Hemolysin II (HlyII) – one of the pathogenic factors of Bacillus cereus, a pore-forming β-barrel toxin – possesses a C-terminal extension of 94 amino acid residues, designated as the C-terminal domain of HlyII (HlyIICTD), which plays an important role in the functioning of the toxin. Our previous work described a monoclonal antibody (HlyIIC-20), capable of strain-specific inhibi-tion of hemolysis caused by HlyII, and demonstrated the dependence of the efficiency of hemol-ysis on the presence of proline at position 324 in HlyII outside the conformational antigenic de-terminant. In this work, we studied 16 mutant forms of HlyIICTD. Each of the mutations, ob-tained by multiple site-directed mutagenesis leading to the replacement of amino acid residues lying on the surface of the 3D structure of HlyIICTD, led to a decrease in the interaction of HlyIIC-20 with the mutant form of the protein. Changes in epitope structure confirm the high conformational mobility of HlyIICTD required for the functioning of HlyII. Comparison of the effect of the introduced mutations on the effectiveness of interaction between HlyIICTD and HlyIIC-20 and a control antibody recognizing a non-overlapping epitope enabled identifying the amino acid residues N339, K340 included in the conformational antigenic determinant rec-ognized by HlyIIC-20.

At 12:28 UTC (8:28 PM, local time) on 15 January 2023, a shallow, moderate earthquake with a magnitude (Mw) of 4.7 and a depth of one (1) kilometer shook the northern part of Leyte Island in the central part of the Philippines. The ground shaking was felt at PHIVOLCS Earthquake Intensity Scale (PEIS) VI, (very strong) equivalent to Modified Mercalli Intensity (MMI) VI in areas along the strike of the Leyte segment of the Philippine Fault. Even though this earthquake is classified as moderate in size, significant geologic, structural, and socio-economic impacts were documented. Most of the earthquake impacts that include surface rupture, liquefaction, and mass movements are limited near the epicentral area. The earthquake produced an 8-km-long surface rupture, the first documented rupture generated by a magnitude <6 along the Philippine Fault, and is unusual in other surface rupturing earthquakes worldwide. It injured 18 persons and damaged 434 residential buildings and 26 public infrastructures which amounts to about 500 thousand US dollars. Structural damages were caused by surface rupture and/or poor engineering construction practices. Analysis of the seismic waveform and recorded peak ground acceleration (PGA) suggests that the subsurface geology influenced the ground motion amplification in an area near the epicenter. Based on the hypocenter, focal mechanism solution, aftershock distribution, and actual field observation, the causative fault for this event is the northern end of the Philippine Fault – Leyte segment. Moderate in size, this earthquake emphasizes the need and importance of documenting moderate-sized earthquakes as a tool and guide for medium and long-term earthquake risk assessment and resiliency.

Plasminogen (Plg) is the inactive form of plasmin (Plm) that exists in two major glycoforms, referred to as glycoforms I and II (GI and GII). In the circulation, Plg assumes an activation-resistant 'closed' conformation via interdomain interactions and is mediated by the lysine binding site (LBS) on the kringle (KR) domains. These inter-domain interactions can be readily disrupted when Plg binds to lysine/arginine residues on protein targets or free L-lysine and analogues. This causes Plg to convert into an 'open' form which is crucial for activation by host activators. In this study, we investigated how various ligands affect the kinetics of Plg conformational change using small-angle X-ray scattering (SAXS). We began by examining the open and closed conformations of Plg using size-exclusion chromatography (SEC) coupled with SAXS. Next, we developed a high throughput (HTP) 96-well SAXS assay setup. This method enables us to determine the Kopen value, which is used to compare ligands’ effect on Plg conformation directly. Based on our analysis using Plg GII, we have found that the Kopen for ε-aminocaproic acid (EACA) is approximately three times greater than that of Tranexamic acid (TXA), which is widely recognized as a highly effective ligand. We demonstrated further that Plg undergoes a conformational change when it binds to the C-terminal peptides of the inhibitor α2-antiplasmin and receptor Plg‑RKT. Our findings suggest that, besides the C-terminal lysine, internal lysine(s) are also necessary for the formation of open Plg. Finally, we compared the conformational changes of Plg GI and GII directly and found that the closed form of GI, which has an N-linked glycosylation, is less stable. To summarize, we have successfully determined the response of Plg to various ligand/receptor peptides by directly measuring the kinetics of its conformational changes.

