Sialome of a Generalist Lepidopteran Herbivore: Identification of Transcripts and Proteins from Helicoverpa armigera Labial Salivary Glands

Although the importance of insect saliva in insect-host plant interactions has been acknowledged, there is very limited information on the nature and complexity of the salivary proteome in lepidopteran herbivores. We inspected the labial salivary transcriptome and proteome of Helicoverpa armigera, an important polyphagous pest species. To identify the majority of the salivary proteins we have randomly sequenced 19,389 expressed sequence tags (ESTs) from a normalized cDNA library of salivary glands. In parallel, a non-cytosolic enriched protein fraction was obtained from labial salivary glands and subjected to two-dimensional gel electrophoresis (2-DE) and de novo peptide sequencing. This procedure allowed comparison of peptides and EST sequences and enabled us to identify 65 protein spots from the secreted labial saliva 2DE proteome. The mass spectrometry analysis revealed ecdysone, glucose oxidase, fructosidase, carboxyl/cholinesterase and an uncharacterized protein previously detected in H. armigera midgut proteome. Consistently, their corresponding transcripts are among the most abundant in our cDNA library. We did find redundancy of sequence identification of saliva-secreted proteins suggesting multiple isoforms. As expected, we found several enzymes responsible for digestion and plant offense. In addition, we identified non-digestive proteins such as an arginine kinase and abundant proteins of unknown function. This identification of secreted salivary gland proteins allows a more comprehensive understanding of insect feeding and poses new challenges for the elucidation of protein function

Ready for new waves: optimizing SARS-CoV-2 variants monitoring in pooled samples with droplet digital PCR

IntroductionThe declaration of the end of the Public Health Emergency for COVID-19 on May 11th, 2023, has shifted the global focus led by WHO and CDC towards monitoring the evolution of SARS-CoV-2. Augmenting these international endeavors with local initiatives becomes crucial to not only track the emergence of new variants but also to understand their spread. We present a cost-effective digital PCR-based pooled sample testing methodology tailored for early variant surveillance.MethodsUsing 1200 retrospective SARS-CoV-2 positive samples, either negative or positive for Delta or Omicron, we assessed the sensitivity and specificity of our detection strategy employing commercial TaqMan variant probes in a 1:9 ratio of variant-positive to variant-negative samples.ResultsThe study achieved 100% sensitivity and 99% specificity in 10-sample pools, with an Area Under the Curve (AUC) exceeding 0.998 in ROC curves, using distinct commercial TaqMan variant probes.DiscussionThe employment of two separate TaqMan probes for both Delta and Omicron establishes dual validation routes, emphasizing the method’s robustness. Although we used known samples to model realistic emergence scenarios of the Delta and Omicron variants, our main objective is to demonstrate the versatility of this strategy to identify future variant appearances. The utilization of two divergent variants and distinct probes for each confirms the method’s independence from specific variants and probes. This flexibility ensures it can be tailored to recognize any subsequent variant emergence, given the availability of its sequence and a specific probe. Consequently, our approach stands as a robust tool for tracking and managing any new variant outbreak, reinforcing our global readiness against possible future SARS-CoV-2 waves

REFLEXO(ÃO)

O cenário atual tem mostrado grande número de pessoas que, após a pandemia, passaram a apresentar problemas psicológicos. Considerando esse elemento, nosso projeto tem como tema principal o "espelho” como um instrumento de reflexão. Nosso objetivo principal é proporcionar ao participante um momento pessoa para com pessoa, onde poderá refletir diante ao espelho, tendo um maior entendimento de quem ela é naquele instante. Importante destacar que em momentos de reflexão é possível perceber que em meio a todo caos existem soluções, que todo labirinto tem uma saída. E que alguns momentos e técnicas podem ajudar as pessoas a entender que para todo problema tem uma solução e com algum esforço é possível conseguir sair de situações difíceis. Um dos métodos bastante utilizados vem da psicanálise, que consiste em fazer com que o/a paciente possa conectar com sua própria mente e assim entender a si mesma/o e poder arrumar soluções para seus problemas. A metodologia adotada será uma dinâmica na qual o espelho será utilizado como meio para podermos realizar o trabalho, onde dentro da cabine feita com panos escuros e uma luminária estará apoiado sobre um banco e a parede, e ali estará sendo reproduzido um áudio no qual será gravado pelos próprios alunos participantes do projeto, citarão pequenos trechos da obra "O Espelho - João Guimarães Rosa", em meio a esse momento a pessoa que estará participando da atividade terá a oportunidade de ter uma maior conexão com seu eu interior. Temos como propósito final o intuito direcionado ao interagente, em que ele possa aproveitar aquele momento consigo mesmo, uma parada no dia para destinar um tempo ao autoconhecimento e consequente dimensionamento dos problemas e situações vivenciados. Além disso podemos considerar a satisfação individual dos próprios integrantes do grupo em estar proporcionando um instante de aprendizado, conhecimento e ponderação de quem estiver participando

