Functional Profiling of Antibody Immune Repertoires in Convalescent Zika Virus Disease Patients

The re-emergence of Zika virus (ZIKV) caused widespread infections that were linked to Guillain-Barré syndrome in adults and congenital malformation in fetuses, and epidemiological data suggest that ZIKV infection can induce protective antibody responses. A more detailed understanding of anti-ZIKV antibody responses may lead to enhanced antibody discovery and improved vaccine designs against ZIKV and related flaviviruses. Here, we applied recently-invented library-scale antibody screening technologies to determine comprehensive functional molecular and genetic profiles of naturally elicited human anti-ZIKV antibodies in three convalescent individuals. We leveraged natively paired antibody yeast display and NGS to predict antibody cross-reactivities and coarse-grain antibody affinities, to perform in-depth immune profiling of IgM, IgG, and IgA antibody repertoires in peripheral blood, and to reveal virus maturation state-dependent antibody interactions. Repertoire-scale comparison of ZIKV VLP-specific and non-specific antibodies in the same individuals also showed that mean antibody somatic hypermutation levels were substantially influenced by donor-intrinsic characteristics. These data provide insights into antiviral antibody responses to ZIKV disease and outline systems-level strategies to track human antibody immune responses to emergent viral infections

IMPACTO DA “MÃE DE LEITE ELETRÔNICA” NO DESEMPENHO ZOOTÉCNICO DE LEITÕES LACTENTES

Com os avanços da genética na suinocultura e a aplicação de tecnologias no manejo o número de leitões nascidos, por parto, tem aumentado significativamente. Por consequência, as leitegadas raramente são uniformes e muitos leitões apresentam baixo peso ao nascimento. Pesquisas indicam que a cada leitão extra na média de nascidos acarreta uma redução de 100g no peso ao nascimento, dobrando o número de leitões que nascem com peso abaixo de 800g. O aumento da taxa de mortalidades entre os leitões com baixo peso, ainda nos primeiros dias de vida, é algo comum. Logo, torna-se essencial o emprego de alternativas que contribuam tanto para a diminuição da mortalidade pré-desmame quanto para o alcance da uniformidade da leitegada. Com o intuito de melhorar esses índices foi avaliado a influência do equipamento “mãe de leite eletrônica” sobre o desempenho zootécnico de leitões no período entre o nascimento e o desmame. O equipamento foi cedido por colaboradores da Empresa STA, de Joinville/SC. Cada lote experimental contou com dois grupos: o controle (GC) e o tratamento (GT). No GC os leitões receberam apenas aleitamento natural, fornecido pela mãe. No GT os leitões foram submetidos ao regime de aleitamento misto (natural, fornecido pela mãe; e o artificial, fornecido pelo equipamento o leite de vaca). Foram avaliadas 6 leitegadas, duas por experimento. As fêmeas eram provenientes de matrizes híbridas Landrace x Large White, entre o 3º e 6º parto. Ao todo realizaram-se 3 procedimentos experimentais. O aleitamento propiciado pelo equipamento beneficiou todas as leitegadas do GT já nos três primeiros dias de vida. Muitos leitões que apresentaram dificuldades de amamentarem-seadequadamente na mãe, pela competição da leitegada pelos tetos, conseguiram sobreviver pela disponibilidade de leite oferecida artificialmente. Por ocasião do desmame, o peso médio e o número de leitões foram similares numericamente entre as 6 leitegadas avaliadas durante o período experimental. No entanto, no GT observou-se maior desuniformidade entre os leitões, uma vez que alguns animais ganharam peso mais rápido pelo acesso também ao leite artificial. Apesar de auxiliar na sobrevivência dos leitões, evitando maiores taxas de mortalidade pré-desmame, principalmente nos primeiros dias após o parto, não houve contribuição do equipamento sobre a uniformidade dos lotes. Concluímos que a “mãe de leite eletrônica” possibilitou não só o interesse como também a ingestão de leite artificial pelos leitões, de modo com que mais tetos ficassem disponíveis para outros leitões durante a mamada. Evidenciamos também que tanto leitões refugos como os mais fracos se beneficiaram do aleitamento misto, mantendo seu desenvolvimento de forma adequada até o desmame. Por fim, a equipe indica a utilização do equipamento, no mínimo, durante os 14 primeiros dias de vida dos leitões

In Vitro and In Vivo Investigation of the Efficacy of Arylimidamide DB1831 and Its Mesylated Salt Form - DB1965 - against Trypanosoma cruzi Infection

