Development of high-yielding white maize hybrids with better chapatti-making quality compared to traditionally used local landraces.

Introduction: The present research focuses on the chapatti making quality of high-yielding white maize hybrids compared to available low-yielding local yellow and white landraces in India. Materials and methods: In this study, the top nine superior hybrids were selected for testing the physical properties of the maize kernels, proximate composition of flours and chapattis, physical parameters of chapatti, textural properties, sensory evaluation of chapattis and pasting properties of maize flour. Results and discussion: The results revealed the superiority of white maize hybrids (WMH), viz., WHM 1, WHM 2, and WHM 8 over the local yellow and white landraces for most of the parameters studied. In sensory analysis, though, the yellow landrace was considered superior by the panellists in terms of colour but the white maize hybrids outperformed in overall sensory analysis and were more acceptable than the yellow and white maize landraces. These high yielding white maize hybrids with good consumer acceptance may cater for the needs of rural and tribal populations in India who prefer white maize as a staple food.

Operational outcomes of propofol sedation versus fentanyl, midazolam and diphenhydramine sedation for endoscopies and colonoscopies at an academic medical center.

BACKGROUND: On July 1st, 2021, the University of Colorado Hospital (UCH) implemented new sedation protocols in the luminal gastrointestinal (GI) suite. GI proceduralist supervised, Nurse Administered Sedation with fentanyl, midazolam, and diphenhydramine (NAS) sedation was transitioned to Monitored Anesthesia Care with propofol under physician anesthesiologist supervision (MAC). OBJECTIVE: To determine if there are statistically significant reductions in Sedation-Start to Scope-In time (SSSI) when using Monitored Anesthesia Care with propofol (MAC) versus Nurse Administered Sedation with fentanyl, midazolam, and diphenhydramine (NAS). Secondary objectives were to determine if statistically significant improvements to other operational times, quality measures, and satisfaction metrics were present. METHOD: This study was a retrospective analysis of a natural experiment resultant of a change from NAS to MAC sedation protocols. Outcomes for NAS protocols from 1/1/21-6/30/21 were compared to outcomes of MAC protocols from the dates 8/1/21-10/31/21. Results were analyzed using Quasi-Poisson regression analysis and stratified based on upper GI, lower GI, and combined procedures. Patient demographic data including age, biological sex, comorbidities, and BMI, were adjusted for in the analysis. ASA matching was not performed as nursing sedation does not use ASA classifications. Pre-anesthesia co-morbidities were assessed via evaluation of a strict set of comorbidities abstracted from the electronic medical record. Perioperative operational outcomes include Sedation Start to Scope-In (SSSI), In-Room to Scope-In Time (IRSI), Scope Out to Out of Room (SOOR), Total Case Length (TCL), and Post Anesthesia Care Unit Length of Stay (PACU LOS). Quality outcomes include PACU Administered Medications (PAM), and Clinician Satisfaction Scores (CSS). RESULTS: A total of 5,582 gastrointestinal (GI) endoscopic cases (upper, lower, and combined endoscopies) were observed. Statistically significant decreases in SSSI of 2.5, 2.1, and 2.2 minutes for upper, lower, and dual GI procedures were observed when using MAC protocols. A statistically significant increase in satisfaction scores of 47.0 and 19.6 points were observed for nurses and proceduralists, respectively, when using MAC. CONCLUSION: MAC protocols for endoscopic GI procedures at UCH led to statistically significant decreases in the time required to complete procedures thus increasing operational efficiency.

Molecular morphology & interactions, functional properties, rheology and in vitro digestibility of ultrasonically modified pearl millet and sorghum starches.

