Dynamic rheological behavior of high-amylose wheat dough during various heating stages: Insight from its starch characteristics.

The objective of this study was to understand how the dynamic rheological behaviors of high-amylose wheat (HAW) dough during various heating stages measured using a mixolab were affected by the starch properties. At the heating stage of 30 °C - 90 °C, low minimum (C2) and peak (C3) torques were observed for HAW doughs, which resulted from their reduced starch granule swelling. During holding at 90 °C, HAW doughs had low minimum (C4) and C3 - C4 torques, indicating a good resistance to mechanical shear and endogenous enzyme degradation. HAW doughs also had low final (C5) and setback (C5 - C4) torques, consistent with their low starch swelling power and solubility. The increased amylose in HAW starch formed long-chain double-helical B-type polymorph and amylose-lipid complex, which resulted in high starch gelatinization-temperatures and enthalpy change, low swelling power and solubility, low pasting viscosity, and high resistance of swollen granules to mechanical shear and enzyme degradation. The overall patterns of dough-rheological behavior of HAW doughs during heating were similar to their respective starch pasting profiles, indicating that starch was the dominant contributor to the dough rheology during heating. This study provides useful information for food applications and manufacturing of HAW-based products, especially none-fermented products requiring firm texture and low viscosity.

Understanding resistant-starch formation during drying high-amylose maize kernels.

Interests in using high-amylose maize (HAM) flour and starch for low glycemic index foods continue to grow. The objective of this work was to understand resistant-starch formation during drying the HAM kernels. Freshly harvested HAM kernels with 28.2 % initial moisture were subjected to sun drying (~30 °C) or hot-air drying at 50 °C, 70 °C, 90 °C, or 110 °C. The enzymatic digestibility of HAM flour decreased from 63.6 % to 41.1 % as the drying temperature increased from 30 °C to 110 °C. The swelling power, solubility, and overall viscosity of HAM flours milled from kernels dried at 110 °C decreased, whereas the peak and conclusion gelatinization temperatures, enthalpy change, and relative crystallinity increased compared to those of flours from kernels dried at 30 °C, 50 °C, 70 °C, and 90 °C. Light microscopic and scanning electron microscopic images showed that starch granule aggregation in HAM flour increased with increasing drying-temperatures. The aggregates remained after 16 h enzymatic hydrolysis of cooked HAM flours. These results suggested that the increase of enzymatic resistance of HAM flour resulted from the formation of high temperature-resistant ordered structures in starch granules and the starch aggregates less accessible to enzymatic hydrolysis.

Miniaturized CO2 Gas Sensor Using 20% ScAlN-Based Pyroelectric Detector.

NDIR CO2 gas sensors using a 10-cm-long gas channel and CMOS-compatible 12% doped ScAlN pyroelectric detector have previously demonstrated detection limits down to 25 ppm and fast response time of ∼2 s. Here, we increase the doping concentration of Sc to 20% in our ScAlN-based pyroelectric detector and miniaturize the gas channel by ∼65× volume with length reduction from 10 to 4 cm and diameter reduction from 5 to 1 mm. The CMOS-compatible 20% ScAlN-based pyroelectric detectors are fabricated over 8-in. wafers, allowing cost reduction leveraging on semiconductor manufacturing. Cross-sectional TEM images show the presence of abnormally oriented grains in the 20% ScAlN sensing layer in the pyroelectric detector stack. Optically, the absorption spectrum of the pyroelectric detector stack across the mid-infrared wavelength region shows ∼50% absorption at the CO2 absorption wavelength of 4.26 µm. The pyroelectric coefficient of these 20% ScAlN with abnormally oriented grains shows, in general, a higher value compared to that for 12% ScAlN. While keeping the temperature variation constant at 2 °C, we note that the pyroelectric coefficient seems to increase with background temperature. CO2 gas responses are measured for 20% ScAlN-based pyroelectric detectors in both 10-cm-long and 4-cm-long gas channels, respectively. The results show that for the miniaturized CO2 gas sensor, we are able to measure the gas response from 5000 ppm down to 100 ppm of CO2 gas concentration with CO2 gas response time of ∼5 s, sufficient for practical applications as the average outdoor CO2 level is ∼400 ppm. The selectivity of this miniaturized CO2 gas sensor is also tested by mixing CO2 with nitrogen and 49% sulfur hexafluoride, respectively. The results show high selectivity to CO2 with nitrogen and 49% sulfur hexafluoride each causing a minimum ∼0.39% and ∼0.36% signal voltage change, respectively. These results bring promise to compact and miniature low cost CO2 gas sensors based on pyroelectric detectors, which could possibly be integrated with consumer electronics for real-time air quality monitoring.

