Preparation and textural properties of white salted noodles made with hard red winter wheat flour partially replaced by different levels of cross-linked phosphorylated RS4 wheat starch.

BACKGROUND: Resistant starch (RS) has health benefits and can be used as a functional ingredient in various food products. Kansas hard red winter (HRW) wheat is conventionally used for bread making and this is attributed to its strong gluten network. To develop Asian white salted noodles with a high RS content, HRW wheat flour was partially replaced with cross-linked phosphorylated RS4 wheat starch. Vital wheat gluten or wheat protein isolate was added to compensate for textural changes due to the addition of RS. RESULTS: The maximum recommended level of RS4 starch to replace HRW wheat flour was 40%. The substitution of 10-40 parts of RS4 for flour did not change hardness in cooked noodles but it did reduce their extensibility, cohesiveness, and springiness, which was probably due to the non-swelling properties of RS4. At 40 parts of RS4 replacement, supplementation of 2-8 parts of vital wheat gluten or wheat protein isolate in the composite flour notably enhanced the hardness and extensibility of cooked noodles, whereas cohesiveness and springiness were minimally affected. Supplemental vital wheat gluten produced a thicker protein network than endogenous protein or added wheat protein isolate, giving cooked noodles greater breaking force and distance. CONCLUSION: RS4 could be used as a functional ingredient to replace up to 40% of hard wheat flour for making Asian noodles while maintaining their hardness after cooking. The extensibility of cooked noodles with high RS4 could be noticeably enhanced by supplementation with vital wheat gluten in the composite flour (RS/flour = 40/60). © 2020 Society of Chemical Industry.

Optimization and characterization of high pressure homogenization produced chemically modified starch nanoparticles.

Chemically modified starch (RS4) nanoparticles were synthesized through homogenization and water-in-oil mini-emulsion cross-linking. Homogenization was optimized with regard to z-average diameter by using a three-factor-three-level Box-Behnken design. Homogenization pressure (X1), oil/water ratio (X2), and surfactant (X3) were selected as independent variables, whereas z-average diameter was considered as a dependent variable. The following optimum preparation conditions were obtained to achieve the minimum average size of these nanoparticles: 50 MPa homogenization pressure, 10:1 oil/water ratio, and 2 g surfactant amount, when the predicted z-average diameter was 303.6 nm. The physicochemical properties of these nanoparticles were also determined. Dynamic light scattering experiments revealed that RS4 nanoparticles measuring a PdI of 0.380 and an average size of approximately 300 nm, which was very close to the predicted z-average diameter (303.6 nm). The absolute value of zeta potential of RS4 nanoparticles (39.7 mV) was higher than RS4 (32.4 mV), with strengthened swelling power. X-ray diffraction results revealed that homogenization induced a disruption in crystalline structure of RS4 nanoparticles led to amorphous or low-crystallinity. Results of stability analysis showed that RS4 nanosuspensions (particle size) had good stability at 30 °C over 24 h.

Carrier-Tumor Cell Membrane Interactions for Optimized Delivery of a Promising Drug, 4(RS)-4-F4t-Neuroprostane.

Nanomedicines engineered to deliver molecules with therapeutic potentials, overcoming drawbacks such as poor solubility, toxicity or a short half-life, are targeted towards their cellular destination either passively or through various elements of cell membranes. The differences in the physicochemical properties of the cell membrane between tumor and nontumor cells have been reported, but they are not systematically used for drug delivery purposes. Thus, in this study, a new approach based on a match between the liposome compositions, i.e., membrane fluidity, to selectively interact with the targeted cell membrane was used. Lipid-based carriers of two different fluidities were designed and used to deliver 4(RS)-4-F4t-Neuroprostane (F4t-NeuroP), a potential antitumor molecule derived from docosahexaenoic acid (DHA). Based on its hydrophobic character, F4t-NeuroP was added to the lipid mixture prior to liposome formation, a protocol that yielded over 80% encapsulation efficiency in both rigid and fluid liposomes. The presence of the active molecule did not modify the liposome size but increased the liposome negative charge and the liposome membrane fluidity, which suggested that the active molecule was accommodated in the lipid membrane. F4t-NeuroP integration in liposomes with a fluid character allowed for the selective targeting of the metastatic prostate cell line PC-3 vs. fibroblast controls. A significant decrease in viability (40%) was observed for the PC-3 cancer line in the presence of F4t-NeuroP fluid liposomes, whereas rigid F4t-NeuroP liposomes did not alter the PC-3 cell viability. These findings demonstrate that liposomes encapsulating F4t-NeuroP or other related molecules may be an interesting model of drug carriers based on membrane fluidity.

