Gossypol mediated DNA degradation

Gossypol, a polyphenolic compound isolated from cotton plant was found to degrade pBR322 DNA Image in a reaction which required the presence of a metal ion, a reducing agent (2-mercaptoethanol) and oxygen as revealed after agarose gel electrophoresis. Fe3+ and Co2+ showed maximum degradation whereas addition of Ca2+ and Mg2+ prevented the gossypol mediated DNA damage. Gossypol caused degradation of rat liver DNA incubated Image even in the absence of added metal ions and 2-mercaptoethanol. Incubation of intact rat liver nuclei with gossypol reveled DNA degradation and nuclei isolated from rats treated with gossypol Image showed higher succestibility to DNA fragmentation when incubated with gossypol Image than control nuclei. EcoRl and AIuI digestion of DNA isolated from gossypol treated rats gave clear cut evidence for DNA degradation. These observations indicate that gossypol is genotoxic and considereable care has to be exercised in its use. SDS, sodium dodecayl sulphate; TE buffer, Tris-HCL-EDTA buffer.

Fusarium wilt of cotton: Population diversity and implications for management

Fusarium wilt of cotton, caused by the fungus Fusarium oxysporum Schlechtend. f. sp. vasinfectum (Atk.) Snyd. & Hans, was first identified in 1892 in cotton growing in sandy acid soils in Alabama (8). Although the disease was soon discovered in other major cotton-producing areas, it did not become global until the end of the next century. After its original discovery, Fusarium wilt of cotton was reported in Egypt (1902) (30), India (1908) (60), Tanzania (1954) (110), California (1959) (33), Sudan (1960) (44), Israel (1970) (27), Brazil (1978) (5), China (1981) (17), and Australia (1993) (56). In addition to a worldwide distribution, Fusarium wilt occurs in all four of the domesticated cottons, Gossypium arboretum L., G. barbadense L., G. herbaceum L., and G. hirsutum L. (4,30). Disease losses in cotton are highly variable within a country or region. In severely infested fields planted with susceptible cultivars, yield losses can be high. In California, complete crop losses in individual fields have been observed (R. M. Davis, unpublished). Disease loss estimates prepared by the National Cotton Disease Council indicate losses of over 109,000 bales (227 kg or 500 lb) in the United States in 2004 (12).

Difference covering arrays and pseudo-orthogonal Latin squares

A pair of Latin squares, A and B, of order n, is said to be pseudo-orthogonal if each symbol in A is paired with every symbol in B precisely once, except for one symbol with which it is paired twice and one symbol with which it is not paired at all. A set of t Latin squares, of order n, are said to be mutually pseudo-orthogonal if they are pairwise pseudo-orthogonal. A special class of pseudo-orthogonal Latin squares are the mutually nearly orthogonal Latin squares (MNOLS) first discussed in 2002, with general constructions given in 2007. In this paper we develop row complete MNOLS from difference covering arrays. We will use this connection to settle the spectrum question for sets of 3 mutually pseudo-orthogonal Latin squares of even order, for all but the order 146.

Soil sorption-desorption of phosphorus from piggery effluent compared with inorganic sources

The leaching of phosphorus (P) within soils can be a limiting consideration for the sustainable operation of intensive livestock enterprises. Sorption curves are widely used to assist estimation of P retention, though the effect of effluent constituents on their accuracy is not well understood. We conducted a series of P-sorption-desorption batch experiments with an Oxic Haplustalf (soil 1), Haplusterts (soils 2 and 3), and a Natrustalf (soil 4). Phosphorus sources included effluent, orthophosphate-P in a matrix replicating the effluent's salt constituents (the reference solution), and an orthophosphate-P solution. Treated soils were incubated for up to 193 days before sequential desorption extraction. Effluent constituents, probably the organic or particulate components, temporarily increased the vulnerability of sorbed-P to desorption. The increase in vulnerability was removed by 2-113 days of incubation (25 degrees C). Despite vigorous extraction for 20 consecutive days, some P sorbed as part of the treatments of soils 1 and 2 was not desorbed. The increased vulnerability due to effluent constituents lasted a maximum of about one cropping season and, for all other treatments, adsorption curves overestimated vulnerability to desorption. Therefore, adsorption curves provide a conservative estimate of vulnerability to desorption where effluent is used in continued crop production in these soils.

