Stealth monoolein-based nanocarriers for delivery of siRNA to cancer cells

While the delivery of small interfering RNAs (siRNAs) is an attractive strategy to treat several clinical con- ditions, siRNA-nanocarriers stability after intravenous administration is still a major obstacle for the development of RNA-interference based therapies. But, although the need for stability is well recognized, the notion that strong stabilization can decrease nanocarriers efficiency is sometimes neglected. In this work we evaluated two stealth functionalization strategies to stabilize the previously validated dioctade- cyldimethylammonium bromide (DODAB):monoolein (MO) siRNA-lipoplexes. The nanocarriers were pre- and post-pegylated, forming vectors with different stabilities in biological fluids. The stealth nanocarriers behavior was tested under biological mimetic conditions, as the production of stable siRNA-lipoplexes is determinant to achieve efficient intravenous siRNA delivery to cancer cells. Upon incubation in human serum for 2 h, by fluorescence Single Particle Tracking microscopy, PEG-coated lipo- plexes were found to have better colloidal stability as they could maintain a relatively stable size. In addi- tion, using fluorescence fluctuation spectroscopy, post-pegylation also proved to avoid siRNA dissociation from the nanocarriers in human serum. Concomitantly it was found that PEG-coated lipoplexes improved cellular uptake and transfection efficiency in H1299 cells, and had the ability to silence BCR-ABL, affecting the survival of K562 cells. Based on an efficient cellular internalization, good silencing effect, good siRNA retention and good col- loidal stability in human serum, DODAB:MO (2:1) siRNA-lipoplexes coated with PEG-Cer are considered promising nanocarriers for further in vivo validation. Statement of Significance This work describes two stealth functionalization strategies for the stabilization of the previously validated dioctadecyldimethylammonium bromide (DODAB):monoolein (MO) siRNA-lipoplexes. These nanocarriers are capable of efficiently incorporating and delivering siRNA molecules to cells in order to silence genes whose expression is implicated in a pathological condition. The main objective was to functionalize these nanocarriers with a coating conferring protection to siRNA in blood without compromising its efficient delivery to cancer cells, validating the potential of DODAB:MO (2:1) siRNA-lipoplexes as therapeutic vec- tors. We show that the stealth strategy is determinant to achieve a stable and efficient nanocarrier, and that DODAB:MO mixtures have a very promising potential for systemic siRNA delivery to leukemic cells.FEDER through POFC-COMPETE and by national funds from FCT I.P. through the strategic funding UID/BIA/04050/2013 (CBMA) and PEst-C/FIS/UI0607/2013 (CFUM) and PTDC/QUI/69795/2006. We thank the support of the Frame Work Program 7 of the European Commission: BIOCAPS (316265, FP7/REGPOT) and Xunta de Galicia, Spain (Agrupamento INBIOMED, Grupo con potencial crecimiento) reference IF/00498/2012, scholarship SFRH/BD/68588/2010. NanoDelivery-I&D em Bionanotecnologia, Lda. for access to their equipment

The evolving SARS-CoV-2 epidemic in Africa: Insights from rapidly expanding genomic surveillance

