of younger fishers.
• 3. These findings contrast markedly with catch and catch per effort patterns from statistics for frigate tuna and bullet tuna (combined) in southern Sri Lanka (1994–2004). No stock decline is evident, and at least one report in the early 1990s advocated increasing exploitation rates by 40% to maximize yields.
• 4. Although not a primary research objective, fisher observations on frigate tuna populations were also analysed to help evaluate possible effects of the 2004 tsunami. Most fishers reported post‐tsunami decline, but mainly from a larger new generation of fishers, rather than extra boats provided by aid money or (direct or indirect) biophysical impacts from the tsunami.
• 5. Reliance on fishery statistics, especially for mixed species and over a limited period, can be risky and easily mask true stock status. Evidence of harvesting effects on frigate tuna in southern Sri Lanka is evident using questionnaire data over a longer time scale.
• 6. This study provides another compelling case of the ‘shifting baseline syndrome’, whereby fishers of different ages have altered perceptions/experiences of their environment. This may be its first reported occurrence in Sri Lanka. Traditional knowledge from this and similar surveys may provide national fishery management with valuable insights and help improve conservation prospects for frigate tuna and other marine resources. Copyright © 2009 John Wiley & Sons, Ltd.

     

Chagos feels the pinch: assessment of holothurian (sea cucumber) abundance,illegal harvesting and conservation prospects in British Indian Ocean Territory

• 1. Data are analysed from visual censuses of shallow‐water holothurians (sea cucumbers) in 72 shallow water transects 100 m×2 m within four atolls of Chagos. Mean holothurian abundance in Diego Garcia, where harvesting is absent, was 18.5 individuals/transect (all transects) and 55.4 individuals/transect (only those containing holothurians). In the three exploited atolls, mean abundance did not exceed 3.5 and 5.2 individuals/transect, respectively.
• 2. Comparison with data collected during this study and an earlier investigation reveals a marked decline over four years in both mean and maximum density of commercially valuable Stichopus chloronotus and Holothuria atra in Salomon and Peros Banhos, both exploited atolls, and also for Holothuria nobilis in the latter.
• 3. Holothurian counts were also made along an extensive transect (21 km×4 m) encircling Salomon atoll. Abundance showed highly significant negative correlation with fishing pressure, the latter estimated using an ordinal (0–3) scale (R_s=?0.605, P?0.01). Harvesting effects were not discernible using data from 200 m² transects.
• 4. While recent studies have shown Chagos is virtually pristine regarding contaminant levels, its holothurian resources are under increasing pressure. Results from this study, and examination of Sri Lanka's fishing activity in distant waters, point to heavy and illegal harvesting.
• 5. Stronger measures are needed to control the illegal fishery, to prevent holothurian abundances falling to the non‐sustainable levels now prevalent across much of the Indo‐Pacific, and to ensure that Chagos remains a biodiversity hotspot and environment of international renown. Use of smaller surveillance vessels would facilitate this.Copyright © 2009 John Wiley & Sons, Ltd.