RNA extraction from self-assembling peptide hydrogels to allow qPCR analysis of encapsulated cells

Self-assembling peptide hydrogels offer a novel 3-dimensional platform for many applications in cell culture and tissue engineering but are not compatible with current methods of RNA isolation; owing to interactions between RNA and the biomaterial. This study investigates the use of two techniques based on two different basic extraction principles: solution-based extraction and direct solid-state binding of RNA respectively, to extract RNA from cells encapsulated in four β-sheet forming self-assembling peptide hydrogels with varying net positive charge. RNA-peptide fibril interactions, rather than RNA-peptide molecular complexing, were found to interfere with the extraction process resulting in low yields. A column-based approach relying on RNA-specific binding was shown to be more suited to extracting RNA with higher purity from these peptide hydrogels owing to its reliance on strong specific RNA binding interactions which compete directly with RNA-peptide fibril interactions. In order to reduce the amount of fibrils present and improve RNA yields a broad spectrum enzyme solution—pronase—was used to partially digest the hydrogels before RNA extraction. This pre-treatment was shown to significantly increase the yield of RNA extracted, allowing downstream RT-qPCR to be performed

TRESK channel contribution to nociceptive sensory neurons excitability: modulation by nerve injury

<p>Abstract</p> <p>Background</p> <p>Neuronal hyperexcitability is a crucial phenomenon underlying spontaneous and evoked pain. In invertebrate nociceptors, the S-type leak K⁺channel (analogous to TREK-1 in mammals) plays a critical role of in determining neuronal excitability following nerve injury. Few data are available on the role of leak K_2Pchannels after peripheral axotomy in mammals.</p> <p>Results</p> <p>Here we describe that rat sciatic nerve axotomy induces hyperexcitability of L4-L5 DRG sensory neurons and decreases TRESK (K2P18.1) expression, a channel with a major contribution to total leak current in DRGs. While the expression of other channels from the same family did not significantly change, injury markers ATF3 and Cacna2d1 were highly upregulated. Similarly, acute sensory neuron dissociation (<it>in vitro </it>axotomy) produced marked hyperexcitability and similar total background currents compared with neurons injured <it>in vivo</it>. In addition, the sanshool derivative IBA, which blocked TRESK currents in transfected HEK293 cells and DRGs, increased intracellular calcium in 49% of DRG neurons in culture. Most IBA-responding neurons (71%) also responded to the TRPV1 agonist capsaicin, indicating that they were nociceptors. Additional evidence of a biological role of TRESK channels was provided by behavioral evidence of pain (flinching and licking), in vivo electrophysiological evidence of C-nociceptor activation following IBA injection in the rat hindpaw, and increased sensitivity to painful pressure after TRESK knockdown in vivo.</p> <p>Conclusions</p> <p>In summary, our results clearly support an important role of TRESK channels in determining neuronal excitability in specific DRG neurons subpopulations, and show that axonal injury down-regulates TRESK channels, therefore contributing to neuronal hyperexcitability.</p

I progetti gestiti dai Coordinatori: analisi del loro profilo e successo

Obiettivo. Descrivere la storia dei progetti affidati/ gestiti dai Coordinatori infermieristici ospedalieri. Metodi. \uc8 stato incluso un campione di 56 Coordinatori in ruolo da almeno un anno nei reparti di 13 Ospedali del nord Italia, contattati con criterio di convenienza. Tramite un\u2019intervista strutturata sono stati raccolti dati sui progetti gestiti nel 2009, tipologia, origine (bottom up; top down), il numero di operatori coinvolti e stato del progetto al momento dell\u2019intervista (concluso, incompleto, abbandonato). Risultati. Nel 2009 i Coordinatori hanno gestito 114 progetti, in media 1.8/ciascuno (\ub11.2): 94 (82.5%) erano progetti di miglioramento, 17 (14.9%) di accreditamento, e 3 (2.6%) di ricerca. I progetti avevano coinvolto complessivamente 2.732 persone (73.7% dei team) con un impegno medio di 84 ore ciascuno; 55 (48.2%) progetti erano ancora in corso, 52 (45.6%) conclusi, 5 (4.4%) incompleti (ovvero mancavano di valutazione) mentre 2 (1.8%) erano stati abbandonati. Conclusioni. Gli infermieri sono coinvolti in numerosi progetti nelle aziende sanitarie. La fase pi\uf9 trascurata \ue8 il monitoraggio dei risultati e il loro consolidamento: i progetti assorbono molte risorse e per questo \ue8 fondamentale che siano correttamente gestiti e partano da reali problemi ed esigenze dei pazienti

