Freezing and thawing in Antarctica: characterization of antifreeze protein (AFP) producing microorganisms isolated from King George Island, Antarctica.

Antarctic temperature variations and long periods of freezing shaped the evolution of microorganisms with unique survival mechanisms. These resilient organisms exhibit several adaptations for life in extreme cold. In such ecosystems, microorganisms endure the absence of liquid water and exhibit resistance to freezing by producing water-binding molecules such as antifreeze proteins (AFP). AFPs modify the ice structure, lower the freezing point, and inhibit recrystallization. The objective of this study was to select and identify microorganisms isolated from different Antarctic ecosystems based on their resistance to temperatures below 0 °C. Furthermore, the study sought to characterize these microorganisms regarding their potential antifreeze adaptive mechanisms. Samples of soil, moss, permafrost, and marine sediment were collected on King George Island, located in the South Shetland archipelago, Antarctica. Bacteria and yeasts were isolated and subjected to freezing-resistance and ice recrystallization inhibition (IR) tests. A total of 215 microorganisms were isolated, out of which 118 were molecularly identified through molecular analysis using the 16S rRNA and ITS regions. Furthermore, our study identified 24 freezing-resistant isolates, including two yeasts and 22 bacteria. A total of 131 protein extracts were subjected to the IR test, revealing 14 isolates positive for AFP production. Finally, four isolates showed both freeze-resistance and IR activity (Arthrobacter sp. BGS04, Pseudomonas sp. BGS05, Cryobacterium sp. P64, and Acinetobacter sp. M1_25C). This study emphasizes the diversity of Antarctic microorganisms with the ability to tolerate freezing conditions. These microorganisms warrant further investigation to conduct a comprehensive analysis of their antifreeze capabilities, with the goal of exploring their potential for future biotechnological applications.

Plant growth regulators and mobilization of reserves in imbibition phases of yellow passion fruit.

The production of seedlings of the passion fruit tree, usually, is sexual, and the seeds are not uniform in the seedling emergence, and soaking treatments of seeds can provide faster and more uniform germination. It was aimed to study the action of plant growth regulators and the mobilization of reserves in the stages of soaking of yellow passion fruit seeds. The seeds were soaked for five hours in solutions containing plant growth regulators, in a completely randomized design, in a factorial 8 x 4, with four replications. The first factor corresponds to eight plant growth regulators: T1 - distilled water (control); T2 - 6-benzylaminepurine ​​500 mg L-1; T3 - 4-(3-indolyl) butyric acid 500 mg L-1; T4 - gibberellic acid 500 mg L-1; T5 - spermine 250 mg L-1; T6 - spermine 750 mg L-1; T7 - spermidine 750 mg L-1; T8 - spermidine 1250 mg L-1; and the second factor, to the four soaking times: zero, four, 72 and 120 hours, corresponding, respectively, to the dry seed, and to phases I, II, and III of the imbibition curve. It was evaluated the biochemical composition of seeds (lipids, soluble sugars and starch). The seeds showed accumulation of lipids in phase III; the content of soluble sugars increased in phase I and decreased in phase II. The starch content increased until the phase II and decreased in phase III. Starch is the main reserve in the seeds and the main source of energy used in phase III; soaking the seeds in polyamines generates an accumulation of lipids in the seeds and soaking in plant growth regulators increases the burning of starch.

Molecular profiling in bee venom allergy: clinical and therapeutic characterization in a Portuguese cohort.

Summary: Background. Bee venom allergy (BVA) can trigger local and systemic allergic reactions, including anaphylaxis. Recently, the molecular sensitization profile has gained importance in the reaction's stratification and venom immunotherapy (VIT). Methods. Retrospective analysis of patients with hypersensitivity to BVA, confirmed by specific sIgE to Apis mellifera ≥0.35 kU/L and/or positive skin tests to bee venom commercial extract, evaluated in specialized consultation. Demographic, clinical, and laboratory data (including molecular Api m 1, 4, and 10) were analyzed, looking for risk factors associated with the severity of the index reaction and reactions during VIT. Results. 93 patients were included (55.9% male; median age of 46 years), 57.3% with atopic comorbidities, and 23.4% with cardiovascular comorbidities. The median specific IgE to Apis mellifera was 6.7 kU/L (IQR 1.0-20.3) kU/L. Regarding the molecular profile, the median IgE to Api m 1 was 0.5 kU/L (57.5% positive out of all measurements); Api m 4 - 0.01 kU/L (11.9% positive), and Api m 10 - 0.3 kU/L (50.0% positive). No patient was monosensitized to Api m 4. The median age of the most severe sting reaction was 36 (IQR 26-48) years, with a median severity (Müeller scale) of 3 (IQR 2-3). Forty-seven patients (50.5%) underwent VIT, with 35.6% of reactions recorded. Allergic reactions during VIT were recorded in 35.6% of cases. The severity of the index reaction correlated positively with older ages (p=0.040; r=0.249), in contrast to monosensitization to Api m 1, which was an independent predictor of milder reactions (p=0.015). Sensitization to Api m 10 was associated with a higher likelihood of reactions during VIT (p=0.038) but potentially less systemic reactions at re-stings (p=0.097). Conclusions. Molecular sensitization profile appears to be relevant not only to the severity of index reactions but also during VIT. Studies of a large cohort of patients with molecular profiles are essential to validate these results and improve the clinical and therapeutic approach to BVA.