This article aims to assess the benefits of floor-slab insulation measures using extruded polystyrene (XPS) and polyisocyanurate (polyiso) insulation materials at various levels of insulation thicknesses for a detached residential building. The EnergyPlus simulation analysis was carried out within the seven energy zones (represented by eight locations) of South Africa in accordance with the South African national code for building energy efficiency (SANS10400-XA). The energy savings and payback periods due to use of the insulation over a lifecycle period of 50 years were assessed. Cape Town (zone 4) behaved differently from other locations and hardly benefitted from the application of floor-slab insulation measures. Generally, polyiso insulation performed better than XPS, for vertical gap insulation. For vertical gap insulation, lower insulation thicknesses required higher insulation depths to maximize energy savings. Similarly, lower insulation thicknesses required higher perimeter insulation widths to maximize energy savings, for the horizontal perimeter insulation method. The locations that benefitted most from vertical gap floor-slab insulation were Pretoria (zone2), Kimberley (zone6), Nelspruit (zone3), Fraserburg (zone7), Welkom (zone1), Mthatha (zone5), Ixopo (zone5H) and Cape Town (zone4) in that order. This order was almost similar with those for the horizontal perimeter floor-slab insulation and horizontal full floor-slab insulation methods.

Genetic engineering technology offers opportunities to improve many important agronomic traits in crops, including insect-resistance. However, genetically modified (GM) exogenous proteins in edible tissues of transgenic crops has become an issue of intense public concern. To advance the application of GM techniques in maize, a Cre/loxP-based strategy was developed for manipulating the transgenes in green tissues while locking it in non-green tissues. In the strategy, the site-specific excision can be used to switch on or off the expression of transgenes at specific tissues. In this work, two basic transgenic maize named KEY carrying the Cre gene and LOCK containing the Vip3A gene with a blocked element, were obtained based on their separate fusion gene cassettes. The expression level and concentration of Vip3A were observed with high specific accumulation in green tissues (leaf and stem), and only a small amount in root and kernel tissues in the KEY × LOCK hybrids. The insect-resistance of transgenic maize against two common lepidopteran pests, Ostrinia furnacalis and Spodoptera frugiperda, was assessed in the laboratory and field. The results indicated that the hybrids possessed high resistance levels against the two pests, with mortality rates above 73.6% and damage scales below 2.4 compared with the control group. Our results suggested that the Cre/loxP-mediated genetic engineering approach has a competitive advantage in GM maize. Overall, findings from this study are significant for providing a feasible strategy for transgenes avoiding expression in edible parts and exploring novel techniques toward the biosafety of GM plants.

Based on the theory that certain geological fault zones surpass the influence of other abiotic and biotic site factors for mound-building red wood ants (Formica rufa group), an area northwest of Tuttlingen with extremely high ant density was systematically surveyed, taking into account official geological maps. Indicators of active fault zones were recorded. These were additionally examined in a model area using a high-resolution digital terrain model (DTM 1-meter). The study area is located in the Swabian Jura at the western edge of the Albstadt shear zone, where recent seismic activity is documented by frequent earthquakes. The earthquakes are considered to be the cause of gas permeability in the strike-slip faults, around which the clustered nesting sites were mapped. More than 2900 nests of the Formica rufa group were mapped and their site conditions analysed in different geomorphological positions. In addition to recording forestry site conditions, a geological classification was carried out, as well as an analysis of soil gases/mofettes in areas with the highest nest densities. The correlation between forestry data and nest sites did not reveal a consistent pattern. In contrast, the relationships with gas-permeable strike-slip faults, though predominantly covered with Quaternary sediments, could be identified through CO₂- and CH₄-rich gas emissions, mofettes, and doline features. The interpretations were complicated due to different mapping ages and signatures of the geological maps. Fortunately, this issue could be analysed and evaluated in a sub-area with mappings from 1936/71 and 1995 regarding presumed or confirmed fault trajectories. New insights were gained regarding the relationships between Formica nest sites, gas-permeable faults, and groundwater levels on narrow mountain ridges with steep relief. Here, the lateral emission of CO₂ depends on the groundwater level and does not occur vertically directly above the fault. C- and H-isotope data of CO₂ and CH₄ do not show a consistent source. While methane is mainly derived from biological production by Archaea in the upper crust and in swamp areas, CO₂ mostly originates from the mantle. The results of the investigations revealed that a multitude of geological and ecological factors must be considered for the interpretation of the occurrences of Formica sites northwest of Tuttlingen. These factors confer a distinct originality to this region, contributing not only to an enhanced understanding of site conditions but also furnishing information regarding environmentally relevant gas emissions.

