Heating-cooling extrusion cycles as a method to improve the physicochemical properties of extruded corn starch.

This work proposed a controlled method to modify the physicochemical properties of corn starch through heating and cooling extrusion (HCE) cycles. It was used native corn starch adjusted to 60% moisture. It was then subjected to 5 HCE cycles at 100 and 125 °C, at 10 rpm. Water absorption index (WAI), water solubility index (WSI), resistant starch (RS), thermal properties, viscosity, FTIR, and X-ray were evaluated. For WAI and WSI, a gradual increase was observed on each HCE cycle. Thermal properties shown that enthalpy decrease with each HCE cycles due to more gelatinization. Viscosity properties shown a thermally stable starch conditions being directly proportional to HCE cycles. The RS increased for each 5 HCE. XRD revealed that HCE cycle changed the starch structure from an orthorhombic structure to V-type crystalline structure. Finally, it was concluded that HCE cycles is a method to produce corn starch with controlled physicochemical properties.

Structural changes in popped sorghum starch and their impact on the rheological behavior.

Sorghum has been used to expand snacks such as pop sorghum. However, it is still unknown how the structural changes during the popping affect its rheological and functional properties. This study evaluated the structural changes of popped sorghum starch (PS) and their impact on rheological behavior. Moisture sorghum was adjusted to 11, 15, and 20% before popped. Morphology, X-ray pattern (XRP), infrared spectra (IR), thermal properties, and rheological behavior before and after popping were evaluated. Micrographs showed a honeycomb-like structure in PS. XRP showed partial damage to the orthorhombic crystals of the sorghum starch after PS, while the growth of crystalline lamellae was also generated (13.08 and 20.01°). IR showed structural damage as the signal at 1045 cm-1 disappeared in PS. The IM increased to gelatinization of the starch. The rheological behavior of PS displayed better thermal stability, with the lowest breakdown (25 ± 3.5 cP), setback (253 ± 11.3 cP), and final (1337 ± 5.7 cP) viscosity. The consistency coefficient k and flow behavior index n increase, meaning a loss of the pseudoplastic character. Viscoelastic properties increased in PS, suggesting the formation of cross-links and a stable matrix. Correlation analysis showed a strong relationship between structural changes and the rheological behavior of PS.

Impact of the popping process on the structural and thermal properties of sorghum grains (Sorghum bicolor L. Moench).

The popping process has been widely used as a technique for obtaining snacks. This study evaluated the effect of the popping process on the structural and thermal properties of sorghum. Seven varieties of sorghum were used. Raw sorghum grains were adjusted to 11% moisture and popped at 210 °C for 90 s with hot air. Microstructure, thermal and viscosity properties, and X-Ray and infrared spectrum were measured in raw and popped sorghum. The popping process produced an ordered honeycomb-like structure in the sorghum. The viscosity profile showed an increase in the thermal stability of popped sorghum. DSC measurements showed a starch gelatinization and a second transition about to 145 °C. XRD diffractograms display a reduction in the amplitude of the crystalline orthorhombic structure peaks. Finally, infrared indicated a change in the short-range structure and protein denaturation due to the popping process.

Effect of the nixtamalization process on the protein bioaccessibility of white and red sorghum flours during in vitro gastrointestinal digestion.

Protein bioaccessibility is a major concern in sorghum (Sorghum bicolor L. Moench) due to potential interactions with tannins affecting its nutritional value. Technological treatments such as boiling or alkaline cooking have been proposed to address this problem by reducing tannin-protein interactions. This research aimed to evaluate the impact of nixtamalization in the protein bioaccessibility from two sorghum varieties (red and white sorghum) during in vitro gastrointestinal digestion. Nixtamalization increased protein bioaccessibility in the non-digestible fraction (NDF) (5.26 and 26.31% for red and white sorghum, respectively). However, cooking showed a higher permeation speed of protein from red sorghum flours at the end of the intestinal incubation (9.42%). The SDS-PAGE profile of the digested fraction (DF) at 90 min of intestinal incubation indicated that, for red sorghum, cooking allows the formation of α and γ-kafirins while nixtamalization increase α-kafirin release. Principal Components Analysis (PCA) showed the association between nixtamalization and dissociation of δα kafirin complexes and increased protein content in the digestible fraction. In silico interactions indicated the highest biding energies for (+)-catechin and kafirin fractions (ß-kafirin: -7.0 kcal/mol; γ-kafirin: -5.8 kcal/mol, and δ-kafirin: -6.8 kcal/mol), suggesting a minor influence of depolymerized proanthocyanidin fractions with sorghum proteins as a result of the nixtamalization process. In conclusion, nixtamalization increased the bioaccessibility of sorghum proteins, depolymerizing condensed tannins, and breaking protein-tannin complexes. Such technological process improves the nutrimental value of sorghum, supporting its inclusion in the human diet.