The present research investigated to study the effect of ultrasound treatment on isolated pearl millet starch (PMS) and sorghum starch (SS). Ultrasonication was applied to PMS and SS for 10, 15, and 20 min. Ultrasonically modified pearl millet and sorghum starches evaluated for their techno-functionality, pasting profile, morphology, in vitro starch digestibility, XRD, and molecular interactions. Ultrasound treatment increased water and oil absorption capacity, swelling power, and solubility with treatment time. For ultrasonicated PMS and SS, a significant increase (p < 0.05) in paste clarity (PC) (70.05 % and 67.23 %), freeze-thawing stability (FTS), gel consistency (GC) (25.05 mm and 32.95 mm), and in vitro starch digestibility were observed (57.70 g/100 g and 50.29 g/100 g), whereas no significant changes were recorded for the color values after the ultrasound treatment. Variations in pasting property were also observed in ultrasonicated starches with treatment duration. SEM images confirmed ultrasonication mainly forms pores and indentations on starch granule surface. FTIR spectra and X-ray diffractogram for ultrasonicated starches revealed a slight decrease in the peak intensity and A-type X-ray pattern with lower relative crystallinity (RC) than the native starches. G' > G″ value, indicating the elastic behavior and lower tan δ value, depicting viscous behavior and high gel strength.

Development of a Dihydroquinoline-Pyrazoline GluN2C/2D-Selective Negative Allosteric Modulator of the N-Methyl-d-aspartate Receptor.

Subunit-selective inhibition of N-methyl-d-aspartate receptors (NMDARs) is a promising therapeutic strategy for several neurological disorders, including epilepsy, Alzheimer's and Parkinson's disease, depression, and acute brain injury. We previously described the dihydroquinoline-pyrazoline (DQP) analogue 2a (DQP-26) as a potent NMDAR negative allosteric modulator with selectivity for GluN2C/D over GluN2A/B. However, moderate (<100-fold) subunit selectivity, inadequate cell-membrane permeability, and poor brain penetration complicated the use of 2a as an in vivo probe. In an effort to improve selectivity and the pharmacokinetic profile of the series, we performed additional structure-activity relationship studies of the succinate side chain and investigated the use of prodrugs to mask the pendant carboxylic acid. These efforts led to discovery of the analogue (S)-(-)-2i, also referred to as (S)-(-)-DQP-997-74, which exhibits >100- and >300-fold selectivity for GluN2C- and GluN2D-containing NMDARs (IC50 0.069 and 0.035 µM, respectively) compared to GluN2A- and GluN2B-containing receptors (IC50 5.2 and 16 µM, respectively) and has no effects on AMPA, kainate, or GluN1/GluN3 receptors. Compound (S)-(-)-2i is 5-fold more potent than (S)-2a. In addition, compound 2i shows a time-dependent enhancement of inhibitory actions at GluN2C- and GluN2D-containing NMDARs in the presence of the agonist glutamate, which could attenuate hypersynchronous activity driven by high-frequency excitatory synaptic transmission. Consistent with this finding, compound 2i significantly reduced the number of epileptic events in a murine model of tuberous sclerosis complex (TSC)-induced epilepsy that is associated with upregulation of the GluN2C subunit. Thus, 2i represents a robust tool for the GluN2C/D target validation. Esterification of the succinate carboxylate improved brain penetration, suggesting a strategy for therapeutic development of this series for NMDAR-associated neurological conditions.

Characterization of bioprocessed white and red sorghum flours: Anti-nutritional and bioactive compounds, functional properties, molecular, and morphological features.

In the current investigation, white and red sorghum grains were exposed to bioprocessing techniques, such as soaking, germination, fermentation, and dual processing (both germination and fermentation). Germination and fermentation resulted in improved bioactive profile attributing to better antioxidant activity, whereas a reduction in antinutrient components was observed. On the other hand, soaking had decreased phenolic components and anti-nutritional factors attributing to their leaching in the soaking water. A significant change in the functional properties and color profile was also observed on bioprocessing. It also caused alterations in the morphological structure of the starch-protein matrix and molecular interactions of certain functional groups that reveal the synthesis of certain new bioactive compounds in the flour. The alterations in the bioprocessed flours occurred due to the structural breakdown attributing to the activity of hydrolytic enzymes that were activated during the processing treatments. Bioprocessing was also responsible for the degradation of the starch granules and unfolding of the protein matrix, thus altering the in vitro nutrient digestibility of the flours. Principal component analysis was used to authenticate the differences between different treatments and observations recorded. These bioprocessed flours could be potential ingredients for several valorized cereal products.

Effect of extrusion processing on techno-functional properties, textural properties, antioxidant activities, in vitro nutrient digestibility and glycemic index of sorghum-chickpea-based extruded snacks.

