Miracle-dz/UniFAR-FTUs huggingface dataset:
Efficient secretory production of large‐size heterologous enzymes in Bacillus subtilis: A secretory partner and directed evolution

Secretory production of recombinant proteins provides a simple approach to the production and purification of target proteins in the enzyme industry. We developed a combined strategy for the secretory production of three large‐size heterologous enzymes with a special focus on 83‐kDa isoamylase (IA) from an archaeon Sulfolobus tokodaii in a bacterium Bacillus subtilis. First, a secretory protein of the B. subtilis family 5 glycoside hydrolase endoglucanase (Cel5) was used as a fusion partner, along with the NprB signal peptide, to facilitate secretory production of IA. This secretory partner strategy was effective for the secretion of two other large enzymes: family 9 glycoside hydrolase from Clostridium phytofermentas and cellodextrin phosphorylase from Clostridium thermocellum. Second, the secretion of Cel5‐IA was improved by directed evolution with two novel double‐layer Petri‐dish‐based high‐throughput screening (HTS) methods. The high‐sensitivity HTS relied on the detection of high‐activity Cel5 on the carboxymethylcellulose/Congo‐red assay. The second modest‐sensitivity HTS focused on the detection of low‐activity IA on the amylodextrin‐I2 assay. After six rounds of HTS, a secretory Cel5‐IA level was increased to 234 mg/L, 155 times the wild‐type IA with the NprB signal peptide only. This combinatory strategy could be useful to enhance the secretory production of large‐size heterologous proteins in B. subtilis.
Xylem Vessel Anatomy of Sweet Cherries Grafted onto Dwarfing and Nondwarfing Rootstocks

Xylem vessel anatomy was examined in tissues surrounding the graft union of sweet cherry (Prunus avium L.) scion (stem) and nondwarfi ng, semi-dwarfi ng, or dwarfi ng rootstock (root) combinations, to characterize potential changes in anatomical features during the initial stages of graft union formation. Vessel element length, frequency, diameter, lumen area (LAV), and mean vessel hydraulic diameter (VDh) were examined in 'Rainier' (P. avium) scion wood grafted onto nondwarfi ng 'Colt' (P. pseudocerasus L. x P. avium) or 'F 12/1' (P. avium) rootstock and semi-dwarfi ng 'Gisela 6' ('Gi 6' (P. cerasus L. x P. canescens L.)), or dwarfi ng 'Gisela 5' ('Gi 5' (P. cerasus x P. canescens)) rootstock systems in: heterograft combinations (commercial scion-rootstock combinations); homografts (scion and rootstock are the same genetic material); and reciprocal heterografts (rootstock tissue grafted onto scion tissue). Vessel element length was not affected by rootstock, but vessel frequency and lumen area in graft union tissues were smaller in 'Rainier'/'Gi 5' (dwarfi ng combination) than in 'Rainier'/'Colt' (nondwarfi ng combination). The heterograft combination of 'Rainier'/'Gi 5' had smaller scion LAV, lower VDh and narrower vessels than homograft or reciprocal heterograft combinations. As callus differentiated into vascular elements, xylem rays in 'Rainer'/'Gi 5' tended to develop at an acute angle to the longitudinal axis of the tree and there was an increase in nonfunctional phloem in 'Rainer'/'Gi 5' compared to 'Rainer'/'F 12/1'. Collectively, the data provides further evidence that a combination of smaller and fewer vessels in the scion and graft union, as well as irregular vessel orientations in the vascular tissue within dwarfi ng combinations could contribute to hydraulic resistance in the graft union resulting in reduced scion growth (dwarfi ng).
DOCKING STUDIES AND SYNTHESIS OF N-MANNICH BASES OF ISATIN DERIVATIVES AND THEIR BIOLOGICAL ACTIVITY

A series of different schiffs bases of 5-substituted-1H indole-2,3 dione were synthesized by refluxing ethanol with different aromatic primary amine and hydrazine.Then a series of different mannich bases were synthesized from schiffs bases of different 5-substituted-1H indole-2,3 dione with different secondary aromatic amine in presence of formaldehyde. In the present study designed for docking studies considered that derivatives of isatin demonstrated protective activity against convulsion. Isatin is a versatile starting material for the synthesis of number of organic compounds. Structures of the synthesized compounds were confirmed by FT-IR, 1 H-NMR, 13 C and mass spectral data.
Meristic and Morphometric Characteristics of Spirlin, Alburnoides bipunctatus Bloch 1782 (Actinopterygii: Cyprinidae) of the Letca Area - Someș River

