We evaluated graft patency by computed tomography and explored the determinants of intraoperative mean graft flow (MGF) and its contribution to predict early graft occlusion.One hundred and forty-eight patients under a single surgeon were prospectively enrolled. Arterial and endoscopically harvested venous conduits were used. Intraoperative graft characteristics and flows were collected. Graft patency was blindly evaluated by a follow-up computed tomography at 11.4 weeks (median).n = 22 of 422; 8% venous and 3% arterial). Thirteen were performed on non-significant proximal stenosis while 9 on occluded or >70% stenosed arteries. Arterial and venous graft MGF were lower in females , with median differences of 10 and 13.5 ml/min, respectively. Arterial and venous MGF were associated positively with target vessel diameter ≥1.75 mm  and negatively with pulsatility index . MGF was an independent predictor of graft occlusion, adjusting for EuroSCORE-II, pulsatility index, graft size and graft type . An MGF cut-off of 26.5 ml/min for arterial  and 36.5 ml/min for venous grafts  performed well in predicting early graft occlusion.Graft occlusion rate was 5.2%  surgery is considered the gold standard for the treatment of patients with advanced coronary artery disease, with randomized clinical trials corroborating its effectiveness [1]. Coronary artery bypass graft (CABG) surgery is considered the gold standard for the treatment of patients with advanced coronary artery disease, with randomized clinical trials corroborating its effectiveness . DespiteIntraoperative assessment of graft flow is believed to be a useful index of graft quality, which may be associated with short- and mid-term graft failure .In this study, we first evaluated potential determinants of intraoperative mean graft flow (MGF) and secondly examined the ability of MGF to predict early graft failure, as assessed by computed tomography (CT)-angiography.This prospective, observational, non-blinded, non-randomized study included 148 consecutively enrolled patients, operated by a single surgeon. All patients had CABG performed and agreed to have postoperative CT-angiography.Study protocols were in agreement with the declaration of Helsinki and all participants provided written informed consent prior to enrolment. Ethics approval was obtained from South-Central, Oxford C Research Ethics Committee, reference MREC 11/SC/0140.Data were prospectively collected and retrospectively analysed. Exclusion criteria included active inflammatory disease, neoplastic, stage-4 renal impairment (eGFR < 30 ml/min) or end-stage liver disease. Demographic characteristics and past medical history were obtained from patients’ notes and from direct interview of the patients by a research investigator Table .™  was used to assess the quality of the grafts during the operation. MGFs and pulsatility indexes (PIs) were recorded using a standardized technique, after protamine administration and with a mean systemic blood pressure between 70 and 80 mmHg. The patency of all grafts was assessed by coronary CT-angiography at a median of 11.4 weeks (range: 6–52 weeks) following CABG [All patients underwent CABG by a single surgeon experienced in both, on- and off-pump techniques. The majority of patients (74%) had their operation on-pump and 18% underwent concomitant valve surgery. Skeletonized internal mammary arteries (IMAs), endoscopically harvested saphenous veins or radial arteries were used as conduits, patients with sequential or Y-grafts were excluded from this cohort. Detailed surgical procedures are presented in the ing CABG . A grafting CABG . The dising CABG  (SupplemU-tests or Student’s t-tests for independent (unpaired) measurements as appropriate. We estimated that 148 patients would allow for detecting an occlusion odds ratio of 0.49 per unit of the continuous predictor MGF, assuming a normal distribution of MGF, with the power of 0.9 and α = 0.05.Continuous variables were tested for normality using the Shapiro–Wilk test, and non-normally distributed variables were analysed with non-parametric tests or log-transformed prior to analysis. Univariate associations between continuous normally distributed variables were evaluated with Pearson’s correlation coefficient; continuous variable differences between 2 groups were assessed by Mann–Whitney Bstand.) are presented for each covariate.A mixed-effects multilevel multiple linear regression analysis was performed, where MGF was used as the dependent variable of interest whilst sex, graft type , PI and target vessel size were used as independent covariates . To provide more insight into the differences between arterial and venous grafts, a secondary multiple linear regression sub-analyses were performed. Continuous variables were log-transformed. Standardized Beta-coefficients . Early graft failure was the dichotomous outcome of interest. Continuous variables were log-transformed. We maintained the continuous nature of continuous variables in our principal analysis, in order to maximize the data information towards addressing our primary statistical question, especially given the moderate sample size (hence odds ratios for continuous variables were defined per unit of each variable). Given the clinical necessity of using cut-off values to robustly select high- versus low-risk grafts, we also conducted a P-values were calculated and P < 0.05 was considered statistically significant. Robust variances were used in the regression models to accommodate possible assumption model violations.All statistical analyses were performed using the SPSS statistical software version 24 . Two-tailed n = 22), 8% (n = 15) of endoscopically harvested venous and 3% (n = 7) of arterial conduits  were performed in 148 participants . There was no intraoperative mortality, periprocedural myocardial infarctions, early recurrent angina, acute heart failure or early postoperative need for ercutaneous coronary intervention or graft revision. Overall early occlusion rate was 5.2%  anastomosed onto native coronary arteries with non-significant narrowing of <70% , 6 (27%) were anastomosed onto totally occluded native coronary arteries (CTO) and 4 grafts (18%) had no clear cause that could contribute or justify their early occlusion.Retrospective 3D-QCA analysis of the preoperative coronary angiogram for the same cohort of occluded grafts showed that 4 of the grafted native coronary arteries had significant proximal stenosis , 13 had <70% stenosis  and 5 vessels were CTOs .rarterial = –0.061, P = 0.51; rvenous = 0.061, P = 0.51), hyperlipidaemia , diabetes  or smoking status . Flow measurements were lower in females, with a median difference of 10 ml/min for arterial (P = 0.01) and 13.5 ml/min for venous grafts . MGF was, however, inversely correlated with PI  and target vessel size  were independently associated with MGF for arterial grafts, whilst venous graft MGF was only independently associated with PI  without any significant association with PI . IMA grafts had a comparable occlusion rate  to that reported in the literature . The obsWe report a high early patency of endoscopically harvested venous conduits (92%), which is similar or better to the reported in the literature for either endoscopically or open technique harvested vein conduits. This finding supports current literature which suggests that endoscopic vein harvesting can give at least equivalent patency rate to open vein harvesting , 19 and The CT angiographic subgroup analysis of the occluded grafts revealed that the majority  were performed on target native coronary arteries without significant proximal disease or anastomosed onto CTO arteries with well-developed collaterals. This supports previous reports which suggest that competitive flow from the native coronaries or aggressive collaterals and advanced diffuse disease of small re-canalized CTO arteries could play an important role in graft occlusion, especially for arterial grafts , 22.Our results support a move towards systematic evaluation of the preoperative angiogram and/or associated intracoronary physiology data and more standardized surgical techniques including the routine use of TTFM to guide contemporary CABG revascularization strategies, especially when multiple arterial grafts are used , 23. CT-In our analysis, we reported that MGF is predominantly influenced by the size of the target vessel ≥1.75 mm and sex, and it correlates with PI for both arterial and venous grafts, while there was no difference of patency rates or MGF when grafts are performed by a surgeon experienced in both, off- and on-pump techniques . These rWe identified that grafts had lower MGF in females, with a median difference of 10 ml/min for arteries and 13.5 ml/min for veins which can be explained by the fact that the size of the coronary arteries and heart muscle is smaller in females and in accordance to similar findings reported by our group, in a different cohort of CABG patients .The multivariable regression analysis in our study demonstrated that MGF was the most reliable and independent predictor of early graft failure, after adjusting for established risk factors. Although the PI was found to correlate inversely with MGF, it did not seem to provide further predictive information, a finding that goes against the already known and widely accepted, predictive value of the PI , 28 and et al. [In a retrospectively analysis of 345 patients, Lehnert reported a direct correlation between flows and graft patency, at 1-year follow-up using conventional coronary angiography. The authors reported an occlusion rate of 6.5% for IMAs and 16.9% for SVG. They recommended that satisfactory patency rates can be achieved when MGF is >20 ml/min for IMAs and >40 ml/min for SVG . Di Giamet al.  proposedet al. , the intet al. .Our study is reporting a very low graft failure rate with a predictive MGF cut-off value of 26.5 ml/min for arterial and 36.5 ml/min for venous grafts. However, the flow alone must be interpreted with caution and the decision to revise a graft (which in our cohort was 4.5%) should be in conjunction with the clinical, anatomical and physiological status of the patient at the time of the measurement. The routine use of intraoperative TTFM is recommended by the 2018 ESC/EACTS guidelines (Class IIa) and National Institute of Clinical Excellence (NICE) and its intraoperative use can influence the decision to revise up to 5% of poor-performing grafts during CABG, leading to better early patency rates .n = 13). The multivariable analysis is based on a small cohort of patients with low event rate and as such, the model is at risk of being over-fitted and further validation of the reported results, including the impact of gender on the MGF values, is required. This dataset is derived from a single-centre, single-surgeon experience, which may comprise a source of selection bias. However, our study is one of the largest that report on the use of TTFM to assess early graft failure with a CT angiogram and it is reporting a very low rate of early graft failure, reinforcing previous publications on the benefits that TTFM can offer to modern coronary artery by-pass surgery.Our study has a number of limitations. It is an observational study with retrospective analyses of collected data; however, this is partially mitigated by prospectively nature of data collection and having the CT-angiograms and 3D-QCA analysis performed by operators blinded to the TTFM measurements. Fractional flow reserve assessment of the proximal stenosis was not routinely performed which could have helped us to limit further the possibility of performing by-pass grafts to arteries with <70% proximal stenosis. We limited our outcome only on totally occluded grafts, thus we did not analyse data regarding partial or flow-limiting stenosis and we did not correlate our results with medium- or long-term postoperative clinical findings. Of all the arterial grafts in our cohort, radial artery conduits displayed a high occlusion rate, however, this should be interpreted with extreme caution given the very small number of radial artery used in this cohort (MGF seems to be the most reliable predictor of early graft patency and is influenced by intrinsic biological factors but not by cardiopulmonary bypass or endoscopic vein harvesting. We demonstrate that MGF is lower in females and that it is mainly influenced by local haemodynamic factors such as vessel size ≥1.75 mm and PI. We estimate that MGF > 26.5 ml/min for arterial and MGF > 36.5 ml/min for venous grafts perform well in predicting early arterial and venous graft patency. Contemporary CABG is associated with very low early graft failure rates and TTFM is an effective tool of intraoperative quality assurance.ICVTS online.Conflict of interest: George Krasopoulos: receives honoraria for being a consultant, advisor and speaker for Medistim ASA and Abbott. Charalambos Antoniadis: founder, director and shareholder of Caristo diagnostic, a University of Oxford spinout company. All relevant data are within the manuscript and its supporting information files.Andrea D'Alessio: Conceptualization; Data curation; Methodology; Resources; Supervision; Visualization; Writing—original draft; Writing—review & editing. Ioannis Akoumianakis: Conceptualization; Data curation; Formal analysis; Methodology; Supervision; Writing—original draft; Writing—review & editing. Andrew Kelion: Conceptualization; Data curation; Investigation; Methodology; Supervision; Writing—review & editing. Dimitrios Terentes-Printzios: Conceptualization; Data curation; Formal analysis; Investigation; Supervision; Writing—review & editing. Andrew Lucking: Conceptualization; Investigation; Methodology; Supervision; Writing—review & editing. Sheena Thomas: Data curation; Investigation. Danilo Verdichizzo: Conceptualization; Resources; Supervision; Writing—review & editing. Amar Keiralla: Conceptualization; Resources. Charalambos Antoniades: Supervision; Validation. George Krasopoulos: Conceptualization; Data curation; Methodology; Supervision; Validation; Writing—original draft; Writing—review & editing.Interactive CardioVascular and Thoracic Surgery thanks the anonymous reviewers for their contribution to the peer review process of this article.ivab298_Supplementary_DataClick here for additional data file.


