Valorization of kiwi waste through composting.

Kiwi waste from the calibration process is a major environmental problem of kiwi production due to landfill deposition. This work aims to contribute to the agronomic use of recycled kiwi waste through composting. With this objective, a composting experiment was carried out with kiwi fruit waste mixed with 5%, 10% and 20% (fresh weight) of wheat straw from bundles used to protect kiwifruit trunks from frost, as abulking agent to increase aeration, in the piles 5S, 10S and 20S, respectively. The highest temperatures for piles 5S and 10S were above 60°C, whereas the temperature did not reach 40°C in the pile with the highest straw content (20S) because the aeration increased heat loss in addition to increased C/N ratio of this pile. Also, the amount of organic matter mineralized decreased with increasing amount of straw because of the high C/N ratio of the straw. The highest total N (29.7 g kg-1) and the lowest C/N ratio (13) of the compost with 5% of straw is important from the agricultural point of view to promote N availability. In contrast, the high electrical conductivity (4.6 dS m-1) of this compost increases the risk of salt accumulation in the soil. Our results show that the compost with 10% straw, with high degree of maturation, absence of poor hygiene indicators as coliforms and pathogens as Salmonella sp., high organic matter content and rich in nutrients, together with the adequate compost pH and low electrical conductivity improves compost quality.

Indexing of Left Atrial Volume by Body Surface Area and Height in a Brazilian Population without Previous Heart Disease and with a Normal Heart on Echocardiography. Behavior in Obese and Overweight Patients.

Background: Left atrial (LA) volume indexing for body surface area (BSA) may underestimate LA size in obese and overweight people. Since LA volume is a risk marker for some cardiovascular events, it is suggested that indexing for height would be an alternative more appropriate method. The aims of this study were to find normal and the best cutoff values for LA volume indexed for height in our population. Methods: Echocardiograms from 2018 to 2021 were reviewed and patients without known cardiac disease and completely normal echocardiograms that had the left atrial volume (LAvol) measured by biplane Simpson's method were included. LAvol was indexed by BSA (ml/m2), by height (LAvol/m), by height raised to exponent 2.7 (ml/ m2.7) and by height squared (ml/h2). Results: A total of 545 patients, 50.5 ± 13.4 y., 335 females (61,5%) were analyzed. There were 145 normal weight (26.6%), 215 overweight (39.4%), 154 obese (28.3%) and 31 low weight (5.7%) patients. To establish normal values we included only the normal weight group and considered normal values from 2SD below to 2SD above the mean. Mean and normal values were: LAvol/h 26.0 ±4.5, 17 - 35 ml/m, LAvol/ht2 16 ± 2.8, 10.4 - 21.6 ml/ ht2 and LAvol/ht2.7 11.4 ± 2.2, 7.0 - 15.8 ml/m2.7. The normal LAvol/ht2.7 differed between male and female (11.4 ± 2.4 and 12.8 ± 2.6, p < 0.001). LA diameter, LAvol, LAvol/h, LAvol/h2 and LAvol/ht2.7 increased progressively from low-weight, normal weight, overweight and obese patients (p< 0.0001), but not LAvol/BSA. When indexing LAvol for height, for height2 and for height2.7 20.8%, 22.7% and 21.4% of the obese patients, respectively, were reclassified as enlarged LA, and 7.4%, 8.8% and 8.4% of the overweight patients as well. Using ROC curve analysis, LAvol/h2 had the highest AUC ant the best predictive value to identify LA enlargement and LAvol/BSA the worst one. Conclusions: Normal values for LAvol indexed for height by three different methods are described in normal individuals. We reinforce that LAvol indexation for BSA underestimates LA size in obese and overweight patients and in these groups, specially, indexing for height2 is probably the best method to evaluate LAvol.

Phytochemical screening and antibacterial assessment of two macroalgae Ulva papenfussi and Ulva nematoidea (Chlorophyta) against the bacterium Vibrio parahaemolyticus.