Abstract Spirlin (Alburnoides bipunctatus) is an important element of the trophic chain of aquatic ecosystems in Romaniaâ€™s hilly areas. To assess the conservation status of this species, 21 specimens were taken from the Letca area, SAƒlaj County. The specimens caught were weighed and analysed from the meristic (8 determinations) and morphometric (36 determinations) points of view. The obtained results reflect the homogeneity of the fish population in terms of morphology (their coefficients of variation showing lower values). This study will continue in the future with the analyses of the spirlin populations from several sectors of the SomeE™ River and its tributaries.
Perinatal risk factors for congenital hypothyroidism

Abstract Congenital hypothyroidism (CH) is one of the most common neonatal endocrine diseases. This retrospective cohort study aimed to identify the potential perinatal risk factors for CH and to differentiate between transient and permanent CH (TCH and PCH, respectively) as well as determine their prevalence in a southeastern Chinese population. This study was based on an 18-year surveillance of a neonatal CH screening program in a large tertiary hospital. A retrospective review of the maternal and neonatal perinatal exposures was conducted. Of the 205,834 newborns screened between 2000 and 2018, 189 were diagnosed with CH (1/1089). Among the 131 CH patients who again underwent thyroid function testing (TFT) after discontinuation of levothyroxine at the age of 3 years, 61 (46.6%) were diagnosed with PCH and 70 (53.4%) were diagnosed with TCH. In the maternal characteristics model, women aged 35 years or older and those who had thyroid disease and/or diabetes mellitus during pregnancy had increased risk of having an offspring with CH (P = .001, .000, and .001, respectively). Significant associations were found with regard to parity and the risk of CH in the offspring (P = .000). In the neonatal characteristics model, infants with female sex, preterm birth, post-term birth, low birth weight, other birth defects, and those born as part of multiple births (P = .011, .034, .001, .000, .000, and .003, respectively) had increased risk of CH. The rate of newborns with other birth defects was higher in the PCH group than that in the TCH group (P = .008), whereas the rate of maternal thyroid disease, newborns with low birth weight, and newborns with preterm birth was higher in the TCH group than that in the PCH group (P = .041, .020, and .013, respectively). The levothyroxine dose (μg/kg/day) at 1 year, 2 years, and 3 years old was significantly lower in the TCH group than that in the PCH group (P = .000, .000, and .000, respectively). Perinatal factors should be considered during the diagnosis and treatment of CH.
Antibacterial and cytotoxic antibacterial potential of ethanol extract and fractions from Aristolochia galeata Mart. ex Zucc

The aim of this investigation was to evaluate the phytochemical, antibacterial and cytotoxic activities of ethanol extract and hexane, dichlorometane, ethyl acetate and hydroethanolic fractions from Aristolochia galeata’s rhizomes. The minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) were evaluated by the broth microdilution assay to investigate the antibacterial activity of various extracts and fractions of A. galeata against Gram-positive and Gram-negative bacteria. The cytotoxicity of plant samples was evaluated in human cervix carcinoma cell line (HeLa) by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. The phytochemical study showed the presence of main secondary metabolites, steroids, flavonoids, coumarins and alkaloids. The ethanol extract and their fractions presented antimicrobial activity against Gram-positive bacteria, and Staphylococcus aureus was regarded the most sensitive strain with MIC of 250 µg/ml for the ethanol extract. The dichlorometane fraction showed bactericidal activity with the value of 1250 µg/ml and moderate cytotoxicity in front of the HeLa cell line tested (CC50 = 90 µg/ml). The results showed that A. galeata had effective antibacterial activity against Gram-positive bacteria and compounds extracted from A. galeata Mart. ex Zucc could be used as possible antimicrobials. The good antimicrobial activity and the low cytotoxicity presented by the hexane fraction can be promised for the new molecules with antibiotic activity.
A Single Load of Fructose Attenuates the Risk of Exercise-Induced Hypoglycemia in Adults With Type 1 Diabetes on Ultra-Long-Acting Basal Insulin: A Randomized, Open-Label, Crossover Proof-of-Principle Study

