Asymmetrical processing of olfactory input in the piriform cortex mediates "activation" of the avian navigation circuitry

The role of odors in the long-distance navigation of birds has elicited intense debate for more than half a century. Failure to resolve many of the issues fueling this debate is due at least in part to the absence of controls for a variety of non-specific effects that odors have on the navigational process. The present experiments were carried out to investigate whether the olfactory inputs are involved only in “activation” of neuronal circuitry involved in navigation or are also playing a role in providing directional information. Experienced adult pigeons were exposed to controlled olfactory stimuli during different segments of the journey (release site vs. displacement + release site). Protein levels of IEGs (immediate early genes used to mark synaptic activity) were analyzed in areas within the olfactory/navigation avian circuitry. The results indicate that 1) exposure to natural odors at the release site (and not before) elicit greater activation across brain regions than exposure to filtered air, artificial odors, and natural odors along the entire outward journey (from home to the release site, inclusive); 2) activation of the piriform cortex in terms of odor discrimination is lateralized; 3) activation of the navigation circuitry is achieved by means of lateralized activation of piriform cortex neurons. Altogether, the findings provide the first direct evidence that activation of the avian navigation circuitry is mediated by asymmetrical processing of olfactory input occurring in the right piriform cortex.

Characterization of a thymidylate synthase (TS)-inducible cell line: a model system for studying sensitivity to TS- and non-TS-targeted chemotherapies

Thymidylate synthase (TS) is responsible for the de novo synthesis of thymidylate, which is required for DNA synthesis and repair and which is an important target for fluoropyrimidines such as 5-fluorouracil (5-FU), and antifolates such as Tomudex (TDX), ZD9331, and multitargeted antifolate (MTA). To study the importance of TS expression in determining resistance to these agents, we have developed an MDA435 breast cancer-derived cell line with tetracycline-regulated expression of TS termed MTS-5. We have demonstrated that inducible expression of TS increased the IC(50) dose of the TS-targeted therapeutic agents 5-FU, TDX, and ZD9331 by 2-, 9- and 24-fold respectively. An IC(50) dose for MTA was unobtainable when TS was overexpressed in these cells, which indicated that MTA toxicity is highly sensitive to increased TS expression levels. The growth inhibitory effects of the chemotherapeutic agents CPT-11, cisplatin, oxaliplatin, and Taxol were unaffected by TS up-regulation. Cell cycle analyses revealed that IC(50) doses of 5-FU, TDX and MTA caused an S-phase arrest in cells that did not overexpress TS, and this arrest was overcome when TS was up-regulated. Furthermore, the S-phase arrest was accompanied by 2- to 4-fold increased expression of the cell cycle regulatory genes cyclin E, cyclin A, and cyclin dependent kinase 2 (cdk2). These results indicate that acute increases in TS expression levels play a key role in determining cellular sensitivity to TS-directed chemotherapeutic drugs by modulating the degree of S-phase arrest caused by these agents. Moreover, CPT-11, cisplatin, oxaliplatin, and Taxol remain highly cytotoxic in cells that overexpress TS.