Concordant molecular and phenotypic data delineate new taxonomy and conservation priorities for the endangered mountain yellow-legged frog

The mountain yellow-legged frog Rana muscosa sensu lato, once abundant in the Sierra Nevada of California and Nevada, and the disjunct Transverse Ranges of southern California, has declined precipitously throughout its range, even though most of its habitat is protected. The species is now extinct in Nevada and reduced to tiny remnants in southern California, where as a distinct population segment, it is classified as Endangered. Introduced predators (trout), air pollution and an infectious disease (chytridiomycosis) threaten remaining populations. A Bayesian analysis of 1901 base pairs of mitochondrial DNA confirms the presence of two deeply divergent clades that come into near contact in the Sierra Nevada. Morphological studies of museum specimens and analysis of acoustic data show that the two major mtDNA clades are readily differentiated phenotypically. Accordingly, we recognize two species, Rana sierrae, in the northern and central Sierra Nevada, and R. muscosa, in the southern Sierra Nevada and southern California. Existing data indicate no range overlap. These results have important implications for the conservation of these two species as they illuminate a profound mismatch between the current delineation of the distinct population segments (southern California vs. Sierra Nevada) and actual species boundaries. For example, our study finds that remnant populations of R. muscosa exist in both the southern Sierra Nevada and the mountains of southern California, which may broaden options for management. In addition, despite the fact that only the southern California populations are listed as Endangered, surveys conducted since 1995 at 225 historic (1899-1994) localities from museum collections show that 93.3% (n=146) of R. sierrae populations and 95.2% (n=79) of R. muscosa populations are extinct. Evidence presented here underscores the need for revision of protected population status to include both species throughout their ranges.

Heritability estimates for growth in the tropical abalone Haliotis asinina using microsatellites to assign parentage

The tropical abalone Haliotis asinina is a wild-caught and cultured species throughout the Indo-Pacific as well as being an emerging model species for the study of haliotids. H. asinina has the fastest recorded natural growth rate of any abalone and reaches sexual maturity within one year. As such, it is a suitable abalone species for selective breeding for commercially important traits such as rapid growth. Estimating the amount of variation in size that is attributable to heritable genetic differences can assist the development of such a selective breeding program. Here we estimated heritability for growth-related traits at 12 months of age by creating a single cohort of 84 families in a full-factorial mating design consisting of 14 sires and 6 dams. Of 500 progeny sampled, 465 were successfully assigned to their parents based on shared alleles at 5 polymorphic microsatellite loci. Using an animal model, heritability estimates were 0.48 ± 0.15 for shell length, 0.38 ± 0.13 for shell width and 0.36 ± 0.13 for weight. Genetic correlations were > 0.98 between shell parameters and weight, indicating that breeding for weight gains could be successfully achieved by selecting for shell length.

Language in Acquisition : Early Lexical Development and Associations between Lexicon and Grammar Findings from Full-Term and Very-Low-Birth-Weight Finnish Children