Pharmacokinetic, pharmacodynamic, and immunogenic rationale for optimal dosing of pegvaliase, a PEGylated bacterial enzyme, in adult patients with phenylketonuria.

Phenylketonuria (PKU), a deficiency in the activity of the enzyme phenylalanine hydroxylase, leads to toxic levels of phenylalanine (Phe) in the blood and brain. Pegvaliase (recombinant Anabaena variabilis phenylalanine ammonia lyase conjugated with polyethylene glycol) is approved to manage PKU in patients aged greater than or equal to 18 years in the United States and in patients aged greater than or equal to 16 years in the European Union. Pharmacokinetic, pharmacodynamic, and immunogenicity results from five open-label pegvaliase trials were assessed. Studies with induction/titration/maintenance (I/T/M) dosing regimens demonstrated pharmacokinetic stabilization and sustained efficacy associated with maintenance doses (20, 40, or 60 mg/day). Immune-mediated pegvaliase clearance was high during induction/titration phases when the early immune response was peaking. The combination of low drug dosage and high drug clearance led to low drug exposure and minimal decreases in blood Phe levels during induction/titration. Higher drug exposure and substantial reductions in blood Phe levels were observed later in treatment as drug clearance was reduced due to the maturation of the immune response, which allowed for increased dosing to target levels. The incidence of hypersensitivity reactions was temporally associated with the peaking of the early antidrug immune response and decreased with time as immune response matured after the first 6 months of treatment. These results support an I/T/M dosing regimen and suggest a strategy for administration of other nonhuman biologics to achieve efficacy and improve tolerability.

Pegvaliase for the treatment of phenylketonuria: A pivotal, double-blind randomized discontinuation Phase 3 clinical trial.

INTRODUCTION: Pegvaliase is a recombinant Anabaena variabilis phenylalanine ammonia lyase (PAL) enzyme under investigation for treatment of adult phenylketonuria (PKU). This manuscript describes results of a randomized discontinuation trial (RDT) designed to evaluate the effects of pegvaliase treatment on blood phenylalanine (Phe) and neuropsychiatric outcomes in adults with PKU. METHODS: PRISM-2 is a 4-part, Phase 3 study that enrolled adults with PKU receiving pegvaliase treatment (initiated in a prior Phase 2 or Phase 3 study). The RDT, Part 2 of PRISM-2, was an 8-week trial that evaluated change in blood Phe concentrations, neuropsychiatric and neurocognitive measures, and safety outcomes in PRISM-2 participants who had achieved at least a 20% blood Phe reduction from pre-treatment baseline with pegvaliase treatment. Participants were randomized 2:1 to either continue pegvaliase (20 mg/day or 40 mg/day) or switch to matching placebo. RESULTS: The pooled pegvaliase group enrolled 66 participants and each placebo group enrolled 14 participants. The primary endpoint of change in blood Phe concentration from RDT entry to RDT Week 8 was met with clinically meaningful and statistically significant differences between the pegvaliase and placebo groups. Mean (SD) blood Phe at the beginning of the RDT when all participants were receiving pegvaliase was 563.9 µM (504.6) in the group assigned to the 20 mg/day placebo group (n = 14), 508.2 µM (363.7) in those assigned to the 40 mg/day placebo group (n = 14), and 503.9 µM (520.3) in those assigned to continue pegvaliase treatment (n = 58). At Week 8 of the RDT, the least squares mean change (95% confidence interval) in blood Phe was 949.8 µM (760.4 to 1139.1) for the 20 mg/day placebo group and 664.8 µM (465.5 to 864.1) for the 40 mg/day placebo group in comparison to 26.5 µM (-68.3 to 121.3) for the pooled (20 mg/day and 40 mg/day) pegvaliase group (P < 0.0001 for pooled pegvaliase group vs each placebo group). Adverse events (AEs) were usually lower in the pooled placebo group when compared to the pooled pegvaliase group. The most common AEs for the pooled pegvaliase and pooled placebo groups were arthralgia (13.6% and 10.3%, respectively), headache (12.1% and 24.1%), anxiety (10.6% and 6.9%), fatigue (10.6% and 10.3%), and upper respiratory tract infection (1.5% and 17.2%). CONCLUSION: Mean blood Phe reduction was sustained in the pegvaliase group, while placebo groups had mean blood Phe concentration increase toward pre-treatment baseline levels. Results from this study confirmed the efficacy of pegvaliase in maintaining reduced blood Phe concentrations with a manageable safety profile for most participants.