Study of pesticide effects on non-target organisms

A utilização insustentável de pesticidas, especialmente em zonas com elevado valor ecológico constitui uma ameaça à integridade dos ecossistemas. Sendo um problema à escala mundial, e também no contexto nacional, o presente trabalho pretende ser um contributo para a avaliação dos efeitos de pesticidas em organismos não alvo terrestres e, principalmente, aquáticos, em contextos de progressiva relevância ecológica. Neste sentido, o estudo foi direccionado para áreas (A1 e A2) integradas numa zona agrícola extensa em Portugal, utilizada para a produção de milho e, principalmente, de arroz (Baixo Mondego), a qual sustenta uma elevada biodiversidade. O estudo teve início na área A1, onde a monitorização físico-química e os ensaios com amostras naturais (ensaios WET - whole effluent tests) provenientes desta área evidenciaram que, apesar da ausência de pesticidas, as amostras de água colhidas no canal que atravessava os arrozais foram as mais nocivas para o crescimento de Pseudokirchneriella subcapitata e Chlorella vulgaris. Uma vez que outras fontes de contaminação (produção de gado) actuavam em A1, o estudo prosseguiu apenas na área A2. Assim, em A2, começou-se por determinar a toxicidade individual e da mistura de dois herbicidas formulados aplicados nos campos de arroz (Viper®) e milho (Mikado®) em condições laboratoriais. Viper® foi o herbicida mais tóxico, tanto para o crescimento de P. subcapitata e C. vulgaris, como para a sobrevivência, reprodução e crescimento de Daphnia longispina e Daphnia magna. Adicionalmente, estimou-se que a mistura Viper®/Mikado® induz efeitos antagonistas no crescimento de P. subcapitata e efeitos sinérgicos no crescimento de C. vulgaris e na sobrevivência dos dafnídeos. A avaliação da toxicidade destes herbicidas formulados e seus ingredientes activos no comportamento de minhocas terrestres (Eisenia andrei), usando solos naturais, demonstrou que Viper® e penoxsulam causaram uma % de evitamento superior nos organismos expostos. Contudo, o risco para E. andrei será à partida reduzido se as taxas de aplicação dos herbicidas forem respeitadas. Ensaios WET foram novamente usados para testar amostras naturais da área A2. Verificou-se que a qualidade do sistema aquático e do arrozal diminuiu durante a estação agrícola, em paralelo com a presença de nutrientes e pesticidas. O crescimento algal foi inibido, apesar dos parâmetros de história de vida dos dafnídeos terem sido estimulados. O resultado desta avaliação subestimou, em certos casos, os impactos reais causados pela aplicação de pesticidas. A avaliação in situ simultânea à aplicação de herbicidas nos arrozais demonstrou que os efeitos registados foram de facto restritos aos pulsos de herbicidas. A inibição das taxas de alimentação de D. longispina e D. magna forneceram um sinal precoce de alterações no sistema, seguido pela diminuição da sua sobrevivência e do crescimento de P. subcapitata. Em suma, as diferentes fases da avaliação efectuada confirmaram a existência de condições desfavoráveis devido às práticas agrícolas, reforçando a necessidade de se conjugar ensaios laboratoriais com avaliações in situ de maior relevância ecológica, para reduzir o grau de incerteza aliado à determinação dos riscos.

Multiple predator effects and native prey responses to two non-native Everglades cichlids

Non-native predators may have negative impacts on native communities, and these effects may be dependent on interactions among multiple non-native predators. Sequential invasions by predators can enhance risk for native prey. Prey have a limited ability to respond to multiple threats since appropriate responses may conflict, and interactions with recent invaders may be novel. We examined predator–prey interactions among two non-native predators, a recent invader, the African jewelfish, and the longer-established Mayan cichlid, and a native Florida Everglades prey assemblage. Using field enclosures and laboratory aquaria, we compared predatory effects and antipredator responses across five prey taxa. Total predation rates were higher for Mayan cichlids, which also targeted more prey types. The cichlid invaders had similar microhabitat use, but varied in foraging styles, with African jewelfish being more active. The three prey species that experienced predation were those that overlapped in habitat use with predators. Flagfish were consumed by both predators, while riverine grass shrimp and bluefin killifish were eaten only by Mayan cichlids. In mixed predator treatments, we saw no evidence of emergent effects, since interactions between the two cichlid predators were low. Prey responded to predator threats by altering activity but not vertical distribution. Results suggest that prey vulnerability is affected by activity and habitat domain overlap with predators and may be lower to newly invading predators, perhaps due to novelty in the interaction.

Evoking non-repertory verbal behavior across operant classes : the effects of motor echoic sign language training within the context of a motivating operation

The individual effects that echoic, mand, and sign language training procedures have on the acquisition of verbal behavior have been widely demonstrated, but more efficient strategies are still needed. This study combined all three treatment strategies into one treatment intervention in order to investigate the joint effects they may have on verbal behavior. Six participants took part in the study. Intervention totaled 1 hour/day for 5 days/week until mastery criterion for motor echoic behavior was achieved. Although motor echoic behavior were solely targeted for acquisition, significant increases in spontaneous motor mands were noted in all treatment participants. Additionally, 4 treatment participants also demonstrated significant gains in vocal echoics and spontaneous vocal mands. No significant increases were noted for the control participant. Results suggest that the aforementioned procedure may provide more efficient results as a first-step to teaching a functional repertoire of verbal behavior to developmentally delayed children.