OBJECTIVE While the adjustment of insulin is an established strategy to reduce the risk of exercise-associated hypoglycemia for individuals with type 1 diabetes, it is not easily feasible for those treated with ultra-long-acting basal insulin. The current study determined whether pre-exercise intake of fructose attenuates the risk of exercise-induced hypoglycemia in individuals with type 1 diabetes using insulin degludec. RESEARCH DESIGN AND METHODS Fourteen male adults with type 1 diabetes completed two 60-min aerobic cycling sessions with or without prior intake (30 min) of 20 g of fructose, in a randomized two-period crossover design. Exercise was performed in the morning in a fasted state without prior insulin reduction and after 48 h of standardized diet. The primary outcome was time to hypoglycemia (plasma glucose ≤3.9 mmol/L) during exercise. RESULTS Intake of fructose resulted in one hypoglycemic event at 60 min compared with six hypoglycemic events at 27.5 ± 9.4 min of exercise in the control condition, translating into a risk reduction of 87.8% (hazard ratio 0.12 [95% CI 0.02, 0.66]; P = 0.015). Mean plasma glucose during exercise was 7.3 ± 1.4 mmol/L with fructose and 5.5 ± 1.1 mmol/L in the control group (P < 0.001). Lactate levels were higher at rest in the 30 min following fructose intake (P < 0.001) but were not significantly different from the control group during exercise (P = 0.32). Substrate oxidation during exercise did not significantly differ between the conditions (P = 0.73 for carbohydrate and P = 0.48 for fat oxidation). Fructose was well tolerated. CONCLUSIONS Pre-exercise intake of fructose is an easily feasible, effective, and well-tolerated strategy to alleviate the risk of exercise-induced hypoglycemia while avoiding hyperglycemia in individuals with type 1 diabetes on ultra-long-acting insulin.
Type VI secretion system effector proteins: effective weapons for bacterial competitiveness.

The Type VI secretion system (T6SS) is a protein translocation nanomachine widespread amongst Gram-negative bacteria and used as a means to deliver effectors directly into target bacterial or eukaryotic cells. These effectors have a wide variety of functions within target cells which ultimately help the secreting cell gain a competitive fitness advantage. Here we discuss the different ways in which these effectors can be delivered by the T6SS and the diverse mechanisms by which they exert their noxious action upon recipient cells. We also highlight the existence of roles for T6SS effectors beyond simply the killing of neighbouring cells. This article is protected by copyright. All rights reserved.
Root exudation (net efflux of amino acids) may increase rhizodeposition under elevated CO2

Increases in atmospheric CO2 concentration ([CO2]) can lead to global climate change and theoretically could enhance carbon (C) deposition in soil, but data on this complex issue are contradictory. One approach for clarifying the diverse forces influencing plant‐derived C in the rhizosphere involves defining how elevated [CO2] alters the fundamental process of C transfer from plant roots to the soil. We examine here how a step increase in [CO2] affects the innate influx and efflux components of root exudation in axenic plants, as one foundation for understanding how climate change may affect rhizodeposition. Increasing [CO2] from 425 to 850 μmol mol−1 during short‐term trials enhanced shoot and root dry weight (P<0.01) of annual rye grass (Lolium multiflorum Lam.) and medic (Medicago truncatula L.) but had no effect on growth of maize (Zea mays L.). Root amino‐acid flux in the same plants changed only in maize, which increased the efflux rate (nmol g root fresh weight−1 h−1) of six amino acids (arginine, alanine, proline, tyrosine, lysine and leucine) significantly (P<0.05) under elevated [CO2]. None of the three plant species altered the steady‐state concentration of 16 amino acids released into a hydroponic solution with changing [CO2], apparently because amino‐acid influx rates, measured at 2.5 μm, consistently exceeded efflux rates. Indeed, plants recovered amino acids at rates 94–374% higher than they were lost from roots regardless of [CO2]. These results indicate that, in theory, any effect of [CO2] doubling on amino‐acid efflux can be offset by innately higher rates of influx. In practice, however, higher rates of amino‐acid cycling (i.e., efflux+influx) for each root segment (in C4 maize) or from more root tissue (in the two C3 species) should increase root exudation by plants exposed to elevated [CO2] as additional amino acids would be adsorbed to soil particles or be taken up by soil microorganisms.
The prediction of body composition in live ewes in early lactation from live weight and estimates of gut contents and total body water