Potential of coffee straw biochal as a substrate conditioner in seed lettuce and sorghum germination and vigority.

The use of residues from coffee production to obtain biochar is a sustainable approach, which aims to minimize the environmental impact of these materials. In this study, the effect of adding coffee straw biochar on the physiological quality of lettuce and sorghum seeds was investigated. Thus, the objective of this work was to study the effect of adding different concentrations of coffee biochar in the substrate composition on the physiological quality of lettuce (Lactuca sativa) and sorghum (Sorghum bicolor) seeds. The experimental design used was completely randomized, with five concentrations of biochar (0; 7.5; 15; 30 and 60%), conducted with four replications of 25 seeds. The use of biochar in the concentrations studied does not provide an increase in the average germination percentage and vigor of lettuce and sorghum seeds. The increase in the concentration of biochar caused less seed vigor, suggesting a toxic effect. For seed germination, there was no significant difference between lettuce and sorghum species, regardless of treatment. For the germination speed index, sorghum seeds have higher means, except for the treatment with the addition of 15% coffee straw biochar. Lettuce seeds have higher shoot length averages, except for treatment with 100% commercial substrate. The sorghum seeds have higher mean root length and dry mass than lettuce, regardless of the treatment.

Tasting to preserve: An educational activity to promote children's positive attitudes towards intraspecific diversity conservation.

On the edge of causing the sixth big mass extinction event, the development of positive attitudes towards the conservation of intraspecific diversity from early ages is essential to overcome the biodiversity crisis we currently face. However, there is no information available on elementary school students' attitudes toward intraspecific diversity conservation nor is there a framework available to perform such analysis. For this study we designed, implemented, and evaluated an educational activity planned for third graders (8 to 13 years old) to explore the intraspecific diversity of vegetables and promote healthy eating habits. Additionally, a framework was developed to evaluate students' attitudes towards intraspecific diversity conservation and applied to semi-structured interviews conducted with students before and after engaging in the educational activity. In this paper we present a reliable framework, developed aligned with the ABC model of attitudes, based on literature, and adapted to elementary school students' responses, to evaluate students' attitudes toward intraspecific diversity. Our results show that, before the educational activity, most students choose a non-biodiverse option, justifying this choice with the affective component of attitudes: mostly emotional factors but also aesthetic and social/cultural factors. After the educational activity, we observed a significant increase in the frequency of students that choose the biodiverse option and that justified it with the cognitive component of attitudes: mainly with biology and health knowledge factors, but also with economic and ethical knowledge factors. Our findings support the positive impact of educational activities that explore vegetable varieties on students' attitudes toward intraspecific diversity conservation. This activity may also be used to foster education for sustainability and address socioscientific issues aligned with diverse sustainable development goals.

Maturation and harvest time of Ateleia glazioveana Baill. seeds.

Ateleia glazioveana Baill. is a pioneer, rustic and can be used for forest recovery. This work aimed to study the process of physiological maturation of this species. The research was carried out in the city of Alegre - ES, the trees were identified in the floral anthesis and accompanied during the filling of the fruits and development of the seeds until the complete maturation. The fruits were harvested at the following stages 7, 14, 21, 28, 35 and 42 days after anthesis, and characterized according to: morphometry, moisture, fresh and dry mass of fruits and seeds, germination, germination speed index, shoot and root length and dry mass of seedlings. The regression equations were adjusted for the main characteristics analyzed as a function of the harvest period. The point of physiological maturity of timbó occurred at 42 days after anthesis.

Multivariate analysis of agronomic, physicochemical, and physiological characters of passion fruit hybrids cultivated at different environments.