INTRODUCTION Investment in Africa over the past year with regard to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequencing has led to a massive increase in the number of sequences, which, to date, exceeds 100,000 sequences generated to track the pandemic on the continent. These sequences have profoundly affected how public health officials in Africa have navigated the COVID-19 pandemic. RATIONALE We demonstrate how the first 100,000 SARS-CoV-2 sequences from Africa have helped monitor the epidemic on the continent, how genomic surveillance expanded over the course of the pandemic, and how we adapted our sequencing methods to deal with an evolving virus. Finally, we also examine how viral lineages have spread across the continent in a phylogeographic framework to gain insights into the underlying temporal and spatial transmission dynamics for several variants of concern (VOCs). RESULTS Our results indicate that the number of countries in Africa that can sequence the virus within their own borders is growing and that this is coupled with a shorter turnaround time from the time of sampling to sequence submission. Ongoing evolution necessitated the continual updating of primer sets, and, as a result, eight primer sets were designed in tandem with viral evolution and used to ensure effective sequencing of the virus. The pandemic unfolded through multiple waves of infection that were each driven by distinct genetic lineages, with B.1-like ancestral strains associated with the first pandemic wave of infections in 2020. Successive waves on the continent were fueled by different VOCs, with Alpha and Beta cocirculating in distinct spatial patterns during the second wave and Delta and Omicron affecting the whole continent during the third and fourth waves, respectively. Phylogeographic reconstruction points toward distinct differences in viral importation and exportation patterns associated with the Alpha, Beta, Delta, and Omicron variants and subvariants, when considering both Africa versus the rest of the world and viral dissemination within the continent. Our epidemiological and phylogenetic inferences therefore underscore the heterogeneous nature of the pandemic on the continent and highlight key insights and challenges, for instance, recognizing the limitations of low testing proportions. We also highlight the early warning capacity that genomic surveillance in Africa has had for the rest of the world with the detection of new lineages and variants, the most recent being the characterization of various Omicron subvariants. CONCLUSION Sustained investment for diagnostics and genomic surveillance in Africa is needed as the virus continues to evolve. This is important not only to help combat SARS-CoV-2 on the continent but also because it can be used as a platform to help address the many emerging and reemerging infectious disease threats in Africa. In particular, capacity building for local sequencing within countries or within the continent should be prioritized because this is generally associated with shorter turnaround times, providing the most benefit to local public health authorities tasked with pandemic response and mitigation and allowing for the fastest reaction to localized outbreaks. These investments are crucial for pandemic preparedness and response and will serve the health of the continent well into the 21st century

Body composition and net energy requirements of Brazilian Somali lambs

The aim of this study was to determine the energy requirements for maintenance (NEm) and growth of 48 Brazilian Somali ram lambs with an average initial body weight of 13.47±1.76 kg. Eight animals were slaughtered at the trials beginning as a reference group to estimate the initial empty body weight (EBW) and body composition. The remaining animals were assigned to a randomised block design with eight replications per block and five diets with increasing metabolisable energy content (4.93, 8.65, 9.41, 10.12 and 11.24 MJ/kg dry matter). The logarithm of heat production was regressed against metabolisable energy intake (MEI), and the NEm (kJ/kg0.75 EBW/day) were estimated by extrapolation, when MEI was set at zero. The NEm was 239.77 kJ/kg0.75 EBW/day. The animal’s energy and EBW fat contents increased from 11.20 MJ/kg and 208.54 g/kg to 13.54 MJ/kg and 274.95 g/kg of EBW, respectively, as the BW increased from 13 to 28.70 kg. The net energy requirements for EBW gain increased from 13.79 to 16.72 MJ/kg EBW gain for body weights of 13 and 28.70 kg. Our study indicated the net energy requirements for maintenance in Brazilian Somali lambs were similar to the values commonly recommended by the United States’ nutritional system, but lower than the values recommended by Agricultural Research Council and Commonwealth Scientific and Industrial Research Organization. Net requirements for weight gain were less compared to the values commonly recommended by nutritional system of the United States

Efficacy of whole-body vibration training on brain-derived neurotrophic factor, clinical and functional outcomes, and quality of life in women with fibromyalgia syndrome: a randomized controlled trial

Whole-body vibration (WBV) is an exercise modality or treatment/prophylaxis method in which subjects (humans, animals, or cells) are exposed to mechanical vibrations through a vibrating platform or device. The vibrations are defined by their direction, frequency, magnitude, duration, and Biology 2021, 10, 965. https://doi.org/10.3390/biology10100965 https://www.mdpi.com/journal/biology Biology 2021, 10, 965 2 of 30 the number of daily bouts. Subjects can be exposed while performing exercises, hold postures, sitting, or lying down. Worldwide, WBV has attracted significant attention, and the number of studies is rising. To interpret, compare, and aggregate studies, the correct, complete, and consistent reporting of WBV-specific data (WBV parameters) is critical. Specific reporting guidelines aid in accomplishing this goal. There was a need to expand existing guidelines because of continuous developments in the field of WBV research, including but not limited to new outcome measures regarding brain function and cognition, modified designs WBV platforms and attachments (e.g., mounting a chair on a platform), and comparisons of animal and cell culture studies with human studies. Based on Delphi studies among experts and using EQUATOR recommendations, we have developed extended reporting guidelines with checklists for human and animal/cell culture research, including information on devices, vibrations, administration, general protocol, and subjects. In addition, we provide explanations and examples of how to report. These new reporting guidelines are specific to WBV variables and do not target research designs in general. Researchers are encouraged to use the new WBV guidelines in addition to general design-specific guideline

