Binding Capacity
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2022 ◽  
Anna Balcarak ◽  
Ewelina Macech-Klicka ◽  
Maciej Wakula ◽  
Rafal Tomecki ◽  
Krzysztof Goryca ◽  

HAX1 is a human protein with no known homologues or structural domains, mutations in which cause severe congenital neutropenia through mechanisms that are poorly understood. Previous studies reported RNA-binding capacity of HAX1, but the role of this binding in physiology and pathology remains unexplained. Here we report transcriptome-wide characterization of HAX1 RNA targets using RIP-seq and CRAC, indicating that HAX1 binds transcripts involved in ribosome biogenesis and rRNA processing. Using CRISPR knockouts we find that RNA targets of HAX1 partially overlap with transcripts downregulated in HAX1 KO, implying a role in mRNA stabilization. Gene ontology analysis demonstrated that genes differentially expressed in HAX1 KO (including genes involved in ribosome biogenesis and translation) are also enriched in a subset of genes whose expression correlates with HAX1 expression in four analyzed neoplasms. Functional connection to ribosome biogenesis was also demonstrated by gradient sedimentation ribosome profiles, which revealed differences in the small subunit:monosome ratio in HAX1 WT/KO. We speculate that changes in HAX1 expression may be important for the etiology of HAX1-linked diseases through dysregulation of translation.

Eric P. Skaar ◽  
Roger Echols ◽  
Yuko Matsunaga ◽  
Anju Menon ◽  
Simon Portsmouth

AbstractCritically ill patients often present with low serum iron levels or anemia. We evaluated the impact of iron levels and iron homeostasis on the efficacy and safety of cefiderocol, an iron-chelator siderophore cephalosporin, in patients with nosocomial pneumonia in a post hoc analysis of the randomized, double-blind, Phase 3 APEKS-NP study (NCT03032380). Patients with Gram-negative nosocomial pneumonia received cefiderocol 2 g, 3-h infusion, q8h, or high-dose, extended-infusion meropenem 2 g, 3-h infusion, q8h, for 7–14 days. Efficacy and safety parameters, including specific iron homeostasis parameters (i.e., hepcidin, iron, total iron binding capacity, transferrin saturation), were analyzed according to baseline iron levels. In the cefiderocol and meropenem arms, 79.1% (117/148) and 83.3% (125/150) randomized patients, respectively, had low baseline serum iron levels. Rates of 14-day (12.3% [14/114] vs 11.6% [14/121]) and 28-day all-cause mortality (20.5% [23/112] vs 19.0% [23/121]), clinical cure (63.2% [72/114] vs 67.2% [82/122]), and microbiological eradication (43.6% [41/94] vs 48.1% [51/106]) at test of cure were similar in cefiderocol vs meropenem arms, respectively. In the overall safety population, rates of anemia-related adverse events were similar (cefiderocol arm 18.2% [27/148], meropenem arm 18.7% [28/150]). Changes from baseline to test of cure in hepcidin, iron, total iron binding capacity, and transferrin saturation were similar between treatment arms. Cefiderocol treatment did not affect iron homeostasis, and its efficacy and safety were not influenced by baseline serum iron levels. registration: NCT03032380. Date of registration: 26 January 2017.

2022 ◽  
Tatiana A. Ignatova ◽  
Nataliya G. Strokova ◽  
Anna N. Roshchina

When agarophytes are processed, 16-89% algal waste (AW) is formed. Due to the high content of minerals in this waste and the presence of proteins that are resistant to the action of proteolytic enzymes, such AW is mainly used in the production of feed for farm animals. The significant content of polysaccharides in dry AW indicates that it could be used as a raw material for the production of dietary fiber (DF). Due to the difference in the chemical composition of AW depending on the type of red algae, different approaches for its deproteinization have been used. Thus, a 3% alkali solution should be used for the deproteinization of algal waste from the red algae Gracilaria, a 0.5% alkali solution for GelidiumAW, and a 1% sodium carbonate solution for A. plicataAW.The duration of the deproteinization process is 30 min at a temperature of 97±2 ∘C for all types of AW. In this study, functional and technological solutions of DF from AW were developed. The results showed that their water binding capacity was 6-22 g of water per 1 g of the preparation, the fat binding capacity was 1.6-3.3 g of fat per 1 g of the preparation, and the swelling capacity was 46-312% depending on the type of red algae.The obtained DF was used in the production of minced fish food products of the ‘fish sticks’ type, based on minced fish and consisting of cod and pink salmon. The study of the chemical composition and calorie content of the developed food product showed the possibility of its use in dietary nutrition. Keywords: red algae, algal waste, deproteinization, dietary fiber

Gels ◽  
2022 ◽  
Vol 8 (1) ◽  
pp. 48
Yuliya Frolova ◽  
Varuzhan Sarkisyan ◽  
Roman Sobolev ◽  
Mariia Makarenko ◽  
Michael Semin ◽  