Physico-chemical, textural, functional, and nutritional properties of the twin screw extruded whole sorghum-chickpea (8:2) snacks was investigated using in vitro procedures. The extruded snacks were analyzed for the effect of variations in extruded conditions on their properties: barrel BT (BT) (130-170°C) and feed moisture (FM) (14%-18%), keeping screw speed constant (400 rpm). The results revealed that specific mechanical energy (SME) decreased (74.4-60.0) in response to rise in both BT and FM, whereas expansion ratio (ER) had shown an alternative relation as it decreased with elevated FM (2.17 at 14%, 130°C to 2.14 at 16%, 130°C) and increased with BT (1.75 at 18%, 130°C to 2.48 at 18%, 170°C). The values of WAI and WSI improved with the surge in BT, which was associated with enhanced disruption of starch granules at higher BT. Raise in FM incremented the total phenolic content (TPC) and hence the antioxidant activity (AA) (FRAP and DPPH) along with the hardness of snacks. As per in vitro starch digestibility is concerned, slowly digestible starch (SDS) content as well as glycemic index (51-53) of the extrudates depressed with increasing BT and FM. Also, lower BT and FM improved the functional properties such as expansion ratio, in-vitro protein digestibility, and overall acceptability of the snacks. A positive correlation was seen among SME and hardness of the snacks, WSI and ER, TPC and AA, SDS and Exp-GI, color and OA, texture and OA.

Discovery of 5'-Substituted 5-Fluoro-2'-deoxyuridine Monophosphate Analogs: A Novel Class of Thymidylate Synthase Inhibitors.

5-Fluorouracil and 5-fluorouracil-based prodrugs have been used clinically for decades to treat cancer. Their anticancer effects are most prominently ascribed to inhibition of thymidylate synthase (TS) by metabolite 5-fluoro-2'-deoxyuridine 5'-monophosphate (FdUMP). However, 5-fluorouracil and FdUMP are subject to numerous unfavorable metabolic events that can drive undesired systemic toxicity. Our previous research on antiviral nucleotides suggested that substitution at the nucleoside 5'-carbon imposes conformational restrictions on the corresponding nucleoside monophosphates, rendering them poor substrates for productive intracellular conversion to viral polymerase-inhibiting triphosphate metabolites. Accordingly, we hypothesized that 5'-substituted analogs of FdUMP, which is uniquely active at the monophosphate stage, would inhibit TS while preventing undesirable metabolism. Free energy perturbation-derived relative binding energy calculations suggested that 5'(R)-CH3 and 5'(S)-CF3 FdUMP analogs would maintain TS potency. Herein, we report our computational design strategy, synthesis of 5'-substituted FdUMP analogs, and pharmacological assessment of TS inhibitory activity.

Neurotherapeutic Potential of Water-Soluble pH-Responsive Prodrugs of EIDD-036 in Traumatic Brain Injury.

The C-20 oxime of progesterone, EIDD-036 (2), demonstrates neuroprotection and improved outcomes in animal models of traumatic brain injury (TBI). However, 2 suffers from poor solubility, which renders it unsuitable for rapid administration. Previous prodrugs of 2 aimed at improving solubility by incorporating enzymatically labile amino acid and phosphate ester promoieties. These approaches were effective but led to limitations with in vivo administration. Herein, we disclose a pH-responsive water-soluble prodrug strategy to improve exposure to 2 through enzyme-independent activation. Compound 13l was identified as a lead that exhibits water-solubility, stability in acidic solutions, and rapid conversion to 2 at physiological pH. Administration of 13l to rats resulted in a twofold increase in exposure to 2 compared to the previous generation phosphate prodrug, EIDD-1723 (6). In a rat model of TBI, treatment with 13l resulted in a significant decrease in cerebral edema when administered postinjury.

