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3.
PeerJ ; 6: e5654, 2018.
Article in English | MEDLINE | ID: mdl-30280031

ABSTRACT

Most pollination ecosystem services studies have focussed on wild pollinators and their dependence on natural floral resources adjacent to crop fields. However, managed pollinators depend on a mixture of floral resources that are spatially separated from the crop field. Here, we consider the supporting role these resources play as an ecosystem services provider to quantify the use and availability of floral resources, and to estimate their relative contribution to support pollination services of managed honeybees. Beekeepers supplying pollination services to the Western Cape deciduous fruit industry were interviewed to obtain information on their use of floral resources. For 120 apiary sites, we also analysed floral resources within a two km radius of each site based on geographic data. The relative availability of floral resources at sites was compared to regional availability. The relative contribution of floral resources-types to sustain managed honeybees was estimated. Beekeepers showed a strong preference for eucalypts and canola. Beekeepers selectively placed more hives at sites with eucalypt and canola and less with natural vegetation. However, at the landscape-scale, eucalypt was the least available resource, whereas natural vegetation was most common. Based on analysis of apiary sites, we estimated that 700,818 ha of natural vegetation, 73,910 ha of canola fields, and 10,485 ha of eucalypt are used to support the managed honeybee industry in the Western Cape. Whereas the Cape managed honeybee system uses a bee native to the region, alien plant species appear disproportionately important among the floral resources being exploited. We suggest that an integrated approach, including evidence from interview and landscape data, and fine-scale biological data is needed to study floral resources supporting managed honeybees.

4.
Proc Biol Sci ; 282(1816): 20152053, 2015 Oct 07.
Article in English | MEDLINE | ID: mdl-26446814

ABSTRACT

Brood-site mutualisms represent extreme levels of reciprocal specialization between plants and insect pollinators, raising questions about whether these mutualisms are mediated by volatile signals and whether these signals and insect responses to them covary geographically in a manner expected from coevolution. Cycads are an ancient plant lineage in which almost all extant species are pollinated through brood-site mutualisms with insects. We investigated whether volatile emissions and insect olfactory responses are matched across the distribution range of the African cycad Encephalartos villosus. This cycad species is pollinated by the same beetle species across its distribution, but cone volatile emissions are dominated by alkenes in northern populations, and by monoterpenes and a pyrazine compound in southern populations. In reciprocal choice experiments, insects chose the scent of cones from the local region over that of cones from the other region. Antennae of beetles from northern populations responded mainly to alkenes, while those of beetles from southern populations responded mainly to pyrazine. In bioassay experiments, beetles were most strongly attracted to alkenes in northern populations and to the pyrazine compound in southern populations. Geographical matching of cone volatiles and pollinator olfactory preference is consistent with coevolution in this specialized mutualism.


Subject(s)
Coleoptera/physiology , Pollination , Symbiosis , Volatile Organic Compounds/metabolism , Zamiaceae/physiology , Animals , Biological Evolution , Geography , Odorants/analysis , Olfactory Perception , South Africa
5.
PLoS One ; 10(8): e0135152, 2015.
Article in English | MEDLINE | ID: mdl-26252495

ABSTRACT

Plants provide fundamental support systems for life on Earth and are the basis for all terrestrial ecosystems; a decline in plant diversity will be detrimental to all other groups of organisms including humans. Decline in plant diversity has been hard to quantify, due to the huge numbers of known and yet to be discovered species and the lack of an adequate baseline assessment of extinction risk against which to track changes. The biodiversity of many remote parts of the world remains poorly known, and the rate of new assessments of extinction risk for individual plant species approximates the rate at which new plant species are described. Thus the question 'How threatened are plants?' is still very difficult to answer accurately. While completing assessments for each species of plant remains a distant prospect, by assessing a randomly selected sample of species the Sampled Red List Index for Plants gives, for the first time, an accurate view of how threatened plants are across the world. It represents the first key phase of ongoing efforts to monitor the status of the world's plants. More than 20% of plant species assessed are threatened with extinction, and the habitat with the most threatened species is overwhelmingly tropical rain forest, where the greatest threat to plants is anthropogenic habitat conversion, for arable and livestock agriculture, and harvesting of natural resources. Gymnosperms (e.g. conifers and cycads) are the most threatened group, while a third of plant species included in this study have yet to receive an assessment or are so poorly known that we cannot yet ascertain whether they are threatened or not. This study provides a baseline assessment from which trends in the status of plant biodiversity can be measured and periodically reassessed.