This study aimed to find relationships between the properties of beeswax-based oleogels and the type of oil used. The influence of linseed, sunflower, olive, and fish oils was studied. For these oils, the fatty acid composition, the content of total polar components, and the iodine value were characterized. Textural and thermodynamic properties were determined for oleogels, the oil-binding capacity was estimated, and the morphology of crystals was studied. The concentration of beeswax in all oleogels was 6.0% w/w. It was shown that the type of oil has a significant influence on all characteristics of the oleogels. The use of different oils at the same technological treatment leads to the formation of crystals of diverse morphology—from platelets to spherulites. At the same time, it was revealed that some characteristics of oils have a varying contribution to the properties of oleogels. The content of total polar materials in oils is associated with a decrease in strength parameters (yield value and elastic modulus) and the oil-binding capacity of oleogels. In its turn, the iodine value of oils has a close positive correlation with the melting and crystallization temperatures of oleogels. The results obtained in this article indicate that the properties of beeswax-based oleogels can be directed by changing the oil composition.

2022 ◽  
Vol 8 (1) ◽  
Biradar Shivaleela ◽  
S. C. Srushti ◽  
S. J. Shreedevi ◽  
R. L. Babu

Abstract Background Inflammatory diseases are the vast array of disorders caused by inflammation. During most inflammatory events, many cytokines expressions were modulated, and one such cytokine is tumor necrosis factor-alpha (TNF-α). TNF-α is mainly secreted by monocytes and macrophages. Notably, it has been proposed as a therapeutic target for several diseases. The anti-TNF biology approach is mainly based on monoclonal antibodies. The fusion protein and biosimilars are prevalent in treating inflammation for decades. Only a few small molecule inhibitors are available to inhibit the expression of TNF-α, and one such promising drug was thalidomide. Therefore, the study was carried out to design thalidomide-based small molecule inhibitors for TNF-α. The main objective of our study is to design thalidomide analogs to inhibit TNF-α using the insilico approach. Results Several thalidomide analogs were designed using chemsketch. After filtration of compounds through ‘Lipinski rule of 5’ by Molinspiration tool, as a result, five compounds were selected. All these compounds were subjected to molecular docking, and the study showed that all five compounds had good binding energy. However, based on ADMET predictions, two compounds (S3 and S5) were eliminated. Conclusions Our preliminary results suggest that S1, S2, S4 compounds showed potential ligand binding capacity with TNF-α and, interestingly, with limited or no toxicity. Our preliminary investigation and obtained results have fashioned more interest for further in vitro studies.

2022 ◽  
Vol 12 ◽  
Luyao Tang ◽  
Enwen Guo ◽  
Lan Zhang ◽  
Ying Wang ◽  
Shan Gao ◽  

Carbohydrate-binding modules (CBMs), as an important auxiliary module, play a key role in degrading soluble alginate by alginate lyase, but the function on alginate gel has not been elucidated. Recently, we reported alginate lyase VxAly7B containing a CBM32 and a polysaccharide lyase family 7 (PL7). To investigate the specific function of CBM32, we characterized the full-length alginate lyase VxAly7B (VxAly7B-FL) and truncated mutants VxAly7B-CM (PL7) and VxAly7B-CBM (CBM32). Both VxAly7B-FL and native VxAly7B can spontaneously cleavage between CBM32 and PL7. The substrate-binding capacity and activity of VxAly7B-CM to soluble alginate were 0.86- and 1.97-fold those of VxAly7B-FL, respectively. Moreover, CBM32 could accelerate the expansion and cleavage of alginate gel beads, and the degradation rate of VxAly7B-FL to alginate gel beads was threefold that of VxAly7B-CM. Results showed that CBM32 is not conducive to the degradation of soluble alginate by VxAly7B but is helpful for binding and degradation of insoluble alginate gel. This study provides new insights into the function of CBM32 on alginate gel, which may inspire the application strategy of CBMs in insoluble substrates.

2022 ◽  
Shuren Guo ◽  
Xiaohuan Mao ◽  
Xiaohua Li ◽  
Huan Ouyang

Abstract Background: Disorders of iron metabolism has been implicated in cardiovascular disease. However, the association of serum ferritin and coronary artery disease (CAD) remains inconsistent. Here, we investigated the associations of serum iron metabolism with the incidence of CAD, the severity of coronary artery stenosis, metabolic biomarkers, and 1-year restenosis after coronary artery revascularization. Methods: A total of 643 CAD patients and 643 healthy controls were enrolled to assess the associations of serum iron status with the presence of CAD, the severity of CAD, and 1-year rehospitalization after revasculation. Serum iron metabolism and other metabolic markers were measured in all subjects. All statistical analyses were analyzed using SPSS22.0 software and STATA statistical package.Results: Serum level of iron metabolism markers, including serum iron, ferritin, unsaturated transferrin iron binding capacity (UIBC), Total iron binding capacity (TIBC) levels, in CAD groups was significantly higher than the control group (P<0.001). UIBC and TIBC were negatively correlated with ferritin in both sexes. Serum level of iron (OR=0.806, 95% CI (0.687-0.944), UIBC (OR=0.919, 95% CI (0.852-0.992), and TIBC (OR=0.864, 95% CI (0.787-0.95) were found to have a protective role for CAD in women (P<0.05, Table 3). The OR for ferritin was significant in the both sexes (OR=1.029, 95% CI (1.002-1.058) in men, OR=1.02, 95% CI (1.005-1.034) in women, P<0.05). Conclusion: Low Serum level of iron, UIBC, TIBC and ferritin levels were found to have a protective role for CAD in women, but not in men. Elevated serum ferritin is independently and positively associated with CAD in men and women.