Bioactivity evaluation of least explored traditionally acclaimed medicinally potent herb Nanorrhinum ramosissimum(Wall. ) Betsche

Abstract Present study analysed the therapeutic potential of traditionally acclaimed medicinal herb Nanorrhinum ramosissimum, using plant parts extracted with different solvents (10 mg/mL). Shoot extracts exhibited comparatively better antimicrobial properties, in comparison to root extracts. Total phenolic content was estimated, to ascertain its dependency on antioxidant properties of plant extracts. Antioxidant assay revealed promising results in comparison to IC50 value of standard ascorbic acid (52.2±0.07 µg/mL), for methanolic extracts of shoot (61.07±0.53 µg/mL and 64.33±0.33 µg/mL) and root (76.705±0.12 µg/mL and 89.73±0.28 µg/ mL) for in vivo and in vitro regenerants respectively. Correlation coefficient R2 values ranged between 0.90-0.95, indicating a positive correlation between phenolic contents and antioxidant activity. Plant extracts were also able to inhibit DNA oxidative damage again indicating their antioxidative potential. Antidiabetic potential was confirmed by alpha amylase inhibition assay where shoot methanolic extracts (invivo, in vitro) exhibited the best IC50 values (54.42±0.16 µg/mL, 66.09±0.12 µg/mL) in comparison to standard metformin (41.92±0.08 µg/mL). Ethanolic extracts of roots (in vitro, invivo) exhibited the relative IC50 values (88.97±0.32µg/mL,96.63±0.44 µg/mL) indicating that shoot parts had a better alpha amylase inhibition property; thus proving the herb's bioactive potential and its prospective therapeutic source for curing various ailments.

Modulation in Techno-Functional, Textural Properties, In Vitro Starch Digestibility and Macromolecular-Structural Interactions of Pasta with Potato (Solanum tuberosum L.).

The replacement of semolina with potato flour (PF) and potato mash (PM) at different levels was assessed for its effects on pasta quality. The results showed that the addition of PF and PM increased the pasting viscosity of the blends; in addition, PF enhanced the functional properties, while PM reduced them. The minimum cooking time decreased with PF and PM, while the PF pasta exhibited a higher cooking loss (5.02 to 10.44%) than the PM pasta, which exhibited a lower cooking loss. The pasta with PF and PM showed an increase in the total phenolic and flavonoid content, with reduced in vitro digestibility as confirmed by Fourier transform infrared spectroscopy. The PF pasta exhibited lower lightness and higher yellowness than the PM pasta, and its firmness and toughness also modulated owing to the complex interaction between potato starches and the gluten protein matrix, as evident from scanning electron microscopy. Sensory data revealed that pasta containing 30% PF and 16% PM was highly acceptable.

Cereal bar functionalised with non-conventional alfalfa and dhaincha protein isolates: quality characteristics, nutritional composition and antioxidant activity.

The utilization of conventional protein sources like gluten, soy, dairy proteins, and nuts in the development of protein-enriched cereal bars presents a challenge for their consumption by the population suffering from celiac and other food protein allergies. In the present investigation, protein-rich cereal bars were developed using non-conventional protein isolates (alfalfa and dhiancha (API & DPI) and were evaluated for their quality attributes, nutritional composition, and bioactive potential. The incorporation of protein isolates increased the weight, density, and non-enzymatic browning and decreased the water activity in the bars. The hardness of the bar increased with the addition of protein isolates; however, reduced hardness was observed at 7.5 and 10% levels of API. Supplementation with protein isolates enhanced the protein content (7.83-16.71%), total phenols (1642-4956 GAE µg/g), total flavonoids (268-984 QE µg/g), DPPH radical scavenging activity (96.38-114.82 TEAC µmol/100 g) and reducing power (1926-3586 AAE µg/g) of the bars. Cereal bars maintained good sensory score and overall acceptability at 10 and 5% level of incorporation of API and DPI respectively.

Effect of extrusion processing on techno-functional, textural and bioactive properties of whole-grain corn flour-based breakfast cereals sweetened with honey.