Citric acid (CA) treatment is a convenient, mild and environmentally friendly strategy to modify the composition, structure and function of starch through hydrolysis and esterification, which expands the application of starch in industry. In this paper, the effects of CA modification on amylose content, amylopectin chain length distribution, microscopic morphology, solubility and swelling ability, thermodynamic properties, gelatinization properties, digestibility properties, texture properties and the film-forming properties of starch were summarized. The application status and development trend of CA modified starch were reviewed, which has important implications for the targeted utilization of CA modified starch in the future. Starch is a polysaccharide composed of glucose units and is the main storage form of carbohydrates in higher plants . Starch Acid hydrolysis and esterification are two common chemical methods in the industrial production of modified starch. The process of acid hydrolysis is relatively simple. The starch is suspended in an organic acid or inorganic acid solution, followed by stirring, filtering, neutralizing, washing and drying steps . The starch retains the granular structure after acid hydrolysis, while its viscosity decreases and fluidity increases. As shown in Citric acid (CA) is an organic acid widely used in the food and daily chemical industries. It is non-toxic with a good solubility, anti-oxidation and mild reaction. Moreover, CA is a licensed additive and is widely used in the production of jams, juices, soft drinks, biscuits and other foods ,8. CA caIn this paper, the effects of CA modification on the amylose content, amylopectin chain length distribution, microscopic morphology, solubility and swelling ability, thermodynamic properties, gelatinization properties, digestibility properties, texture properties and film-forming properties in starch were described  sedimentation method. The iodine binding method is based on the binding capacity of amylose to iodine, and its content is determined by colorimetry. Since the long side chains of amylopectin can also bind to iodine, the assay results tend to exceed the true value. In addition, the ConA sedimentation method indirectly determines the content of amylose by enzymatic hydrolysis, based on the specific binding property of ConA and amylopectin. Since ConA binds to the less branched amylopectin moiety, its assay results also exceed the true value . In the In natural starch, amylopectin exhibits a highly branched structure, and α-D-glucose is linearly connected by α-1,4 glycosidic bonds to form the main chain and each the short branch of amylopectin. The chain and each short branch are linked through α-1,6 glycosidic bonds. Generally, enzymes (isoamylase and pullulanase) are used to debranch amylopectin into short linear structures, and then high-performance size-exclusion chromatography (HPSEC) and other methods are used to separate the branched starch. The chain length distribution of chain starch was determined. Li et al. found that with the prolongation of the starch hydrolysis time by HCl, the long-chain starch and medium-chain starch in the amorphous region preferentially underwent hydrolysis, and the long-chain starch (degree of polymerization (DP) ≥ 37), medium-chain starch (DP 25–36) and short-chain starch were hydrolyzed preferentially. The relative proportions of (DP 6–24) decreased sequentially . Kim et The microscopic morphology of starch from different sources includes round , oval , polyhedron , etc. A scanning electron microscope (SEM), a light microscope , a polarized light microscope  and a laser confocal microscope  and other equipment are commonly used for detection. Huo et al. found that CA modification had no significant effect on the polyhedral structure of rice starch granules, while as the concentration increased, slight erosion marks appeared on the surface of some granules . Similar−1 and (1022/995) cm−1 peak intensities in the spectrum is regarded as an indicator of the ordered structure of the starch granules, in which the (1047/1022) cm−1 peak intensity ratio reflects the order degree of the starch molecules. The larger the ratio, the higher the order degree [Starch crystal is composed of a short-range ordered structure and long-range ordered structure. The short-range ordered structure refers to the double helical structure composed of short chain parts in amylose and amylopectin, while the long-range ordered structure refers to double helical molecules. The chains form different polymorphs in certain regions of starch granules through intermolecular interactions . The shor degree . The lonr degree ,27. Diffr degree .In the process of starch hydrolysis, the amorphous region degrades faster. As the acid molecules penetrate into the crystalline region, the surface layer of the crystalline region will also be degraded and the phenomenon of erosion will occur. However, the hydrolysis rate of the crystalline region is much slower than that of the amorphous region, so that the starch granules can retain the crystalline structure . At presA high temperature increases the vibration of starch molecules in water and breaks the intermolecular hydrogen bonds, resulting in more sites for starch molecules to interact with water molecules. At the same time, water molecules enter the crystallization zone, the crystalline state is transformed into an amorphous state, and the starch undergoes irreversible swelling . Most st0), peak gelatinization temperature (TC), termination gelatinization temperature (TP) and gelatinization enthalpy (ΔH) characterized in the DSC curve. It is generally believed that starch hydrolysis results in the destruction of the amorphous region, requiring more energy to melt the crystalline structure during the heating process, and increasing ΔH [The thermodynamic properties of starch can be measured by differential scanning calorimetry (DSC) through the initial gelatinization temperature  is used to measure, through the peak viscosity (peak viscosity), valley value in the RVA curve trough viscosity, holding strength, final viscosity, break down viscosity, setback, peak time and pasting temperature and other indicators to characterize. Studies have found that starch hydrolysis leads to a significant reduction in gelatinization viscosity, which is mainly related to the destruction of the amorphous region and the reduction of the solubility and swelling ability of starch granules ,45,47. Lp ≤ 0.05) from 31.71% to 39.43% and 37.55% to 53.38%, respectively [According to the digestibility, starch can be divided into rapidly digestible starch (RDS), slowly digestible starch (SDS) and resistant starch (RS). Fast-digestible starch refers to starch that is rapidly digested and absorbed in the small intestine (<20 min). Slow-digestible starch refers to starch that is completely digested and absorbed in the small intestine but at a slower rate (20–120 min). Resistant starch refers to starch that cannot be digested and absorbed in the small intestine, but can be partially fermented in the large intestine by intestinal microorganisms . The digectively . On the ectively .The texture characteristics of starch can be detected by a texture profile analysis (TPA), using a physical property analyzer. Parameters, such as gumminess, springiness, and chewiness are characterized. Studies have found that with the increase of the degree of hydrolysis, the gel strength of starch gradually decreases, which is mainly related to the depolymerization of starch granules and the degradation of amylose molecules ,45. MoinUtilizing its retrogradation property, starch can be made into films through gelatinization, cooling, retrogradation, drying and other processes. Starch film has the characteristics of a good stretchability, transparency, folding resistance, water insolubility and low air permeability, which can be applied in the field of green packaging. Studies have shown that the hydrolysis reaction can improve the mechanical strength and water vapor transmission rate of starch films and reduce the elongation at break. Retrogradation of starch is mainly related to the binding of amylose molecules. Amylopectin molecules are not prone to retrogradation because of the branched structure, which have an inhibitory effect on the retrogradation of amylose. The hydrolysis reaction leads to a decrease in the molecular weight of amylopectin in starch granules and a decrease in the degree of branching. In the film forming solution, the small molecular weight starch molecular chains are dispersed in the gaps of the amylopectin molecules, which are conducive to the formation of an orderly arrangement of the starch molecular chains through hydrogen bonding during the film forming process. The water vapor transmission rate is mainly related to the polar groups in the starch film. The starch molecular chain produced by hydrolysis has a large number of hydroxyl groups, which can improve the dissolution and diffusion of water vapor in the starch film . SimilarCA has the characteristics of a good solubility, pure acidity, strong chelating ability and low toxicity, and has been certified by the US FDA as a “generally recognized as safe” (GRAS) food additive, can be used in food processing as a sour agent, antioxidant, buffer, color retention agent, etc. Rice is one of the most commonly consumed staple food around the world and is an excellent source of starch. Cooked rice generally has a high blood sugar response, which is not good for the blood sugar control of diabetic patients . Lee et Porous starch is a modified starch, which can be used as a catalyst, adsorbent, slow-release agent, etc. in different fields, and has the advantages of a high efficiency, non-toxicity, safety, and biodegradability. Porous starch can be obtained by hydrolyzing starch with enzymes or acids below the gelatinization temperature. The use of CA as a hydrolysis catalyst has the advantages of safety and controllability, low price, and mild hydrolysis. Kim et al. found that the hydrolysis of CA resulted in the partial breakage of the crystal structure of corn starch and an increase in the area of the central cavity of starch granules, resulting in a significant increase in its adsorption capacity in water and oil . SimilarPickering emulsion is a stable emulsion that replaces traditional surfactants with solid particles, which has important application value in the fields of food, medicine and cosmetics. Ordinary starch has a large particle size (>10 μm) and weak hydrophobicity, making it difficult to adsorb on the oil-water interface to form a