Induction of male sex behavior in pony mares with testosterone propionate

Two pony mares were administered 150 mg of testosterone propionate every other day for 20 days (ten injections) and every ten days there-after. An additional two mares and one stallion were not treated and served as controls. Testosterone propionate was dissolved in absolute ethanol and administered subcutaneously. Sex behavior tests were conducted 26 and 40 days after the first injection. Control mares exhibited very little male sex behavior. Both testosterone propionatetreated mares, however, exhibited mounting, sniffing, flehmen, biting and vocalization behavior in the presence of an estrous mare. The testosterone propionate-treated mares mounted and bit estrous mares more frequently than the stallion but exhibited less sniffing, flehmen and vocalization behavior in the presence of an estrous mare than the stallion. Testosterone propionate-treated mares and the stallion mounted an estrous mare 23.3 +/- 9.7 seconds and 172.5 +/- 22.5 seconds, respectively, after being introduced into the pen. Mares in estrus were mounted by the testosterone propionate-treated mares and the stallion an average of 4.0 +/- 1.3 and 1.0 +/- 0 times, respectively, during a ten-minute test. None of the non-estrous mares was ever mounted by the testosterone propionate-treated mares. In summary, testosterone propionate induced male sex behavior in intact mares and the testosterone propionate-treated mares effectively detected estrous mares.     

ARP101, a selective MMP-2 inhibitor, induces autophagy-associated cell death in cancer cells

Autophagy is a catabolic cellular process involving self-digestion and turnover of macromolecules and entire organelles. Autophagy is primarily a protective process in response to cellular stress, but it can be associated with cell death. Genetic evidence also supports autophagy function as a tumor suppressor mechanism. To identify specific regulators to autophagy, we screened the Lopac 1280 and the Prestwick chemical libraries using a cell-based screening system with autophagy marker (green fluorescence protein conjugated LC3 protein (GFP-LC3)). We identified ARP101, a selective matrix metalloproteinase-2 (MMP-2) inhibitor as one of the most potent inducer of autophagy. ARP101 treatment was highly effective in inducing the formation of autophagosome and conversion of LC3I into LC3II. Moreover, ARP101-induced autophagy was completely blocked in mouse embryo fibroblasts that lacked autophagy related gene 5 (ATG5^−/− MEF). Interestingly, cell death induced by ARP101 was not inhibited by zVAD, a pan caspase inhibitor, whereas, it was efficiently suppressed by addition of 3-methyladenine, an autophagy inhibitor. These results suggest that the selective MMP-2 inhibitor, ARP101, induces autophagy and autophagy-associated cell death.     

Key biocultural values to guide restoration action and planning in New Zealand

A pluralist and cross‐cultural approach that accommodates differing values while encouraging the collaboration and social cohesion necessary for the complex task of ecological restoration is needed. We used qualitative and quantitative analyses to investigate value assigned to biocultural restoration of coastal forests in northern New Zealand by 26 interviewees from three groups (environmental managers, Māori community members, and community project leaders). Māori community members primarily emphasized the importance of Cultural Stewardship and Use in the restoration process, while placing less emphasis on Ecological Integrity. Otherwise, all participants shared common trends, culminating in three interrelated value sets: (1) Personal Engagement, (2) Connection, and (3) the generation and transfer of Knowledge & Wisdom. These values demonstrate that restoration's benefits to people and community are as significant as its reparations of ecological components. Despite differences, all stakeholders were united in a broadly common goal to restore socio‐ecological systems. Their knowledge and shared passion for conservation signal enormous promise for accelerated and effective restoration of coastal forests, if it is conducted using a pluralistic approach. Because some values expressed were intangible and complex, with cross‐cultural dimensions, current valuation tools used by ecological economists to guide management investment fail to adequately account for, in particular, Māori values of ecological restoration.     

禾谷镰刀菌降解毒素基因Tri101的克隆和序列分析

禾谷镰刀菌Tri101基因编码的单端孢酶烯3-O-乙酰转移酶可通过加乙酰基的形式使禾谷镰刀菌产生的单族毒素(如DON)转变为较低的毒性。本研究利用RT-PCR技术从禾谷镰刀菌0623中扩增并克隆了Tri101基因的cDNA片段,测序结果表明,Tri101基因核苷酸序列阅读框架全长1356bp(GenBank序列号:GQ907236),编码451个氨基酸的多肽,推测分子量为49.45kD,等电点为5.14。氨基酸序列同源性比对结果表明,它与Kimura报道的禾谷镰刀菌Tri101氨基酸序列同源性最高,为99.56%,与其它13种镰刀菌的Tri101氨基酸序列的同源性分别为97.91%-75.68%。系统进化树分析结果表明,Fusarium graminearium0623与Fusarium sporotrichioides属于同一进化枝且与Fusarium asiaticum有较近的亲缘关系,而与F.oxysporum、F.moniliforme、F.nygamai、F.nisikadoi和F.decemcellulare的亲缘关系较远。     