The aim was to analyse the growth and compositional development of the receptive and expressive lexicons between the ages 0,9 and 2;0 in the full-term (FT) and the very-low-birth-weight (VLBW) children who are acquiring Finnish. The associations between the expressive lexicon and grammar at 1;6 and 2;0 in the FT children were also studied. In addition, the language skills of the VLBW children at 2;0 were analysed, as well as the predictive value of early lexicon to the later language performance. Four groups took part in the studies: the longitudinal (N = 35) and cross-sectional (N = 146) samples of the FT children, and the longitudinal (N = 32) and cross-sectional (N = 66) samples of VLBW children. The data was gathered by applying of the structured parental rating method (the Finnish version of the Communicative Development Inventory), through analysis of the children´s spontaneous speech and by administering a a formal test (Reynell Developmental Language Scales). The FT children acquired their receptive lexicons earlier, at a faster rate and with larger individual variation than their expressive lexicons. The acquisition rate of the expressive lexicon increased from slow to faster in most children (91%). Highly parallel developmental paths for lexical semantic categories were detected in the receptive and expressive lexicons of the Finnish children when they were analysed in relation to the growth of the lexicon size, as described in the literature for children acquiring other languages. The emergence of grammar was closely associated with expressive lexical growth. The VLBW children acquired their receptive lexicons at a slower rate and had weaker language skills at 2;0 than the full-term children. The compositional development of both lexicons happened at a slower rate in the VLBW children when compared to the FT controls. However, when the compositional development was analysed in relation to the growth of lexicon size, this development occurred qualitatively in a nearly parallel manner in the VLBW children as in the FT children. Early receptive and expressive lexicon sizes were significantly associated with later language skills in both groups. The effect of the background variables (gender, length of the mother s basic education, birth weight) on the language development in the FT and the VLBW children differed. The results provide new information of early language acquisition by the Finnish FT and VLBW children. The results support the view that the early acquisition of the semantic lexical categories is related to lexicon growth. The current findings also propose that the early grammatical acquisition is closely related to the growth of expressive vocabulary size. The language development of the VLBW children should be followed in clinical work.

Reduced efficacy of macrocyclic lactone treatments in controlling gastrointestinal nematode infections of weaner dairy calves in subtropical eastern Australia.

Faecal Egg Count Reduction Tests (FECRTs) for macrocyclic lactone (ML) and levamisole (LEV) drenches were conducted on two dairy farms in the subtropical, summer rainfall region of eastern Australia to determine if anthelmintic failure contributed to severe gastrointestinal nematode infections observed in weaner calves. Subtropical Cooperia spp. were the dominant nematodes on both farms although significant numbers of Haemonchus placei were also present on Farm 2. On Farm 1, moxidectin pour-on (MXD) drenched at 0.5 mg kg-1 liveweight (LW) reduced the overall Cooperia burden by 82% (95% confidence limits, 37-95%) at day 7 post-drench. As worm burdens increased rapidly in younger animals in the control group (n = 4), levamisole was used as a salvage drench and these calves withdrawn from the trial on animal welfare grounds after sample collection at day 7. Levamisole (LEV) dosed at 6.8 mg kg-1 LW reduced the worm burden in these calves by 100%, 7 days after drenching. On Farm 2, MXD given at 0.5 mg kg-1 LW reduced the faecal worm egg count of cooperioids at day 8 by 96% (71-99%), ivermectin oral (IVM) at 0.2 mg kg-1 LW by 1.6% (-224 to 70%) and LEV oral at 7.1 mg kg-1 LW by 100%. For H. placei the reductions were 98% (85-99.7%) for MXD, 0.7% (-226 to 70%) for IVM and 100% for LEV. This is the first report in Australia of the failure of macrocyclic lactone treatments to control subtropical Cooperia spp. and suspected failure to control H. placei in cattle.

Chemical changes during the aging and decomposition of composite solid propellants

During the thermal decomposition of orthorhombic ammonium perchlorate (AP) at 230°C, where the decomposition is only up to 30 wt %, there is an accumulation in the solid of acids, the concentration of which increases up to 15% decomposition, after which it decreases till it reaches the original value. Similar observations have been made in the polystyrene (PS)/AP propellant systems. Aging studies of PS/AP propellants have been carried out earlier [1], where it has been shown that for the aged propellants the thermal decomposition (TD) rate at 230°C and 260°C and ambient pressure burning rate (Image ) both increase and this increase is due to the formation of reactive intermediate “polystyrene peroxide (PSP).” In the present studies it has been observed that during the aging of the propellant at 150°C, the acid is formed and gets accumulated in the propellant, which may also be responsible for the increase in TD rate and perhaps may be more effective than PSP.

Adsorption of CO and N2 on modified transition-metal surfaces

Electron spectroscopic studies clearly demonstrate that modification of the surfaces of Mn, Fe and Ni metals by chlorine significantly decreases the strength of interaction between the metal and adsorbed molecules such as CO and N2. This is in contrast to the effect of electropositive additives such as Ba and Al which increase the adsorption bond strength significantly.

