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.

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.

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).

Phytoscreening of Vochysiaceae species: Molecular identification by HPLC-ESI-MS/MS and evaluating of their antioxidant activity and inhibitory potential against human α-amylase and protein glycation.

Scientific research based on medicinal plants has been highlighted as a complementary treatment to T2DM, stand out the Vochysiaceae family, which have been widely used in folk medicine by traditional South American communities to treat some diseases. Our study aimed to investigate the antioxidant and antiglycation activities of ethanol extracts of leaves (LF) and stem barks (SB) of Vochysiaceae species, evaluated their capacities to inhibit glycoside and lipid hydrolases related to T2DM and molecular identification by HPLC-ESI-MS/MS. Our main findings indicate that the ethanolic extract of four of eight analyzed plants such as LF and SB of Q. grandiflora, Q. parviflora, V. elliptica and Calisthene major exhibited, respectively, potential of α-amylase inhibition (IC50 of LF: 5.7 ±â€¯0.6, 4.1 ±â€¯0.5, 5.8 ±â€¯0.5, 3.2 ±â€¯0.6 and IC50 of SB: 3.3 ±â€¯0.7, 6.2 ±â€¯2.0, 121.0 ±â€¯8.6 and 11.2 ±â€¯2.8 µg/mL), capacities of antioxidant (ORAC of LF: 516.2 ±â€¯0.1, 547.6 ±â€¯4.9, 544.3 ±â€¯6.1, 442.6 ±â€¯2.4 and ORAC of SB: 593.6 ±â€¯22.3, 497.7 ±â€¯0.8, 578 ±â€¯12.3, 593.6 ±â€¯19.5 µmol trolox eq/g; FRAP of LF: 796.1 ±â€¯0.9, 427.7 ±â€¯22.0, 81.0 ±â€¯1.9, 685 ±â€¯37.9 and FRAP of SB: 947.4 ±â€¯24.9, 738.6 ±â€¯24.3, 98.8 ±â€¯7.9, 970.8 ±â€¯13.9 µmol trolox eq/g; DPPH IC50 of LF: 14.2 ±â€¯1.8, 36.3 ±â€¯6.9, 11.8 ±â€¯1.9, 13.3 ±â€¯1.2 and DPPH IC50 of SB: 16.0 ±â€¯3.0, 15.5 ±â€¯1.9, 126.1 ±â€¯23. 6, 5.3 ±â€¯0.3 µg/mL, respectively) and antiglycation (BSA/Frutose IC50 of LF: 43.1 ±â€¯3.4, 52.1 ±â€¯6.0, 175.5 ±â€¯32, 8, 111.8 ±â€¯14.7 and BSA/Frutose IC50 of SB:, 40.1 ±â€¯11.9, 51.2 ±â€¯16. 7, 46.6 ±â€¯5.7, 53.5 ±â€¯13.6 µg/mL) and presence of polyphenols, such as flavonoids and condensed tannins. The extracts presented low ability to inhibit α-glycosidase and lipase enzymes in the initial assays, with values below 40% of inhibition. In BSA/methylglyoxal, only Q. grandiflora SB, V. eliptica LF and V. tucanorum LF showed activity (IC50: 655.5 ±â€¯208.5, 401.9 ±â€¯135.2 and 617.1 ±â€¯80.6 µg/mL, respectively) and only C. major LF and SB, in Arg/methylglyoxal (IC50: 485.1 ±â€¯130.8 and 468.0 ±â€¯150.5 µg/ml, respectively). This study presented new findings about the biological and pharmacological potential of some species of Vochysiaceae family, contributing to the understanding of the action and efficacy in use of these plants, in their management of postprandial hyperglycemia and in glycation and oxidative processes that contribute to managing diabetes mellitus.

Lack of mutagenic effect by multi-walled functionalized carbon nanotubes in the somatic cells of Drosophila melanogaster.

Carbon nanotubes (CNTs) are formed by rolling up a single graphite sheet into a tube. Among the different types of CNTs, the multi-walled carbon nanotubes (MWCNTs) comprise a set of concentric nanotubes with perfect structures. Several uses for MWCNTs have been suggested to be included in biological applications such as manufacturing of biosensors, carriers of drugs. However, before these materials can be put on the market, it is necessary to know their genotoxic effects. Thus, this study aims to evaluate the mutagenicity of multi-walled carbon nanotubes (MWCNTs) functionalized in somatic cells of Drosophila melanogaster, using the somatic mutation and recombination test (SMART). This assay detects the loss of heterozygosity of marker genes expressed phenotypically on the wings of the fly. Larvae of three days were used, resulting from ST cross, with basal levels of the cytochrome P450 and larvae of high metabolic bioactivity capacity (HB cross). They were treated with different concentrations of MWCNTs functionalized. The MH descendants, analyzed in both ST and HB crosses, had no significant effects on the frequency of mutant. Based on the results and on the experimental conditions mentioned in this study, it was concluded that MWCNTs were not mutagenic in D. melanogaster.

On the characterization of energy networks of proteins.

The construction of a realistic theoretical model of proteins is determinant for improving the computational simulations of their structural and functional aspects. Modeling proteins as a network of non-covalent connections between the atoms of amino acid residues has shown valuable insights into these macromolecules. The energy-related properties of protein structures are known to be very important in molecular dynamics. However, these same properties have been neglected when the protein structures are modeled as networks of atoms and amino acid residues. A new approach for the construction of protein models based on a network of atoms is presented. This method, based on interatomic interaction, takes into account the energy and geometric aspects of the protein structures that were not employed before, such as atomic occlusion inside the protein, the use of solvation, protein modeling and analysis, and the use of energy potentials to estimate the energies of interatomic non-covalent contacts. As a result, we achieved a more realistic network model of proteins. This model has the virtue of being more robust in face of different unknown variables that usually are arbitrarily estimated. We were able to determine the most connected residues of all the proteins studied, so that we are now in a better condition to study their structural role.