Epitope interactions of monoclonal antibodies targeting CD20 and their relationship to functional properties

Several novel anti-CD20 monoclonal antibodies are currently in development with the aim of improving the treatment of B cell malignancies. Mutagenesis and epitope mapping studies have revealed differences between the CD20 epitopes recognized by these antibodies. Recently, X-ray crystallography studies confirmed that the Type I CD20 antibody rituximab and the Type II CD20 antibody obinutuzumab (GA101) differ fundamentally in their interaction with CD20 despite recognizing a partially overlapping epitope on CD20. The Type I CD20 antibodies rituximab and ofatumumab are known to bind to different epitopes. The differences suggest that the biological properties of these antibodies are not solely determined by their core epitope sequences, but also depend on other factors, such as the elbow hinge angle, the orientation of the bound antibody and differential effects mediated by the Fc region of the antibody. Taken together, these factors may explain differences in the preclinical properties and clinical efficacy of anti-CD20 antibodies.     

Analysis of functions in plasmid pHZ1358 influencing its genetic and structural stability in <Emphasis Type="Italic">Streptomyces lividans</Emphasis> 1326

The complete DNA sequence of plasmid pHZ1358, a widely used vector for targeted gene disruption and replacement experiments in many Streptomyces hosts, has been determined. This has allowed a detailed analysis of the basis of its structural and segregational instability, compared to the high copy number plasmid pIJ101 of Streptomyces lividans 1326 from which it was derived. A 574-bp DNA region containing sti (strong incompatibility locus) was found to be a determinant for segregational instability in its original S. lividans 1326 host, while the structural instability was found to be related to the facile deletion of the entire Escherichia coli-derived part of pHZ1358, mediated by recombination between 36-bp direct repeats. A point mutation removing the BamHI site inside the rep gene encoding a replication protein (rep*) and/or a spontaneous deletion of the 694-bp region located between rep and sti including the uncharacterized ORF85 (orf85 ⁻) produced little or no effect on stability. A pHZ1358 derivative (pJTU412, sti ⁻ , rep*, orf85 ⁻) was then constructed which additionally lacked one of the 36-bp direct repeats. pJTU412 was demonstrated to be structurally stable but segregationally unstable and, in contrast to sti ⁺ pHZ1358, allowed efficient targeted gene replacement in S. lividans 1326.     

In vitro and in vivo manipulations of bacteriophage Mu DNA: cloning of Mu ends and construction of mini-Mu's carrying selectable markers

Recombinant plasmids carrying one or both ends of the bacteriophage Mu genome were constructed by molecular cloning. Transposable mini-Mu's with selectable markers (ampicillin resistance, kanamycin resistance or the entire lac operon of Escherichia coli) inserted between the Mu ends were also constructed. As a source of lac operon DNA, a pBR322 derivative with a 27 kb insert containing the lac operon was constructed. The plasmids with both ends of Mu (mini-Mu's) conferred full Mu immunity upon the host cells. However, the same mini-Mu's containing kan or lac inserts were defective in immunity. A summary of the construction and physical characterization, including restriction endonuclease cleavage maps and some of the biological properties of the plasmids, is presented.     

Dmt-Tic-NH-CH2-Bid (UFP-502), a potent DOP receptor agonist: in vitro and in vivo studies

Knockout and pharmacological studies demonstrated that the activation of delta opioid peptide (DOP) receptors produces antidepressant-like effects in rodents. Here we report the results obtained with the novel DOP ligand H-Dmt-Tic-NH-CH₂-Bid (UFP-502). UFP-502 bound with high affinity (pK_i 9.43) to recombinant DOP receptors displaying moderate selectivity over MOP and KOP. In CHO_hDOP [³⁵S]GTPγS binding and mouse vas deferens experiments, UFP-502 behaved as a potent (pEC₅₀ 10.09 and 10.70, respectively) full agonist. In these preparations, naloxone, naltrindole and N,N(CH₃)₂Dmt-Tic-OH showed similar pA₂ values against UFP-502 and DPDPE and the same rank order of potency. In vivo in mice, UFP-502 mimicked DPDPE actions, producing a significant reduction of immobility time after intracerebroventricular administration in the forced swimming test and a clear antinociceptive effect after intrathecal injection in the tail withdrawal assay. However, while the effects of DPDPE were fully prevented by naltrindole those evoked by UFP-502 were unaffected (tail withdrawal assay) or only partially reversed (forced swimming test). In conclusion, UFP-502 represents a novel and useful chemical template for the design of selective agonists for the DOP receptor.