The present study investigated the effect of honey and extrusion processing parameters on techno-functional and bioactive properties of whole-grain corn flour breakfast cereals. The central composite rotatable design (CCRD) was used to plan the experiments using feed moisture (FM), extrusion temperature (ET), and honey level as process variables and sectional expansion ratio (SER), bulk density (BD), water absorption index (WAI), water solubility index (WSI), and textural hardness as response variables. The feed containing honey and whole-grain corn flour was extruded through a twin-screw extruder according to CCRD. The data were fit to the appropriate regression models based on model significance and insignificant lack of fit. The selected experiments from CCRD were considered for studying the bioactive properties, and the effect of ET and honey level on bioactive properties was determined. The results of the study indicated that FM and honey adversely affected the SER, BD, and textural hardness, while ET augmented these properties of breakfast cereals. The WAI decreased with an increase in honey level and ET during extrusion. The incremental addition of honey in the mix as well extrusion at elevated temperature led to higher WSI. The whole-grain corn flour added with 10.22% honey and extruded at 16.06% FM, and 138.07°C ET produced optimum quality breakfast cereals with 0.70 desirability. The total phenolic content of breakfast cereals decreased, and antioxidant activity and hydroxymethylfurfural content increased upon extrusion at the higher temperatures. Honey addition lowers the loss of phenolic content during extrusion and improves the antioxidant activity of breakfast cereals.

The Toxoplasma glucan phosphatase TgLaforin utilizes a distinct functional mechanism that can be exploited by therapeutic inhibitors.

Toxoplasma gondii is an intracellular parasite that generates amylopectin granules (AGs), a polysaccharide associated with bradyzoites that define chronic T. gondii infection. AGs are postulated to act as an essential energy storage molecule that enable bradyzoite persistence, transmission, and reactivation. Importantly, reactivation can result in the life-threatening symptoms of toxoplasmosis. T. gondii encodes glucan dikinase and glucan phosphatase enzymes that are homologous to the plant and animal enzymes involved in reversible glucan phosphorylation and which are required for efficient polysaccharide degradation and utilization. However, the structural determinants that regulate reversible glucan phosphorylation in T. gondii are unclear. Herein, we define key functional aspects of the T. gondii glucan phosphatase TgLaforin (TGME49_205290). We demonstrate that TgLaforin possesses an atypical split carbohydrate-binding-module domain. AlphaFold2 modeling combined with hydrogen-deuterium exchange mass spectrometry and differential scanning fluorimetry also demonstrate the unique structural dynamics of TgLaforin with regard to glucan binding. Moreover, we show that TgLaforin forms a dual specificity phosphatase domain-mediated dimer. Finally, the distinct properties of the glucan phosphatase catalytic domain were exploited to identify a small molecule inhibitor of TgLaforin catalytic activity. Together, these studies define a distinct mechanism of TgLaforin activity, opening up a new avenue of T. gondii bradyzoite biology as a therapeutic target.

Assessment of physicochemical, rheological, and thermal properties of Indian rice cultivars: Implications on the extrusion characteristics.

The implications of physicochemical, rheological, and thermal properties of seven eminent Indian rice cultivars (PR 114, 121, 122, 123, 124, 126, and 127) on the extrusion behavior and physico-functionalities of the extrudates were investigated. The amylose and amylopectin content of the cultivars ranged between 12.72 to 28.86% and 71.14 to 87.28% in addition with protein and crude fat content that varied from 7.05 to 9.15% and 0.49 to 1.17%, respectively. The onset (r = 0.98), peak (r = 0.95), and conclusion (r = 0.98) temperatures of the cultivars were in positive correlation with amylose. Likewise, pasting temperature (r = 0.979), final viscosity (r = 0.91), set back viscosity (r = 0.89), and stability ratio (r = 0.90) of the cultivars demonstrated a significant positive correlation with the amylose content. However, peak (r = - 0.879) and hold viscosity (r = - 0.89) were negatively correlated. The cultivars were extruded at feed moisture of 15%, screw speed of 500 rpm and barrel temperature of 150°C. The extrudates characteristics viz., expansion ratio-1.82 (PR 123); bulk density-184 g/cc (PR 123); specific mechanical energy-262.35 Wh/kg; water absorption index (WAI)-6.26 (PR 122); water solubility index-48.52% (PR 123); hardness-148.63 N (PR 122); and hydration power-284% (PR 122) were viably hyphenated with the physicochemical and rheological behavior of cultivars. The physico-functional characterization of the extrudates in terms of their starch and protein structural indexes, α-amylase susceptibility; water soluble carbohydrates and proteins revealed the possibility of exploring these cultivars as a functionally viable and diverse ingredient for the production of ready-to-eat extrudates.