SUMMARY The efficacy of estimates of gut contents and total body water in increasing the precision with which the chemical composition of the body could be estimated in early lactation was evaluated in 36 Finnish Landrace × Dorset Horn ewes. The ewes were fed at two levels in pregnancy, and, in lactation, given diets of two metabolizable energy concentrations. The allometric relationships relating weight of chemical fat and protein to emptybody weight were not affected by treatment or stage of lactation. Inclusion of an index of gut contents, based on dry-matter intake, indigestibility and retention time of food residues, together with live weight in a regression equation predicting weight of body fat, only slightly increased the precision of estimate compared with equations using live weight alone. There was a close negative relationship between the proportions of water and fat in live weight. Inclusion of weight of body water with live weight in a regression equation predicting weight of body fat markedly increased the precision of estimate and the residual error (0·81 kg) was similar at different stages of lactation. However, when deuterium oxide space was used instead of body water there was only a small increase in precision of estimate and the residual error varied from 5·3 kg in early lactation to 2·1 kg in mid-lactation. The relationship between deuterium oxide space and body water was shown to be variable and altered by stage of lactation, and these differences were associated with differences in rate of water turnover in the animal's body. It is concluded that estimates of body water are unsuitable for estimating weight of body fat in early lactation.
Dual brain stimulation enhances interpersonal learning through spontaneous movement synchrony

Abstract Social interactive learning denotes the ability to acquire new information from a conspecific—a prerequisite for cultural evolution and survival. As inspired by recent neurophysiological research, here we tested whether social interactive learning can be augmented by exogenously synchronizing oscillatory brain activity across an instructor and a learner engaged in a naturalistic song-learning task. We used a dual brain stimulation protocol entailing the trans-cranial delivery of synchronized electric currents in two individuals simultaneously. When we stimulated inferior frontal brain regions, with 6 Hz alternating currents being in-phase between the instructor and the learner, the dyad exhibited spontaneous and synchronized body movement. Remarkably, this stimulation also led to enhanced learning performance. These effects were both phase- and frequency-specific: 6 Hz anti-phase stimulation or 10 Hz in-phase stimulation, did not yield comparable results. Furthermore, a mediation analysis disclosed that interpersonal movement synchrony acted as a partial mediator of the effect of dual brain stimulation on learning performance, i.e. possibly facilitating the effect of dual brain stimulation on learning. Our results provide a causal demonstration that inter-brain synchronization is a sufficient condition to improve real-time information transfer between pairs of individuals.
Raised plasma vitellogenin in male wild brown trout (Salmo trutta) near a wastewater treatment plant in ireland

Ovotestis (intersex) and raised plasma vitellogenin in male fish are widely employed biomarkers of estrogencontamination in the aquatic environment. In the present study, these biomarkers were used to determine whether Irish rivers contain estrogenic chemicals at levels capable of affecting the reproductive health and success of exposed fish populations. A number of aquatic ecosystems were investigated (the rivers Liffey, Lee, and Bandon and the Killarney lakes). A survey of male wild brown trout (Salmo trutta) was carried out to assess the incidence of endocrine disruption in the feral fish population; no evidence of intersex was found in any of the wild fish sampled. Raised plasma vitellogenin was detected, however, in the wild brown trout downstream of a major municipal wastewater treatment plant on the river Liffey.
TCR β-Chain Sharing in Human CD8+ T Cell Responses to Cytomegalovirus and EBV1