Simultaneous analysis studies of several agronomic traits in cultivated plants make it possible to identify phenotypic and genotypic differences due to environmental variations, such as altitude. Therefore, the objective was to evaluate, through multivariate analysis of agronomic, physicochemical and physiological characters, passion fruit hybrids cultivated in different environments. The hybrids used were Gigante Amarelo, Rubi do Cerrado and Sol do Cerrado, cultivated in the southern region of Espírito Santo in four municipalities/environments: Marataízes (41 m), Jerônimo Monteiro (104 m), Alegre (711 m), and Ibitirama (1016 m). The agronomic characters of the plants, the physical-chemical characteristics of the fruits and the physical, biochemical and physiological qualities of the seeds were analyzed. The Singh method was used to determine the most important differentiating characters between hybrids growing in different environments. Based on these characters, a dissimilarity matrix was generated and a principal coordinate analysis was performed. It was observed that the pulp yield was influenced by altitude. The three hybrids showed greater performance in terms of agronomic characters at altitude (41 m) than at altitude (104 m). The Sol do Cerrado hybrid showed high performance in the physical-chemical characteristics of the fruits at altitude (104 m).

COVID-19 vaccine confidence in the post-vaccination era: Perceptions among adults with immune-mediated inflammatory diseases.

Objective: Assess the perceived protection afforded by a range of COVID-19 vaccines in immune-mediated inflammatory diseases patients previously vaccinated against SARS-CoV-2. Study design: Survey. Methods: On-line cross-sectional survey aimed at evaluating the perceived protection (and its determinants) afforded by a range of COVID-19 vaccines among immune-mediated inflammatory diseases previously vaccinated for COVID-19. Results: Out of 493 eligible respondents who lived in Brazil, 397 (80.5%) were confident that their primary vaccination series would protect them against severe COVID-19. In multivariate analysis, only overlapping immune-mediated inflammatory diseases remained (negatively) associated with the perception of protection. Conclusions: No influence was found between COVID-19 vaccine types and the perception of protection after initial vaccinations.

Bothrops jararacussu snake venom decreases CD1d, CD83, and CD86 expression on bone marrow-derived dendritic cells.

This study was designed to characterize mice bone marrow (BM) and bone marrow-derived dendritic cells (BMDC) and to compare the surface markers expression and inflammatory cytokine liberation in response to LPS and Bothrops jararacussu venom (BjV) stimulation. Typical morphology was observed in BM and BMDCs from the 4th up to the 8th day of culture using recombinant mouse GM-CSF and IL-4. A high basal level of MHC-II, CD1d, CD83, CD11c, CD80, and low CD86 was expressed by BM cells. After stimulation with GM-CSF/IL-4 for BMDCs differentiation, the BM cells differentiated into BMDCs presented MHC-II, CD1d, CD83, CD11c, CD86, and CD80 expression on the 4th - 8th day accompanied with high levels of TNF-α liberated. The difference between the surface markers' expression was observed in this time course in which CD1d, CD11c, and CD80 remained in high levels of expression, while MHC-II and CD83 showed moderate expression during the differentiation period. Also, cytokines liberation was monitored over the period of the BMDCs culture, and on the 6th day, low levels of IL-6 and IL-1ß were found, while high levels of TNF-α on the 4th and 8th days, both of which contributed to the maturity of the BMDCs. Maturation of DCs with LPS showed significant upregulation of surface markers (MHC-II, CD1d, CD83, CD86, CD80) and pro-inflammatory cytokines (IL-1ß, IL-6, TNF-α) liberation. On the other hand, BjV induced a decrease in CD1d, CD11c, CD83, and CD86 expression in mature BMDCs which was not observed when LPS was used to stimulate BMDCs which probably induces impairment in T-cell activation.

Relevance of the diagnosis of hypersensitivity reactions to antineoplastic and biological agents: experience with drug provocation test.