stable emulsion. Therefore, CA treatment can improve the hydrophobicity of starch, preparing a good starch based solid particle emulsifier. Pickering emulsion, prepared with CA modified starch as emulsifier has a good emulsifying ability and storage stability. The diameter of oil droplets in the emulsion is small and uniform, and the oil droplets are not easy to coalesce. The emulsion prepared from natural starch depolymerized after being stored at 25 ℃ for 3 h, while the emulsion prepared from CA modified starch still maintained a good stability under the same conditions for 12 h, and its emulsification index reached more than 90%. The esterification reaction can improve the hydrophobicity of starch molecules and reduce the surface tension of the oil-water interface . Lv et aStarch-based fat substitutes have texture, taste and stability similar to those of fat-containing foods, and are recognized as safe fat substitutes, which have attracted more and more attention. However, most natural starches have the characteristics of easy aging, a low shear resistance, and poor thermal stability, and are not suitable as fat substitutes. CA modified starch has been prepared as starch-based fat substitutes in different food matrices, such as mayonnaise. Park et al. compared the physicochemical properties of octenyl succinic anhydride modified starch, toughened modified starch, CA modified starch, acetylated modified starch and moist heat modified starch, and found that CA modification can improve the solubility, water holding capacity and swelling capacity of starch. Moreover, the mayonnaise product obtained by replacing oil and fat with CA-modified starch has a good color, viscoelasticity and emulsion stability .Starch-based biodegradable materials have an excellent biocompatibility and biodegradability, and can be used in packaging, construction, medicine and other fields. Starch-based film materials have the characteristics of being colorless, odorless, good flexibility, high transparency and impermeability to oxygen, etc., but also have disadvantages, such as poor mechanical properties and poor water vapor permeability. Related studies have found that CA modification can improve the mechanical strength, barrier properties, tensile strength and thermal stability of starch films, and reduce the water vapor transmission rate, which endowed its application in the bioactive packaging field ,66. LikeCA treatment is an convenient, mild and environmentally friendly strategy to modify the composition, structure and function of starch through hydrolysis and esterification, which expands the application of starch in the fields of food, cosmetics, chemical industry and medicine. In recent years, scholars have made progress in the study of modified starch with CA, while there are still many challenges in the maturity and commercial application of this technology. First of all, the mechanism of CA modification on the crystal structure of starch granules is not yet clear. The changes in the degradation of the amorphous region, the depolymerization and repolymerization of starch molecules, and the formation of amylose-lipid complexes during the starch modification process can be further explored. Secondly, in order to deeply understand the mechanism of CA-modified starch on the nutritional quality and texture properties of food, the interaction of esterified-modified starch with protein, dietary fiber, fat and other components in a high temperature and high pressure environment need to be investigated. Thirdly, in order to further improve the physiochemical properties and producing efficiency of CA-modified starch, CA modification can be combined with other methods, such as enzymatic hydrolysis, extrusion, microwave, ultrasonication and radiation ,73. Fina


BRCA2) mutations are associated with tumorigenesis in humans and dogs. BRCA2 is a DNA damage repair protein that interacts with RAD51, another DNA-damage-repair protein, to preserve DNA stability. Tumors, such as mammary tumors, are developed when their function or interaction is lost. It is currently unclear how the interaction activity of BRCA2 and RAD51 is regulated. We focused on a highly conserved region (HCR) near the BRC repeats, which are RAD51-interacting domains in BRCA2. We demonstrated that HCR regulated the RAD51-interaction activity of BRC repeats. In addition, we looked for HCR mutations in canine mammary tumors and discovered I1110M mutations. The effect of four HCR mutations, including I1110M, on the RAD51-interaction activity of BRC repeats was investigated. Three HCR mutations  found in canine mammary tumors or human breast cancers influenced the RAD51-interaction activity of BRC repeats. According to this study, HCR regulated the RAD51-interacting activity of BRC repeats, and HCR mutations may be related to tumorigenesis in both dogs and humans.Mammary tumors are the most common tumor type in female dogs and one of the major problems in veterinary oncology. Breast cancer 2, early onset (BRCA2) malignant mutations are associated with tumorigenesis in humans and dogs. BRCA2 plays a pivotal role in homologous recombination repair by recruiting RAD51 recombinase to DNA damage sites to maintain genome stability. To recruit RAD51, BRCA2 must interact with RAD51 via BRC repeats, but the regulation of this interaction has been unclear. In this study, we focused on a highly conserved region (HCR) near BRC repeats. Using co-immunoprecipitation and mammalian two-hybrid assay, we found that HCR suppressed the RAD51-interaction activity of BRC repeats and that substitutions of HCR phosphorylation sites affected it. In canine tumor samples, we found ten mutations, including a novel HCR mutation (I1110M) from canine tumor samples. The effect of four HCR mutations, including I1110M, on the RAD51-interaction activity of BRC repeats was tested. One of the HCR mutations found in canine mammary tumors increased the interaction, but the two mutations found in human breast cancers decreased it. This study suggested that the HCR regulated the RAD51-interacting activity of BRC repeats through HCR phosphorylation and that mutations in HCR may be related to tumorigenesis in both dogs and humans.Mammary tumors are the most prevalent type of tumors in female dogs. Breast cancer 2, early onset ( Canis lupus familiaris) . W. W23]. WIn humans, three phosphorylation sites, S1106, S1123, and T1128, are phosphorylated by ATM/ATR [BRCA2 exon 11, which encodes the HCR and BRC repeats, in the mammary tumor samples , 2213A>G (N669D), 2329T>C (synonymous), 2609A>C (K801Q), 2696A>G (I830V), 3538T>G (I1110M), 4481A>C (T1425P), 4512A>G (K1435R), 5788G>A (synonymous), and 6886A>G (I2226M) (We identified 10 mutations in (I2226M) . We also(I2226M) . Homozyg(I2226M) . We analhttps://brcaexchange.org (accessed on 21 July 2021)) [We identified a novel mutation, I1110M, in the HCR. Then, we speculated that mutations in the HCR could contribute to tumorigenesis through BRCA2 dysfunction. Thus, we investigated the effect of mutations in the HCR on the RAD51-interaction activity of BRC1 and BRC2. Some mutations were also reported in HCRs from canine mammary tumors and human breast cancer samples ) . The A11y 2021)) . Almost HeLa cells stably expressing canine cBRC1-HCR-BRC2 harboring A1108G, I1110M, S1114P, or T1115P mutants were generated to evaluate these mutations using co-immunoprecipitation. The expression of exogenous BRCA2 peptide and endogenous RAD51 or PP2A-C subunit (α and β) was assessed via Western blotting B. All cBIn this study, we identified the HCR as a RAD51-interaction regulatory region for BRC repeats located between BRC1 and BRC2 in BRCA2. The BRC repeats–RAD51 interaction is ordinarily attenuated by the presence of the HCR. A previous study found that the DNA-damage-signaling kinases ATM and ATR phosphorylated the HCR following DNA damage . ConsistIt was reported that BRCA2 interacted with the phosphatase PP2A through the HCR and that the interaction between PP2A and HCR is critical for HR . We alsoIt was also suggested that the HCR mutation disrupting the interaction with PP2A did not influence the RAD51-interaction activity of BRCA2 . The PP2BRCA2 exon 11, which encodes the HCR and BRC repeats, from canine mammary tumor and tumor-free samples. We identified 10 mutations. The mutations 2135A>G (E643K), 2213A>G (N669D), 2609A>C (K801Q), 2696A>G (I830V), 4481A>C (T1425P), 4512A>G (K1435R), and 6886A>G (I2226M) have been previously reported by ourselves and other groups; however, the others were novel (2329T>C (synonymous); accession No. LC687404, 3538T>G (I1110M); accession No. LC687405, and 5788G>A (synonymous); accession No. LC687406) [According to our findings and those of others, the HCR plays an important role in BRCA2 function . Mutatio696A>G I8V, 4481A>C687406) ,34,35. AC687406) . In thisBecause the I1110M (3538T>G) mutation is located in the HCR, we also examined the effect of mutations in HCR on the BRC repeats–RAD51 interaction activity. We also examined three other mutations: the A1108G mutation found in canine mammary tumor samples in the previous report, and S1114P and T1115P, corresponding to human BRCA2 S1115P and T1116P mutations, respectively, in breast cancer samples . The pepDuring the analysis of the RAD51-interacting activity, we realized that HeLa cells expressing the peptide containing BRC2 decreased endogenous RAD51 expression. This amount of reduced RAD51 expression seems to be related to the expression level of the peptide containing BRC2. We did not investigate how the BRC2 peptide decreased the expression level of endogenous RAD51, but this interesting issue should be elucidated in the future.In this study, we identified a novel function of the HCR in BRCA2 located between BRC1 and BRC2 in canine BRCA2. The HCR regulated the RAD51-interaction activity of BRC repeats. We also confirmed the phosphorylation of the HCR, which could regulate the RAD51-interaction activity of BRC repeats. Additionally, we identified the novel homozygosity mutation, I1110M (3538T>G), in canine mammary tumor samples. Although this mutation did not have any effect on RAD51-interacting activity, the reported HCR mutation, A1108G from canine mammary tumors, increased the RAD51-interacting activity, and the two HCR mutations derived from human breast cancer, S1114P and T1115P, reduced the RAD51-interacting activity of BRC repeats. Thus, this study provides new insights into how BRCA2 regulates the interaction with RAD51 using the BRC repeats and the HCR, as well as offers possible future directions for research into the tumorigenic role of BRCA2 mutations in dogs and humans.