Expanding the toolbox of metabolically stable lipid prodrug strategies.

Nucleoside- and nucleotide-based therapeutics are indispensable treatment options for patients suffering from malignant and viral diseases. These agents are most commonly administered to patients as prodrugs to maximize bioavailability and efficacy. While the literature provides a practical prodrug playbook to facilitate the delivery of nucleoside and nucleotide therapeutics, small context-dependent amendments to these popular prodrug strategies can drive dramatic improvements in pharmacokinetic (PK) profiles. Herein we offer a brief overview of current prodrug strategies, as well as a case study involving the fine-tuning of lipid prodrugs of acyclic nucleoside phosphonate tenofovir (TFV), an approved nucleotide HIV reverse transcriptase inhibitor (NtRTI) and the cornerstone of combination antiretroviral therapy (cART). Installation of novel lipid terminal motifs significantly reduced fatty acid hepatic ω-oxidation while maintaining potent antiviral activity. This work contributes important insights to the expanding repertoire of lipid prodrug strategies in general, but particularly for the delivery and distribution of acyclic nucleoside phosphonates.

Anti-nutrient & bioactive profile, in vitro nutrient digestibility, techno-functionality, molecular and structural interactions of foxtail millet (Setaria italica L.) as influenced by biological processing techniques.

The present investigation aimed at assessing the impact of biological processing techniques on bio-and techno-functional characteristics of foxtail millet (Setaria italica L.). Grains were exposed to soaking, germination, fermentation and combination of aforesaid treatments and significant variation (p < 0.05) in anti-nutritional factors, in vitro starch and protein digestibility, bioactive constituents and associated antioxidant potential was noted. Bioprocessed flours were characterized by altered functional properties due to hydrolytic action of activated enzymes. ATR-FTIR spectra and X-ray diffraction patterns revealed structural variation in macromolecular arrangement, synthesis of bioactive compounds in bioprocessed flours and slight reduction in the crystallinity of starch molecules. Bioprocessed flours exhibited degraded protein matrix; however, only fermentation and combination treatments caused hydrolysis of granular starch. Principal component analysis was employed to validate the differences in processing treatments and observations. The results are suggestive that bioprocessed flours could serve as potential ingredients with improved techno-and bio-functionality in valorized cereal products.

ω-Functionalized Lipid Prodrugs of HIV NtRTI Tenofovir with Enhanced Pharmacokinetic Properties.

Tenofovir (TFV) is the cornerstone nucleotide reverse transcriptase inhibitor (NtRTI) in many combination antiretroviral therapies prescribed to patients living with HIV/AIDS. Due to poor cell permeability and oral bioavailability, TFV is administered as one of two FDA-approved prodrugs, both of which metabolize prematurely in the liver and/or plasma. This premature prodrug processing depletes significant fractions of each oral dose and causes toxicity in kidney, bone, and liver with chronic administration. Although TFV exalidex (TXL), a phospholipid-derived prodrug of TFV, was designed to address this issue, clinical pharmacokinetic studies indicated substantial hepatic extraction, redirecting clinical development of TXL toward HBV. To circumvent this metabolic liability, we synthesized and evaluated ω-functionalized TXL analogues with dramatically improved hepatic stability. This effort led to the identification of compounds 21 and 23, which exhibited substantially longer t1/2 values than TXL in human liver microsomes, potent anti-HIV activity in vitro, and enhanced pharmacokinetic properties in vivo.

Cooperative Kinetics of the Glucan Phosphatase Starch Excess4.

Glucan phosphatases are members of a functionally diverse family of dual-specificity phosphatase (DSP) enzymes. The plant glucan phosphatase Starch Excess4 (SEX4) binds and dephosphorylates glucans, contributing to processive starch degradation in the chloroplast at night. Little is known about the complex kinetics of SEX4 when acting on its complex physiologically relevant glucan substrate. Therefore, we explored the kinetics of SEX4 against both insoluble starch and soluble amylopectin glucan substrates. SEX4 displays robust activity and a unique sigmoidal kinetic response to amylopectin, characterized by a Hill coefficient of 2.77 ± 0.63, a signature feature of cooperativity. We investigated the basis for this positive kinetic cooperativity and determined that the SEX4 carbohydrate-binding module (CBM) dramatically influences the binding cooperativity and substrate transformation rates. These findings provide insights into a previously unknown but important regulatory role for SEX4 in reversible starch phosphorylation and further advances our understanding of atypical kinetic mechanisms.

