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Enzymatic degumming using phospholipase C (PLC) enzymes may be used in environmentally friendly processes with improved oil recovery yields. In this work, phosphatidylinositol-specific phospholipase C (PIPLC) candidates obtained from an in silico analysis were evaluated for oil degumming. A PIPLC from Lysinibacillus sphaericus was shown to efficiently remove phosphatidylinositol from crude oil, and when combined with a second phosphatidylcholine and phosphatidylethanolamine-specific phospholipase C, the three major phospholipids were completely hydrolyzed, providing an extra yield of oil greater than 2.1%, compared to standard methods. A remarkably efficient fed-batch Escherichia coli fermentation process producing ∼14 g/L of the recombinant PIPLC enzyme was developed, which may facilitate the adoption of this cost-effective oil-refining process.

The growing demand for food and biofuels urges the vegetable oil processing industry to adopt cleaner technologies to mitigate the environmental pollution caused by chemical refining processes. Over the past decade, several enzymatic methods have proven to be efficient at reducing the generated waste, but improving the benefit-cost ratio is still necessary for the widespread adoption of this technology. In this work, we show that lecithin:cholesterol acyltransferase from Aeromonas enteropelogenes (LCATAE) provides a higher extra-yield of soybean oil than a type A1 phospholipase (PLA) enzyme currently commercialized for soybean oil deep degumming. Our model indicates that crude soybean oil treated with the new enzyme generates 87% more neutral oil from phospholipids than the widely used PLA, with the corresponding reduction in waste and byproducts generation. The refined oil retains the phytosterols naturally present in crude oil, enriching its nutritional value. The results presented here position LCATAE as a promising candidate to provide the green solutions needed by the industrial oil processing sector.Key points• Selected LCAT gene candidates were expressed in E. coli.• Aeromonas enteropelogenes LCAT hydrolyzes all the phospholipids present in crude soybean oil.• The LCAT enzyme provides a higher yield of neutral oil than commercial PLA enzymes and generates less waste.• The degummed oil retains sterols with high nutritional value.

The range of vaccines developed against severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) provides a unique opportunity to study immunization across different platforms. In a single-center cohort, we analyzed the humoral and cellular immune compartments following five coronavirus disease 2019 (COVID-19) vaccines spanning three technologies (adenoviral, mRNA and inactivated virus) administered in 16 combinations. For adenoviral and inactivated-virus vaccines, heterologous combinations were generally more immunogenic compared to homologous regimens. The mRNA vaccine as the second dose resulted in the strongest antibody response and induced the highest frequency of spike-binding memory B cells irrespective of the priming vaccine. Priming with the inactivated-virus vaccine increased the SARS-CoV-2-specific T cell response, whereas boosting did not. Distinct immune signatures were elicited by the different vaccine combinations, demonstrating that the immune response is shaped by the type of vaccines applied and the order in which they are delivered. These data provide a framework for improving future vaccine strategies against pathogens and cancer. Becher et al. perform a head-to-head comparison of multiple severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) vaccine prime-boost combinations and analyze the ensuing humoral and cellular responses in a large randomized cohort.

On 21 February 2020, a resident of the municipality of Vo’, a small town near Padua (Italy), died of pneumonia due to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection 1 . This was the first coronavirus disease 19 (COVID-19)-related death detected in Italy since the detection of SARS-CoV-2 in the Chinese city of Wuhan, Hubei province 2 . In response, the regional authorities imposed the lockdown of the whole municipality for 14 days 3 . Here we collected information on the demography, clinical presentation, hospitalization, contact network and the presence of SARS-CoV-2 infection in nasopharyngeal swabs for 85.9% and 71.5% of the population of Vo’ at two consecutive time points. From the first survey, which was conducted around the time the town lockdown started, we found a prevalence of infection of 2.6% (95% confidence interval (CI): 2.1–3.3%). From the second survey, which was conducted at the end of the lockdown, we found a prevalence of 1.2% (95% CI: 0.8–1.8%). Notably, 42.5% (95% CI: 31.5–54.6%) of the confirmed SARS-CoV-2 infections detected across the two surveys were asymptomatic (that is, did not have symptoms at the time of swab testing and did not develop symptoms afterwards). The mean serial interval was 7.2 days (95% CI: 5.9–9.6). We found no statistically significant difference in the viral load of symptomatic versus asymptomatic infections ( P = 0.62 and 0.74 for E and RdRp genes, respectively, exact Wilcoxon–Mann–Whitney test). This study sheds light on the frequency of asymptomatic SARS-CoV-2 infection, their infectivity (as measured by the viral load) and provides insights into its transmission dynamics and the efficacy of the implemented control measures. The authors describe the prevalence of SARS-CoV-2 infection, viral load and the frequency of symptomatic versus asymptomatic and presymptomatic infection in an Italian town, before and after a strict 14-day lockdown.