On the characterization of energy networks of proteins

The construction of a realistic theoretical model of proteins is determinant for improving the computational simulations of their structural and functional aspects. Modeling proteins as a network of non-covalent connections between the atoms of amino acid residues has shown valuable insights into these macromolecules. The energy-related properties of protein structures are known to be very important in molecular dynamics. However, these same properties have been neglected when the protein structures are modeled as networks of atoms and amino acid residues. A new approach for the construction of protein models based on a network of atoms is presented. This method, based on interatomic interaction, takes into account the energy and geometric aspects of the protein structures that were not employed before, such as atomic occlusion inside the protein, the use of solvation, protein modeling and analysis, and the use of energy potentials to estimate the energies of interatomic non-covalent contacts. As a result, we achieved a more realistic network model of proteins. This model has the virtue of being more robust in face of different unknown variables that usually are arbitrarily estimated. We were able to determine the most connected residues of all the proteins studied, so that we are now in a better condition to study their structural role.

Thermodynamic evaluation and modeling of proton and water exchange associated with benzamidine and berenil binding to beta-trypsin.

Serine-proteases are involved in vital processes in virtually all species. They are important targets for researchers studying the relationships between protein structure and activity, for the rational design of new pharmaceuticals. Trypsin was used as a model to assess a possible differential contribution of hydration water to the binding of two synthetic inhibitors. Thermodynamic parameters for the association of bovine beta-trypsin (homogeneous material, observed 23,294.4 +/- 0.2 Da, theoretical 23,292.5 Da) with the inhibitors benzamidine and berenil at pH 8.0, 25 degrees C and with 25 mM CaCl2, were determined using isothermal titration calorimetry and the osmotic stress method. The association constant for berenil was about 12 times higher compared to the one for benzamidine (binding constants are K = 596,599 +/- 25,057 and 49,513 +/- 2,732 M(-1), respectively; the number of binding sites is the same for both ligands, N = 0.99 +/- 0.05). Apparently the driving force responsible for this large difference of affinity is not due to hydrophobic interactions because the variation in heat capacity (DeltaCp), a characteristic signature of these interactions, was similar in both systems tested (-464.7 +/- 23.9 and -477.1 +/- 86.8 J K(-1) mol(-1) for berenil and benzamidine, respectively). The results also indicated that the enzyme has a net gain of about 21 water molecules regardless of the inhibitor tested. It was shown that the difference in affinity could be due to a larger number of interactions between berenil and the enzyme based on computational modeling. The data support the view that pharmaceuticals derived from benzamidine that enable hydrogen bond formation outside the catalytic binding pocket of beta-trypsin may result in more effective inhibitors.

Thermodynamic evaluation and modeling of proton and water exchange associated with benzamidine and berenil binding to ß-trypsin

Serine-proteases are involved in vital processes in virtually all species. They are important targets for researchers studying the relationships between protein structure and activity, for the rational design of new pharmaceuticals. Trypsin was used as a model to assess a possible differential contribution of hydration water to the binding of two synthetic inhibitors. Thermodynamic parameters for the association of bovine ß-trypsin (homogeneous material, observed 23,294.4 ± 0.2 Da, theoretical 23,292.5 Da) with the inhibitors benzamidine and berenil at pH 8.0, 25°C and with 25 mM CaCl2, were determined using isothermal titration calorimetry and the osmotic stress method. The association constant for berenil was about 12 times higher compared to the one for benzamidine (binding constants are K = 596,599 ± 25,057 and 49,513 ± 2,732 M-1, respectively; the number of binding sites is the same for both ligands, N = 0.99 ± 0.05). Apparently the driving force responsible for this large difference of affinity is not due to hydrophobic interactions because the variation in heat capacity (DCp), a characteristic signature of these interactions, was similar in both systems tested (-464.7 ± 23.9 and -477.1 ± 86.8 J K-1 mol-1 for berenil and benzamidine, respectively). The results also indicated that the enzyme has a net gain of about 21 water molecules regardless of the inhibitor tested. It was shown that the difference in affinity could be due to a larger number of interactions between berenil and the enzyme based on computational modeling. The data support the view that pharmaceuticals derived from benzamidine that enable hydrogen bond formation outside the catalytic binding pocket of ß-trypsin may result in more effective inhibitors.

Physico-chemical characterization of meglumine antimoniate.

The leishmanicidal drug, meglumine antimoniate (MA), has been synthesized by the reaction of antimony oxyhydrated and N-methyl glucamine. Infrared and solid state NMR 13C analysis of MA and the ligand strongly suggests that antimony binds to N-methyl glucamine through the oxygen of C-3 carbon. Potentiometric titration indicated that, between pH 4.5 and 7.5, MA exists in the zwitterionic form.

Reparacao de ferimento cardiaco:ocorrencia de dois episodios no mesmo paciente. / Repair of cardiac injury: ocurrence of two episodes in the same patient

Os autores relatam o caso de um paciente que por duas vezes foi vitima de ferimentos cardiacos com perfuracao de ventriculo esquerdo, tendo em ambas as ocasioes sido submetido a cirurgia de urgencia em um hospital de interior, sem equipamentos especializados ou medicos treinados em cirurgia cardiaca. Houve pleno sucesso em ambas as cirurgias e o paciente manteve-se vivendo normalmente apesar das sequelas remanescentes que, mais tarde, foram reveladas pela cineangiocoronariografia (obstrucao da coronaria descendente anterior esquerdo a aneurisma de ventriculo esquerdo)