Pegvaliase for the treatment of phenylketonuria: Results of a long-term phase 3 clinical trial program (PRISM).

BACKGROUND: Phenylketonuria (PKU) is caused by phenylalanine hydroxylase (PAH) deficiency that results in phenylalanine (Phe) accumulation. Pegvaliase, PEGylated recombinant Anabaena variabilis phenylalanine ammonia lyase (PAL), converts Phe to trans-cinnamic acid and ammonia, and is a potential enzyme substitution therapy to lower blood Phe in adults with PKU. METHODS: Two Phase 3 studies, PRISM-1 and PRISM-2, evaluated the efficacy and safety of pegvaliase treatment using an induction, titration, and maintenance dosing regimen in adults with PKU. In PRISM-1, pegvaliase-naïve participants with blood Phe >600 µmol/L were randomized 1:1 to a maintenance dose of 20 mg/day or 40 mg/day of pegvaliase. Participants in PRISM-1 continued pegvaliase treatment in PRISM-2, a 4-part clinical trial that includes an ongoing, open-label, long-term extension study of pegvaliase doses of 5 mg/day to 60 mg/day. RESULTS: Of 261 participants who received pegvaliase treatment, 72.0% and 32.6% reached ≥12 months and ≥ 24 months of study treatment, respectively, and 65% are still actively receiving treatment. Mean (SD) blood Phe was 1232.7 (386.4) µmol/L at baseline, 564.5 (531.2) µmol/L at 12 months, and 311.4 (427) µmol/L at 24 months, a decrease from baseline of 51.1% and 68.7%, respectively. Within 24 months, 68.4% of participants achieved blood Phe ≤600 µmol/L, 60.7% of participants achieved blood Phe ≤360 µmol/L, below the upper limit recommended in the American College of Medical Genetics and Genomics PKU management guidelines, and 51.2% achieved blood Phe ≤120 µmol/L, below the upper limit of normal in the unaffected population. Improvements in neuropsychiatric outcomes were associated with reductions in blood Phe and were sustained with long-term pegvaliase treatment. Adverse events (AEs) were more frequent in the first 6 months of exposure (early treatment phase) than after 6 months of exposure (late treatment phase); 99% of AEs were mild or moderate in severity and 96% resolved without dose interruption or reduction. The most common AEs were arthralgia (70.5%), injection-site reaction (62.1%), injection-site erythema (47.9%), and headache (47.1%). Acute systemic hypersensitivity events consistent with clinical National Institute of Allergy and Infectious Diseases and the Food Allergy and Anaphylaxis Network anaphylaxis criteria were observed in 12 participants (17 events); of these, 6 participants remained on treatment. Acute systemic hypersensitivity events including potential events of anaphylaxis were not associated with immunoglobulin E, and all events resolved without sequelae. CONCLUSION: Results from the PRISM Phase 3 program support the efficacy of pegvaliase for the treatment of adults with PKU, with a manageable safety profile in most participants. The PRISM-2 extension study will continue to assess the long-term effects of pegvaliase treatment.

Experimental demonstration of highly sensitive optical sensor based on grating-assisted light coupling between strip and slot waveguides.

An optical sensor based on grating-assisted light coupling between a strip waveguide and a slot waveguide is demonstrated (the sensor was proposed and analyzed in [Opt. Express21, 5897-5909 (2013)]. The wavelength at which the light is strongly coupled between two waveguides is used to the measure the external medium's refractive index. The sensor was fabricated with silicon nitride waveguides and obvious grating induced band-rejection and band-pass characteristics were observed. The measured sensitivity of the fabricated sensor was -756.1 nm/RIU. Furthermore, by covering the strip waveguide with the silicon dioxide cladding, the sensitivity was measured to be as large as -1970 nm/RIU, which was 2.6 times enhanced. The experimental results agreed well with the calculated sensitivity values.

Rapid evolving into acute myeloid leukemia in a patient with multiple myeloma and concurrent myelodysplasia after VTD therapy.

The association of Myelodysplasia (MDS) and multiple myeloma (MM) has been usually described not only as a complication of chemotherapy but also in the absence of preceding chemotherapy or together at the time of diagnosis. Optimal therapies of a coexisting MM and MDS have not been well established up to now. We report a case of MDS diagnosed simultaneously with MM. After treatment with VTD (bortezomib, thalidomide, dexamethasone) marked anti-myeloma activity was observed, but it was associated with rapid progression of the MDS to acute myeloid leukemia (AML). The leukemic transformation in our case most probably reflects the natural progression of MDS, though it clearly demonstrates that VTD is ineffective in controlling blast proliferation in MDS. To our knowledge, this is the first case report on MDS in the setting of MM with rapid evolution to AML to VTD therapy. More data from more cases are needed, to find the potential utility of VTD therapy in coexisting MDS and MM patients.