Drought impacts are increasingly recognized as interdisciplinary, socially influenced processes often resulting in uneven outcomes across different social groups, rather than as mere hydro-climatic events. Yet, many drought impact studies do not fully integrate the knowledge and perspectives of those who directly experience the impacts of droughts. While knowledge co-creation represents a promising avenue to address this challenge, there still remains an ample margin of improvement in the depth and breadth of transdisciplinary approaches in drought impact studies: most studies either limit co-creation to specific phases of the research process (breadth) or fail to meaningfully incorporate non-academic knowledge within those phases (depth). Drawing from a diverse body of literature on transdisciplinarity in sustainability science, integrated water resources management, socio-hydrology, science and technology studies, and critical water studies, we delineate five key dimensions which can support broader and deeper knowledge co-creation processes in drought impact studies, including (1) setting up a collaborative space, (2) framing the co-modelling process, (3) shaping a shared knowledge of drought, (4) co-selecting and co-developing models to understand drought impacts, and (5) being aware of power biases and knowledge imbalances. Incorporating all five dimensions promotes broader and more comprehensive studies, while exploring each dimension in greater detail enhances their depth. Together, these dimensions provide conceptual guidance for developing transdisciplinary approaches that are more integrated, power-sensitive, inclusive, situated, and reflexive.

OBJECTIVE: To determine the pharmacokinetic and pharmacodynamic properties of an oral insulin (OI) formulation compared with subcutaneously injected regular human insulin (RHI). RESEARCH DESIGN AND METHODS: Ten male patients with type 2 diabetes (means ± SD; A1C 7.0 ± 1.1%; BMI 28.3 ± 2.7 kg/m²) received either 300 units of insulin combined with 400 mg of delivery agent orally or 15 units RHI subcutaneously under isoglycemic clamp conditions. RESULTS: Maximum insulin concentration was greater and onset of action was faster with OI (Cmax 93 ± 71 vs. 33 ± 11 μU/ml; AUCGIR⁽⁰⁻¹h⁾ 173 ± 86 vs. 27 ± 32 mg/kg; P < 0.05). Mean insulin concentration and glucose infusion rate returned to baseline within 3 h after OI administration. Relative bioavailability of OI was 7 ± 4% (1st 2 h). CONCLUSIONS: This proof-of-concept study demonstrated that absorption of OI is feasible under fasting conditions. OI has a fast onset and a short duration of action but also shows a rather high between-subject variability in absorption.

Elevated surface concentrations of nitrogen dioxide (NO2) are associated with poor air quality, making its detection and monitoring important for human health and the environment. Existing instruments such as the TROPOMI satellite currently deliver daily global maps of NO2 tropospheric columns, and the future Sentinel-4 instrument will return hourly maps. While areas of strong concentrations (cities, large industries) can be detected in these satellite observations, their spatiotemporal resolution remains too coarse to capture local hot spots and quick variations. In the context of urban air quality monitoring, we present a new type of remote sensing instrument capable of observing spatial and temporal gradients in the NO2 field, which is not currently possible with either space instruments or from the routine operations of conventional diffraction grating and other ground-based remote sensing instruments. This novel instrument is based on an acousto-optical tunable filter (AOTF) located at the heart of a telecentric imaging system. The instrument acquires spectral images in the region 430–455 nm, where NO2 exhibits strong absorption features. A dense spectral sampling was commanded in order to enable the application of the DOAS (differential optical absorption spectroscopy) method in the processing of the spectra measured by each detector pixel. In March 2024, the instrument was deployed at the BAQUNIN supersite for atmospheric research, located in the center of Rome. In order to validate the NO2 camera measurements, coincident acquisitions by a MAX-DOAS and a Pandora spectrometer were performed. The results show very good agreement among the three instruments. They also illustrate the additional capabilities of the NO2 camera in observing the spatial and temporal variability of the urban NO2 field.