Chagas disease is caused by infection with the intracellular protozoan parasite Trypanosoma cruzi. At present, nifurtimox and benznidazole, both compounds developed empirically over four decades ago, represent the chemotherapeutic arsenal for treating this highly neglected disease. However, both drugs present variable efficacy depending on the geographical area and the occurrence of natural resistance, and are poorly effective against the later chronic stage. As a part of a search for new therapeutic opportunities to treat chagasic patients, pre-clinical studies were performed to characterize the activity of a novel arylimidamide (AIA - DB1831 (hydrochloride salt) and DB1965 (mesylate salt)) against T.cruzi. These AIAs displayed a high trypanocidal effect in vitro against both relevant forms in mammalian hosts, exhibiting a high selectivity index and a very high efficacy (IC50 value/48 h of 5–40 nM) against intracellular parasites. DB1965 shows high activity in vivo in acute experimental models (mouse) of T.cruzi, showing a similar effect to benznidazole (Bz) when compared under a scheme of 10 daily consecutive doses with 12.5 mg/kg. Although no parasitological cure was observed after treating with 20 daily consecutive doses, a combined dosage of DB1965 (5 mg/kg) with Bz (50 mg/kg) resulted in parasitaemia clearance and 100% animal survival. In summary, our present data confirmed that aryimidamides represent promising new chemical entities against T.cruzi in therapeutic schemes using the AIA alone or in combination with other drugs, like benznidazole

Setting research priorities to improve global newborn health and prevent stillbirths by 2025

Acknowledgements: The authors thank the expert group for their time and effort in this priority–setting exercise, and the contribution of anonymous reviewers to the final version of this report. We acknowledge inputs from Dr Diane Morof from Centers for Disease Control and Prevention, USA. Funding: This work was supported by Save the Children.Peer reviewedPublisher PD

Genomics and proteomics approaches to the study of cancer-stroma interactions

<p>Abstract</p> <p>Background</p> <p>The development and progression of cancer depend on its genetic characteristics as well as on the interactions with its microenvironment. Understanding these interactions may contribute to diagnostic and prognostic evaluations and to the development of new cancer therapies. Aiming to investigate potential mechanisms by which the tumor microenvironment might contribute to a cancer phenotype, we evaluated soluble paracrine factors produced by stromal and neoplastic cells which may influence proliferation and gene and protein expression.</p> <p>Methods</p> <p>The study was carried out on the epithelial cancer cell line (Hep-2) and fibroblasts isolated from a primary oral cancer. We combined a conditioned-medium technique with subtraction hybridization approach, quantitative PCR and proteomics, in order to evaluate gene and protein expression influenced by soluble paracrine factors produced by stromal and neoplastic cells.</p> <p>Results</p> <p>We observed that conditioned medium from fibroblast cultures (FCM) inhibited proliferation and induced apoptosis in Hep-2 cells. In neoplastic cells, 41 genes and 5 proteins exhibited changes in expression levels in response to FCM and, in fibroblasts, 17 genes and 2 proteins showed down-regulation in response to conditioned medium from Hep-2 cells (HCM). Nine genes were selected and the expression results of 6 down-regulated genes (<it>ARID4A</it>, <it>CALR</it>, <it>GNB2L1</it>, <it>RNF10</it>, <it>SQSTM1</it>, <it>USP9X</it>) were validated by real time PCR.</p> <p>Conclusions</p> <p>A significant and common denominator in the results was the potential induction of signaling changes associated with immune or inflammatory response in the absence of a specific protein.</p

Setting research priorities to improve global newborn health and prevent stillbirths by 2025.

BACKGROUND: In 2013, an estimated 2.8 million newborns died and 2.7 million were stillborn. A much greater number suffer from long term impairment associated with preterm birth, intrauterine growth restriction, congenital anomalies, and perinatal or infectious causes. With the approaching deadline for the achievement of the Millennium Development Goals (MDGs) in 2015, there was a need to set the new research priorities on newborns and stillbirth with a focus not only on survival but also on health, growth and development. We therefore carried out a systematic exercise to set newborn health research priorities for 2013-2025. METHODS: We used adapted Child Health and Nutrition Research Initiative (CHNRI) methods for this prioritization exercise. We identified and approached the 200 most productive researchers and 400 program experts, and 132 of them submitted research questions online. These were collated into a set of 205 research questions, sent for scoring to the 600 identified experts, and were assessed and scored by 91 experts. RESULTS: Nine out of top ten identified priorities were in the domain of research on improving delivery of known interventions, with simplified neonatal resuscitation program and clinical algorithms and improved skills of community health workers leading the list. The top 10 priorities in the domain of development were led by ideas on improved Kangaroo Mother Care at community level, how to improve the accuracy of diagnosis by community health workers, and perinatal audits. The 10 leading priorities for discovery research focused on stable surfactant with novel modes of administration for preterm babies, ability to diagnose fetal distress and novel tocolytic agents to delay or stop preterm labour. CONCLUSION: These findings will assist both donors and researchers in supporting and conducting research to close the knowledge gaps for reducing neonatal mortality, morbidity and long term impairment. WHO, SNL and other partners will work to generate interest among key national stakeholders, governments, NGOs, and research institutes in these priorities, while encouraging research funders to support them. We will track research funding, relevant requests for proposals and trial registers to monitor if the priorities identified by this exercise are being addressed