Food systems ◽  
2022 ◽  
Vol 4 (4) ◽  
pp. 269-277
A. A. Papakhin ◽  
Z. M. Borodina

The use of debranching enzymes in starch hydrolysis is a topical direction for obtaining new types of starch products with controlled properties and a potential for the further use. The aim of the work was to study an effect of pullulanase (EC3.2.1.41) on maize amylopectin starch in the native and gelatinized state. The objects of the research were maize amylopectin starch and enzyme preparation Promozyme D6 (Novozymes, Denmark). High-performance liquid chromatography (HPLC) was used to determine the carbohydrate composition of hydrolysates. The mass fraction of reducing substances (RS) was determined by the Lane and Eynon method. A rotational viscometer was used to measure dynamic viscosity of the starch hydrolysis products. It was found that analyzed starch in the native state showed low enzymatic sensitivity to the action of pullulanase with insignificant changes in viscosity, solubility and iodine binding capacity of the samples. Pullulanase showed the highest effect on gelatinized starch during the first eight hours of incubation. After eight hours, the maximum degree of starch hydrolysis by pullulanase at a dose of 10 units/g dry matter (DM) was 4.7% on DM basis, iodine binding capacity of the hydrolysate was D600 = 0.343 (in the control experiment D600 = 0.154), and the viscosity of the hydrolysate decreased from 7887 mPa · s to 4.3 mPa · s. Hydrolysates cooled to 8 °C and held for 20 hours along with hydrolysates that were not cooled showed high susceptibility to attack by glucoamilase (97–98%) at 60 °C and 24 hours of saccharification, which suggested the absence of their resistance to the action of glucoamilase in the conditions of the experiment. The use of pullulanase in dextrinization of the analyzed starch, which was gelatinized and partly hydrolyzed by α-amylase (RS6.1%), enabled obtaining hydrolysates with the mass fraction of reducing substances in a range of 10–24% on DM basis with the process duration of 2 to 24 hours and the enzyme dose of 2–10 units, which contained mainly maltotriose, maltohexose and maltoheptose with their total amount of 45–60% on DM basis. The results indicate a need for further research of the biocatalytic action of pullulanase to develop new methods for enzymatic modification of starch.

Gels ◽  
2022 ◽  
Vol 8 (1) ◽  
pp. 30
Sheah Yee Ghan ◽  
Lee Fong Siow ◽  
Chin Ping Tan ◽  
Kok Whye Cheong ◽  
Yin Yin Thoo

The present work investigated the interaction between soy lecithin (SL), glyceryl monostearate (GMS), and water in structuring palm olein (PO) to create an organogel having similar mechanical properties to commercial spread. Extreme vertices mixture design was used to optimize the composition of PO-based organogel. The resulting model showed a good fit to the predicted data with R2 ≥ 0.89. The optimum composition was 8% SL, 22% GMS, 28% water, and 42% PO (w/w) to produce a mean firmness of 1.91 N, spreadability of 15.28 N s−1, and oil binding capacity (OBC) of 83.83%. The OBC of optimized organogel was 10% higher than commercial spread product, and no significant difference was observed in the mechanical properties (p > 0.05). The microstructure, as well as the rheological and thermal properties of the optimized organogel were characterized. Fourier transform infrared analysis indicated that hydrogen bonding and van der Waals interactions were the key driving forces for organogelation. The mixture of SL and GMS favored the formation of β′ + β form crystals with a predominance of the β′ form. These results have important implications for the development of PO-based organogel as a potential fat replacer in the production of low-fat spread.

2022 ◽  
Vol 2022 (1) ◽  
pp. pdb.prot103127
Edward A. Greenfield

In an antigen capture assay for hybridoma screening, the detection method identifies the presence of the antigen. Often this is achieved by labeling the antigen directly. In this assay, the polyvinyl chloride (PVC) wells of a high-binding-capacity ELISA plate are first coated with an affinity-purified rabbit anti-mouse immunoglobulin and then incubated with hybridoma tissue culture supernatant. Monoclonal antibodies in the supernatant are “captured” on the coated PVC surface and detected by screening with biotin- or histidine (His)–tagged antigen. The antigen can be labeled to a high specific activity and thus very little antigen is required for this procedure.

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