Bracts of maize (Zea mays, L.), a large agro-industrial residue and a source of functional biopolymers, were evaluated as a source of additives in bread-making. After drying and milling, the dried powder obtained (BP), its alcohol insoluble residue (AIR: ~70% yield on BP; composed by 26% arabinoxylans, 26% cellulose, 7% lignin, 5.5% pectins), and the ferulate-crosslinked arabinoxylans (AX-OH) isolated by 4%-KOH treatment from AIR, were evaluated. Bread loaves were made by replacing the wheat flour in 1.5% (w/w) by each fraction, applying two elaboration methodologies: required and constant hydration. Doughs’ thermomechanical behavior during kneading showed that the rheological properties and water availability were affected within both methodologies of bread making. The major effect was noted in the formulation with the fiber enriched in arabinoxylans (AX-OH) when added at required hydration. The hydration percentage tended to increase with the enrichment in arabinoxylans, varying from 58.7% in the dough made without wheat flour replacement, to 62% in AX-OH dough, with the highest content of hemicellulose. Dough stability and protein weakening were the most influenced parameters, mainly owing to AX-OH fibers, suggesting that they interfered the water distribution and hence the gluten network formation. As expected, fibers’ addition decreased the specific volume of bread, while firmness analysis showed that all maize fibers also delayed bread aging at required hydration, probably by hindering the diffusion of water and, hence, leading to starch retrogradation delay. Fibers from maize bracts can be a source of biopolymers useful as natural additives in bakery products.

HLA-G is a promiscuous immune checkpoint molecule. The HLA-G gene presents substantial nucleotide variability in its regulatory regions. However, it encodes a limited number of proteins compared to classical HLA class I genes. We characterized the HLA-G genetic variability in 4640 individuals from 88 different population samples across the globe by using a state-of-the-art method to characterize polymorphisms and haplotypes from high-coverage next-generation sequencing data. We also provide insights regarding the HLA-G genetic diversity and a resource for future studies evaluating HLA-G polymorphisms in different populations and association studies. Despite the great haplotype variability, we demonstrated that: (1) most of the HLA-G polymorphisms are in introns and regulatory sequences, and these are the sites with evidence of balancing selection, (2) linkage disequilibrium is high throughout the gene, extending up to HLA-A, (3) there are few proteins frequently observed in worldwide populations, with lack of variation in residues associated with major HLA-G biological properties (dimer formation, interaction with leukocyte receptors). These observations corroborate the role of HLA-G as an immune checkpoint molecule rather than as an antigen-presenting molecule. Understanding HLA-G variability across populations is relevant for disease association and functional studies.

Although non-small cell lung cancer (NSCLC) remains a deadly disease, new predictive biomarkers have emerged to assist in managing the disease, of which one of the most promising is the programmed death-ligand 1 (PD-L1). Each, variant seem to modulate the function of immune checkpoints differently and affect response to adjuvant treatment and outcome in NSCLC patients. We thus investigated the influence of these PD-L1 genetic variations in genetically admixed NSCLC tissue samples, and correlated these values with clinicopathological characteristics, including prognosis. We evaluated PD-L1 non-coding genetic variants and protein expression in lung adenocarcinomas (ADC), squamous cell carcinomas (SqCC), and large cell carcinomas (LCC) in silico. Microarray paraffin blocks from 70 samples of ADC (N=33), SqCC (N=24), and LCC (N=13) were used to create PD-L1 multiplex immunofluorescence assays with a Cell Signaling E1L3N clone. Fifteen polymorphisms of the gene were investigated by targeted sequencing and evaluated in silico using dedicated tools. Although polymorphisms seemed not to interfere with protein expression, PD-L1 expression varied among different histological subtypes, as did clinical outcomes, with the rs4742098A>G, rs4143815G>C, and rs7041009G>A variants being associated with relapse ( =0.01; =0.05; =0.02, respectively). The rs7041009 GG genotype showed a significant correlation with younger and alive patients compared to carriers of the A allele ( =0.02 and 0.01, respectively). The Cox regression model showed that the rs7041009 GG genotype may influence OS ( <0.01) as a co-dependent factor associated with radiotherapy and recurrence in NSCLC patients. Furthermore, the Kaplan-Meier survival curves showed that rs7041009 and rs4742098 might impact PPS in relapsed patients. In silico approaches identified the variants as benign. non-coding variants play an important role in modulating immune checkpoint function and may be explored as immunotherapy biomarkers. We highlight the rs7041009 variant, which impacts OS and PPS in NSCLC patients.