Demanda mediana por serviços públicos e desempenho eleitoral: evidências do modelo do eleitor mediano para os municípios brasileiros

O objetivo deste trabalho é testar empiricamente a validade do modelo do eleitor mediano para o caso brasileiro. O teste foi viabilizado pela possibilidade de reeleição, estabelecida em 1997 por meio da Emenda da Reeleição. Com informações municipais do Censo Populacional e do TSE para o ano de 2000, identificaram-se as cestas ofertadas por candidatos à reeleição como os gastos efetuados durante seus primeiros mandatos. Em seguida, estimou-se a demanda por serviços públicos locais. Relaxando a hipótese de informação perfeita dos candidatos em relação à demanda mediana, e impondo uma hipótese menos forte, de expectativas racionais, justificou-se a estimação de uma medida de erro de percepção dos candidatos à reeleição em relação à demanda mediana. Em seguida, estimou-se o impacto do módulo dessa medida sobre o desempenho eleitoral dos candidatos à reeleição usando modelos de seleção. O resultado obtido sugere que o modelo do eleitor mediano é válido para os municípios brasileiros

Distribution map of Ambrosia artemisiifolia L. (Asteraceae) in Italy

The spread of the invasive and allergenic Ambrosia artemisiifolia L. in Italy was analysed and mapped using distribution data from a wide range of sources. Ambrosia artemisiifolia occupies 1057 floristic quadrants which are mostly distributed in the Po plain. The distribution obtained represents the basis to implement urgent management strategies

Trust in scientists and their role in society across 68 countries

Science is crucial for evidence-based decision-making. Public trust in scientists can help decision makers act on the basis of the best available evidence, especially during crises. However, in recent years the epistemic authority of science has been challenged, causing concerns about low public trust in scientists. We interrogated these concerns with a preregistered 68-country survey of 71,922 respondents and found that in most countries, most people trust scientists and agree that scientists should engage more in society and policymaking. We found variations between and within countries, which we explain with individual- and country-level variables, including political orientation. While there is no widespread lack of trust in scientists, we cannot discount the concern that lack of trust in scientists by even a small minority may affect considerations of scientific evidence in policymaking. These findings have implications for scientists and policymakers seeking to maintain and increase trust in scientists

Perceptions of science, science communication, and climate change attitudes in 68 countries – the TISP dataset

Science is integral to society because it can inform individual, government, corporate, and civil society decision-making on issues such as public health, new technologies or climate change. Yet, public distrust and populist sentiment challenge the relationship between science and society. To help researchers analyse the science-society nexus across different geographical and cultural contexts, we undertook a cross-sectional population survey resulting in a dataset of 71,922 participants in 68 countries. The data were collected between November 2022 and August 2023 as part of the global Many Labs study “Trust in Science and Science-Related Populism” (TISP). The questionnaire contained comprehensive measures for individuals’ trust in scientists, science-related populist attitudes, perceptions of the role of science in society, science media use and communication behaviour, attitudes to climate change and support for environmental policies, personality traits, political and religious views and demographic characteristics. Here, we describe the dataset, survey materials and psychometric properties of key variables. We encourage researchers to use this unique dataset for global comparative analyses on public perceptions of science and its role in society and policy-making