This study examines the capacity of a Site-Specific Fodder Management Decision Support System (SSFM-DSS) model to assist in successful cultivation of sericea lespedeza [SL; Lespedeza cuneata (Dum.-Cours.) G. Don.], with a specific focus on small-scale agricultural systems in the southeastern United States (U.S.). The study emphasizes incorporating advanced geospatial technologies, such as Geographic Information Systems (GIS), remote sensing, and Global Navigation Satellite Systems (GNSS), to improve fodder production. The research showcases the versatility of SL under varying environmental conditions, underscoring its significance in promoting sustainable livestock production. The methodology integrates empirical field data, geospatial analysis, and predictive modeling to formulate an SSFM strategy customized to address the distinct difficulties climate change presents, such as sudden and severe drought conditions. The study introduces an automated geospatial model for assessing the appropriateness of SL production throughout Alabama, Georgia, and South Carolina. The model uses many environmental criteria, including soil properties, climate fluctuations, and terrain. Furthermore, the research was instrumental in development of a WebGIS Dashboard, which offers farmers a decision assistance system to enhance the sustainable production of SL. The results highlight the significant impact that a SSFM-DSS can have on promoting sustainable agricultural practices, providing a solution to address the issues of food security and environmental concerns in agriculture.

Introduction: Infection of a wound during the thirty days after surgery is considered a surgical site infection (SSI). Because of lack of large studies or documented guidelines for SSI after surgery, we aimed to study risk factors and comorbidities that may lead to SSI.Methods: A cross-sectional study was conducted in one month (May, 2023) included 111 patients. All patients underwent to either abdominal or urological surgeries. Inclusion criteria were: patients aged 18 or older, any type of abdominal or urological open surgery, and no documented history of infection at surgical site infection.Results: we found 24 (21.6 %) patients out of 111 developed SSI after surgery. Male predominance was clear in this study. The number of patients with age 40 years or less was 60 (54 %) and 10 of them developed SSI (14.2 %).Patients who stayed more than 24 hour of preoperative at hospital were less in number than patients who did not (36.9% vs. 63%; p=0.002) respectively. Patients with emergency procedure were 12 patients (10.8 %) in contrast to 99 patients (89.1 %) who underwent a planned surgery (p-value= 0.003). Conclusion: our study revealed a higher incidence of SSI after surgery. Older patients with comorbidities, preoperative stay, emergency surgery, male sex are all correlated to the developing SSI.

In the world's first pig-to-human cardiac xenotransplant, the elevated levels of porcine cytomeg-alovirus (PCMV) are considered key factors contributing to patient mortality. This has renewed attention to PCMV, a virus widely prevalent in pigs. Currently, there are no effective drugs or vaccines targeting PCMV, and its high detection difficulty poses challenges for prevention and control research. In this study, antiviral small hairpin RNA (shRNA) was selected and inserted into the Rosa26 and miR-17-92 loci of pigs via a CRISPR/Cas9-mediated knock-in strategy. Further in vitro viral challenge experiments demonstrated that these genetically edited pig cells could effec-tively limit PCMV replication. Through this process, we constructed a PCMV-infected cell model, validated partial viral interference sites, enhanced gene knock-in efficiency, performed gene editing at two different gene loci, and ultimately demonstrated that RNA interference (RNAi) technology combined with CRISPR/Cas9 has the potential to generate pig cells with enhanced antiviral infec-tion capabilities. This opens up possibilities for the future production of pig populations with anti-viral functionalities.