Summary: Background. Evidence regarding drug provocation test (DPT) with chemotherapeutic agents is scarce. The aim of our study is to describe the experience of DPT in patients with a history of hypersensitivity reactions (HSRs) to antineoplastic and biological agents. Methods. This was an eight-year retrospective, observational, descriptive study of patients with a history of HSRs to chemotherapy who were submitted to DPT. Anamnesis, skin tests (ST) and DPT were analyzed. Patients with a negative DPT were submitted to at least one regular supervised administration (RSA). Patients with positive DPT or HSR during RSA were offered rapid drug desensitization (RDD). Results. A total of 54 patients were submitted to DPT. The most common suspected drugs were platins (n = 36), followed by taxanes (n = 11). Most initial reactions were classified as grade II (n = 39) according to Brown's grading system. ST with platinum (n = 35), taxanes (n = 10) and biological agents (n = 4) were negative, except for one intradermal test with paclitaxel, which was positive. A total of 64 DPTs were performed. Eleven percent of all DPTs were positive (platins (n = 6), doxorubicin (n = 1)). Of the 57 RSA with the culprit drugs, 2 were positive (platins). The diagnosis of hypersensitivity was confirmed by DPT/RSA in 9 patients. All patients with positive DPT/RSA presented HSRs of equal or less severity than the initial one. Conclusions. DPT followed by RSA allowed to exclude HSRs in 45 patients (55 culprit drugs). DPT before desensitization prevents non-hypersensitivity patients from undergoing RDD. In our study DPT was safe, all reactions were managed by an allergist.

Allergic emergencies in the prehospital setting: a 5 year retrospective study.

Summary: Background. Patients with severe allergic conditions often request support from the prehospital emergency services given the rapid, unexpected and potentially life-threatening nature of the reactions, such as anaphylaxis. Studies regarding prehospital incidents for allergic conditions are scarce. This study aimed to characterize prehospitalar medical requesting assistance due to suspected hypersensitivity reactions (HSR). Methods. Retrospective study of allergic-related requesting assistances between 2017-2022 of a Portuguese emergency dispatch centre - Emergency and Resuscitation Medical Vehicle (VMER), in Coimbra University Hospital. Demographic and clinical variables were analysed, including clinical manifestations, anaphylaxis severity grading, therapeutic interventions, and post-incident allergic work-up. Regarding anaphylactic events, three diagnosis timings were compared: on-site, hospital emergency department and Investigator-diagnosis based on data reviewed. Results. Out of 12689 VMER requesting assistances, 210 (1.7%) were classified as suspected HSR reactions. After on-site medical evaluation, 127 (60.5%) cases maintained the HSR classification (median age 53 years; 56% males) and the main diagnoses included HSR to Hymenoptera venom (29.9%), food allergy (29.1%), and pharmaceutical drugs (25.5%). Anaphylaxis was assumed on-site in 44 (34.7%) cases, in the hospital emergency department in 53 cases (41.7%) and by investigators in 76 (59.8%) cases. Regarding management, epinephrine was administered on-site in 50 cases (39.4%). Conclusions. The main reason for prehospital requesting assistance was HSR to Hymenoptera venom. A high proportion of incidents met the criteria for anaphylaxis and despite the inherent difficulties of the prehospital setting, many of the on-site diagnoses agreed with the criteria. Regarding management, epinephrine was underused in this setting. Referral to specialized consultation is crucial for the management of prehospital incidents.

Use of granules and free salts for Fe and Zn fortification of leafy vegetables: Improvements in trace element bioaccessibility and fulfillment of Dietary Reference Intakes.

BACKGROUND: Leafy vegetables represent an excellent dietary source of trace elements such as Fe and Zn. Nevertheless, Fe and Zn bioaccessibility can lessen due to a high concentration of anti-nutritional compounds. The encapsulation of Fe and Zn salts as granules could be used to fortify these leafy vegetables. METHOD: Three leafy vegetables, spinach, Swiss chard and Ethiopian mustard were fortified with iron sulfate and zinc sulfate as granules and free salts in order to test the improvements in the bioaccessibility and fulfillments of DRIs. Fe and Zn granules were prepared in a fluidized bed granulator. A probabilistic analysis was performed, using experimental data, to assess bioaccessible intake and fulfillments of DRIs in European populations. RESULTS: Fe contents ranged between 4.8 mg/100 g of Ethiopian mustard to 157.4 mg/100 g of spinach. Fe and Zn bioaccessibility percentages were low for Swiss chard and spinach without fortification. Fortification with granules improved Fe bioaccessibility of these latter vegetables (196 and 223 mg/100 g). Zn contents in samples without fortification ranged between 2.3 mg/100 g for Ethiopian mustard and 7.4 mg/100 g for spinach. Zn fortification as granules improved Zn bioaccessibility for the three vegetables studied. Thus, Zn bioccessible concentrations ranged between 17.4 and 108 mg/100 g for the solubility assay and between 5.9 and 31.1 mg/100 g for the dialyzability assay. Besides, the probability analysis showed that fortification had a better performance in meeting DRIs for those populations with higher consumption levels of leafy vegetables. CONCLUSIONS: The probability analysis demonstrated that fortification can be a suitable strategy to meet DRIs for both trace elements, which was especially remarkable for Fe. Fortification with granule was more effective in most the cases, although for Ethiopian mustard, free salt of Fe showed a better performance.