Pea starch and some legume starches are the side streams of plant-based protein production. Structural modification toward moderate digestibility and desirable functionality is a way to increase the economic values of these side-stream starches. We applied an innovative and sustainable technique, high-pressure homogenization, to alter pea starch structure, which resulted in a high level of complexation with the small phenolic acid molecule, gallic acid, to alter starch digestibility. This study showed a great level of disruption of the compact starch structure represented by the decrease in gelatinization temperature, enthalpy change, and relative crystallinity. The addition of a high concentration (10%) of gallic acid contributed to a typical V-type X-ray diffractometry pattern. Data demonstrated a significant decrease (~23%) in the susceptibility to α-amylase and an increase in resistant starch (~13%). In addition, starch functionality was improved with a reduced retrogradation rate. Pea starch responded to the high-pressure homogenization process well. Compared with the rice and maize starch reported in the literature, pea starch required a reduced amount of gallic acid to form a high level of complexation with a significant delay in starch digestion. Pisum sativum) is a preferred crop as it is allergen-free and is typically not genetically modified [w/w), depending on the variety [Legumes are a primary source of plant protein, and pea , the short processing time with HPH (~12 min in this study) may prevent the heat degradation of phenolic acids. In this study, we modified pea starch digestibility through complexation with GA via HPH, and the modification via HPH processing was characterized.Aspergillus niger (319 U/mL), invertase from S. cerevisiae (334 U/mg), and α-amylase from porcine pancreas  were from Sigma-Aldrich . All reagents and chemicals were of analytical grade and used without further purification. Enzyme activity was defined and determined by the manufacturers, and newly purchased enzymes were used in this study.Commercial pea starch was received from Roquette , and as noted by the manufacturer, the native pea starch contained 0.1% of crude fat and 0.5% of protein. Gallic acid  was from Sigma-Aldrich . D-Glucose Assay Kit was purchased from Megazyme International Ltd. . Pepsin from porcine gastric mucosa (371 U/mg), pancreatin from porcine pancreas (8x USP), amyloglucosidase from w/w) were prepared using deionized water at approximately 27 ± 5 °C. In sequence, GA (5% and 10% weight based on pea starch) was added into the starch slurry with constant stirring for 3 min. Then, the mixture was homogenized via a high-pressure homogenizer  at 180 MPa for four cycles, which was 3 min for each cycle and a total of 12 min. A control treatment without GA was prepared with the same procedure. Due to the smooth warming of the mixtures promoted by the high-pressure homogenization process, the mixtures were cooled to approximately 30 °C and then washed twice with 50% (v/v) ethanol solution. The suspension was centrifugated (2800× g for 5 min) to remove free (uncomplexed) GA. The precipitates were dehydrated in an oven at 37 °C for approximately 14 h. Dehydrated samples were ground into powder using a coffee grinder . Powders were passed through a 100-mesh screen before further analyses.Pea starch slurries (10% TS/ΔHNS) × 100; where ΔHTS is the enthalpy change of HPH-treated starches and ΔHNS is the enthalpy change of native pea starch.The thermal properties of the pea starch–GA complexes and of the control treatment were determined by differential scanning calorimetry . Samples (6 mg) were weighted in aluminum pans, mixed with 18 μL of deionized water, and then equilibrated at ambient temperature for 24 h. The pans were scanned from 10 °C to 150 °C at 10 °C/min, and onset (To), peak (Tp), and conclusion (Tc) temperatures, enthalpy change (ΔH), and the range of gelatinization temperature  were recorded. The starch gelatinization degree (GD) was obtained following the equation: DSG (%) =  equipped with an IμSCu microfocus sealed tube, with a step size of 0.02°, a target voltage of 50 kV, a current of 0.8 mA, and a scan time of 300 s, and the samples were scanned from 3° to 35°. The relative crystallinity (RC) of the starches was obtained by using the software Origin 8.0 , and using the following equation as described by Vanier et al. (2019): RC (%) = (Ac/(Ac + Aa)) × 100; where Ac is the crystalline area and Aa is the amorphous area on the X-ray diffractograms .g for 10 min. An aliquot of the mixture (15 μL) was diluted with deionized water (240 μL) in a 96-well microplate, followed by adding the Folin–Ciocalteu reagent . The mixture was kept on the bench for 3 min, and then 30 μL of sodium carbonate (1N) was added into the mixture. The final mixtures were kept at proximately 27 °C for 90 min in the dark. The absorbance at 760 nm was recorded using a plate reader and compared against a GA standard curve. The complex index was defined as the percentage of the detected GA amount in treatments to the amount of GA added in the modification.The GA content in the complexes was estimated by the Folin–Ciocalteu procedure as described by Chi et al. (2019)  and SancStarch–GA samples (500 mg) were mixed with a 25 mL solution (pH 6.9), which contained calcium chloride (0.1 mM), glycerol (0.2 mg/mL), and sodium azide (0.2 mg/mL). The substrate mixtures were prewarmed at 37 °C for 5 min, and then 5 mL of α-amylase suspension (0.33 U/mg starch) was added into the mixture. The hydrolysis was conducted in a water bath controlled at 37 °C. An aliquot of 0.5 mL was taken after 0, 5, 10, 15, 20, 40, 60, 80, 100, and 120 min and heated in a boiling water bath for 10 min to inactivate enzyme activity. The concentration of reducing sugars released by α-amylase hydrolysis was determined by the miniaturized Somogyi–Nelson assay .g for 10 min. Subsequently, amyloglucosidase  and invertase  were added to working solution B.To determine the rapidly digestible starch (RDS), slowly digestible starch (SDS), and resistant starch (RS) fractions, in vitro digestibility of pea starch–GA complexes was carried out following the method of Englyst and Cummings (1992) . Pepsin g for 20 min, and glucose content present in the supernatant was analyzed using a GOPOD reagent. The released glucose after 20 and 120 min was labeled as G20 and G120, respectively, by which RDS, SDS, and RS fractions were calculated as described in the formulas below:For the gastric phase, the starch–GA complexes (0.5 g) were dispersed in 5 mL of sodium acetate buffer 0.05 mol/L (pH 5.2) and equilibrated at 37 °C for 5 min. Then, 10 mL of enzyme working solution A and three glass marbles were added to the samples, and the mixture was incubated in a shaking water bath (150 rpm) at 37 °C for 30 min. In sequence, to start the intestinal phase, 5 mL of enzyme solution B was added to the mixture, and it was further incubated at 37 °C for 120 min. Aliquots of the hydrolysates (1 mL) were collected at times of 0, 20, and 120 min of intestinal phase simulation and placed in a boiling water bath (10 min) to inactivate the enzymes. The samples were centrifuged at 10,800× TS means the total starch (TS) content of the complexes used for digestibility measurement, equal to 0.5 g.® software .Data analysis was carried out by analysis of variance (ANOVA). Tukey’s test compared the means at the 5% level of significance using SASThe thermal properties of the pea starch, HPH-treated pea starch (HPH-PS), and HPH-treated pea starch–gallic acid complex (HPH-PS-GA) were examined with DSC, and the results are presented in p < 0.05), indicating a lower thermostability of the starch structure. HPH-treated samples had a narrower gelatinization temperature range than native pea starch, suggesting a higher homogeneity of amylopectin crystals in the HPH-treated samples compared to the native pea starch. With the presence of GA, HPH-PS-GA had a higher gelatinization degree than HPH-PS.HPH-treated samples had a reduction in onset (To), peak (Tp), and conclusion (Tc) temperatures and in enthalpy change (ΔH) compared to native pea starch, indicating that HPH facilitated starch gelatinization. The addition of GA, both at 5% and 10%, did not significantly change gelatinization temperature, although both treatments had a slightly higher peak temperature (Tp) than HPH-treated starch without GA . The preAfter cold storage for seven days, HPH-PS and HPH-PS-GA-5% samples had a higher retrogradation percentage  than untreated pea starch 84.7%; . Samplesp < 0.05).Results of the α-amylase hydrolysis kinetic study showed HPH-PS-GA-10% reduced approximately 23% of the digestion rate compared with HPH-PS . The addHPH modification with GA greatly disrupted the highly organized starch molecular structure. The structural disruption was caused by the mechanical change, including high pressure and high shear force, during HPH processing. The outcome of such structural disruption was the enhancement of hydration during heating , which pp < 0.05) might be due to the molecular leaching promoted by HPH. However, the complexation with GA reduced retrogradation, and this desirable outcome was achieved only with a high concentration of GA. The bonding between GA and starch molecules may hinder the interaction between starch molecules and delay retrogradation.HPH modification with GA improved pea starch functionality with a lower retrogradation rate. HPH alone increased the retrogradation rate from 85% to 99%, and such a significant change  [Conventionally, HPH has neither generated a high degree of complexation ,15,23 now/w) .HPH was selected for this study to overcome a few disadvantages in conventional heat-based treatments, such as heat damage to phenolic compounds and higher energy demand for a long heating process. Findings of the change of digestibility and functionalities in this study suggest HPH is an effective method for the industry to utilize pea starch. Researchers may consider extending the study to other legume starches with similar granular structures.In modern diets, many processed starchy foods are rapidly digested into glucose. Much research has demonstrated various modifications of starch digestibility. This study aimed to demonstrate an innovative technique and “clean-label” approach  utilizing side-stream starch to alter starch digestibility. We believe that the selection of initial material (starch) is critical because each starch from different botanical sources responds to technology in different manners. Pea starch and many other legume starches that primarily originate from the massive production of plant-sourced proteins have a distinguished granular architecture that varies from commercial cereal  and tuber starch . This study demonstrated a successful and effective complexation of pea starch with GA through an innovative and sustainable manufacturing technique, high-pressure homogenization. A V-type inclusion complex was formed, the susceptibility to α-amylase was greatly reduced, and the retrogradation rate was significantly hindered with a relatively low amount of gallic acid compared with the complexation of cereal starches. Utilizing the natural starch structure with a suitable technique is an effective way to produce desirable functionality.