Pharmacogenetic Variation at CYP2D6, CYP2C9, and CYP2C19: Population Genetic and Forensic Aspects

Pharmacogenetics deals with genetically determined variation in drug response. In this context, three phase I drug-metabolizing enzymes, CYP2D6, CYP2C9, and CYP2C19, have a central role, affecting the metabolism of about 20-30% of clinically used drugs. Since genes coding for these enzymes in human populations exhibit high genetic polymorphism, they are of major pharmacogenetic importance. The aims of this study were to develop new genotyping methods for CYP2D6, CYP2C9, and CYP2C19 that would cover the most important genetic variants altering the enzyme activity, and, for the first time, to describe the distribution of genetic variation at these loci on global and microgeographic scales. In addition, pharmacogenetics was applied to a postmortem forensic setting to elucidate the role of genetic variation in drug intoxications, focusing mainly on cases related to tricyclic antidepressants, which are commonly involved in fatal drug poisonings in Finland. Genetic variability data were obtained by genotyping new population samples by the methods developed based on PCR and multiplex single-nucleotide primer extension reaction, as well as by collecting data from the literature. Data consisted of 138, 129, and 146 population samples for CYP2D6, CYP2C9, and CYP2C19, respectively. In addition, over 200 postmortem forensic cases were examined with respect to drug and metabolite concentrations and genotypic variation at CYP2D6 and CYP2C19. The distribution of genetic variation within and among human populations was analyzed by descriptive statistics and variance analysis and by correlating the genetic and geographic distances using Mantel tests and spatial autocorrelation. The correlation between phenotypic and genotypic variation in drug metabolism observed in postmortem cases was also analyzed statistically. The genotyping methods developed proved to be informative, technically feasible, and cost-effective. Detailed molecular analysis of CYP2D6 genetic variation in a global survey of human populations revealed that the pattern of variation was similar to those of neutral genomic markers. Most of the CYP2D6 diversity was observed within populations, and the spatial pattern of variation was best described as clinal. On the other hand, genetic variants of CYP2D6, CYP2C9, and CYP2C19 associated with altered enzymatic activity could reach extremely high frequencies in certain geographic regions. Pharmacogenetic variation may also be significantly affected by population-specific demographic histories, as seen within the Finnish population. When pharmacogenetics was applied to a postmortem forensic setting, a correlation between amitriptyline metabolic ratios and genetic variation at CYP2D6 and CYP2C19 was observed in the sample material, even in the presence of confounding factors typical for these cases. In addition, a case of doxepin-related fatal poisoning was shown to be associated with a genetic defect at CYP2D6. Each of the genes studied showed a distinct variation pattern in human populations and high frequencies of altered activity variants, which may reflect the neutral evolution and/or selective pressures caused by dietary or environmental exposure. The results are relevant also from the clinical point of view since the genetic variation at CYP2D6, CYP2C9, and CYP2C19 already has a range of clinical applications, e.g. in cancer treatment and oral anticoagulation therapy. This study revealed that pharmacogenetics may also contribute valuable information to the medicolegal investigation of sudden, unexpected deaths.

Who did pharmacists vaccinate in Australia’s first Pharmacist Immunisation Pilot?

Introduction: The Queensland Pharmacist Immunisation Pilot (QPIP) began in April 2014, and was Australia’s first to allow pharmacists vaccination. An aim of QPIP was to investigate participants’ satisfaction with the service, and their overall experience with the service. Method: Patient demographics and previous influenza vaccination experiences were recorded using GuildCare software. After receiving the influenza vaccine from the pharmacist, participants were asked to complete a ‘post-vaccination satisfaction questionnaire’. Results: A total of 10,889 participants received influenza vaccinations from a pharmacist, and >8000 participants completed the post-vaccination survey. Males accounted for 37% of participants, with the majority of participants aged between 45-64 years (53%). Almost half of the participants had been vaccinated before, the majority at a GP clinic (60%), and most participants reported receiving their previous influenza vaccination from a nurse (61%). Interestingly, 7% were unsure which healthcare professional had vaccinated them, and 1% thought a pharmacist had administered their previous vaccination. It was also noteworthy that approximately 10% of all participants were eligible to receive a free vaccination under the National Immunisation Program, but opted to receive their vaccine in a pharmacy. Overall, 95% were happy to receive their vaccination from a pharmacy in the future and 97% would recommend this service to other people. Conclusion: Participants were overwhelmingly positive in their response to the pharmacist vaccination pilot. These findings have paved the way for expanding the scope of practice for pharmacists with the aim to increase vaccination rates across the country. The pilot has now been expanded to include the administration of vaccinations for measles and pertussis.