Nanoscale friction on MoS2/graphene heterostructures.

Stacked hetero-structures of two-dimensional materials allow for a design of interactions with corresponding electronic and mechanical properties. We report structure, work function, and frictional properties of 1 to 4 layers of MoS2 grown by chemical vapor deposition on epitaxial graphene on SiC(0001). Experiments were performed by atomic force microscopy in ultra-high vacuum. Friction is dominated by adhesion which is mediated by a deformation of the layers to adapt the shape of the tip apex. Friction decreases with increasing number of MoS2 layers as the bending rigidity leads to less deformation. The dependence of friction on applied load and bias voltage can be attributed to variations in the atomic potential corrugation of the interface, which is enhanced by both load and applied bias. Minimal friction is obtained when work function differences are compensated.

Supramolecular porphyrin as an improved photocatalyst for chloroform decomposition.

In this work, the outlying decoration of the free-base meso-(4-tetra) pyridyl porphyrin (H2TPyP) with the RuCl(dppb)(5,5'-Me-bipy) ruthenium complex (here named Supra-H2TPyP) is observed as an improved molecular photocatalyst for dye-mediated chloroform (CHCl3) decomposition via one-photon absorption operating in the visible spectral range (532 nm and 645 nm). Supra-H2TPyP offers a better option for CHCl3 photodecomposition when compared to the same process mediated by pristine H2TPyP, which requires either excited-state- or UV absorption. The chloroform photodecomposition rates for Supra-H2TPyP as well as its excitation mechanisms are explored as a function of distinct laser irradiation conditions.

Poly(ethylene terephthalate), Poly(butylene terephthalate), and Polystyrene Oligomers: Occurrence and Analysis in Food Contact Materials and Food.

Polyesters (PES) and polystyrene (PS) are among the most used plastics in the production of food contact materials (FCM). The existence of compounds that could migrate from these materials into food requires a constant analytical control to ensure the safety of consumers due to consumption. It also implies a significant research challenge for their identification and quantification. One of the most important groups of known FCM migrants are the substances known as oligomers. PES and PS oligomers have long been suspected to possess some toxicological effects. The International Agency for Research on Cancer and the European Food Safety Authority alerted recently to the potential carcinogenicity of styrene, with its oligomers consequently being also in the spotlight. At the same time, PES cyclic oligomers are categorized as having Cramer III toxicity. Many recent works on the occurrence of poly(ethylene terephthalate) (PET), poly(butylene terephthalate) (PBT), and PS oligomers in FCM and food have been published. The oligomeric chemical analysis requires the use of demanding analytical strategies to address their different physicochemical characteristics (melting points, octanol/water partition coefficients, and solubility properties). Chromatographic methods are normally preferred due to the intrinsic complexity of the target matrices, but the reduced amount of reliable analytical standards still hinders the widespread screening analysis of oligomers in food. This work presents the most relevant recent studies and analytical methodologies used in the analysis of PET, PBT, and PS oligomers in food and FCM, as well as current and future challenges.

Allergy to lipid transfer proteins (LTP) in a pediatric population.

Summary: Background. Lipid transfer proteins (LTP) are considered important plant food allergens in the Mediterranean area, but little is known about LTP allergy in pediatric age. Our aim was to characterize LTP allergy in children.Methods. We reviewed the clinical data from all children evaluated in our department with LTP allergy. From the 76 patients with LTP allergy, 26c hildren were included, 50% female, median age 10 years (1-17). Symptoms included urticaria in 58% (n = 15), anaphylaxis in 46% (n = 12) and OAS in 42% (n = 11). Results. Multiple reactions with different foods occurredin 69%. Cofactors were reported in 27% (n = 7). All patients had positive SPT to peach LTP extract and sIgE Pru p 3. No association between the occurrence of severe reactions and sIgE to Pru p 3 (p = 0.462), sIgE to Cor a 8(p = 0.896), SPT to peach LTP extract (p = 0.846) or the number of positive SPT to fruits/tree nuts (p = 0.972; p = 0.676) was found. Ninety-two percent of the patients tolerated fruits from Rosacea family without peel. Twelve percent reported reactions to new LTP containing foods during follow-up. LTP allergy can occur since early childhood. Conclusions. Since anaphylaxisis common and cofactors act as severity enhancers, it is fundamental to recognizeLTP allergy in children. Currently available diagnostic tests (SPT and sIgE) cannot accurately predict food tolerance or anticipate reaction severity.