There was no significant difference in biliary complications between NMP and SCS . Graft and patient survival were significantly better in HOPE than in SCS  and . Graft and patient survival were not significantly different between NMP and SCS. Conclusion: HOPE/D-HOPE and NMP are promising alternatives to SCS for donor liver preservation. They may help address the widening gap between the demand for and availability of donor livers by enabling the rescue and transplantation of marginal livers.Background: The gap between the demand and supply of donor livers is still a considerable challenge. Since static cold storage is not sufficient in marginal livers, machine perfusion is being explored as an alternative. The objective of this study was to assess  hypothermic oxygenated machine perfusion (HOPE/D-HOPE) and normothermic machine perfusion (NMP) in contrast to static cold storage (SCS). Methods: Three databases were searched to identify studies about machine perfusion. Graft and patient survival and postoperative complications were evaluated using the random effects model. Results: the incidence of biliary complications was lower in HOPE vs. SCS (OR: 0.59, 95% CI: 0.36–0.98,  Although the number of liver transplants performed globally has increased yearly, the availability of donor organs is overshadowed by the demand. More and more centers have optimized and adopted the use of extended criteria donor (ECD) organs to narrow the gap ,2. HowevAlthough numerous studies have explored the dynamic preservation of livers over the past two decades using machine perfusion (NMP or HOPE/D-HOPE) compared to SCS in clinical settings, the majority are small sample-size studies. Based on current literature, it is not very clear which may be comparatively better between HOPE/D-HOPE and NMP when compared with SCS, which is the standard method for preserving donor livers. Ischemia re-perfusion injury is one of the main concerns in SCS. Ischemia re-perfusion injury affects graft survival, which influences patient survival. Machine perfusion aims to address this problem. The occurrence of postoperative complications also has an impact on patient survival. Therefore, in this systematic review and meta-analysis, our primary objective is to assess and compare patient and graft survival in liver transplant patients after ex situ machine perfusion compared to SCS. The secondary objective is to evaluate the occurrence of postoperative complications after liver transplantation.The PubMed, Web of Science and Scopus databases were queried for studies reporting on normothermic and hypothermic machine perfusion in liver transplantation through September 2022. The full search syntax for each database is documented in the Two independent reviewers performed the quality assessment of all the studies included in the meta-analysis. The evaluation was according to the Downs and Black checklist . We usedData were extracted independently by the two reviewers using standardized forms. Baseline and outcome data were extracted for the research (NMP/HOPE) and control (CSC) groups. Baseline data includes sample size in each group, age, donor type and BMI. Outcome data includes graft survival, patient survival, biliary complications, hospital stay, vascular complications and primary non-function. Data were collected, aggregated and reported. For studies that did not report survival data, the data were extracted from Kaplan–Meier survival curves using methods described by Tierney et al. .2 statistic. p-value < 0.05 was considered significant.Pooling of available outcome data  was performed using “Review Manager 5.3” using the random effects model. Study heterogeneity was quantified using the DerSimonian–Laird method. The pooled data were presented with their corresponding 95% confidence intervals (CI). The graft and patient survival between the groups were compared using generic inverse variance described by Tierney et al. . The hazA text search was performed on 13 September 2022. A PRISMA flow chart of the search process is presented in p < 0.001, I2: 0%, p = 0.04, I2: 0%, p = 0.38, I2: 55%, p = 0.35, I2: 0%, p = 0.16, I2: 0%, p = 0.84, I2: 0%, p = 0.37, I2: 50%, p = 0.21, I2: 38%, p = 0.49, I2: 0%, Biliary and vascular complications and primary non-function are summarized in p = 0.01, I2: 0%, p = 0.03, I2: 74%, p = 0.002, I2: 0%, p = 0.47, I2: 89%, P = 0.20, I2: 4%, p = 0.03, I2: 0%, p = 0.98, I2: 0%, The graft and patient survival rates for each of the studies are summarized in Given the increasing demand for donor livers, the gap between supply and demand has kept widening. Several approaches have been taken to try to address this issue. One of which has been the use of ECD organs ,34. HoweI2: 0%).Both graft and patient survival in liver transplant recipients of grafts that underwent HOPE/D-HOPE instead of SCS were significantly better. The improvement in graft survival may be associated with reduced ischemia-reperfusion injury in grafts that are preserved using HOPE ,31,36. TI2-statistic, these studies were heterogenous (I2: 89%). On further investigation, we found graft survival in Mergental et al. [I2: 0%). This heterogeneity may be a result of the much smaller sample size in this study compared to the other three studies in the analysis. The NPM and SCS group sample sizes were 22 and 44, respectively, in Mergental et al. [However, in the studies that compared NMP to SCS, there was no significant difference in graft and patient survival, although graft survival slightly favored NMP. We do note though that based on l et al.  to be thl et al. ; 67 and l et al. ; 170 andl et al. ; and 142l et al. .Based on these results, HOPE/D-HOPE may provide more favorable graft and patient survival outcomes than NMP. However, we cannot provide concrete backing for this deduction. As such, it should be interpreted as a bird’s-eye-view takeaway from the findings, which merit further investigation.p = 0.03). However, the studies were significantly heterogeneous (I2: 74%). The heterogeneity here is most likely a result of the different methods of machine perfusion used in the different studies (HOPE vs. NMP). The patient survival was not significantly better in the machine perfusion group than in SCS, although machine perfusion was slightly favored (p = 0.2). Unlike the graft-survival analysis, in this case, the studies were homogenous (I2: 4%). The patient outcome was mostly affected by the survival results in the studies that used NMP. This is perhaps expected since HOPE and NMP are considered to be distinct graft-preservation techniques. HOPE has been reported to promote mitochondrial functional recovery, increase adenosine triphosphate levels and reduce the donor liver injuring the rewarming phase [In a pooled analysis of machine perfusion (NMP and HOPE/D-HOPE) vs. SCS, graft survival was significantly better in the machine perfusion group . However, the difference was not significant for NPM vs. SCS, and yet again, the studies were heterogeneous (I2: 55%). As with graft survival in the studies that compared NPM to SCS, Mergental et al. [I2 of 0%. We did, however, find biliary complications to be lower in machine perfusion as a whole vs. SCS . For the other postoperative outcomes we analyzed (PNF and vascular complications), there were no significant differences between HOPE and SCS, nor between NMP and SCS.We found a similar situation with respect to biliary complications. HOPE had a significantly lower incidence of biliary complications than SCS (l et al.  seems toWe could not conduct a detailed analysis of the potential mediating and confounding factors that may have impacted graft and patient survival in the included studies due to patient data availability limitations. However, this is an important aspect of survival analysis. Graft survival in liver transplant patients may be affected by male recipient–female donor sex mismatch, recipient blood group, number of transplantations, advanced donor age, pre-existing portal vein thrombosis and prolonged cold ischemia time ,42,43. PThe limitations of this study include relatively small sample sizes in some of the studies included. However, since the transplantation of machine-perfused livers is currently being investigated at a limited number of centers, the sample size limitation is still unavoidable. This will undoubtedly change as more liver transplantation centers adopt machine perfusion. Heterogeneity may also have had some impact on the results, especially in the NMP subgroup. In either case, the source of the heterogeneity was a single study whose sample size was much smaller compared to the other studies in the analysis. There may also be limitations due to the inclusion or exclusion bias. There may also be differences in surgical experience at different centers and protocols for HOPE and NMP in the different studies. For all the studies, survival data and hazard ratios were extracted and calculated using the method described by Tierney et al.  in theirMachine perfusion is gaining more interest in donor liver preservation and viability testing for marginal/suboptimal organs. In reported studies, HOPE/D-HOPE has been associated with improved graft and patient survival and reduced biliary complications. NMP has been reported to be helpful in the viability evaluation and rescue of marginal livers. Therefore, HOPE/D-HOPE and NMP are promising alternatives to SCS for donor liver preservation. They may help address the widening gap between the demand for and availability of donor livers by enabling the rescue and transplantation of marginal livers.


Journal of Comparative Physiology A, passed away. To commemorate his life and work, the Editor-in-Chief, Günther K. H. Zupanc, invited Carsten Duch and Ansgar Büschges to write an obituary for the Journal. The submitted manuscript underwent peer and editorial review before a revised version was published online on April 25, 2022 (Duch and Büschges Neuroforum (Duch and Büschges Neurowissenschaftliche Gesellschaft e.V. and published by Walter de Gruyter GmbH. Neither the Editor-in-Chief nor the publisher of the Journal of Comparative Physiology A, Springer Nature, had obtained prior knowledge of, or given consent for, the intended dual publication.On January 25, 2022, Hans-Joachim Pflüger, a former Associate Editor of the The authors acknowledged receipt of this Editorial Expression of Concern but did not comment on its content.


BackgroundSubarachnoid hemorrhage (SAH) grading scales typically evaluate a limited number of cisterns on the axial plane. The goal of our study is to apply a simple quantitative yet comprehensive SAH grading scale to all major intracranial cisterns, including the infratentorial cisterns, with multiplanar computed tomography (CT) reformations.MethodologyWe performed a retrospective review of 94 consecutive cases of spontaneous SAH presenting within 72 hours of onset. SAH was categorized into five grades based on the short-axis thickness of SAH in 20 intracranial cisterns measured on the axial, coronal, and sagittal planes. Statistical analysis was performed for inter-rater agreement with kappa statistics, for inter-plane agreement by Spearman correlation statistics, and for inter-rater and inter-plane agreement by Pearson correlation statistics.ResultsThe extended kappa coefficient for the three reviewers across all 20 cisterns varied from 0.38  to 0.59  on the axial plane. The kappa coefficient for two reviewers varied from 0.46  to 0.70  on the coronal plane and from 0.35  to 0.87  on the sagittal plane. The average grade of cisterns per case demonstrated mostly excellent correlation between the imaging planes with Spearman correlation statistics (≥0.70). Pairwise concordance correlation coefficient of the total SAH score revealed agreement ranging from 0.81 to 0.90 in all three planes. Pearson correlation statistics of the average total SAH scores revealed excellent correlation among the three planes (≥0.91).ConclusionA simple quantitative SAH grading scale can be successfully applied to the supratentorial and infratentorial cisterns in three standard CT imaging planes. Several studies have found a correlation between the risk of vasospasm and the severity of subarachnoid hemorrhage (SAH) . When moParticipantsThis retrospective study was conducted in compliance with the 1996 Health Insurance Portability and