Novel rare variants in congenital cardiac arrhythmia genes are frequent in drug-induced torsades de pointes.

Marked prolongation of the QT interval and polymorphic ventricular tachycardia following medication (drug-induced long QT syndrome, diLQTS) is a severe adverse drug reaction (ADR) that phenocopies congenital long QT syndrome (cLQTS) and is one of the leading causes for drug withdrawal and relabeling. We evaluated the frequency of rare non-synonymous variants in genes contributing to the maintenance of heart rhythm in cases of diLQTS using targeted capture coupled to next-generation sequencing. Eleven of 31 diLQTS subjects (36%) carried a novel missense mutation in genes with known congenital arrhythmia associations or with a known cLQTS mutation. In the 26 Caucasian subjects, 23% carried a highly conserved rare variant predicted to be deleterious to protein function in these genes compared with only 2-4% in public databases (P<0.003). We conclude that the rare variation in genes responsible for congenital arrhythmia syndromes is frequent in diLQTS. Our findings demonstrate that diLQTS is a pharmacogenomic syndrome predisposed by rare genetic variants.

The effect of novel promoter variants in MATE1 and MATE2 on the pharmacokinetics and pharmacodynamics of metformin.

Interindividual variation in response to metformin, first-line therapy for type 2 diabetes, is substantial. Given that transporters are determinants of metformin pharmacokinetics, we examined the effects of promoter variants in both multidrug and toxin extrusion protein 1 (MATE1) (g.-66T → C, rs2252281) and MATE2 (g.-130G → A, rs12943590) on variation in metformin disposition and response. The pharmacokinetics and glucose-lowering effects of metformin were assessed in healthy volunteers (n = 57) receiving metformin. The renal and secretory clearances of metformin were higher (22% and 26%, respectively) in carriers of variant MATE2 who were also MATE1 reference (P < 0.05). Both MATE genotypes were associated with altered post-metformin glucose tolerance, with variant carriers of MATE1 and MATE2 having an enhanced (P < 0.01) and reduced (P < 0.05) response, respectively. Consistent with these results, patients with diabetes (n = 145) carrying the MATE1 variant showed enhanced metformin response. These findings suggest that promoter variants of MATE1 and MATE2 are important determinants of metformin disposition and response in healthy volunteers and diabetic patients.

Optimizing drug outcomes through pharmacogenetics: a case for preemptive genotyping.

Routine integration of genotype data into drug decision making could improve patient safety, particularly if many relevant genetic variants can be assayed simultaneously before prescribing the target drug. The frequency of opportunities for pharmacogenetic prescribing and the potential adverse events (AEs) mitigated are unknown. We examined the frequency with which 56 medications with known outcomes influenced by variant alleles were prescribed in a cohort of 52,942 medical home patients at Vanderbilt University Medical Center (VUMC). Within a 5-year window, we estimated that 64.8% (95% confidence interval (CI): 64.4-65.2%) of individuals were exposed to at least one medication with an established pharmacogenetic association. Using previously published results for six medications with severe, well-characterized, genetically linked AEs, we estimated that 383 events (95% CI, 212-552) could have been prevented with an effective preemptive genotyping program. Our results suggest that multiplexed, preemptive genotyping may represent an efficient alternative approach to current single-use ("reactive") methods and may also improve safety.

Operational implementation of prospective genotyping for personalized medicine: the design of the Vanderbilt PREDICT project.

The promise of "personalized medicine" guided by an understanding of each individual's genome has been fostered by increasingly powerful and economical methods to acquire clinically relevant information. We describe the operational implementation of prospective genotyping linked to an advanced clinical decision-support system to guide individualized health care in a large academic health center. This approach to personalized medicine entails engagement between patient and health-care provider, identification of relevant genetic variations for implementation, assay reliability, point-of-care decision support, and necessary institutional investments. In one year, approximately 3,000 patients, most of whom were scheduled for cardiac catheterization, were genotyped on a multiplexed platform that included genotyping for CYP2C19 variants that modulate response to the widely used antiplatelet drug clopidogrel. These data are deposited into the electronic medical record (EMR), and point-of-care decision support is deployed when clopidogrel is prescribed for those with variant genotypes. The establishment of programs such as this is a first step toward implementing and evaluating strategies for personalized medicine.