Subject(s)
Biodiversity , Conservation of Natural Resources , Endangered Species , Viridiplantae/classification , Databases, Factual , Ecosystem , Extinction, Biological , Geography , Rainforest , Tropical Climate
6.
Mol Biol Cell ; 24(24): 3881-95, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24131997

ABSTRACT

Src interactions with the plasma membrane are an important determinant of its activity. In turn, Src activity modulates its association with the membrane through binding of activated Src to phosphotyrosylated proteins. Caveolin-1 (Cav-1), a major component of caveolae, is a known Src phosphorylation target, and both were reported to regulate cell transformation. However, the nature of Src-Cav-1 interactions, a potential mechanism of their coregulation, remained unclear. Here we used fluorescence recovery after photobleaching beam-size analysis, coimmunoprecipitation, quantitative imaging, and far-Western studies with cells expressing wild type, as well as structural and activity mutants of Src-green fluorescent protein and Cav-1-monomeric red fluorescent protein, to measure their interactions with the membrane and with each other. We show dynamic Src-plasma membrane interactions, which are augmented and stabilized by Cav-1. The mechanism involves phosphorylation of Cav-1 at Tyr-14 by Src and subsequent binding of the Src SH2 domain to phospho-Cav-1, leading to accumulation of activated Src in focal adhesions. This novel Cav-1 function potentially modulates focal adhesion dynamics.


Subject(s)
Caveolin 1/metabolism , Cell Membrane/metabolism , src-Family Kinases/metabolism , Animals , COS Cells , Cell Line , Chlorocebus aethiops , Cholesterol/biosynthesis , Focal Adhesions , Green Fluorescent Proteins/genetics , Luminescent Proteins/genetics , Phosphorylation , Protein Binding , Protein Structure, Tertiary , RNA Interference , RNA, Small Interfering , Rats , Red Fluorescent Protein
7.
Ann Bot ; 112(5): 891-902, 2013 Sep.
Article in English | MEDLINE | ID: mdl-23887092

ABSTRACT

BACKGROUND AND AIMS: Ontogenetic patterns of odour emissions and heating associated with plant reproductive structures may have profound effects on insect behaviour, and consequently on pollination. In some cycads, notably Macrozamia, temporal changes in emission of specific odour compounds and temperature have been interpreted as a 'push-pull' interaction in which pollinators are either attracted or repelled according to the concentration of the emitted volatiles. To establish which mechanisms occur in the large Encephalartos cycad clade, the temporal patterns of volatile emissions, heating and pollinator activity of cones of Encephalartos villosus in the Eastern Cape (EC) and KwaZulu Natal (KZN) of South Africa were investigated. METHODS AND KEY RESULTS: Gas chromatography-mass spectrometry (GC-MS) analyses of Encephalartos villosus cone volatiles showed that emissions, dominated by eucalyptol and 2-isopropyl-3-methoxypyrazine in EC populations and (3E)-1,3-octadiene and (3E,5Z)-1,3,5-octatriene in the KZN populations, varied across developmental stages but did not vary significantly on a daily cycle. Heating in male cones was higher at dehiscence than during pre- and post-dehiscence, and reached a maximum at about 1830 h when temperatures were between 7·0 and 12·0 °C above ambient. Daily heating of female cones was less pronounced and reached a maximum at about 1345 h when it was on average between 0·9 and 3·0 °C above ambient. Insect abundance on male cones was higher at dehiscence than at the other stages and significantly higher in the afternoon than in the morning and evening. CONCLUSIONS: There are pronounced developmental changes in volatile emissions and heating in E. villosus cones, as well as strong daily changes in thermogenesis. Daily patterns of volatile emissions and pollinator abundance in E. villosus are different from those observed in some Macrozamia cycads and not consistent with the push-pull pattern as periods of peak odour emission do not coincide with mass exodus of insects from male cones.