This study was conducted to determine whether the macroalgae Ulva papenfussi and Ulva nematoidea could be alternatives for preventing Litopenaeus vannamei vibriosis caused by the bacterium Vibrio parahaemolyticus. Phytochemical screening was performed on methanolic extracts to qualitatively determine the main groups of bioactive compounds, previous to an in vitro antibacterial test against V. parahaemolitycus. Phenols, polyphenols, flavonoids, and the high presence of carbohydrates were found in both macroalgae. U. papenfussi showed more presence of lipids and alkaloids than U. nematoidea . Macroalgae extracts prepared (v:v) with a 1:1 methanol: dichloromethane solvent was used for the in vitro test using the disc diffusion method (MDD). Filter paper discs impregnated with 1.0, 1.5, 2.0, 3.0, and 4.0 mg of the extracts showed antibacterial activity against V. Parahaemolitycus in a dose-dependent manner in both macroalgae. The inhibition zone varied significantly (p < 0.05) from 8.33 ± 0.12 to 11.41 ± 0.73 mm for 1 to 3 mg of extract levels, respectively. In conclusion, both macroalgae have antibacterial activity in their crude extracts against this bacteria. It is suggested to evaluate it as a feed additive for L. vannamei. This study is the first report on a phytochemical screening and antibacterial activity of these macroalgae against V. parahaemolyticus.

Performance of an interferon-γ release assay-based test for cell-mediated immunity to SARS-CoV-2.

In search for immunological correlates of protection against acute coronavirus disease 2019 (COVID-19) there is a need for high through-put assays for cell-mediated immunity (CMI) to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. We established an interferon-γ release assay -based test for detection of CMI against SARS-CoV-2 spike (S) or nucleocapsid (NC) peptides. Blood samples obtained from 549 healthy or convalescent individuals were measured for interferon-γ (IFN-γ) production after peptide stimulation using a certified chemiluminescence immunoassay. Test performance was calculated applying cutoff values with the highest Youden indices in receiver-operating-characteristics curve analysis and compared to a commercially available serologic test. Potential confounders and clinical correlates were assessed for all test systems. 522 samples obtained from 378 convalescent in median 298 days after PCR-confirmed SARS-CoV-2 infection and 144 healthy control individuals were included in the final analysis. CMI testing had a sensitivity and specificity of up to 89% and 74% for S peptides and 89% and 91% for NC peptides, respectively. High white blood cell counts correlated negatively with IFN-γ responses but there was no CMI decay in samples obtained up to one year after recovery. Severe clinical symptoms at time of acute infection were associated with higher measures of adaptive immunity and reported hair loss at time of examination. This laboratory-developed test for CMI to SARS-CoV-2 NC peptides exhibits excellent test performance, is suitable for high through-put routine diagnostics, and should be evaluated for clinical outcome prediction in prospective pathogen re-exposure.

Predominantly antibody deficiency: case report / Deficiência predominantemente de anticorpos: relato de caso

Abstract Objectives: present a case of Inborn errors of immunity (IEI) as a potential diagnosis in pediatric patients with recurrent infections. Description: male patient, 13 years old, since he was eight months old had recurrent diarrhea, sinusitis, otitis, abscesses and urinary tract infections. At the age of ten, he presented mastoiditis progressing to meningitis, he was admitted to a tertiary hospital, where an immunological evaluation was performed, which led to the diagnosis of Predominantly Antibody Deficiency (PAD), with suspected X-linked Agammaglobulinemia (XLA). Treatment was initiated with administration of intravenous gamma globulin 400 mg/kg every four weeks, with a significant improvement of the condition. Discussion: usually, the diagnosis of XLA tends to be made in the first three years of life. However, in this report, although the first manifestations started at eight months of age, there was a delay of ten years before starting the treatment. In fact, the diagnosis of children and adults with IEI can be delayed if healthcare professionals are unable to find the true cause of recurrent infections. Therefore, the relevance of considering such pathologies in the presence of risk signs is highlighted, as early diagnosis being essential in treating and preventing morbidities.