Balsam fir trees are a specialty agricultural species that are largely used as Christmas trees. Soil or tissue sampling is seldom performed to assess balsam fir nutrition needs, which can contribute to over or under fertilization and subsequent environmental challenges. Remote sensing to determine the normalized difference vegetation index (NDVI) has had strong relationships with nutrition in other plants. This research aimed to determine whether there is a relationship between NDVI and nutrition in balsam fir. A random sample of 45 trees was selected in autumn of 2021 and then another 70 trees selected in spring of 2022. Soil and needle tissue samples were analyzed for N, P, K, Ca, Mg, S, Fe, Zn, Mn, B, and Cu. An uncrewed aerial vehicle equipped with a multispectral camera was flown over the orchard to determine the NDVI for each tree. Nutrition and NDVI were analyzed for correlations through all sampled trees. Separate correlation analyses were repeated for mature trees (greater than 5 years old) and young trees (less than 5 years old). Soil nutrients N, P, K had a significant relationship with NDVI in autumn sampling. Only N was correlated consistently in both autumn and spring, with R2 of 37% and 33% respectively. Relationships were much stronger when only mature trees were considered, increasing to 60% and 51% respectively. NDVI was found to offer predictive power for tissue N that can be useful to producers to help optimize fertilizer input. However, further work is needed to refine these relationships.

Soil, geochemical, microbiological and archaeological studies were conducted at eight settlements dating from the Paleolithic to Late Medieval and New Times near the Southern Trans-Urals mountains, Russia. The forest-steppe landscapes, rivers and abundance of mineral resources have attracted people to region since ancient times. Properties of cultural layer (CLs): close to neutral pH, well structured, absence of salinity, enrichment with exchangeable calcium and anthropogenic phosphorus (0.2–0.4%). The majority of CLs start from 3–25 cm and last up to 40–60 cm and contain 6–10% organic carbon (Corg), in a layer of 0–20 cm because modern processes are developing here, at Ishkulovo site (around 14th century) Corg decreases to 1.3%, because CL is below 80 cm and in the absence of fresh organic material it is mineralized. The content of microbial biomass of the 0–10 cm layer is 520–680 μg С/g of soil, microbes cause the emission of 0.2–1.0 μg C–CO2/g soil per h. The proximity of the studied sites to deposits of copper, chromium, zinc, manganese and Magnitogorsk Metallurgical and Uchaly processing plants creates geochemical anomalies of soils. Pollutants are also released into the CLs from bronze products, ore, ceramic fragments and raw materials from their manufacture. It leads to the concentration of pollutants of the 1st (arsenic, lead, zinc) and 2nd (cobalt, copper, nickel, chromium) hazard classes in the studied CLs, often in quantities higher than the clarke and less often – more than the allowable content for human health.

Celiac disease is an autoimmune disease triggered by oral ingestion of gluten, with certain gluten residues resistant to digestive tract enzymes. Within the duodenum, the remaining peptides incite immunogenic responses, including the generation of autoantibodies and inflammation, leading to irreversible damage. Our previous exploration unveiled a glutenase called Bga1903 derived from the Gram-negative bacterium Burkholderia gladioli. The cleavage pattern of Bga1903 indicated its moderate ability to mitigate the toxicity of pro-immunogenic peptides. The crystal structure of Bga1903, along with the identification of subsites within its active site, was determined. To improve its substrate specificity toward prevalent motifs like QPQ within gluten peptides, the active site of Bga1903 underwent site-directed mutagenesis according to structural insights and enzymatic kinetics. Among the double-site mutants, E380Q/S387L exhibits an approximately 38-fold increase in its specificity constant toward the QPQ sequence, favoring glutamines at the P1 and P3 positions compared to the wild type. Not only does the heightened specificity of E380Q/S387L improve the hydrolytic activity against pro-immunogenic peptides, but also holds this version of the enzyme as a promising candidate as an oral therapy enzyme for celiac disease.

Small island developing states (SIDS) traditionally face a set of challenges like the narrow economic structure, environmental issues, and dependence on a few ranges of economic activities forcing them to open the economy to the exterior. Therefore, their development model, like in Cape Verde depends on migration, remittances, dependence on aid, tourism, and state employment. The current research offers an insight about the nature of Cape Verde’s economy as a SIDS and the degree to which its economy depends on tourism and government services, both of which are supported through aid programs. Understanding Cape Verde’s development model is important to clarify the challenges the country faces, and its development needs to gather a long-term resilience and to understand if it is changing from a MIRAB (Migrations, Remittances, Aid and Bureaucracy) model into another one.