Acid-treated high-amylose corn starch suppresses high-fat diet-induced steatosis.

Resistant starch (RS) has been reported to improve steatosis as well as obesity. Type 4 resistant starch (RS4), a chemically modified starch, is particularly hard to digest and suggesting higher efficacy. However, because the effects of RS4 on steatosis are not yet fully understood, the effects of RS4 on steatosis were examined using a murine high-fat diet model. Seven-week-old male mice were divided into three groups and fed a normal diet, a high-fat diet (HFD), or a high-fat diet with added RS (HFD + RS). Amylofiber SH® produced from acid-treated corn starch was used as the dietary RS. At 22 weeks old, hepatic steatosis and short chain fatty acid (SCFA) content and gut microbiota in cecum stool samples were analyzed. The ratio of body weight to 7 weeks was significantly suppressed in the HFD + RS group compared to the HFD group (132.2 ± 1.4% vs. 167.2 ± 3.9%, p = 0.0076). Macroscopic and microscopic steatosis was also suppressed in the HFD + RS group. Analysis of cecum stool samples revealed elevated SCFA levels in the HFD + RS group compared with the HFD group. Metagenome analysis revealed that Bifidobacterium (17.9 ± 1.9% vs. 3.6 ± 0.7%, p = 0.0019) and Lactobacillus (14.8 ± 3.4% vs. 0.72 ± 0.23%, p = 0.0045), which degrade RS to SCFA, were more prevalent in the HFD + RS group than the HFD group. In conclusion, RS4 suppressed steatosis, and increased Bifidobacterium and Lactobacillus, and SCFAs. RS4 may prevent steatosis by modulating the intestinal environment.

Glycemic and Insulinemic Responses of Healthy Humans to a Nutrition Bar with or without Added Fibersym® RW, a Cross-Linked Phosphorylated RS4-Type Resistant Wheat Starch.

The current study compared postprandial glycemic and insulinemic responses to four nutrition bars containing two different doses of resistant starch type-4. Normoglycemic adults (n = 17) completed six treatments, consuming either 50 g or 30 g digestible carbohydrate as: dextrose beverages (DEX), control puffed wheat bars (PWB), or RS4 test bars (RS4). Glucose (mg/dL) and insulin (µIU/mL) were measured at baseline and 10, 20, 30, 60, 90, and 120 min. There was a main effect of dose and treatment on glucose incremental area under the curve (iAUC, ps < 0.001), such that RS4 (50 g: 941, 95% confidence interval (CI): 501, 1519; 30 g: 481, 95% CI: 186, 914) was lower than PWB (50 g: 1746, 95% CI: 1109, 2528; 30 g: 693, 95% CI: 331, 1188) and DEX (50 g: 1940, 95% CI: 1249, 2783; 30 g:1432, 95% CI: 883, 2114). There was a main effect of dose and treatment on insulin iAUC (ps < 0.001), such that RS4 (50 g: 1993, 95% CI: 1347, 2764; 30 g: 943, 95% CI: 519, 1493) was lower than PWB (50 g: 3501, 95% CI: 2625, 4502; 30 g: 1789, 95% CI: 1193, 256) and DEX (50 g: 3143, 95% CI: 2317, 4095; 30 g: 2184, 95% CI: 1519, 2970). Results demonstrate significantly lower glycemic and insulinemic responses following consumption of nutrition bars containing RS4, regardless of dose, when compared with puffed wheat bars and dextrose.

Neuroprotective effects of DHA-derived peroxidation product 4(RS)-4-F4t-neuroprostane on microglia.

The abundance of docosahexaenoic acid (DHA) in brain membrane phospholipids has stimulated studies to explore its role in neurological functions. Upon released from phospholipids, DHA undergoes enzymatic reactions resulting in synthesis of bioactive docosanoids and prostanoids. However, these phospholipids are also prone to non-enzymatic reactions leading to more complex pattern of metabolites. A non-enzymatic oxidized product of DHA, 4(RS)-4-F4t-Neuroprostane (44FNP), has been identified in cardiac and brain tissues. In this study, we examined effects of the 44FNP on oxidative and inflammatory responses in microglial cells treated with lipopolysaccharide (LPS). The 44FNP attenuated LPS-induced production of reactive oxygen species (ROS) in both primary and immortalized microglia (BV2). It also attenuated LPS-induced inflammation through suppressing NFκB-p65 and levels of iNOS and TNFα. In addition, 44FNP also suppressed LPS-induced mitochondrial dysfunction and upregulated the Nrf2/HO-1 antioxidative pathway. In sum, these findings with microglial cells demonstrated neuroprotective effects of this 44FNP and shed light into the potential of nutraceutical therapy for neurodegenerative diseases.