Resumo Objetivos: apresentar um caso de Erro Inato da Imunidade (EII) como diagnóstico em potencial de pacientes pediátricos com infecções de repetição. Descrição: paciente masculino, 13 anos, desde os oito meses de idade apresentou quadros repetidos de diarreias, sinusites, otites, abscessos e infecções do trato urinário; destacando-se a otite, sinusite e diarreia pela maior recorrência. Aos dez anos, quando apresentou mastoidite evoluindo para meningite, foi internado em um hospital terciário, onde foi realizada avaliação imunológica, a qual levou ao diagnóstico de Deficiência Predominantemente de Anticorpos (DPAs), tendo como suspeita a agamaglobulinemia ligada ao cromossomo X (ALX). Foi iniciado tratamento com administração de gamaglobulina endovenosa 400 mg/kg a cada quatro semanas, ocorrendo melhora significativa do quadro. Discussão: normalmente, o diagnóstico da ALX tende a ser feito nos primeiros três anos de vida. Neste relato, entretanto, embora as primeiras manifestações tenham iniciado aos oito meses de idade, ocorreu um atraso de dez anos até o início do tratamento. De fato, o diagnóstico de crianças e adultos com EII pode ser retardado se os profissionais de saúde não conseguirem encontrar a causa das infecções recorrentes. Destaca-se, portanto, a relevância de se considerar tais patologias na vigência de sinais de riscos, pois o diagnóstico precoce é fundamental para tratar e prevenir morbidades.

Clinical Development of mRNA Vaccines: Challenges and Opportunities.

The emergence of safe and effective mRNA platform-based COVID-19 vaccines from the recent pandemic has changed the face of vaccine development. Compared with conventional technologies used historically, mRNA-based vaccines offer a rapid flexible and robust approach to preventing disease caused by transient viral strains such as SAR2-CoV-2 variants of concern and seasonal influenza. Adaptations in the formulation of the mRNA delivery systems such as with lipid nanoparticle delivery (LNP) used in mRNA-1273 and BNT16b2b have enabled this technology to flourish under the urgent collective response and collaborative regulatory understanding derived from COVID-19 vaccine development. The application of mRNA-based therapeutics in other areas holds potential promise including combination vaccines that might deliver protections against multiple infectious diseases. Future studies and further advances in mRNA-based technologies will provide insight into the clinical efficacy and real-world effectiveness of vaccines as well as provisions with respect to the impact of reactogenicity profiles. Overall, the success of mRNA-based COVID-19 vaccines has helped unlock a platform likely to result in many more candidate vaccines entering clinical evaluation to address the unmet medical needs of other diseases including viral respiratory diseases, herpesviruses, and historically challenging vaccine targets such as HIV.

Subcutaneous delivery of FGF21 mRNA therapy reverses obesity, insulin resistance, and hepatic steatosis in diet-induced obese mice.

Fibroblast growth factor 21 (FGF21) is a promising therapeutic agent for treatment of type 2 diabetes (T2D) and non-alcoholic steatohepatitis (NASH). We show that therapeutic levels of FGF21 were achieved following subcutaneous (s.c.) administration of mRNA encoding human FGF21 proteins. The efficacy of mRNA was assessed following 2-weeks repeated s.c. dosing in diet-induced obese (DIO), mice which resulted in marked decreases in body weight, plasma insulin levels, and hepatic steatosis. Pharmacokinetic/pharmacodynamic (PK/PD) modelling of several studies in both lean and DIO mice showed that mRNA encoding human proteins provided improved therapeutic coverage over recombinant dosed proteins in vivo. This study is the first example of s.c. mRNA therapy showing pre-clinical efficacy in a disease-relevant model, thus, showing the potential for this modality in the treatment of chronic diseases, including T2D and NASH.

Inefficient Placental Virus Replication and Absence of Neonatal Cell-Specific Immunity Upon Sars-CoV-2 Infection During Pregnancy.