The CD8+ TCR repertoires specific for many immunogenic epitopes of CMV and EBV are dominated by a few TCR clonotypes and involve public TCRs that are shared between many MHC-matched individuals. In previous studies, we demonstrated that the observed sharing of epitope-specific TCRβ chains between individuals is strongly associated with TCRβ production frequency, and that a process of convergent recombination facilitates the more efficient production of some TCRβ sequences. In this study, we analyzed a total of 2836 TCRβ sequences from 23 CMV-infected and 10 EBV-infected individuals to investigate the factors that influence the sharing of TCRβ sequences in the CD8+ T cell responses to two immunodominant HLA-A*0201-restricted epitopes from these viruses. The most shared TCRβ amino acid sequences were found to have two features that indicate efficient TCRβ production, as follows: 1) they required fewer nucleotide additions, and 2) they were encoded by a greater variety of nucleotide sequences. We used simulations of random V(D)J recombination to demonstrate that the in silico TCRβ production frequency was predictive of the extent to which both TCRβ nucleotide and amino acid sequences were shared in vivo. These results suggest that TCRβ production frequency plays an important role in the interindividual sharing of TCRβ sequences within CD8+ T cell responses specific for CMV and EBV.
CENTRO INTERDISCIPLINARIO DE INVESTIGACIÓN PARA EL DESARROLLO INTEGRAL REGIONAL

In this research work, the effect on capillary water absorption, moisture loss, and chloride diffusion coefficient, resulting from the use of a 3% cactus mucilage solution to replace water in concrete mixtures was evaluated. Also, the effect of the time dependant chloride diffusion coefficient obtained on the initiation period for steel corrosion in a hypothetical concrete structure under specific environmental conditions and exposure to chlorides was evaluated using the software Life 365. The materials used were white Portland cement, cactus mucilage (Opuntia ficus indica), river aggregates and distilled water. Three control concrete mixtures were designed and prepared with water/cement ratios of 0.30, 0.45 and 0.60, and three mixtures with the same water/cement ratios containing a 3% cactus mucilage solution. Specimens measuring 100 mm in diameter and 200 mm in height were cast and cured for zero and seven days at 23 ± 3 o C. Results from drying in a natural environment indicate that the cactus mucilage reduces moisture loss only at a ratio of 0.30. At the ratios of 0.45 and 0.60 the specimens prepared with cactus mucilage lost a higher amount of water compared to control specimens. This is explained by the higher capillary porosity observed in the specimens containing cactus mucilage and the hydration retarding effect of the mucilage. Results of capillary water absorption indicate that the cactus mucilage had a positive effect in all ratios (0.30, 0.45 and 0.60), showing a lower water absorption with respect to control mixtures. With respect to the chloride diffusion, specimens at a ratio of 0.30 and containing cactus mucilage showed a lower diffusion coefficient at all ages for both moist curing regimes. In the ratio 0.45, the cactus mucilage reduces the diffusion coefficient only at 120 days for both moist curing regimes. At a ratio of 0.60, the cactus mucilage reduces the chloride diffusion only when moist curing for seven days. In the prediction of the initiation period for the steel corrosion, the cactus mucilage increased this time by approximately 15 years with respect to the control specimens. This was only observed at a ratio of 0.30 and moist curing for seven days. The increase in other ratios was only marginal.
Reconstruction of Tertiary Metasequoia forests. I. Test of a method for biomass determination based on stem dimensions

Abstract Accurate reconstruction of the biomass, structure, and productivity of ancient forests from their fossilized remnants remains an interesting challenge in paleoecology. In well-preserved Tertiary fossil Metasequoia forests of Canada's Arctic, in situ stumps and fragments of stems, treetops, and branches contain substantial information about tree dimensions that can be used to determine tree height, stand biomass, and other characteristics such as canopy depth and structure, and the history of stand development. To validate a method for reconstructing the biomass of the Eocene floodplain Metasequoia forests of Axel Heiberg Island, we measured stump diameters and spacing, and stem, branch, and treetop characteristics in living Metasequoia glyptostroboides and Chamaecyparis thyoides stands in ways that simulate the limited measurements that can be made in well-preserved fossil forests in Canada and probably elsewhere. We used those limited measurements to estimate tree height and volume, branch and foliar dry weights, and tree biomass. The estimates derived from the limited data set are usually within 15% of the estimates derived from the methods currently used in forest ecology for determining those metrics in modern forests. Under appropriate conditions, the biomass of ancient forests can be estimated with reasonable confidence.