Accountability Act and approved by the Institutional Review Board  that waived the patient consent requirement. Relevant consecutive cases were identified on our institutional database through our picture archiving and communication system (PACS) using the Primordial search engine . We used the search terms “subarachnoid” and “SAH” and set the date parameters as January 2007 to February 2018. The search yielded 215 consecutive cases of spontaneous SAH admitted to our emergency department. Inclusion criteria were patients diagnosed with SAH on CT within three days of onset and multiplanar coronal and sagittal CT reformations. In total, 94 consecutive cases were identified for inclusion in our study.Image acquisitionMultidetector CT machines from different vendors were used in the cases studied. Images were acquired from the vertex to C2 vertebral level on the axial plane, with 3 to 5 mm slice thickness at zero spacing, 240 mm field of view, and a 512 × 512-pixel image matrix. Coronal and sagittal reformations were performed with 2 to 3 mm slice thickness at zero spacing.Image interpretationCT images were independently analyzed on the PACS by three fellowship-trained neuroradiologists (two neuroradiologists with more than 10 years of experience and a graduate neuroradiology fellow) who were blinded to the clinical data, radiology reports, and all angiographic images, including CT angiography and digital subtraction angiography. The images on the axial plane were analyzed by all three reviewers  and in the sagittal and coronal planes by two reviewers (R1 and R2). The maximum thickness of SAH in the shortest axis of a cistern in each imaging plane was measured and categorized into one of five grades: 0, no SAH; I, ≤3 mm; II, >3 to ≤6 mm; III, >6 to ≤11 mm; and IV, >11 mm  on the grades was assessed by estimating the kappa and extended kappa coefficients between the reviewers. Kappa <0 was rated as below-chance agreement, 0.01-0.20 as slight agreement, 0.21-0.40 as fair agreement, 0.41-0.60 as moderate agreement, 0.61-0.80 as substantial agreement, and 0.81-0.99 as almost perfect agreement . AgreemeA total of 94 cases  with an average age of 54.6  were evaluated. In total, 1,880 cisterns were assessed per imaging plane. Overall, 13,160 cisterns were assessed by the reviewers, 827 (6%) of which were documented as DA. The prepontine cistern was documented as DA on the coronal plane 82 and 71 times by R1 and R2, respectively. On the coronal plane, the premedullary cistern was documented as DA 74 and 46 times by R1 and R2, respectively, and the C1-C2 cistern was documented as DA 60 times by R1. This resulted in insufficient data for these three cisterns on the coronal plane. On the sagittal plane, the right cerebellomedullary cistern was scored as DA in 50 cases and the left in 46 cases by R1. On the same plane, the anterior suprasellar cistern was scored as DA in 62 cases and the interhemispheric fissure in 79 cases by R2. The axial plane allowed grading of all cisterns by the three reviewers without difficulty. Statistical analyses of the cerebellomedullary cisterns, anterior suprasellar cistern, and interhemispheric fissure on the sagittal plane were obtained with sparse data.The results of the IRA on the axial plane between the three reviewers R1, R2, and R3) and on the coronal and sagittal planes between the two reviewers (R1 and R2) are shown in Table , and R3 The risk of cerebral vasospasm is related to the severity of SAH ,13. TradThe Hijdra scale has produced considerable interobserver variation when used to grade individual cisterns . HoweverCONSCIOUS 1 to 3 trials evaluating the efficacy of the endothelin receptor antagonist, clazosentan, in the treatment of cerebral vasospasm, rely on simple quantitative measurements of variables such as the thickness and length of SAH in a cistern rather than using qualitative grading scales -17. MeasThere were a few challenges to grading SAH with our scale. Difficulties occurred in measuring the short thickness of SAH in a very small number of cisterns on certain planes. This limitation was seen when grading the prepontine, premedullary, and C1-C2 cisterns on the coronal plane because their long axes precluded short-axis measurement of the SAH thickness. Similarly, on the sagittal plane, sparse grading data was available for the anterior suprasellar, interhemispheric, and bilateral cerebellomedullary cisterns. However, in all these challenging instances, grading was accomplished on at least one plane.Some studies have categorized SAH with a thickness of >4 mm as “thick SAH clots.” Observation of “thick SAH clots” in three or more cisterns was used to distinguish diffuse SAH and successfully predict the risk of vasospasm-induced morbidity and outcomes ,18. UnliThe retrospective design was a major limitation of our study. Artificial intelligence (AI) is increasingly used in imaging and, in the future, may minimize or eliminate the need for manual grading of SAH. However, current automated techniques incorporate several manual steps in the evaluation of SAH, and none is yet well accepted in clinical practice ,20. To dWe have presented a simple quantitative SAH grading scale using multiplanar CT scan images that can be successfully applied to the supratentorial and infratentorial cisterns on the three standard CT imaging planes. The proposed grading scale addresses a major shortcoming of previous scales with its inclusion of infratentorial cisterns. Our quantitative method of grading SAHs uses axial, coronal, and sagittal multiplanar CT reformations and is practical to apply in clinical settings. It offers a strong IRA for the overall grading of the extent of SAHs.


Globally, 296 million people are infected with chronic hepatitis B (CHB) and at risk for cirrhosis and hepatocellular carcinoma. Suppressive therapy with a nucleos(t)ide analogue (NUC) such as tenofovir alafenamide (TAF) reduces risk of cirrhosis and cancer. Transient elastography measures liver stiffness and is used to monitor liver fibrosis in CHB. This study evaluates liver stiffness change among people taking NUC and determines correlates with improvement in liver stiffness.Participants enrolled at the University of Maryland, Baltimore were treated with TAF 25mg daily for 2 years. Liver stiffness (FibroScan©) was measured at baseline and year 2. Mild (F0/F1) and advanced (≥ F2) fibrosis, was defined as ≤ 7.4kPa and ≥ 7.5kPa, respectively. Correlates of liver stiffness change from baseline to year 2 was evaluated with multiple linear regression. P-value of < 0.05 was considered statistically significant.Since 2017, 60 have completed the study. Twenty-nine were switched from another NUC to TAF and 31 were off NUC treatment at baseline . Baseline alanine transaminase (ALT) was higher in the "No NUC" group . Most (67.2%) participants had mild fibrosis at baseline. Nineteen had advanced fibrosis. Among those with advanced fibrosis, fibrosis stage improved by at least 1 stage in 14 with no change in 5. Liver stiffness improved in the No NUC group , but not in the switch group. Subgroup analysis of those with advanced fibrosis also showed reduction in liver stiffness in the No NUC group only. Increased baseline ALT and baseline liver stiffness were predictive of reduction in liver stiffness .Baseline liver stiffness predicts change in liver stiffness with NUC therapyIncreased liver stiffness, and therefore fibrosis, at baseline was associated with greater reductions in liver stiffness, and therefore liver stiffness, after 2 years of NUC therapy .Liver stiffness improved with 2 years of NUC treatment in people with CHB, including those with advanced liver fibrosis. Either baseline ALT or liver stiffness can predict reduction in liver stiffness. This may reflect an overestimation of fibrosis stage in the presence of liver inflammation at NUC initiation. Repeating FibroScan after initiation of NUC treatment would therefore be useful, especially in people with elevated ALT, for guiding CHB care.Lydia Tang, MBChB, Gilead Sciences: Grant/Research Support Shyam Kottilil, MD, PhD, Arbutus Pharmaceuticals: Grant/Research Support|Gilead: Grant/Research Support|Merck: Grant/Research Support|Regeneron Pharmaceuticals: Advisor/Consultant|Silverback Therapeutics: Advisor/Consultant|The Liver Company: Advisor/Consultant|Yufan Biotechnologies: Advisor/Consultant.


O), peak GT (TP), and conclusion GT (TC). Among the thermodynamic properties, ΣDP 12–24 was significantly positively correlated with To, Tp, and Tc, ΣDP 25–36 was significantly negatively correlated with To, Tp, and Tc, and ΣDP ≥ 37 had no correlation with the thermodynamic properties. The results of correlation analysis with RVA spectrum characteristic values showed that ΣDP 6–11 was significantly negatively correlated with hot paste viscosity (HPV), cool paste viscosity (CPV), consistency viscosity (CSV), peak time (PeT), and pasting temperature (PaT) among the Rapid Visco Analyzer (RVA) profile characteristics, ΣDP 12–24 was significantly positively correlated with HPV, CPV, CSV, PeT, and PaT, and ΣDP ≥ 25 had no correlation with the viscosity characteristics. Therefore, we concluded that the amylopectin structure had a greater effect on the TO, TP, TC, ΔH and peak viscosity, HPV, CPV, CSV, PeT, and PaT. The glutinous rice varieties with a higher distribution of short chains and a lower distribution of medium and long chains in the amylopectin structure resulted in lower GT and RVA spectrum characteristic values.Glutinous rice has very low amylose content and is a good material for determining the structure and physicochemical properties of amylopectin. We selected 29 glutinous rice varieties and determined the amylopectin structure by high-performance anion exchange chromatography with the pulsed amperometric detection method. We also determined the correlation between amylopectin structure and the physicochemical properties of starch extracted from these varieties. The results showed that the amylopectin chain ratio Σdegree of polymerization (DP) ≤ 11/ΣDP ≤ 24 of 29 glutinous rice varieties was greater than 0.26, signifying that these varieties contained type II amylopectin. The results of the correlation analysis with gelatinization temperature showed that ΣDP 6–11 was significantly negatively correlated with the onset gelatinization temperature (GT) (T The structure of amylopectin has an important effect on the physicochemical properties of starch ,2, whichIn this study, 29 glutinous rice varieties of different qualities were used as study materials, including 17 varieties of indica rice and 12 varieties of japonica rice . Test vaThe following experimental instruments were used in this study: TU-1810D ultraviolet-visible spectrophotometer: Beijing Puxi Company, Beijing, China; Thermo ICS5000+ ion chromatography system: Thermo Fisher Scientific, Waltham, MA, USA; Matersizer 3000 Laser Particle Size Analyzer: Malvern Instruments Ltd., Worcestershire, UK; X’Pert Pro X-ray diffractometer: PANalytical, Almelo, The Netherlands; RVA-TecMaster viscometer: Perten, Stockholm, Sweden; and DSC 4000 differential scanning calorimeter (DSC): Perkin-Elmer, Waltham, MA, USA.Wx) gene to detect glutinous rice varieties. The polymerase chain reaction (PCR) was performed under the following conditions: 5 min at 94 °C, followed by 30 s at 94 °C, 30 s at 55 °C, and 30 s at 72 °C for 35 cycles, and 10 min at 72 °C for a final extension. Only waxy varieties have 492 bp PCR products, while non waxy varieties have no PCR products [DNA was extracted according to the cetrimonium bromide method , and seqproducts . PCR ampAC was determined according to the Chinese national standard GB/T15683-2008 .Starch was extracted and purified according to the method of Wei et al. .According to the method of Zhang et al. , the amyX’Pert Pro X-ray diffractometer  was used to analyze the X-ray diffraction patterns of the crystallographic structure of starch. The relative crystallinity of starch was calculated using the MDI Jade software.The starch viscosity was determined using the RVA-TecMaster viscometer  and its supporting software, Thermal Cycle for Windows, according to the American Association of Cereal Chemists operating procedures (1995 61-02) . The obtO), peak GT (TP), conclusion GT (TC), and gelatinization enthalpy (ΔH) were recorded. The specific methods are: weigh 5.0 mg of rice