Subject(s)
Flowers/chemistry , Insecta/physiology , Monoterpenes/metabolism , Oils, Volatile/metabolism , Zamiaceae/chemistry , Animals , Flowers/physiology , Host-Parasite Interactions , Hydrocarbons/metabolism , Odorants , Plant Oils/metabolism , Pollen/chemistry , Pollen/physiology , Pollination , Reproduction , Temperature , Thermogenesis , Zamiaceae/physiology
8.
PLoS One ; 8(5): e63570, 2013.
Article in English | MEDLINE | ID: mdl-23717448

ABSTRACT

We aggregated data on butterfly-host plant associations from existing sources in order to address the following questions: (1) is there a general correlation between host diversity and butterfly species richness?, (2) has the evolution of host plant use followed consistent patterns across butterfly lineages?, (3) what is the common ancestral host plant for all butterfly lineages? The compilation included 44,148 records from 5,152 butterfly species (28.6% of worldwide species of Papilionoidea) and 1,193 genera (66.3%). The overwhelming majority of butterflies use angiosperms as host plants. Fabales is used by most species (1,007 spp.) from all seven butterfly families and most subfamilies, Poales is the second most frequently used order, but is mostly restricted to two species-rich subfamilies: Hesperiinae (56.5% of all Hesperiidae), and Satyrinae (42.6% of all Nymphalidae). We found a significant and strong correlation between host plant diversity and butterfly species richness. A global test for congruence (Parafit test) was sensitive to uncertainty in the butterfly cladogram, and suggests a mixed system with congruent associations between Papilionidae and magnoliids, Hesperiidae and monocots, and the remaining subfamilies with the eudicots (fabids and malvids), but also numerous random associations. The congruent associations are also recovered as the most probable ancestral states in each node using maximum likelihood methods. The shift from basal groups to eudicots appears to be more likely than the other way around, with the only exception being a Satyrine-clade within the Nymphalidae that feed on monocots. Our analysis contributes to the visualization of the complex pattern of interactions at superfamily level and provides a context to discuss the timing of changes in host plant utilization that might have promoted diversification in some butterfly lineages.


Subject(s)
Butterflies/classification , Magnoliopsida , Animals , Biological Evolution , Feeding Behavior , Herbivory , Likelihood Functions , Models, Biological , Phylogeny , Species Specificity
9.
Phytochemistry ; 85: 82-91, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23098901

ABSTRACT

Volatiles play a key role in attraction of pollinators to cycad cones, but the extent to which volatile chemistry varies among cycad species is still poorly documented. Volatile composition of male and female cones of nineteen African cycad species (Encephalartos; Zamiaceae) was analysed using headspace technique and gas chromatography-mass spectrometry (GC-MS). A total of 152 compounds were identified among the species included in this study, the most common of which were monoterpenes, nitrogen-containing compounds and unsaturated hydrocarbons. Male and female cones emitted similar volatile compounds which varied in relative amounts with two unsaturated hydrocarbons (3E)-1,3-octadiene and (3E,5Z)-1,3,5-octatriene present in the volatile profile of most species. In a multivariate analysis of volatile profiles using non-metric multidimensional scaling (NMDS), a number of species clusters were identified according to shared emission of unsaturated hydrocarbons, pyrazines, benzenoids, aldehydes, alkanes and terpenoids. In comparison, terpenoids are common in Zamia and dominant in Macrozamia species (both in the family Zamiaceae) while benzenoids, esters, and alcohols are dominant in Cycas (Cycadaceae) and in Stangeria (Stangeriaceae). It is likely that volatile variation among Encephalartos species reflects both phylogeny and adaptations to specific beetle pollinators.


Subject(s)
Zamiaceae/chemistry , Animals , Gas Chromatography-Mass Spectrometry , Hydrocarbons/metabolism , Phylogeny , Pollination/physiology , Terpenes/metabolism , Zamiaceae/classification , Zamiaceae/physiology
10.
Int J Gynecol Cancer ; 22(6): 1044-9, 2012 Jul.
Article in English | MEDLINE | ID: mdl-22622952