Rising organic charge in northern freshwaters is attributed to increasing levels of Dissolved Natural Organic Matter (DNOM) and shifts in water chemistry. Organic charge concentration may be determined by charge balance calculations (Org.-) or modelled (OAN-) using the conceptual Oliver and Hruška models based on the density of weak acid functional sites (SD) present in DNOM. The charge density (CD) is governed by the SD and protonation and complexation reactions on the functional groups. The models use this SD as a key parameter in empirically fitting the model to Org.-. Utilizing extensive datasets of water chemistry, this study shows that spatial and temporal differences in SD and CD are influenced by variations in the humic-to-fulvic ratio of DNOM and the organic aluminum (Al) complexation, as well as the mole fraction of CD to SD governed by the acidity. Site median SD obtained for 44 long-term monitored acid-sensitive lakes was 11.1 and 13.9 µEq/mg C for the Oliver and the Hruška model, respectively. During the 34 years of monitoring the CD increased by 70%, likely due to rising pH and declining Al complexation with DNOM. Present-day median SD values for the Oliver and Hruška models in 16 low-order streams are 13.8 and 15.8 µEq/mg C, respectively, and 10.8 and 12.5 µEq/mg C, respectively, in 10 high order rivers.

The human landscape transforming activity contributes to the global change of vegetation in different forms. Hunting is one of the most ancient human landscape-shaping activities. Feeders for hunting are particularly disruptive to vegetation. In the present study, we conducted a vegetation survey in these highly disturbed places. We are investigating the vegetation dynamics over several years in in the turkey oak–sessile oak zone, in two areas with different moisture and shade conditions (forest and clearing). One important background factor is changes in precipitation and temperature. The novelty of our present study lies in the fact that the observation takes place with continuous disturbance, where the human influence is very large. Our results confirm that the weed infestation be detectable at bait sites over a long period, and the interannual changes were typical in every year, according to the fact that in August, more weed species are usually present with a greater cover at the bait sites. The main meteorological factors played a role in the degree of weed infestation in each year. There was a higher proportion of vegetation regeneration in wetter years, while dry periods do not favor successional processes. With the drying of the climate, the disturbed areas are constantly losing their natural value, but wetter weather is not an automatic solution either. A more precise understanding of vulnerable systems and careful planning of treatment interventions is necessary.

In implantology, among the key choices, to obtain predictable results, it’s essential to establish, using CBCT, the bone site where to insert the implants; these sites must be identified on the mucosa and secondarily, during the surgical phase, on the cortical bone. Surgical templates are a valid aid especially in complex cases which require sev-eral implants. In cases of single implant, on the other hand, surgical guides are rarely used and the implant is inserted freehand and the identification of the implant site on the oral mucosa is more difficult; for this reason the clinician uses the teeth in the arch as a ref-erence. This experimental study is based on the use of a thermo-printed mask, con-taining radiopaque references which, worn by edentulous patients during the execu-tion of the CBCT, helps to evaluate the identification of the implant sites on the patient when surgical guides are not used and the free hand technique is adopted. This method is certainly reserved for experienced operators capable of identifying the correct angle of the implants with the freehand technique. The level of experience remains fundamental in the clinician's decision whether or not to use surgical guides; in fact, doctors with little experience should use surgical guides even in the simplest cases to reduce the risk of error.

