Colocalization of different neurotransmitter transporters on synaptic vesicles is sparse except for VGLUT1 and ZnT3

Vesicular transporters (VTs) define the type of neurotransmitter that synaptic vesicles (SVs) store and release. While certain mammalian neurons release multiple transmitters, it is not clear whether the release occurs from the same or distinct vesicle pools at the synapse. Using quantitative single-vesicle imaging, we show that a vast majority of SVs in the rodent brain contain only one type of VT, indicating specificity for a single neurotransmitter. Interestingly, SVs containing dual transporters are highly diverse (27 types) but small in proportion (2% of all SVs), excluding the largest pool that carries VGLUT1 and ZnT3 (34%). Using VGLUT1-ZnT3 SVs, we demonstrate that the transporter colocalization influences the SV content and synaptic quantal size. Thus, the presence of diverse transporters on the same vesicle is bona fide, and depending on the VT types, this may act to regulate neurotransmitter type, content, and release in space and time

Inhibition of resistance-refractory P. falciparum kinase PKG delivers prophylactic, blood stage, and transmission-blocking antiplasmodial activity

The search for antimalarial chemotypes with modes of action unrelated to existing drugs has intensified with the recent failure of first-line therapies across Southeast Asia. Here, we show that the trisubstituted imidazole MMV030084 potently inhibits hepatocyte invasion by Plasmodium sporozoites, merozoite egress from asexual blood stage schizonts, and male gamete exflagellation. Metabolomic, phosphoproteomic, and chemoproteomic studies, validated with conditional knockdown parasites, molecular docking, and recombinant kinase assays, identified cGMP-dependent protein kinase (PKG) as the primary target of MMV030084. PKG is known to play essential roles in Plasmodium invasion of and egress from host cells, matching MMV030084's activity profile. Resistance selections and gene editing identified tyrosine kinase-like protein 3 as a low-level resistance mediator for PKG inhibitors, while PKG itself never mutated under pressure. These studies highlight PKG as a resistance-refractory antimalarial target throughout the Plasmodium life cycle and promote MMV030084 as a promising Plasmodium PKG-targeting chemotype

Chromatin Immunoprecipitation to Analyze DNA Binding Sites of HMGA2

BACKGROUND: HMGA2 is an architectonic transcription factor abundantly expressed during embryonic and fetal development and it is associated with the progression of malignant tumors. The protein harbours three basically charged DNA binding domains and an acidic protein binding C-terminal domain. DNA binding induces changes of DNA conformation and hence results in global overall change of gene expression patterns. Recently, using a PCR-based SELEX (Systematic Evolution of Ligands by Exponential Enrichment) procedure two consensus sequences for HMGA2 binding have been identified. METHODOLOGY/PRINCIPAL FINDINGS: In this investigation chromatin immunoprecipitation (ChIP) experiments and bioinformatic methods were used to analyze if these binding sequences can be verified on chromatin of living cells as well. CONCLUSION: After quantification of HMGA2 protein in different cell lines the colon cancer derived cell line HCT116 was chosen for further ChIP experiments because of its 3.4-fold higher HMGA2 protein level. 49 DNA fragments were obtained by ChIP. These fragments containing HMGA2 binding sites have been analyzed for their AT-content, location in the human genome and similarities to sequences generated by a SELEX study. The sequences show a significantly higher AT-content than the average of the human genome. The artificially generated SELEX sequences and short BLAST alignments (11 and 12 bp) of the ChIP fragments from living cells show similarities in their organization. The flanking regions are AT-rich, whereas a lower conservation is present in the center of the sequences

An epitaph to Section 28? Telling tales out of school about changes and challenges to discourses of sexuality

This is a postprint of an article whose final and definitive form has been published in the International Journal of Qualitative Studies in Education,© 2007 Copyright Taylor & Francis; International Journal of Qualitative Studies in Education, is available online at http://www.informaworld.comThis article seeks to develop an understanding of the professional and personal lives of LGBT teachers in relation to the discriminatory statute Section 28, which prohibited 'promotion' of 'the acceptability of homosexuality as a pretended family relationship' by local education authorities in the UK (except Northern Ireland). Interviews with a small sample of serving teachers are analysed using a feminist poststructuralist methodology to discover whether the removal of this legislation marks a shift in theorization, policy or practice. Findings are arranged to focus on the workings of official policy, on informal or unofficial classroom and staffroom practices, and on relations with a local community. Analysis and discussion reveal a complex matrix of constituents (space, relationships and other variables) only some of which respond to the (perhaps) superficial stimulus of legislative change. Such change goes only a small way to challenge a deeply embedded discourse of inequality, which may respond only to a more profound epistemological transformation

CD4陽性T細胞に発現するA20(tnfaip3) はTh2型アレルギー性気道炎症を抑制する

研究科: 千葉大学大学院医学薬学府（先端医学薬学専攻）学位記番号: 千大院医薬博甲第医1412号博士（医学）千葉大学 = Chiba Universit

Die Aufklärung der Noludarzwischenfälle im Hamburger Hafenviertel (St. Pauli) mit Hilfe kombinierter Analysenmethoden

Arnold W, Grützmacher H-F. Die Aufklärung der Noludarzwischenfälle im Hamburger Hafenviertel (St. Pauli) mit Hilfe kombinierter Analysenmethoden. Deutsche Zeitschrift für die gesamte gerichtliche Medizin. 1969;65(1):44-60.In den Jahren 1965/66 kam es vermehrt zu Beraubungsdelikten im Hamburger Hafenviertel (St. Pauli). Nach den Tatumständen war anzunehmen, daß wahrscheinlich den meist mehr oder weniger alkoholisierten Opfern unbeobachtet ein schnell wirkendes Schlaf- oder Betäubungsmittel beigebracht wurde. Die chemische Untersuchung polizeilich sichergestellter Urinproben der Geschädigten führte zur Isolierung eines Fremdstoffes aus einzelnen dieser Harnasservate, der mit Hilfe kombinierter Analysenmethoden (präparative Dünnschichtchromatographie, IR- und Massenspektrographie) als ein neuer Noludar-Metabolit (2,4-Dioxo-3,3-diäthyl-5-methyl-6-hydroxypiperidin) identifiziert wurde