ABSTRACT

OBJECTIVE: To establish the accuracy of sentinel lymph node (SLN) detection in early cervical cancer. MATERIALS AND METHODS: Sentinel lymph node detection was performed prospectively over a 6-year period in 86 women undergoing surgery for cervical carcinoma by the combined method (Tc-99m and methylene blue dye). Further ultrastaging was performed on a subgroup of 26 patients who had benign SLNs on initial routine histological examination. RESULTS: The SLN was detected in 84 (97.7%) of 86 women by the combined method. Blue dye uptake was not seen in 8 women (90.7%). Sentinel lymph nodes were detected bilaterally in 63 women (73.3%), and the external iliac region was the most common anatomic location (48.8%). The median SLN count was 3 nodes (range, 1-7). Of the 84 women with sentinel node detection, 65 also underwent bilateral pelvic lymph node dissection, and in none of these cases was a benign SLN associated with a malignant non-SLN (100% negative predictive value). The median non-SLN count for all patients was 19 nodes (range, 8-35). Eighteen patients underwent removal of the SLN without bilateral pelvic lymph node dissection. Nine women (10.5%) had positive lymph nodes on final histology. One patient had bulky pelvic nodes on preoperative imaging and underwent removal of the negative bulky malignant lymph nodes and a benign SLN on the contralateral side. This latter case confirms the unreliability of the SLN method with bulky nodes. The remaining 8 patients had positive SLNs with negative nonsentinel lymph nodes. Fifty-nine SLNs from 26 patients, which were benign on initial routine histology, underwent ultrastaging, but no further disease was identified. Four patients (5%) relapsed after a median follow-up of 28 months (range, 8-80 months). CONCLUSION: Sentinel lymph node detection is an accurate and safe method in the assessment of nodal status in early cervical carcinoma.


Subject(s)
Carcinoma/pathology , Lymph Nodes/pathology , Sentinel Lymph Node Biopsy , Uterine Cervical Neoplasms/pathology , Adult , Aged , Female , Humans , Lymphatic Metastasis , Middle Aged , Neoplasm Staging , Prospective Studies , Young Adult
12.
Int J Gynecol Cancer ; 21(3): 559-64, 2011 Apr.
Article in English | MEDLINE | ID: mdl-21430459

ABSTRACT

OBJECTIVES: To determine the accuracy of sentinel lymph node (SLN) detection in vulval carcinoma and to report the reliability and safety of this procedure. METHODS/MATERIALS: For a period of 6 years, we recruited women undergoing surgery for vulval carcinoma. All women had a preoperative biopsy confirming the depth of invasion greater than 1 mm. Sentinel lymph node detection was performed using the combined method (Tc-99m and methylene-blue dye). The standard management included complete inguinofemoral lymphadenectomy. When inguinofemoral lymph nodes were found grossly to be enlarged, these nodes were debulked, and the women subsequently treated with radiotherapy with or without chemotherapy. During the last 2 years of the study, a selected group of women had an SLN dissection alone. The SLNs were ultrastaged when they were negative on routine hematoxylin and eosin examination. RESULTS: Among 60 women undergoing SLN detection, SLN was detected in 59 women (98.3%) with combined method. Blue dye did not detect an SLN in 3 women resulting in a 93.3% detection rate. The median SLN count was 2 nodes (range, 1-9). Of the 60 women, 41 had inguinofemoral lymphadenectomy, 4 had only enlarged inguinofemoral nodes debulked, and 15 had the SLN only removed. The non-SLN count was 9 nodes (range, 3-17). There were no false-negative SLNs. Twenty-one women (35%) had positive nodes on final histology. Ultrastaging increased detection of metastases in 6.9% of nodes relative to routine hematoxylin and eosin examination and upstaged 12% of women. The median follow-up was 24 months (range, 2-66 months). CONCLUSIONS: Sentinel lymph node detection is safe and accurate in assessing lymph node status in women with vulval cancer undergoing staging. The combined method using Tc-99m and methylene blue dye injection for SLN detection has the best detection rate. Routine ultrastaging of negative SLN improves the detection of nodal metastases.