Dynamics of Germination Behaviour, Protein Secondary Structure, Technofunctional Properties, Antinutrients, Antioxidant Capacity and Mineral Elements in Germinated Dhaincha.

RESEARCH BACKGROUND: Dhaincha (Sesbania aculeata) is a forage legume primarily used for green manuring and animal feed. Good nutritional profile of dhaincha makes it a potential alternative legume in human nutrition. However, the presence of high amount of antinutrients poses a problem in its utilisation for food applications. The present investigation intends to germinate dhaincha seeds at different time-temperature regimes and to evaluate the process of germination to ascertain optimal conditions and improve its potential for utilisation. EXPERIMENTAL APPROACH: Dhaincha seeds were germinated at 24, 28 and 32 °C for 24, 48 and 72 h. Germination characteristics and germination loss, spectral characteristics, technofunctionality, antinutrients, bioactive constituents, antioxidant capacity and mineral element content of germinated dhaincha were evaluated. Optimal balance of technobiofunctionality of germinated dhaincha seeds was validated by principal component analysis. RESULTS AND CONCLUSIONS: Sprout length and germination loss increased with the higher germination temperature and prolonged germination time. Seeds showed similar germination rate at 28 and 32 °C and it was markedly higher than at 24 °C. Germination for 24 h resulted in mild conformational changes in the secondary structure of proteins, whereas germination for 48 and 72 h exhibited major conformational changes in the ß-sheets, resulting in the improvement in the hydration and foaming properties. Progression of germination (72 h) caused the decrease of tannin (24.47%), phytic acid (16.38%) and saponin (24.58%) mass fractions, and of trypsin inhibitor (40.33%) and lectin activity (62.50%). Slight decrease of DPPH˙ (3.7%) and ABTS˙+ (18.5%) values was also observed, whereas total flavonoid content (36.14%) and metal chelating activity (26.76%) increased. Total phenolics, FRAP, and reducing power decreased after 24 h, followed by a gradual increase. Zinc extractability increased drastically with germination. Germination at 28 °C for 72 h resulted in higher reduction of antinutrients with optimal retention of antioxidant activity and better functional characteristics, as validated by principal component analysis. NOVELTY AND SCIENTIFIC CONTRIBUTION: Dhaincha is an unknown crop in Europe, and even in Asia it is predominantly used as green manure and animal feed. This research demonstrated that the intervention in germination can transform dhaincha into a promising crop for food industry. Germinated dhaincha exhibited enhanced technobiofunctionality for utilisation in various food formulations.

Generation and characterization of a laforin nanobody inhibitor.

OBJECTIVES: Mutations in the gene encoding the glycogen phosphatase laforin result in the fatal childhood dementia Lafora disease (LD). A cellular hallmark of LD is cytoplasmic, hyper-phosphorylated, glycogen-like aggregates called Lafora bodies (LBs) that form in nearly all tissues and drive disease progression. Additional tools are needed to define the cellular function of laforin, understand the pathological role of laforin in LD, and determine the role of glycogen phosphate in glycogen metabolism. In this work, we present the generation and characterization of laforin nanobodies, with one being a laforin inhibitor. DESIGN AND METHODS: We identify multiple classes of specific laforin-binding nanobodies and determine their binding epitopes using hydrogen deuterium exchange (HDX) mass spectrometry. Using para-nitrophenyl phosphate (pNPP) and a malachite gold-based assay specific for glucan phosphatase activity, we assess the inhibitory effect of one nanobody on laforin's catalytic activity. RESULTS: Six families of laforin nanobodies are characterized and their epitopes mapped. One nanobody is identified and characterized that serves as an inhibitor of laforin's phosphatase activity. CONCLUSIONS: The six generated and characterized laforin nanobodies, with one being a laforin inhibitor, are an important set of tools that open new avenues to define unresolved glycogen metabolism questions.