RNA interference in Lepidoptera: an overview of successful and unsuccessful studies and implications for experimental design

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Identifying the need for specialized palliative care in adult cancer patients – development and validation of a screening procedure based on proxy assessment by physicians and filter questions

Background One challenge in caring for cancer patients with incurable disease is the adequate identification of those in need for specialized palliative care (SPC). The study’s aim was to validate an easy to use phenomenological screening tool. Methods The German tool is based on the National Comprehensive Cancer Network (NCCN) Palliative Care guidelines and contains ten items in five domains that focus e.g. on diagnosis, functional status, complications, comorbidities, and palliative care relevant problems such as symptom management, distress, and support of family and team members. Sum score ranges from 0 to 14 (no need to great need). Assessment to identify SPC needs was done in university hospital wards between 1 and 08/2017 by health care professionals on admission of the patient if the disease was incurable and expected prognosis < 12 months. The Integrated Palliative Outcome Scale (IPOS, staff version), an outcome assessment instrument for palliative care that consists of ten items, served as external criterion; in sub samples inter-rater/test-retest were performed. Results Data from 208 patients with incurable disease and life expectancy < 12 months (54.8% female; average age 63.5 years, range 21–96) were assessed using the tool. The tool has good convergent validity; the correlation between the sum scores of IPOS and our tool showed a significant and substantial effect. The sum score was independent of the patient’s age, gender and primary diagnosis. Patients who already were in contact with SPC had significantly higher screening scores than patients without. With a cut point of  ≥ 5, 80.8% of the screened patients were in need for SPC. Cronbach’s alpha was α = .600. Rater agreement (inter-rater, test-retest) varied between single items. Correlation coefficients showed significant substantial effects. Conclusions This is the first validation of a screening procedure in German language identifying SPC needs of adult patients with advanced cancer and the first using filter questions as a pre-screening. Proxy assessment of SPC needs by physicians in cancer care settings is feasible and the suggested tool presents a valid instrument to trigger a PC consultation. Trial registration The study was not registered

Geographical Distribution and Genetic Diversity of Bank Vole Hepaciviruses in Europe

The development of new diagnostic methods resulted in the discovery of novel hepaciviruses in wild populations of the bank vole (Myodes glareolus, syn. Clethrionomys glareolus). The naturally infected voles demonstrate signs of hepatitis similar to those induced by hepatitis C virus (HCV) in humans. The aim of the present research was to investigate the geographical distribution of bank vole-associated hepaciviruses (BvHVs) and their genetic diversity in Europe. Real-time reverse transcription polymerase chain reaction (RT-qPCR) screening revealed BvHV RNA in 442 out of 1838 (24.0%) bank voles from nine European countries and in one of seven northern red-backed voles (Myodes rutilus, syn. Clethrionomys rutilus). BvHV RNA was not found in any other small mammal species (n = 23) tested here. Phylogenetic and isolation-by-distance analyses confirmed the occurrence of both BvHV species (Hepacivirus F and Hepacivirus J) and their sympatric occurrence at several trapping sites in two countries. The broad geographical distribution of BvHVs across Europe was associated with their presence in bank voles of different evolutionary lineages. The extensive geographical distribution and high levels of genetic diversity of BvHVs, as well as the high population fluctuations of bank voles and occasional commensalism in some parts of Europe warrant future studies on the zoonotic potential of BvHVs.Peer reviewe

UTILIZAÇÃO DE COLEÇÕES ENTOMOLÓGICAS NO IFC-CAMPUS CAMBORIÚ: Uma proposta didática

As aulas práticas são essenciais na aprendizagem na área de entomologia, pois aboa formação dos estudantes passa por experiências que transcendem o campoteórico e despertam a curiosidade. Na disciplina de Defesa Sanitária Vegetal doCurso Técnico em Agropecuária do IFC-Camboriú os alunos dos segundos anos, sãoestimulados a elaborar coleções didáticas entomológicas. O objetivo principal éproporcionar uma vivência prática dos conhecimentos repassados em sala, dentreeles, reconhecimento do habitat, ciclo biológico, comportamento, além das principaisordens e famílias, causadoras de danos irreparáveis às culturas agrícolas. Todas ascoleções foram incorporadas no Museu Entomológico