flour sample with 14.0% water content into an aluminum crucible and add 10 μL deionized water, after mixing, use a matching sample press to seal the crucible, and leave it in a 4 °C refrigerator overnight; before the test, take out the crucible stably, place it at room temperature and balance it for 1 h, and then go on the machine for measurement; take the empty disk as the control, and conduct 10 °C/min heating at 30~110 °C; analyze the sample curve and record TO, TP, TC, and ΔH.The thermodynamic properties were measured using a DSC , and the sample curves were analyzed using the supporting software Pyris Manager. The onset GT , Duncan’s multiple comparisons, and Pearson’s correlation. The means of duplicated measurements were used for the analysis. Significant differences in the mean values were determined at wx is an allelic variation of the Wx gene, which has a recessive mutation due to 23-bp fragment deletion in the second exon of the Wx gene. The appearance of glutinous rice greatly differs from that of non-glutinous rice, but distinguishing this appearance among the heterozygous genotypes is difficult using conventional methods [Wx gene, and the amplified fragment length was 492 bp. According to AC determination and the Wx genotyping results of the tested varieties, AC had little effect on the physicochemical properties of starch in glutinous rice, and the amylopectin structure was the determining factor affecting the physicochemical properties of starch.The glutinous gene  methods . Therefo methods  to detecIn this study, the amylopectin chain length and its chain-length distribution in the 29 test varieties were determined according to the method of He Xiaopeng . The amyThe results of the physicochemical properties of starch in the test varieties are shown in O, TP, and TC values were 62.54–76.91 °C, 70.25–80.66 °C, and 77.92–88.18 °C, respectively. In the heating process of glutinous rice starch, due to the complexity of amylopectin structure, the pasting temperature of individual varieties is higher. Compared with ordinary rice, most glutinous rice varieties had lower GTs. The ΔH was between 6.33–12.10 J/g, with a mean value of 9.30 J/g. The large difference in ΔH among the test varieties indicated that during the heating process of the glutinous rice starch, due to the complexity of the amylopectin structure, the enthalpy change values among the varieties were different.Among the RVA spectrum characteristic values of the test varieties, PKV ranged from 1523 centipoise (cP) to 3425.33 cP, which was generally lower than that of the common rice. HPV ranged from 882 cP to 2111.33 cP, CPV ranged from 1085 cP to 2563 cP, BDV ranged from 641 cP to 1529 cP, SBV ranged from −1222.67 cP to −363.67 cP, CSV ranged from 203 cP to 484 cP, PeT ranged from 3.76 min to 4.78 min, and PaT ranged from 70.95 °C to 83.00 °C. The measurement results of the thermodynamic characteristics showed that the ranges of variation of TFurther correlation analysis between the thermodynamic and physicochemical properties of starch in different glutinous rice varieties  revealedO, TC, and ΔH, the other characteristic values were significantly correlated with the thermodynamic parameters. Relative crystallinity and ΔH were significantly positively correlated. To and Tc were significantly positively correlated with HPV, CPV, CSV, PeT, and PaT. Tp was significantly positively correlated with PKV, HPV, CPV, CSV, PeT, and PaT. ΔH was significantly positively correlated with PaT, HPV, CPV, CSV, and PeT. Among these, the correlation coefficients between PaT and TO, TP, TC, and ΔH were the highest, which were 0.965, 0.944, 0.689, and 0.591, respectively, indicating a great correlation among each of the physicochemical properties of starch. PaT in the RVA spectrum can differ in GTs of different varieties to some extent.Except that BDV and SBV had no correlation with thermodynamic parameters, and PKV had no correlation with TO, TP, TC, and ΔH in the thermodynamic properties. ΣDP 12–24 was significantly positively correlated with HPV, CPV, CSV, PeT, and PaT in the RVA spectrum characteristic values and TO, TP, TC, and ΔH in the thermodynamic properties. ΣDP 25–36 was significantly negatively correlated with PaT in the RVA spectrum characteristic values and TO, TP, and TC in the thermodynamic properties. No correlation was present between ΣDP ≥ 37 and the physicochemical and thermodynamic properties of each starch sample. The average DP was only significantly positively correlated with PeT and PaT in the RVA spectrum characteristic values, indicating that the lesser the short-chain distribution of glutinous rice amylopectin (DP 6–11), the more the medium-long-chain distribution (DP 12–24), and the lesser the long-chain distribution (DP 25–36), the higher the RVA spectrum characteristic values, this is consistent with Zhou’s study [The correlation analysis results of the amylopectin chain length distribution and physicochemical properties of starch are shown in ’s study . The higAmylopectin is the main component of the rice endosperm. With the development of science and technology, the methods for amylopectin structure determination are also gradually developing. Presently, these methods can be divided into two categories: electrophoresis and chromatography, with each category consisting of various submethods and having its own advantages and disadvantages. The methods can be divided into FACE, enzymatic method, gel chromatography , and speHe et al.  used theae rice mutant and a significant increase in the short chains, resulting in a significant decrease in TO of starch. He et al. [O, TP, and TC of the test varieties were significantly negatively correlated with ΣDP 6–11 and ΣDP 25–36 and significantly positively correlated with ΣDP 12–24. These results were consistent with those of previously reported studies, indicating that the amylopectin structure plays similar and important role in the GT of glutinous rice and non-glutinous rice. The relative number of short chains with DP 6–11 and long chains with DP 25–36 should be increased, whereas the relative number of medium-length chains with DP 12–24 should be reduced to improve the rice amylopectin structure for reducing GT.GT refers to the temperature at which a large number of water-absorbing starch granules undergo irreversible expansion, birefringence, and crystallinity disappearance after being heated in a suspension aqueous solution . Vandepue et al.  found thThe RVA spectrum characteristic values are closely related to the eating quality of rice. The rice varieties with lower HPV, CPV, SBV, CSV, PeT, and PaT and a higher BDV are considered to have better grain quality, softer texture, better viscosity, and cold rice texture . By perfO, TP, and TC among the thermodynamic properties, ΣDP 12–24 was significantly positively correlated with To, Tp, and Tc, ΣDP 25–36 was significantly negatively correlated with To, Tp, and Tc. ΣDP 6–11 was significantly negatively correlated with HPV, CPV, CSV, PeT, and PaT among the RVA profile characteristics, ΣDP 12–24 was significantly positively correlated with HPV, CPV, CSV, PeT, and PaT. Therefore, we concluded that the amylopectin structure had a greater effect on the TO, TP, TC, ΔH and peak viscosity, HPV, CPV, CSV, PeT, and PaT. The glutinous rice varieties with a higher distribution of short chains and a lower distribution of medium and long chains in the amylopectin structure resulted in lower GT and RVA spectrum characteristic values. These new findings will significantly assist in revealing the genetic basis of the formation of amylopectin structure in glutinous rice and provide a theoretical basis for the breeding and improvement of glutinous rice starch quality.In this study, the amylopectin chain ratio ΣDP ≤ 11/ΣDP ≤ 24 of 29 glutinous rice varieties was greater than 0.26, signifying that these varieties contained type II amylopectin. The results of the correlation analysis showed that ΣDP 6–11 was significantly negatively correlated with T


Transportation, logistics, storage, and many other sectors provide a wide space for applying Industry 4.0. This era, with its components, represents the equipment necessary to obtain a unique competitive advantage. Being smart through sensors, big data, and digitalization corresponds not only to evolution but also provides protection for businesses in the face of depression. The COVID-19 pandemic caused collapses and defects for very large enterprises and large enterprises, especially for small and medium-sized enterprises (SMEs). This article focuses on SMEs and their profits from using smart sensors. Thus, the aim was to expose the striking effect of Industry 4.0 on earnings during the crisis in the Visegrad Four. The Mann–Kendall trend was used to map the consequences contrasting the period of 2016–2021. The investigation involved samples from 1221 Slovak, 259 Czech, 855 Polish, and 2156 Hungarian enterprises. The results showed that more than 80% of businesses did not have a negative trend in how their earnings changed over time. This fact was confirmed by a z-test for the comparison of one proportion for each analyzed country. The adaptation to Industry 4.0 strengthened the muscle for bankruptcy resilience during the crisis. In addition, it may encourage enterprises to be smart in the same or different sectors. Smart sensors are applied in sectors and their systems to indicate changes or monitor trends in controlling parameters during the Industry 4.0 era . Small aTijani et al.  advocateIn general, Industry 4.0 is a fundamental requirement for process optimization, waste reduction, innovation introduction, and acquiring competitive advantages . Its polThus, the aim of this research was to expose the striking effect of Industry 4.0 on earnings during the crisis in the Visegrad Four. Many studies have confirmed the dependence between the derivatives of Industry 4.0 and the maintenance of earnings for enterprises in general. The contribution of this study compared to previous studies is a comprehensive assessment of SMEs for the V4 region on the issue based on robust samples and confirmation of the significant effect of smart sensors against crises as evidenced by the COVID-19 pandemic example. The novelty consisted of using methodologies that combined trend analysis, heterogeneity occurrence, and the z-test. This article especially focuses on smart sensors; the last incentives have been as follows: Blake and Frajtova Michalikova  providedMarinov et al.  showed tValaskova et al.  linked sHa et al.  enumeratShirmohammadli and Bahreyni  preparedSmart sensors lead to an increase in the accumulation of big data ,69. GibsThis article is structured as follows: A literature review of studies pertaining to the evaluation of smart sensors and the most recent incentives is highlighted first. The statistical techniques employed in the provided study are then demonstrated, together with the dataset that was used. The This study targeted SME enterprises. The choice was limited to businesses that were oriented toward smart sensors because this issue has been scarcely documented in the literature . The perThe earnings were chosen because owners, managers, or other external entities set the greatest momentousness for financial quantification ,75. It hThe financial situations of SMEs were expressed in this investigation as earnings before interest, tax, depreciation, and amortization (EBITDA). This indicator is recommended for use in the comparison of different sectors ,82 becauCreation of a sample.The following methodology was used in this study:The outliers were not excluded because the sub-samples were not balanced to obtain a relevant and robust sample from V4, reflecting the actual situation in the sector ,84,85,86Each country was labeled with a number: 1 for Slovakia, 2 for Czechia, 3 for Poland, and 4 for Hungary. These numbers were used to name the countries in the tested hypotheses.2.Detection of a trend.There was a premise that smart sensors supported enterprises. Then, we could not reject the occurrence of no trend within EBITDA during 2016–2021. The existence of a monotonic trend was found by the Mann–Kendall trend test. The time series for Slovak, Czech, Polish, and Hungarian SMEs were checked for a negative trend against a distribution that was identical. Kliestik et al.  noted