Efeito do óleo essencial de pimenta longa (Piper hispidinervum C. DC) e do emulsificante Tween® 80 sobre o crescimento micelial de Alternaria alternata (Fungi: Hyphomycetes) The effect of Long-pepper essential oil (Piper hispidinervum C. DC.) and of Tween®; 80 emulsifier on the mycelial growth of Alternaria alternate (Fungi: Hyphomycetes)

Este trabalho objetivou avaliar o efeito do óleo essencial de folhas de pimenta longa (Piper hispidinervum C. DC), sobre o crescimento micelial de Alternaria alternata e a análise da influência da concentração do emulsificante Tween® 80 no controle deste fitopatógeno. O óleo essencial foi obtido pela técnica "arraste a vapor d'água", utilizando-se aparelho de Clevenger modificado, e posteriormente submetido, à análise por cromatografia em fase gasosa acoplada a espectrômetro de massas CG-EM a CG. Para os ensaios biológicos, o método foi o bioanalítico in vitro observando-se o crescimento ou inibição do micélio de A. alternata no meio de cultura BDA na presença de diferentes concentrações do óleo essencial (0, 100, 250, 500 e 1000 mgL-1 ) sob diferentes concentrações de Tween® 80. Adotou-se esquema fatorial com quatro repetições, em delineamento inteiramente casualizado (DIC). Observou-se que o óleo essencial de pimenta longa apresentou inibição sobre o crescimento micelial do fungo A. alternataem todas as concentrações analisadas, sendo que na concentração de 1000 mgL-1 esta inibição foi de 100% , e a porcentagem de emulsificante (Tween® 80), influenciou na atividade fungitóxica das concentrações de 250 mgL -1 e 500 mgL -1 do óleo essencial.<br>The object of this work was to evaluate the effectof essential oils from Long-pepper leaves Piper hispidinervum on the mycelial growth of Alternaria alternata and to analyze the influence of emulsifier Tween® 80 concentration in the control of this phytopathogen. The essential oil was obtained by steam stream distillation, using a modified Clevenger apparatus, and the chemical composition was analyzed by gas chromatography coupled to mass spectroscopy GC-MS and GC. For biological tests, the bioanalytical in vitro method was used, observing the growth or inhibition of such phytopathogen in BDA culture media, in the presence of different essential oil concentrations (0, 100, 250, 500, e 1000 mgL -1) under different Tween® 80 concentrations. Factorial scheme with four repetitions, in entirely randomized outline was adopted. The essential oil of Long-pepper inhibited the micellial growth of the A. alternata fungus at all of the concentrations tested. A 100% inhibition at the 1000 mgL -1 concentration was observed, and the percentage of emulsifier (Tween® 80) influenced the fungitoxic activity at the 250 mgL-1 and 500 mgL -1 concentrations of the essential oil

Dermatologist-like explainable AI enhances trust and confidence in diagnosing melanoma

Artificial intelligence (AI) systems have been shown to help dermatologists diagnose melanoma more accurately, however they lack transparency, hindering user acceptance. Explainable AI (XAI) methods can help to increase transparency, yet often lack precise, domain-specific explanations. Moreover, the impact of XAI methods on dermatologists’ decisions has not yet been evaluated. Building upon previous research, we introduce an XAI system that provides precise and domain-specific explanations alongside its differential diagnoses of melanomas and nevi. Through a three-phase study, we assess its impact on dermatologists’ diagnostic accuracy, diagnostic confidence, and trust in the XAI-support. Our results show strong alignment between XAI and dermatologist explanations. We also show that dermatologists’ confidence in their diagnoses, and their trust in the support system significantly increase with XAI compared to conventional AI. This study highlights dermatologists’ willingness to adopt such XAI systems, promoting future use in the clinic. © 2024, The Author(s).53 − 5400.1-007/