Effects of hook and bait on targeted and bycatch fishes in an equatorial atlantic pelagic longline fishery

We examined the effects of different hook style and bait type combinations on the catches of targeted, bycatch, and discarded fishes in equatorial Atlantic waters. In total, 221 longline sets (>305,000 hooks) were deployed from Portuguese pelagic longline vessels (SELECT -PAL Project) during the February–October fishing season. Three different hook styles and two bait types were tested: the traditional J-hook was compared to two circle hooks (one non-offset and one with 10° offset), and squid bait was compared to mackerel. Catch per unit effort (CPU Es) were calculated and compared between the different hook style and bait type combinations, which indicated that the effects of hook style and bait on the CPU Es were species-specific. For example, swordfish CPU Es were higher with J-hooks baited with squid, while for targeted tunas and blue shark only the bait effect was significant, but with opposite effect (i.e., higher catches of tuna with squid bait and higher catches of blue shark with mackerel bait). For the discarded species, at-haulback mortality was also species-specific. Proportions of alive vs dead specimens at time of fishing gear retrieval did not vary significantly by hook style or bait type combinations. The total retained catch was analyzed in value per unit effort (VPU E), and indicated losses in fishery revenue when mackerel was used instead of squid, but not when circle hooks were used instead of J-hooks.

Resisting contradictions : non-Indigenous pre-service teacher responses to critical Indigenous studies

The study examines non-Indigenous pre-service teacher responses to the authorisation of Indigenous knowledge perspectives in compulsory Indigenous studies with a primary focus on exploring the nature and effects of resistance. It draws on the philosophies of the Japanangka teaching and research paradigm (West, 2000), relationship theory (Graham, 1999), Indigenist methodologies and decolonisation approaches to examine this resistance. A Critical Indigenist Study was employed to investigate how non-Indigenous pre-service teachers managed their learning, and how they articulated shifts in resistance as they progressed through their studies. This study explains resistance to compulsory Indigenous and how it can be targeted by Indigenist Standpoint Pedagogy. The beginning transformations in pre-service teacher positioning in relation to Australian history, contemporary educational practice, and professional identity was also explored.

IL-23 is pro-proliferative, epigenetically regulated and modulated by chemotherapy in non-small cell lung cancer

Background IL-23 is a member of the IL-6 super-family and plays key roles in cancer. Very little is currently known about the role of IL-23 in non-small cell lung cancer (NSCLC). Methods RT-PCR and chromatin immunopreciptiation (ChIP) were used to examine the levels, epigenetic regulation and effects of various drugs (DNA methyltransferase inhibitors, Histone Deacetylase inhibitors and Gemcitabine) on IL-23 expression in NSCLC cells and macrophages. The effects of recombinant IL-23 protein on cellular proliferation were examined by MTT assay. Statistical analysis consisted of Student's t-test or one way analysis of variance (ANOVA) where groups in the experiment were three or more. Results In a cohort of primary non-small cell lung cancer (NSCLC) tumours, IL-23A expression was significantly elevated in patient tumour samples (p<0.05). IL-23A expression is epigenetically regulated through histone post-translational modifications and DNA CpG methylation. Gemcitabine, a chemotherapy drug indicated for first-line treatment of NSCLC also induced IL-23A expression. Recombinant IL-23 significantly increased cellular proliferation in NSCLC cell lines. Conclusions These results may therefore have important implications for treating NSCLC patients with either epigenetic targeted therapies or Gemcitabine. © 2012 Elsevier Ireland Ltd.