thS) is defined as:S is E[S] = 0 and the variance p is the number of the tied groups in the data set and jt is the number of data points in the j-th tied group. The statistic S is approximately normally distributed provided that the following Z-transformation is employed:S and more well-known, such as Kendall’s τ, which is given by:The nonparametric statistic  SMEs. There was a negative trend confirmed for the period of 2016–2021.3.Disclosure of a change.Pettitt’s test was run after the trend test for SMEs; it confirmed the negative development. It was done to trace heterogeneity and the year that changed the decrease in EBITDA as based on the study by Kanovsky . There wValaskova et al.  noted thTK provided that the statistic is significant. The significance probability of TK is approximated for a p-value ≤ 0.05 as:The nonparametric statistic is defined as:The following theory was developed and tested separately for each nation:b.H There was a phenomenon of heterogeneity in earnings for smart Slovak, Czech, Polish, and Hungarian SMEs. The pandemic years caused a negative disruption in the period of 2016–2021.4.Confirmation of a proportion.The pandemic years were determined as the years of change within EBITDA. Then, the z-test for the comparison of one proportion was applied to confirm the percentage of SMEs that were protected during a crisis via smart sensors. The findings could be generalized after running this test based on the used samples.Svabova et al.  noted thAn assumption of the The test statistic The test statistic may be greater than the standard normal distribution earchers ,100,101 The following theory was developed and tested separately for each nation:c.H More than 80% of Slovak  SMEs balanced earnings during the crisis.The results assessed the nexus between smart sensors and the retention of earnings by SMEs. The sample included 4491 units from the V4 region and covered a 6-year period. Firstly, the occurrence of a negative trend in earnings in smart enterprises was checked during the period 2016–2021. The significance level alpha was set at level 0.05 for all the investigations. Each country of the Visegrad Four was tested in turn.The following hypotheses were mapped to the analysis in Slovakia:0a:H Smart sensors balanced earnings for Slovak SMEs. There was no trend in the period of 2016–2021.1a.H Smart sensors did not balance earnings for Slovak SMEs. There was a negative trend confirmed for the period of 2016–2021.The Mann–Kendall trend test was run for each enterprise in the sample of 1221 SMEs. If the computed p-value is lower than the significance level alpha, one should reject the null hypothesis and accept the alternative hypothesis based on Based on The following hypotheses were mapped to the analysis in Czechia:0a.H Smart sensors balanced earnings for Czech SMEs. There was no trend in the period of 2016–2021.2a.H Smart sensors did not balance earnings for Czech SMEs. There was a negative trend confirmed for the period of 2016–2021.p-value is lower than the significance level alpha, one should reject the null hypothesis and accept the alternative hypothesis based on The Mann–Kendall trend test was run for each enterprise in the sample of 259 SMEs. If the computed Based on The following hypotheses were mapped to the analysis in Poland:0a.H Smart sensors balanced earnings for Polish SMEs. There was no trend in the period of 2016–2021.3a.H Smart sensors did not balance earnings for Polish SMEs. There was a negative trend confirmed for the period of 2016–2021.p-value is lower than the significance level alpha, one should reject the null hypothesis and accept the alternative hypothesis based on The Mann–Kendall trend test was run for each enterprise in the sample of 855 SMEs. If the computed Based on The following hypotheses were mapped to the analysis in Hungary:0a.H Smart sensors balanced earnings for Hungarian SMEs. There was no trend in the period of 2016–2021.4a.H Smart sensors did not balance earnings for Hungarian SMEs. There was a negative trend confirmed for the period of 2016–2021.p-value is lower than the significance level alpha, one should reject the null hypothesis and accept the alternative hypothesis based on The Mann–Kendall trend test was run for each enterprise in the sample of 2156 SMEs. If the computed Based on Secondly, the negative trend was explored in detail for 460 SMEs. The test for heterogeneity was applied individually in every enterprise with this kind of trend. The pandemic years of 2020 and 2021 were supposed to be the cause of decreased development in the time series. The significance level alpha was set at level 0.05 for all the investigations. Each country of the Visegrad Four was tested in turn.The following hypotheses corresponded to the analysis of Slovakia:0b.H There was no phenomenon of heterogeneity in earnings for smart Slovak SMEs. The pandemic years did not cause a negative disruption in the period of 2016–2021.1b.H There was a phenomenon of heterogeneity in earnings for smart Slovak SMEs. The pandemic years caused a negative disruption in the period of 2016–2021.p-value is lower than the significance level alpha, one should reject the null hypothesis and accept the alternative hypothesis. Thus, the phenomenon of heterogeneity in earnings was accepted for each unit in the sample. Based on Pettitt’s test was run for each enterprise in the sample of 154 Slovak SMEs. If the computed The following hypotheses corresponded to the analysis of Czechia:0b.H There was no phenomenon of heterogeneity in earnings for smart Czech SMEs. The pandemic years did not cause a negative disruption in the period of 2016–2021.2b.H There was a phenomenon of heterogeneity in earnings for smart Czech SMEs. The pandemic years caused a negative disruption in the period of 2016–2021.p-value is lower than the significance level alpha, one should reject the null hypothesis and accept the alternative hypothesis. Thus, the phenomenon of heterogeneity in earnings was accepted for each unit in the sample. Based on Pettitt’s test was run for each enterprise in the sample of 31 Czech SMEs. If the computed The following hypotheses corresponded to the analysis of Poland:0b.H There was no phenomenon of heterogeneity in earnings for smart Polish SMEs. The pandemic years did not cause a negative disruption in the period of 2016–2021.3b.H There was a phenomenon of heterogeneity in earnings for smart Polish SMEs. The pandemic years caused a negative disruption in the period of 2016–2021.p-value is lower than the significance level alpha, one should reject the null hypothesis and accept the alternative hypothesis. Thus, the phenomenon of heterogeneity in earnings was accepted for each unit in the sample. Based on Pettitt’s test was run for each enterprise in the sample of 63 Polish SMEs. If the computed The following hypotheses corresponded to the analysis of Hungary:0b.H There was no phenomenon of heterogeneity in earnings for smart Hungarian SMEs. The pandemic years did not cause a negative disruption in the period of 2016–2021.4b.H There was a phenomenon of heterogeneity in earnings for smart Hungarian SMEs. The pandemic years caused a negative disruption in the period of 2016–2021.p-value is lower than the significance level alpha, one should reject the null hypothesis and accept the alternative hypothesis. Thus, the phenomenon of heterogeneity in earnings was accepted for each unit in the sample. Based on Pettitt’s test was run for each enterprise in the sample of 212 Hungarian SMEs. If the computed A negative trend was proven due to the pandemic and its consequences. Finally, the study exposed the striking effect of Industry 4.0 on earnings. The tests for one proportion disclosed the percentage of SMEs that protected their earnings using smart sensors during a crisis. The findings were generalized based on the analyzed samples for the entire population. The significance level alpha was set at level 0.05 for all the investigations. Each country of the Visegrad Four was tested in turn.The hypotheses supporting the analysis of Slovakia were:0c.H 80% of Slovak SMEs balanced earnings during the crisis via Industry 4.0.1c.H More than 80% of Slovak SMEs balanced earnings during the crisis via Industry 4.0.p-value is lower than the significance level alpha, one should reject the null hypothesis and accept the alternative hypothesis. Thus, more than 80% of Slovak SMEs balanced earnings during the crisis via Industry 4.0. This was generalized based on the p-value < 0.0001 from The z-test for comparison of one proportion tested if the difference between the proportions was equal to 0 or not. If the computed The hypotheses supporting the analysis in Czechia were:0c.H 80% of Czech SMEs balanced earnings during the crisis via Industry 4.0.2c.H More than 80% of Czech SMEs balanced earnings during the crisis via Industry 4.0.p-value is lower than the significance level alpha, one should reject the null hypothesis and accept the alternative hypothesis. Thus, more than 80% of Czech SMEs balanced earnings during the crisis via Industry 4.0. This was generalized based on the p-value = 0.0008 from The z-test for comparison of one proportion tested if the difference between the proportions was equal to 0 or not. If the computed The hypotheses supporting the analysis in Poland were:0c.H 80% of Polish SMEs balanced earnings during the crisis via Industry 4.0.3c.H More than 80% of Polish SMEs balanced earnings during the crisis via Industry 4.0.p-value is lower than the significance level alpha, one should reject the null hypothesis and accept the alternative hypothesis. Thus, more than 80% of Polish SMEs balanced earnings during the crisis via Industry 4.0. This was generalized based on the p-value < 0.0001 from The z-test for comparison of one proportion tested if the difference between the proportions was equal to 0 or not. If the computed The hypotheses supporting the analysis in Hungary were:0c.H 80% of Hungarian SMEs balanced earnings during the crisis via Industry 4.0.4c.H More than 80% of Hungarian SMEs balanced earnings during the crisis via Industry 4.0.p-value is lower than the significance level alpha, one should reject the null hypothesis and accept the alternative hypothesis. Thus, more than 80% of Hungarian SMEs balanced earnings during the crisis via Industry 4.0. This was generalized based on the p-value < 0.0001 from The z-test for comparison of one proportion tested if the difference between the proportions was equal to 0 or not. If the computed The results were discussed to determine whether they were a close match with the last studies from the V4 region. The findings for SMEs (primarily from the transportation and automotive sectors) may be expressed through the juxtaposition of similar ones.Gasiorek  did not Karmanska  exploredKliestik et al.  assessedThe exploration by Michalkova et al.  added 26The aim of this study was to expose the striking effect of Industry 4.0 on earnings during the crisis in the Visegrad Four. Specifically, the advantages of using smart sensors for the retention of earnings as represented by EBITDA were shown. A decrease in earnings may support even the bankruptcy of SMEs. It was proven that COVID-19 caused a negative trend in earnings. During the pandemic crisis (2020–2021), however, smart sensors the balanced earnings of more than 80% of Slovak  SMEs. Based on these results, Industry 4.0 and its components mean not only progress, but also the protection business in which they have been involved. This positive fact, which was based on verifiable samples, is the next reason for governments and authorities to support and speed up programs that apply smart manufacturing systems and technology.The first limitation was that there was not an equal number of enterprises in the samples for each country. The SMEs declared the dominant use of smart sensors as a possible progression of tools. However, we did not check whether there were positive effects of other parts of Industry 4.0 that were implemented in SMEs. Further, the orientation was for all sectors and only SMEs. Thus, future research should be extended to the investigation of large and very large enterprises. In addition, the orientation per sector included balanced samples that involved Industry 4.0. The other parts of Industry 4.0 may also be tested while their effects are being examined.