Pregnant women have been carefully observed during the COVID-19 pandemic, as the pregnancy-specific immune adaptation is known to increase the risk for infections. Recent evidence indicates that even though most pregnant have a mild or asymptomatic course, a severe course of COVID-19 and a higher risk of progression to diseases have also been described, along with a heightened risk for pregnancy complications. Yet, vertical transmission of the virus is rare and the possibility of placental SARS-CoV-2 infection as a prerequisite for vertical transmission requires further studies. We here assessed the severity of COVID-19 and onset of neonatal infections in an observational study of women infected with SARS-CoV-2 during pregnancy. Our placental analyses showed a paucity of SARS-CoV-2 viral expression ex vivo in term placentae under acute infection. No viral placental expression was detectable in convalescent pregnant women. Inoculation of placental explants generated from placentas of non-infected women at birth with SARS-CoV-2 in vitro revealed inefficient SARS-CoV-2 replication in different types of placental tissues, which provides a rationale for the low ex vivo viral expression. We further detected specific SARS-CoV-2 T cell responses in pregnant women within a few days upon infection, which was undetectable in cord blood. Our present findings confirm that vertical transmission of SARS-CoV-2 is rare, likely due to the inefficient virus replication in placental tissues. Despite the predominantly benign course of infection in most mothers and negligible risk of vertical transmission, continuous vigilance on the consequences of COVID-19 during pregnancy is required, since the maternal immune activation in response to the SARS-CoV2 infection may have long-term consequences for children's health.

Impact of lipid nanoparticle size on mRNA vaccine immunogenicity.

Lipid nanoparticles (LNP) are effective delivery vehicles for messenger RNA (mRNA) and have shown promise for vaccine applications. Yet there are no published reports detailing how LNP biophysical properties can impact vaccine performance. In our hands, a retrospective analysis of mRNA LNP vaccine in vivo studies revealed a relationship between LNP particle size and immunogenicity in mice using LNPs of various compositions. To further investigate this, we designed a series of studies to systematically change LNP particle size without altering lipid composition and evaluated biophysical properties and immunogenicity of the resulting LNPs. While small diameter LNPs were substantially less immunogenic in mice, all particle sizes tested yielded a robust immune response in non-human primates (NHP).

Novel approaches for vaccine development.

Vaccines are critical tools for maintaining global health. Traditional vaccine technologies have been used across a wide range of bacterial and viral pathogens, yet there are a number of examples where they have not been successful, such as for persistent infections, rapidly evolving pathogens with high sequence variability, complex viral antigens, and emerging pathogens. Novel technologies such as nucleic acid and viral vector vaccines offer the potential to revolutionize vaccine development as they are well-suited to address existing technology limitations. In this review, we discuss the current state of RNA vaccines, recombinant adenovirus vector-based vaccines, and advances from biomaterials and engineering that address these important public health challenges.

Cytomegalovirus (CMV) Pneumonitis: Cell Tropism, Inflammation, and Immunity.

Human cytomegalovirus (HCMV) is an opportunistic pathogen causing disease mainly in immunocompromised patients or after congenital infection. HCMV infection of the respiratory tract leads to pneumonitis in the immunocompromised host, which is often associated with a bad clinical course. The related mouse cytomegalovirus (MCMV) likewise exhibits a distinct tropism for the lung and thus provides an elegant model to study host-pathogen interaction. Accordingly, fundamental features of cytomegalovirus (CMV) pneumonitis have been discovered in mice that correlate with clinical data obtained from humans. Recent studies have provided insight into MCMV cell tropism and localized inflammation after infection of the respiratory tract. Accordingly, the nodular inflammatory focus (NIF) has been identified as the anatomical correlate of immune control in lungs. Several hematopoietic cells involved in antiviral immunity reside in NIFs and their key effector molecules have been deciphered. Here, we review what has been learned from the mouse model with focus on the microanatomy of infection sites and antiviral immunity in MCMV pneumonitis.

The droplet size of emulsion adjuvants has significant impact on their potency, due to differences in immune cell-recruitment and -activation.

Self-emulsification is routinely used for oral delivery of lipophilic drugs in vivo, with the emulsion forming in vivo. We modified this technique to prepare novel oil-in-water emulsions of varying droplet size and composition on bench to enable adjuvanted vaccine delivery. We used these formulations to show that smaller droplets (20 nm) were much less effective as adjuvants for an influenza vaccine in mice than the emulsion droplet size of commercial influenza vaccine adjuvants (~160 nm). This was unexpected, given the many claims in the literature of the advantages of smaller particulates. We also undertook cell-recruitment mechanistic studies at site of injection and draining lymph nodes to directly address the question of why the smaller droplets were less effective. We discovered that emulsion droplet size and composition have a considerable impact on the ability to recruit immune cells to the injection site. We believe that further work is warranted to more extensively explore the question of whether, the smaller is not 'better', is a more common observation for particulate adjuvants.