The VVBA method for thoracoscopic right middle lobe segmentectomy.

The right middle lobe segments (RS4 and RS5) are very small and the technique for segmentectomy is rarely reported. Here, we report 3 cases of thoracoscopic RS4 segmentectomy with VVBA (V, vein; V, ventilation; B, bronchus; A, artery) method. The VVBA method for thoracoscopic segmentectomy is a simple, safe, and effective procedure for RS4 segmentectomy with clear procedural steps. It overcomes the difficulty in manipulation of incomplete lung fissures and simplifies the surgical procedures.

Metabolic Responses to Native Wheat Starch (MidsolTM 50) versus Resistant Wheat Starch Type 4 (Fibersym® RW): Standard versus Marketplace Testing Protocols.

BACKGROUND: To investigate the effect of resistant starch (RS) on acute glycemic or insulinemic responses, the FDA indicates that control and RS-enriched foods must contain equivalent amounts of digestible carbohydrate. However, RS-containing foods typically contain less digestible carbohydrate per serving than control foods. Thus, controlling for digestible carbohydrate may yield different responses as compared with controlling for serving size. OBJECTIVE: The aim was to compare the postprandial metabolic responses to native wheat starch (NWS) versus RS type 4 (RS4) using digestible carbohydrate-matched portions compared with weight-matched portions. METHODS: A single-blind, randomized-controlled crossover trial examined glycemic and insulinemic responses over 2 h following consumption of 4 cracker conditions and a dextrose beverage in apparently healthy participants (n = 14). Crackers provided 50 g of digestible carbohydrate using the FDA's meal-intervention protocol or 35 g of carbohydrate by weight for the marketplace substitution method. Crackers differed only by the type of starch additive: NWS (MidsolTM 50; MGP Ingredient, Inc.) or RS4 (Fibersym® RW; MGP Ingredients, Inc.). Glucose concentrations were assessed at baseline and at 15, 30, 45, 60, 90, and 120 min; insulin concentrations were measured at baseline and 30, 60, and 120 min. RESULTS: There were no significant differences between 50 g digestible carbohydrate cracker conditions for glucose or insulin incremental AUC (iAUC). The 35 g carbohydrate by weight conditions were not different for glucose iAUC [mean (95% CI): 35 g NWS: 1317 (677, 2169); 35 g RS4: 701 (262, 1351); P > 0.05]. However, insulin iAUC was lower following 35 g RS4 compared with 35 g NWS [35 g RS4: 92 (1, 259); 35 g NWS: 697 (397, 1080); P < 0.01]. CONCLUSIONS: In healthy adults, consumption of RS4 crackers decreased postprandial insulin responses compared with NWS crackers when using the marketplace substitution method compared with the FDA standard testing method, with similar postprandial glucose responses. Comparisons of the FDA standard testing method and the marketplace substitution method should be investigated further to elucidate differential physiological impacts on consumers.

The RS4;11 cell line as a model for leukaemia with t(4;11)(q21;q23): Revised characterisation of cytogenetic features.