Pharmacist vaccination – an Australian first, but what was the impact?

Introduction/background/issues The Queensland Pharmacist Immunisation Pilot is Australia’s first to allow pharmacists vaccination. The pilot ran between April 1st 2014 and August 31st 2014, with pharmacists administering influenza vaccination during the flu season. The aim of this work was to investigate the benefits of trained registered pharmacists administering vaccinations in a community pharmacy setting. Methods Participant demographics and previous influenza vaccination experiences were recorded using GuildCare software. Participants also completed a ‘post-vaccination satisfaction survey’ following their influenza vaccination. Results/discussions A total of 10,889 participant records were analysed. Females accounted for 63% of participants, with the majority of participants aged between 45-64 years (53%). Overall, 49% of participants had been vaccinated before, the majority at a GP clinic (60%). Most participants reported receiving their previous influenza vaccination from a nurse (61%). Interestingly, 1% thought a pharmacist had administered their previous vaccination, while 7% were unsure which health professional had administered it. It was also of note that approximately 10% of all participants were eligible to receive a free vaccination from the National Immunisation Program, but still opted to receive their vaccine in a pharmacy. Over 8,000 participants took part in the post-vaccination survey, 93% were happy to receive their vaccination from a pharmacy in the future while 94% would recommend this service to other people. The remaining 7% and 6% respectively had omitted to fill in those questions. Conclusions/implications These findings have helped pave the way for expanding the scope of practice for pharmacists with the aim to increase vaccination rates across Australia. Key message • Scope of practice and ability for health providers like pharmacists to provide services such as vaccination in primary care. • New service delivery to improve access to service, and increase immunisation rates.

The health professional behind the syringe: Benefits of a pharmacist vaccination program

Background: The Queensland Pharmacist Immunisation Pilot (QPIP) which ran in 2014 was Australia’s first to allow pharmacists to administer vaccinations. An aim of QPIP was to investigate the benefits of trained pharmacists administering vaccinations in a community pharmacy setting. Methods: Participant demographics and previous influenza vaccination experiences were recorded using GuildCare software. Participants also completed a ‘post-vaccination satisfaction survey’ following their influenza vaccination. Results: A total of 10,889 participant records and 8,737 satisfaction surveys were analysed. Overall, 1.9% of the participants reported living with a chronic illness, and 22.5% were taking concomitant medications. As part of the consultation before receiving the vaccine, participants acknowledged the opportunity to discuss other aspects of their health with the pharmacist, including concerns about their general health, allergies, and other medications they were taking. It was worth noting that 17.5% of people would not have received an influenza vaccination if the QPIP service was unavailable. Additionally, approximately 10% of all participants were eligible to receive a free vaccination from the National Immunisation Program, but still opted to receive their vaccine from a pharmacist. Conclusion: The findings from this pilot demonstrate the benefit of a pharmacist vaccination program in increasing vaccination rates, and have helped pave the way for expanding the scope of practice for pharmacists.

Positive results in a real-time PCR for type A influenza associated with the use of an inactivated vaccine.

A real-time reverse transcription polymerase chain reaction (qRT-PCR) test for the matrix gene of type A influenza viruses was used during the 2007 Australian equine influenza (EI) outbreak in order to confirm diagnosis and, later, eradication of the virus. During the EI outbreak, horses being exported required vaccination and individual proof of freedom from EI. At the end of the outbreak, positive results were obtained from four horses destined for export, because of contamination of the samples with the vaccine. This report highlights the need for EI testing and vaccination to occur on separate days and with the collection of swabs for testing to precede vaccination.