The RNA-binding landscape of HAX1 protein indicates its involvement in ribosome biogenesis and translation.

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.

Iron serum levels and iron homeostasis parameters in patients with nosocomial pneumonia treated with cefiderocol: post hoc analysis of the APEKS-NP study

Critically 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. Clinicaltrials.gov registration: NCT03032380. Date of registration: 26 January 2017.

Chestnut peels and wheat bran at different water level influence the physical properties of pan bread

In breadmaking, dietary fibres are used to improve the nutritional quality of the final products; on the other hand, they may affect the physical and sensory properties. This work aimed to the evaluate, on pan breads, the effect of substituting 3 g of wheat flour with an equivalent amount of fibre rich ingredients: chestnut peels (CP) or wheat bran (WB), in comparison to a traditional wheat bread formulation (C). The effect of four levels of added water (54, 60, 66, 71 g/100 of flour) was also tested. The fibre content of CP (33%) and WB (42%) affected their water binding capacity and, consequently, the quality of the final loaves, according to the different water addition levels. In bread crumb, water content and water activity increased proportionally to the water addition levels, being instead in the crust also affected by the presence of fibres: lower water retention capacity was observed for CP, in comparison to WB and C. The loaf volume resulted higher for C in comparison to WB and CP, in relation to the larger dimensions of the crumb pores, probably due to the interfering effect of fibres during the development of the gluten network. Crumb hardness resulted higher for C at low water addition levels, being instead higher for CP at high water addition levels. CP showed a darker and redder colour, than both WB and C bread, for the presence of the brown pigments carried by chestnut peels. PCA analysis confirmed that more water is required for both the fibre-enriched breads to show characteristics similar to the control loaves.

The Feasibility of Using Agarophyte Processing Waste

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

In-Depth Understanding of Granule Compression Behavior under Variable Raw Material and Processing Conditions

Tablet manufacturing involves the processing of raw materials through several unit operations. Thus, the mitigation of input-induced variability should also consider the downstream processability of intermediary products. The objective of the present work was to study the effect of variable raw materials and processing conditions on the compression properties of granules containing two active pharmaceutical ingredients (APIs) and microcrystalline cellulose. Differences in compressibility and tabletability of granules were highlighted in function of the initial particle size of the first API, granule polydispersity and fragmentation. Moreover, interactions were underlined with the atomizing pressure. Changing the supplier of the second API was efficiently controlled by adapting the binder addition rate and atomizing pressure during granulation, considering the starting crystal size. By fitting mathematical models on the available compression data, the influence of diluent source on granule compactibility and tabletability was identified. These differences resumed to the ease of compaction, tableting capacity and pressure sensitivity index due to variable water binding capacity of microcrystalline cellulose. Building the design space enabled the identification of suitable API types and the appropriate processing conditions (spray rate, atomizing pressure, compression force) required to ensure the desired tableting performance.

Prevalence and associated risk factors of iron deficiency without anemia among school adolescents in Mbouda, Western Cameroon

Background: Adolescents are a particularly vulnerable age group to iron deficiency without anemia (IDWA). This study aimed at determining the prevalence and associated risks factors of IDWA among apparently healthy school teenagers in Mbouda, West-Cameroon.Methods: The 778 blood samples were randomly collected from adolescents aged 10-19 years of four schools in Mbouda. Appropriate indicators including haemoglobin and haematocrit, serum iron, ferritin, transferrin, and total iron binding capacity, transferrin saturation were determined using standard methods.Results: The overall prevalence of IDWA was 40.4%. High significant difference in the mean values of serum iron (p≤0.001), serum ferritin (p≤0.001), TIBC (p=0.007), CST (p≤0.001) were observed in participants with IDWA compared to those without IDWA. The sociodemographic risk factors of IDWA in school adolescents were school site (p=0.022; 0.14 (0.90-1.43) and period of menstruation (p=0.015; 1.48 (1.083-2.03). Consumption of roots-tubers [OR: 0.92 (0.48-1.77)] legumes [OR: 2.65 (1.91-3.67)], cereals [OR: 0.90 (0.67-1.22)], increased the risk of IDWA while consumption of vegetables [OR: 0.29 (0.22-0.39)], eggs [OR: 0.66 (0.49-0.90)], and fish-shrimp [OR: 0.172 (0.10-0.295)] decreased the risk of IDWA. Meal consumption frequency had a statistically significant association with IDWA (p≤0.001).Conclusions: This study identified that IDWA was a major public health in adolescents. This study will help in planning and implementation of the policy for prevention of IDWA in adolescents.

A novel mouse model of type 2N VWD was developed by CRISPR/Cas9 gene editing and recapitulates human type 2N VWD

Type 2N von Willebrand disease is caused by mutations in the factor VIII (FVIII) binding site of von Willibrand factor (VWF), resulting in dysfunctional VWF with defective binding capacity for FVIII. Here we developed a novel type 2N mouse model using CRISPR/Cas9 technology. In homozygous VWF2N/2N mice, plasma VWF levels were normal (1167±257 mU/ml) but the VWF was completely incapable of binding FVIII, resulting in 53±23 mU/ml of plasma FVIII levels that were similar to those in VWF deficient (VWF-/-) mice. When wild-type human or mouse VWF was infused into VWF2N/2N mice, endogenous plasma FVIII was restored, peaking at 4-6 hours post-infusion, demonstrating that FVIII expressed in VWF2N mice is viable, but short-lived unprotected in plasma due to dysfunctional 2N-VWF. The whole blood clotting time and thrombin generation were impaired in VWF2N/2N but not in VWF-/- mice. The bleeding time and blood loss in VWF2N/2N mice were similar to wild-type mice in the lateral tail vein or ventral artery injury model. However, VWF2N/2N, but not VWF-/- mice, lost a significant amount of blood during the primary bleeding phase after a tail tip amputation injury model, indicating that there are other alternative pathway(s) that can at least partially restore hemostasis when VWF is absent. In summary, we have developed a novel mouse model by gene editing with both the pathophysiology and clinical phenotype found in severe type 2N patients. This unique model can be used to investigate the biological properties of VWF/FVIII association in hemostasis and beyond.

The Influence of Edible Oils’ Composition on the Properties of Beeswax-Based Oleogels

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.

Thalidomide-based inhibitor for TNF-α: designing and Insilico evaluation

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.

The Function of CBM32 in Alginate Lyase VxAly7B on the Activity on Both Soluble Sodium Alginate and Alginate Gel

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.