Predicting clopidogrel response using DNA samples linked to an electronic health record.

Variants in ABCB1 and CYP2C19 have been identified as predictors of cardiac events during clopidogrel therapy initiated after myocardial infarction (MI) or percutaneous coronary intervention (PCI). In addition, PON1 has recently been associated with stent thrombosis. The reported effects of these variants have not yet been replicated in a real-world setting. We used BioVU, the Vanderbilt DNA repository linked to de-identified electronic health records (EHRs), to find data on patients who were on clopidogrel treatment after an MI and/or a PCI; among these, we identified those who had experienced one or more recurrent cardiac events while on treatment (cases, n = 225) and those who had not experienced any cardiac event while on treatment (controls, n = 468). We found that CYP2C19*2 (hazard ratio (HR) 1.54, 95% confidence interval (CI) 1.16-2.06, P = 0.003) and ABCB1 (HR 1.28, 95% CI 1.04-1.57, P = 0.018), but not PON1 (HR 0.91, 95% CI 0.73-1.12, P = 0.370), were associated with recurrent events. In this population, genetic signals for clopidogrel resistance in ABCB1 and CYP2C19 were replicated, supporting the use of EHRs for pharmacogenomic studies. Our data do not show an association between PON1 and recurrent cardiovascular events.

A common 5'-UTR variant in MATE2-K is associated with poor response to metformin.

Multidrug and toxin extrusion 2 (MATE2-K (SLC47A2)), a polyspecific organic cation exporter, facilitates the renal elimination of the antidiabetes drug metformin. In this study, we characterized genetic variants of MATE2-K, determined their association with metformin response, and elucidated their impact by means of a comparative protein structure model. Four nonsynonymous variants and four variants in the MATE2-K basal promoter region were identified from ethnically diverse populations. Two nonsynonymous variants-c.485C>T and c.1177G>A-were shown to be associated with significantly lower metformin uptake and reduction in protein expression levels. MATE2-K basal promoter haplotypes containing the most common variant, g.-130G>A (>26% allele frequency), were associated with a significant increase in luciferase activities and reduced binding to the transcriptional repressor myeloid zinc finger 1 (MZF-1). Patients with diabetes who were homozygous for g.-130A had a significantly poorer response to metformin treatment, assessed as relative change in glycated hemoglobin (HbA1c) (-0.027 (-0.076, 0.033)), as compared with carriers of the reference allele, g.-130G (-0.15 (-0.17, -0.13)) (P=0.002). Our study showed that MATE2-K plays a role in the antidiabetes response to metformin.

Studies of the photooxidant properties of antibacterial fluoroquinolones and their naphthalene derivatives.

We synthesized and determined the production of reactive oxygen species (ROS) as 1O2, *-O2, *OH, H2O2 during the photolysis with UV-A light of three antibacterial quinolones and their naphthyl ester derivatives. Singlet oxygen and ROS dose-dependant generation from norfloxacin (1), enoxacin (2), ciprofloxacin (3) and their respective naphthyl ester derivatives 4-6 were detecting in cell-free systems by the histidine assay and by luminol-enhanced chemiluminescence (LCL). Both the electronic absorption and emission spectra were quantified and their photostability determined. The antibacterial activity in darkness and under irradiation of compounds 4, 5 and 6 was tested on E. coli and compared with their parent drugs.

Nuclear DNA sequence specific to Leishmania (Viannia) subgenus: a molecular marker for species identification.

As shown by RFLP analysis, there is a high variability in the beta-tubulin gene region of Leishmania sp. Such variability has been used in the identification of these parasites, establishing differences between subgenera of New World Leishmania. We have found a region of 500 bp (beta500) upstream of the coding region of the beta-tubulin gene that is present in all strains tested belonging to the L. (Viannia) subgenus. This region apparently is a repetitive sequence and we have shown that it is specific to the Leishmania (Viannia) subgenus. This sequence has no homology with the genomic DNA isolated from either the species belonging to the L. (Leishmania) subgenus or other Kinetoplastida, such as Trypanosoma cruzi, T. brucei, Leptomonas samueli, or Crithidia fasciciulata. The beta500 sequence showed sufficient variation to be used as a molecular marker in the identification of parasites. We established inter- and intrasubgenus differentiation and were able to discriminate at the species level in the Vianna subgenus. A PCR assay confirmed the specificity of the beta500 sequence.