Subject(s)
Carcinoma, Squamous Cell/diagnosis , Methylene Blue , Practice Patterns, Physicians' , Radiopharmaceuticals , Technetium Tc 99m Sulfur Colloid , Vulvar Neoplasms/diagnosis , Adult , Aged , Aged, 80 and over , Carcinoma, Squamous Cell/surgery , Female , Humans , Lymph Node Excision , Lymphatic Metastasis , Middle Aged , Neoplasm Staging , Prospective Studies , Sentinel Lymph Node Biopsy , Survival Rate , Treatment Outcome , Vulvar Neoplasms/surgery
13.
Ecol Lett ; 14(3): 251-9, 2011 Mar.
Article in English | MEDLINE | ID: mdl-21244594

ABSTRACT

Ongoing expansion of large-scale agriculture critically threatens natural habitats and the pollination services they offer. Creating patches with high plant diversity within farmland is commonly suggested as a measure to benefit pollinators. However, farmers rarely adopt such practice, instead removing naturally occurring plants (weeds). By combining pollinator exclusion experiments with analysis of honeybee behaviour and flower-visitation webs, we found that the presence of weeds allowed pollinators to persist within sunflower fields, maximizing the benefits of the remaining patches of natural habitat to productivity of this large-scale crop. Weed diversity increased flower visitor diversity, hence ameliorating the measured negative effects of isolation from natural habitat. Although honeybees were the most abundant visitors, diversity of flower visitors enhanced honeybee movement, being the main factor influencing productivity. Conservation of natural patches combined with promoting flowering plants within crops can maximize productivity and, therefore, reduce the need for cropland expansion, contributing towards sustainable agriculture.


Subject(s)
Bees/physiology , Biodiversity , Helianthus/physiology , Plant Weeds/physiology , Seeds/growth & development , Agriculture/methods , Animals , Conservation of Natural Resources , Ecosystem , Flowers/physiology , Helianthus/growth & development , Pollination , South Africa
14.
PDA J Pharm Sci Technol ; 65(6): 692, 2011.
Article in English | MEDLINE | ID: mdl-22294601

ABSTRACT

CONFERENCE PROCEEDING Proceedings of the PDA/FDA Adventitious Viruses in Biologics: Detection and Mitigation Strategies Workshop in Bethesda, MD, USA; December 1-3, 2010 Guest Editors: Arifa Khan (Bethesda, MD), Patricia Hughes (Bethesda, MD) and Michael Wiebe (San Francisco, CA).


Subject(s)
Biological Products , Technology , Gene Expression Profiling , Humans , San Francisco , Technology, Pharmaceutical
15.
Anticancer Res ; 29(10): 3845-55, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19846918

ABSTRACT

BACKGROUND: DNA-damaging agents are widely used for the treatment of human malignancies. Agents containing the multifunctional alkylating moiety tetrakis(2-chloroethyl)phosphorodiamidic acid are currently under development as cancer therapeutics. MATERIALS AND METHODS: TLK58747, a phophorodiamidate-based prodrug, was tested in vivo for antitumor efficacy and safety. The in vitro responses of tumor cells to TLK58747 were examined by cytotoxicity assays, cell cycle analysis, immunoblots and microscopy. RESULTS: TLK58747 was efficacious in xenograft models of human breast, pancreas, and prostate cancer, as well as in leukemia and glioma. It caused less bone marrow suppression in rats than did cyclophosphamide. In vitro, TLK58747 inhibited the growth of a wide variety of cancer cells and activated the DNA damage-response pathway, leading to G(2)/M cell cycle arrest and subsequent premature senescence or apoptosis. CONCLUSION: TLK58747 is a promising new alkylating agent with broad antitumor activity and superior safety that warrants further development.


Subject(s)
Antineoplastic Agents, Alkylating/pharmacology , DNA Damage , DNA, Neoplasm/drug effects , Organophosphorus Compounds/pharmacology , Prodrugs/pharmacology , Animals , Antineoplastic Agents, Alkylating/toxicity , Cell Division/drug effects , Cell Growth Processes/drug effects , Cell Line, Tumor , DNA, Neoplasm/metabolism , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , G2 Phase/drug effects , HL-60 Cells , Humans , Male , Mice , Neoplasms/drug therapy , Neoplasms/genetics , Neoplasms/pathology , Organophosphorus Compounds/toxicity , Prodrugs/toxicity , Rats , Rats, Sprague-Dawley , Xenograft Model Antitumor Assays
16.
Trends Plant Sci ; 14(11): 608-13, 2009 Nov.
Article in English | MEDLINE | ID: mdl-19773195

ABSTRACT

The contributions of botanic gardens to conservation biology and global-change research need to be understood within the context of the traditional strengths of such gardens in herbarium collections, living collections and interactions with the public. Here, I propose that research in conservation planning, modelling species responses to climate change, conservation of threatened species and experimental tests of global change build on the core strengths of botanic gardens. However, there are limits to what can be achieved through traditional gardens-based programs, and some botanic gardens have adapted their research to include studies of threatening processes and to monitor and verify global-change impacts. There is an opportunity for botanic gardens to use their living collections more effectively in global-change research and for them to have a role in linking biodiversity conservation with benefits derived from ecosystem services.