Corrigendum: Induction of Robust B Cell Responses After Influenza mRNA Vaccination Is Accompanied by Circulating Hemagglutinin-Specific ICOS+ PD-1+ CXCR3+ T Follicular Helper Cells.

[This corrects the article DOI: 10.3389/fimmu.2017.01539.].

Optimization of Lipid Nanoparticles for Intramuscular Administration of mRNA Vaccines.

mRNA vaccines have the potential to tackle many unmet medical needs that are unable to be addressed with conventional vaccine technologies. A potent and well-tolerated delivery technology is integral to fully realizing the potential of mRNA vaccines. Pre-clinical and clinical studies have demonstrated that mRNA delivered intramuscularly (IM) with first-generation lipid nanoparticles (LNPs) generates robust immune responses. Despite progress made over the past several years, there remains significant opportunity for improvement, as the most advanced LNPs were designed for intravenous (IV) delivery of siRNA to the liver. Here, we screened a panel of proprietary biodegradable ionizable lipids for both expression and immunogenicity in a rodent model when administered IM. A subset of compounds was selected and further evaluated for tolerability, immunogenicity, and expression in rodents and non-human primates (NHPs). A lead formulation was identified that yielded a robust immune response with improved tolerability. More importantly for vaccines, increased innate immune stimulation driven by LNPs does not equate to increased immunogenicity, illustrating that mRNA vaccine tolerability can be improved without affecting potency.

Improved phylogeny of brown algae Cystoseira (Fucales) from the Atlantic-Mediterranean region based on mitochondrial sequences.

Cystoseira is a common brown algal genus widely distributed throughout the Atlantic and Mediterranean regions whose taxonomical assignment of specimens is often hampered by intra- and interspecific morphological variability. In this study, three mitochondrial regions, namely cytochrome oxidase subunit 1 (COI), 23S rDNA (23S), and 23S-tRNAVal intergenic spacer (mt-spacer) were used to analyse the phylogenetic relationships of 22 Cystoseira taxa (n = 93 samples). A total of 135 sequences (48 from COI, 43 from 23S and 44 from mt-spacer) were newly generated and analysed together with Cystoseira sequences (9 COI, 31 23S and 35 mt-spacer) from other authors. Phylogenetic analysis of these three markers identified 3 well-resolved clades and also corroborated the polyphyletic nature of the genus. The resolution of Cystoseira taxa within the three clades improves significantly when the inclusion of specimens of related genera was minimized. COI and mt-spacer markers resolved the phylogeny of some of the Cystoseira taxa, such as the C. baccata, C. foeniculacea and C. usneoides. Furthermore, trends between phylogeny, embryonic development and available chemotaxonomic classifications were identified, showing that phylogenetic, chemical and morphological data should be taken into account to study the evolutionary relationships among the algae currently classified as Cystoseira. The resolution of Cystoseira macroalgae into three well supported clades achieved here is relevant for a more accurate isolation and identification of natural compounds and the implementation of conservation measures for target species.

Report of in vitro antileishmanial properties of Iberian macroalgae.

Here is reported the anti Leishmania infantum activity of 48 hexane, CH2Cl2 and MeOH extracts from 16 macroalgae collected on the Iberian Coast. Seven hexane and CH2Cl2 Cystoseira baccata, Cystoseira barbata, Cystoseira tamariscifolia, Cystoseira usneoides, Dictyota spiralis and Plocamium cartilagineum extracts were active towards promastigotes (IC50 29.8-101.8 µg/mL) inducing strong morphological alterations in the parasites. Hexane extracts of C. baccata and C. barbata were also active against intracellular amastigotes (IC50 5.1 and 6.8 µg/mL, respectively). Fatty acids, triacylglycerols, carotenoids, steroids and meroterpenoids were detected by nuclear magnetic resonance (NMR), and gas chromatography in the Cystoseira extracts. These results suggest that Cystoseira macroalgae contain compounds with antileishmanial activity, which could be explored as scaffolds to the development of novel sources of antiparasitic derivatives.