The beneficial effects of specialist thoracic surgery on the resection rate for non-small-cell lung cancer

We aimed to evaluate the effect of the appointment of a dedicated specialist thoracic surgeon on surgical practice for lung cancer previously served by cardio-thoracic surgeons. Outcomes were compared for the 240 patients undergoing surgical resection for lung cancer in two distinct 3-year periods: Group A: 65 patients, 1994-1996 (pre-specialist); Group B: 175 patients, 1997-1999 (post-specialist). The changes implemented resulted in a significant increase in resection rate (from 12.2 to 23.4%, P<0.001), operations in the elderly (over 75 years) and extended resections. There were no significant differences in stage distribution, in-hospital mortality or stage-specific survival after surgery. Lung cancer surgery provided by specialists within a multidisciplinary team resulted in increased surgical resection rates without compromising outcome. Our results strengthen the case for disease-specific specialists in the treatment of lung cancer. © 2004 Published by Elsevier Ireland Ltd.

Procaspase 8 overexpression in non-small-cell lung cancer promotes apoptosis induced by FLIP silencing

We found that procaspase 8 was overexpressed in non-small-cell lung cancers (NSCLCs) compared with matched normal tissues. The caspase 8 inhibitor FLICE-inhibitory protein (FLIP) was also overexpressed in the majority of NSCLCs. Silencing FLIP induced caspase 8 activation and apoptosis in NSCLC cell lines, but not in normal lung cell lines. Apoptosis induced by FLIP silencing was mediated by the TRAIL death receptors DR4 and DR5, but was not dependent on ligation of the receptors by TRAIL. Furthermore, the apoptosis induced by FLIP silencing was dependent on the overexpression of procaspase 8 in NSCLC cells. Moreover, in NSCLC cells, but not in normal cells, FLIP silencing induced co-localization of DR5 and ceramide, and disruption of this co-localization abrogated apoptosis. FLIP silencing supra-additively increased TRAIL-induced apoptosis of NSCLC cells; however, normal lung cells were resistant to TRAIL, even when FLIP was silenced. Importantly, FLIP silencing sensitized NSCLC cells but not normal cells to chemotherapy in vitro, and silencing FLIP in vivo retarded NSCLC xenograft growth and enhanced the anti-tumour effects of cisplatin. Collectively, our results suggest that due to frequent procaspase 8 overexpression, NSCLCs may be particularly sensitive to FLIP-targeted therapies.

Targeted therapies in non-small cell lung cancer

The majority of non-small cell lung cancer (NSCLC) patients present with advanced disease and with a 5 year survival rate of <15% for these patients, treatment outcomes are considered extremely disappointing. Standard chemotherapy regimens provide some improvement to ~40% of patients. However, intrinsic and acquired chemoresistance are a significant problem and hinder sustained long term benefits of such treatments. Advances in proteomic and genomic profiling have increased our understanding of the aberrant molecular mechanisms that are driving an individual's tumour. The increased sensitivity of these technologies has enabled molecular profiling at the stage of initial biopsy thus paving the way for a more personalised approach to the treatment of cancer patients. Improvements in diagnostics together with a wave of new targeted small molecule inhibitors and monoclonal antibodies have revolutionised the treatment of cancer. To date there are essentially three targeted agents approved for clinical use in NSCLC. The tyrosine kinase inhibitor (TKI) erlotinib, which targets the epidermal growth factor receptor (EGFR) TK domain, has proven to be an effective treatment strategy in patients who harbour activating mutations in the EGFR TK domain. Bevacizumab a monoclonal antibody targeting the vascular endothelial growth factor (VEGF) can improve survival, response rates, and progression-free survival when used in combination with chemotherapy. Crizotinib, a small-molecule drug, inhibits the tyrosine kinase activity of the echinoderm microtubule-associated protein-like 4 anaplastic lymphoma kinase (EML4-ALK) fusion protein, resulting in decreased tumour cell growth, migration, and invasiveness in patients with locally advanced or metastatic NSCLC. The clinical relevance of several other targeted agents are under investigation in distinct molecular subsets of patients with key "driver" mutations including: KRAS, HER2, BRAF, MET, PIK3CA, AKT1,MAP2K1, ROS1 and RET. Often several pathways are activated simultaneously and crosstalk between pathways allows tumour cells to escape the inhibition of a single targeted agent. This chapter will explore the clinical development of currently available targeted therapies for NSCLC as well as those in clinical trials and will examine the synergy between cytotoxic therapies.