Subject(s)
Botany/methods , Climate Change , Conservation of Natural Resources/methods , Ecosystem , Botany/trends , Conservation of Natural Resources/trends , Endangered Species , Environment, Controlled , Environmental Monitoring , Plant Development , Population Dynamics , Research/trends , Research Design
18.
Ann Bot ; 100(7): 1483-9, 2007 Dec.
Article in English | MEDLINE | ID: mdl-17951585

ABSTRACT

BACKGROUND AND AIMS: A deviation from the classical beetle pollination syndrome of dull-coloured flowers with an unpleasant scent is found in the Greater Cape Floral Region of South Africa. Here, monkey beetles (Scarabaeidae) visit brightly coloured, odourless flowers with conspicuous dark spots and centres (beetle marks). The role of flower colour and markings in attracting monkey beetles is still poorly understood. METHODS: Artificial model flowers with different marking patterns were used to test the effect of beetle marks on visitation by monkey beetles. To test whether monkey beetles are conditioned to the colour of the local matrix species, model flowers of different colours were placed in populations of three differently coloured species of Iridaceae. KEY RESULTS: Among all three matrix species the presence of dark markings of some kind (either centres or spots) increased visitation rates but the different matrix species differed in whether the effect was due to a dark centre or to dark spots. Monkey beetles were not conditioned for the colour of the matrix species: model colour was not significant in the Hesperantha vaginata and in the Romulea monadelpha matrices, whereas yellow model flowers were preferred over orange ones in the orange-flowered Sparaxis elegans matrix. CONCLUSIONS: This study is the first to demonstrate that beetle marks attract pollinating monkey beetles in the Greater Cape Floral Region. In contrast to plants with the classical beetle pollination syndrome that use floral scent as the most important attractant of pollinating beetles, plants with the monkey beetle pollination syndrome rely on visual signals, and, in some areas at least, monkey beetles favour flowers with dark beetle markings over unmarked flowers.


Subject(s)
Coleoptera/physiology , Flowers/physiology , Pigmentation/physiology , Predatory Behavior/physiology , Animals , Color Perception/physiology , Contrast Sensitivity/physiology , Flowers/chemistry , Odorants , Pollination/physiology , South Africa
19.
J Cell Biol ; 178(4): 675-86, 2007 Aug 13.
Article in English | MEDLINE | ID: mdl-17698610

ABSTRACT

Src functions depend on its association with the plasma membrane and with specific membrane-associated assemblies. Many aspects of these interactions are unclear. We investigated the functions of kinase, SH2, and SH3 domains in Src membrane interactions. We used FRAP beam-size analysis in live cells expressing a series of c-Src-GFP proteins with targeted mutations in specific domains together with biochemical experiments to determine whether the mutants can generate and bind to phosphotyrosyl proteins. Wild-type Src displays lipid-like membrane association, whereas constitutively active Src-Y527F interacts transiently with slower-diffusing membrane-associated proteins. These interactions require Src kinase activity and SH2 binding, but not SH3 binding. Furthermore, overexpression of paxillin, an Src substrate with a high cytoplasmic population, competes with membrane phosphotyrosyl protein targets for binding to activated Src. Our observations indicate that the interactions of Src with lipid and protein targets are dynamic and that the kinase and SH2 domain cooperate in the membrane targeting of Src.


Subject(s)
Membranes/metabolism , Protein-Tyrosine Kinases/metabolism , Animals , COS Cells , CSK Tyrosine-Protein Kinase , Chlorocebus aethiops , Fluorescence Recovery After Photobleaching , Green Fluorescent Proteins/metabolism , Membranes/chemistry , Paxillin/metabolism , Protein-Tyrosine Kinases/chemistry , src Homology Domains , src-Family Kinases
20.
Menopause ; 14(4): 808; author reply 808-9, 2007.
Article in English | MEDLINE | ID: mdl-17549041
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