The Giant Babax, an endemic bird in China and a nationally protected wildlife species of Class Ⅱ, is primarily distributed in southern and southeastern Tibet. To investigate the factors influencing nesting site selection, this study conducted research on nest characteristics and nesting site preferences at four sample sites around Lhasa from March to October 2023y, using systematic search methods, tracking methods, and sampling methods. The research findings are as follows: (1) The average nest depth was (7.70±1.47)cm, nest height was (19.58±2.97)cm, inner diameter was (10.64±1.73)cm, outer diameter was (24.65±4.8)cm, average weight was (162.3±13.5)g, and average nest spacing within the plot was 396.93m. (2) The nests of Giant Babax were predominantly found on vegetation such as Salix longistamina (38%), Hippophae gyantsensis (20%), Rosa sericea Lindl. (14%), and Populus szechuanica Schneid(11%). They showed a preference for artificial secondary forests as nesting sites. (3) Giant Babax exhibited a preference for habitats with robust trees that had high canopies while being distant from houses and roads, but close to water sources; moreover, the canopy of the nesting tree played a crucial role in their choice of nesting sites. (4) The reproductive success rate reached 76%, with key factors affecting it being the canopy coverage of the nesting tree and the number of helpers. These research results provide fundamental data support for population conservation efforts and ecological resource management strategies concerning the Giant Babax.

This research aimed to determine the most viable locations for establishing solar energy facilities in Yemen through the utilization of the Analytical Hierarchy Process (AHP), Best-Worst Method (BWM), and Geographic Information System (GIS). A total of twelve key criteria were meticulously selected and evaluated based on their impact on the appropriateness of sites for solar power plants. These criteria were amalgamated using a weighted overlay tool, resulting in a map that classified regions as either optimal, highly suitable, or suitable. The findings of the study indicated that the eastern, southern, and northern parts of Yemen are prime candidates for solar energy installations due to their moderate climate and flat topography. On the other hand, the western regions were deemed less favorable. The breakdown of site classifications revealed that 38% of areas were optimal, 61% were highly suitable, and only 1% were considered suitable, based on the assigned weights from the AHP and BWM methodologies. Moreover, a sensitivity analysis was carried out to evaluate the impact of adjusting the weights of Global Horizontal Irradiance (GHI) and slope on site classifications. A minor increase in the weights of GHI (from 30% to 31%) and slope (from 10% to 11%) resulted in a significant alteration in the suitability categories, highlighting the importance of weight adjustments in multi-criteria decision-making processes. In conclusion, the study emphasizes the effectiveness of AHP, BWM, and GIS in assessing site suitability for solar energy facilities, as well as the significance of sensitivity analyses. These approaches provide valuable insights for decision-makers, aiding in strategic planning and fostering sustainable energy development in Yemen.

Insulin therapy is a cornerstone in the management of diabetes, yet the pursuit of optimizing its pharmacokinetic profile remains a focal point in diabetes research. For instance, insulin icodec of Novo Nordisk is a novel long-acting insulin analogue that exhibits an extended duration of action, providing a promising once-weekly treatment option for diabetic patients. However, designing insulin analogues with lower receptor affinity than that of insulin icodec could still produce a further extended duration of action than that of insulin icodec through the suppression of receptor-mediated internalization of insulin analogues. In this study, therefore, I present the design of a novel series of insulin analogues engineered towards lower binding affinity to the insulin receptor compared to that of insulin icodec. Through computational structural biophysics-based rational design strategies, this article aims to further extend the duration of action while maintaining therapeutic efficacy of insulin analogues. Utilizing homology molecular modeling and structural biophysics-based binding affinity calculations, this article puts forward a set of insulin analogues with lower binding affinity to insulin receptor than that of insulin icodec from a structural and biophysical point of view. Overall, this article calls for subsequent in vitro and in vivo evaluations of the efficacy and prolonged action of the engineered insulin analogues to pharmacokinetically test whether these analogues acutally surpass that of insulin icodec as hopeful candidates for next-generation insulin analogue therapies with improved duration of action and enhanced control of blood glucose levels for diabetic patients in future.

Depression marsh intra- and interspecific facilitation in Southeast Florida, dominated by Hypericum fasciculatum, Stillingia aquatica, and Rhynchospora tracyi were studied as a third installment to the author’s ongoing interest in “Hypericum-Stillingia (HS) marsh” vegetation. The project includes a contextual overview in relation to the HS marsh environmental filter, stress-gradient-hypothesis, and harsh-habitat trait convergence vs. divergence. Using field measurements, transects, and quadrats, the effort focused mainly on two research questions: 1. Does the Hypericum fasciculatum microdistribution relate to marsh microtopography? 2. Does H. fasciculatum have positive or negative associations with other species? Tempered by “cause and effect” concerns, the results supported positive feedback autogenic site alteration by H. fasciculatum along with interspecific facilitation.