Vía aérea difícil no prevista secundaria a consumo de inhibidores de la enzima convertidora de angiotensina. Reporte de caso / Unexpected difficult airway management due to the use of ACE inhibitors: case report

Abstract Angioedema induced by the use of angiotensin converting enzyme (ACE) inhibitors is an uncommon but life-threatening complication, especially when the airway is affected, creating unexpected difficult airway management. A prompt differential diagnosis with anaphylactic shock is critical, given that adrenaline treatment does not improve angioedema. We report a case of angioedema induced by ACE inhibitor following in-hospital administration of captopril, with almost impossible intubation, and secondary aspiration during airway management. Angioedema was erroneously treated, because it was mistakenly considered to be an anaphylactic reaction, and it could have ended in death.

Resumen El angioedema es una complicación poco frecuente relacionada con el uso de inhibidores de la enzima convertidora de angiotensina, pero potencialmente mortal, especialmente en el caso de afectar a la vía aérea, generando vías aéreas difíciles no previstas. Es de vital importancia realizar un rápido diagnóstico diferencial del cuadro con el shock anafiláctico, dado que el tratamiento con adrenalina, no mejora el angioedema. Presentamos un caso de angioedema tras administración intrahospitalaria de Captopril a un paciente sano, sin vía aérea difícil prevista, generando una intubación casi imposible y broncoaspiración secundaria durante el manejo de la vía aérea. El cuadro clínico se desencadenó por la confusión del angioedema, con una reacción anafiláctica, realizándose un tratamiento inapropiado. Las consecuencias del mismo pudieron ser mortales.

Neutralization of the Plasmodium-encoded MIF ortholog confers protective immunity against malaria infection.

Plasmodium species produce an ortholog of the cytokine macrophage migration inhibitory factor, PMIF, which modulates the host inflammatory response to malaria. Using a novel RNA replicon-based vaccine, we show the impact of PMIF immunoneutralization on the host response and observed improved control of liver and blood-stage Plasmodium infection, and complete protection from re-infection. Vaccination against PMIF delayed blood-stage patency after sporozoite infection, reduced the expression of the Th1-associated inflammatory markers TNF-α, IL-12, and IFN-γ during blood-stage infection, augmented Tfh cell and germinal center responses, increased anti-Plasmodium antibody titers, and enhanced the differentiation of antigen-experienced memory CD4 T cells and liver-resident CD8 T cells. Protection from re-infection was recapitulated by the adoptive transfer of CD8 or CD4 T cells from PMIF RNA immunized hosts. Parasite MIF inhibition may be a useful approach to promote immunity to Plasmodium and potentially other parasite genera that produce MIF orthologous proteins.

Stable Nanoemulsions for the Delivery of Small Molecule Immune Potentiators.

Adjuvants are required to enhance immune responses to typically poorly immunogenic recombinant antigens. Toll-like receptor agonists (TLRa) have been widely evaluated as adjuvants because they activate the innate immune system. Currently, licensed vaccines adjuvanted with TLRa include the TLR4 agonist monophosphoryl lipid, while additional TLRa are in clinical development. Unfortunately, naturally derived TLRa are often complex and heterogeneous entities, which brings formulation challenges. Consequently, the use of synthetic small-molecule TLRa has significant advantages because they are well-defined discrete molecules, which can be chemically modified to modulate their physicochemical properties. We previously described the discovery of a family of TLR7 agonists based on a benzonaphthyridine scaffold. In addition, we described how Alum could be used to deliver these synthetic TLRa. An alternative adjuvant approach with enhanced potency over Alum are squalene containing oil-in-water emulsions, which have been included in licensed influenza vaccines, including Fluad (MF59 adjuvanted) and Pandemrix (AS03 adjuvanted). Here, we describe how to enable the co-delivery of a TLR7 agonist in a squalene-based oil-in-water emulsion, for adjuvant evaluation.