BACKGROUND: Haematological malignancies harbouring rearrangements of the KMT2A gene represent a unique subtype of leukaemia, with biphenotypic clinical manifestations, a rapid and aggressive onset, and a generally poor prognosis. Chromosomal translocations involving KMT2A often cause the formation of oncogenic fusion genes, such as the most common translocation t(4;11)(q21;q23) producing the KMT2A-AFF1 chimera. AIM: The aim of this study was to confirm and review the cytogenetic and molecular features of the KMT2A-rearranged RS4;11 cell line and put those in context with other reports of cell lines also harbouring a t(4;11) rearrangement. METHODS AND RESULTS: The main chromosomal rearrangements t(4;11)(q21;q23) and i(7q), described when the cell line was first established, were confirmed by fluorescence in situ hybridisation (FISH) and 24-colour karyotyping by M-FISH. Additional cytogenetic abnormalities were investigated by further FISH experiments, including the presence of trisomy 18 as a clonal abnormality and the discovery of one chromosome 8 being an i(8q), which indicates a duplication of the oncogene MYC. A homozygous deletion of 9p21 containing the tumour-suppressor genes CDKN2A and CDKN2B was also revealed by FISH. The production of the fusion transcript KMT2A-AFF1 arising from the der(11)t(4;11) was confirmed by RT-PCR, but sequencing of the amplified fragment revealed the presence of multiple isoforms. Two transcript variants, resulting from alternative splicing, were identified differing in one glutamine residue in the translated protein. CONCLUSION: As karyotype evolution is a common issue in cell lines, we highlight the need to monitor cell lines in order to re-confirm their characteristics over time. We also reviewed the literature to provide a comparison of key features of several cell lines harbouring a t(4;11). This would guide scientists in selecting the most suitable research model for this particular type of KMT2A-leukaemia.

Production of RS4 from rice starch and its utilization as an encapsulating agent for targeted delivery of probiotics.

The research reported in this article is based on the hypothesis that crosslinking of starch can make it a potential wall material for targeted delivery of probiotics by altering its digestion. Three probiotic strains namely Lactobacillus casei, Lactobacillus brevis and Lactobacillus plantarum were microencapsulated with resistant starch. Encapsulation yield (%) of resistant starch microspheres was in the range of 43.01-48.46. The average diameter of resistant starch microparticles was in the range of 45.53-49.29µm. Fourier transform infrared (FT-IR) spectroscopy of microcapsules showed peaks in the region of 900-1300cm-1 and 2918-2925cm-1 which corresponds to the presence of bacteria. Differential Scanning Calorimeter (DSC) showed better thermal stability of resistant starch microcapsules. Microencapsulated probiotics survived well in simulated gastrointestinal conditions and adverse heat conditions. The viability of the microcapsulated lactobacilli also remained high (>7 log cfu g-1) for 2months at 4°C. The results revealed that resistant starch is the potential new delivery carrier for oral administration of probiotics.

Preparation and characterization of resistant starch type IV nanoparticles through ultrasonication and miniemulsion cross-linking.

This study aimed to assess the properties of resistant starch type IV (chemically modified starch, RS4) prepared from a new and convenient synthesis route by using ultrasonication combined with water-in-oil miniemulsion cross-linking technique. A three-factor Box-Behnken design and optimization was used to minimize particle size through the developed RS4 nanoparticles. The predicted minimized Z-Avel (576.1nm) under the optimum conditions of the process variables (ultrasonic power, 214.57W; sonication time, 114.73min; and oil/water ratio, 10.54:1) was very close to the experimental value (651.0nm) determined in a batch experiment. After preparing the RS4 nanoparticles, morphological, physical, chemical, and functional properties were assessed. Results revealed that RS4 nanoparticle size reached about 600nm. Scanning electron microscopy images showed that ultrasonication induced notches and grooves on the surface. Under polarized light, the polarized cross was impaired. X-ray diffraction results revealed that the crystalline structure was disrupted. Smaller or no endotherms were exhibited in DSC analysis. In the FTIR graph, new peaks at 1532.91 and 1451.50cm(-1) were observed, and pasting properties were reduced. Amylose content, solubility, and SP increased, but RS content decreased. Anti-digestibility remained after ultrasonication. The prepared RS4 nanoparticles could be extensively used in biomedical applications and in the development of new medical materials.

Optimisation of the reaction conditions for the production of cross-linked starch with high resistant starch content.

The optimum reaction conditions (temperature and pH) for the preparation of cross-linked (CL) corn and wheat starches with maximum resistant starch (RS) content were investigated by using response surface methodology (RSM). According to the preliminary results, five levels were selected for reaction temperature (38-70 °C) and pH (10-12) in the main study. RS contents of the CL corn and wheat starch samples increased with increasing temperature and pH, and pH had a greater influence on RS content than had temperature. The maximum RS content (with a maximum p value of 0.4%) was obtained in wheat starch cross-linked at 38 °C and pH 12. In the case of CL corn starch, the optimum condition was 70 °C and pH 12. CL corn and wheat starch samples were also produced separately under the optimum conditions and their RS contents were 80.4% and 83.9%, respectively. These results were also in agreement with the values predicted by RSM.