A rapid amplification/detection assay for analysis of Mycobacterium tuberculosis using an isothermal and silicon bio-photonic sensor complex.

Global tuberculosis (TB) control is hampered by cost and slow or insensitive diagnostic methods to be used for TB diagnosis in clinic. Thus, TB still remains a major global health problem. The failure to rapidly and accurately diagnose of TB has posed significant challenges with consequent secondary resistance and ongoing transmission. We developed a rapid Mycobacterium tuberculosis (MTB) amplification/detection method, called MTB isothermal solid-phase amplification/detection (MTB-ISAD), that couples isothermal solid-phase amplification and a silicon biophotonics-based detection sensor to allow the simultaneous amplification and detection of MTB in a label-free and real-time manner. We validated the clinical utility of the MTB-ISAD assay by detecting MTB nucleic acid in sputum samples from 42 patients. We showed the ability of the MTB-ISAD assay to detect MTB in 42 clinical specimens, confirming that the MTB-ISAD assay is fast (<20 min), highly sensitive, accurate (>90%, 38/42), and cost-effective because it is a label-free method and does not involve thermal cycling. The MTB-ISAD assay has improved time-efficiency, affordability, and sensitivity compared with many existing methods. Therefore, it is potentially adaptable for better diagnosis across various clinical applications.

Single-channel Mach-Zehnder interferometric biochemical sensor based on two-lateral-mode spiral waveguide.

We propose and demonstrate a single-channel Mach-Zehnder interferometric (MZI) biochemical sensor consisting of two single-mode waveguides connected by a two-lateral-mode spiral sensing waveguide through two discontinuous junctions. The use of a two-lateral-mode waveguide offers the advantage of simple fabrication using single-step lithography and etching process. Meanwhile, the two-mode waveguide folded in a spiral layout can achieve high sensitivity of a long sensing waveguide while providing a compact sensing area compatible with commercial spotting machine and requiring small volume of sample. The sensor is demonstrated in silicon waveguides and the effect of the discontinuity offset distance on the interference visibility is studied. The bulk and surface sensitivity of a fabricated sensor with a 4582-µm-long two-mode spiral waveguide folded within a 185 µm diameter spot are characterized to be 461.6 π/RIU (refractive index unit) and 1.135 π/ng mm(-2), respectively. The biosensing capability of the sensor is verified by the measurement of biotin-streptavidin interaction of different concentrations.

Single-dose, subcutaneous recombinant phenylalanine ammonia lyase conjugated with polyethylene glycol in adult patients with phenylketonuria: an open-label, multicentre, phase 1 dose-escalation trial.

BACKGROUND: Phenylketonuria is an inherited disease caused by impaired activity of phenylalanine hydroxylase, the enzyme that converts phenylalanine to tyrosine, leading to accumulation of phenylalanine and subsequent neurocognitive dysfunction. Phenylalanine ammonia lyase is a prokaryotic enzyme that converts phenylalanine to ammonia and trans-cinnamic acid. We aimed to assess the safety, tolerability, pharmacokinetic characteristics, and efficacy of recombinant Anabaena variabilis phenylalanine ammonia lyase (produced in Escherichia coli) conjugated with polyethylene glycol (rAvPAL-PEG) in reducing phenylalanine concentrations in adult patients with phenylketonuria. METHODS: In this open-label, phase 1, multicentre trial, single subcutaneous injections of rAvPAL-PEG were given in escalating doses (0·001, 0·003, 0·010, 0·030, and 0·100 mg/kg) to adults with phenylketonuria. Participants aged 18 years or older with blood phenylalanine concentrations of 600 µmol/L or higher were recruited from among patients attending metabolic disease clinics in the USA. The primary endpoints were safety and tolerability of rAvPAL-PEG. Secondary endpoints were the pharmacokinetic characteristics of the drug and its effect on concentrations of phenylalanine. Participants and investigators were not masked to assigned dose group. This study is registered with ClinicalTrials.gov, number NCT00925054. FINDINGS: 25 participants were recruited from seven centres between May 6, 2008, and April 15, 2009, with five participants assigned to each escalating dose group. All participants were included in the safety population. The most frequently reported adverse events were injection-site reactions and dizziness, which were self-limited and without sequelae. Two participants had serious adverse reactions to intramuscular medroxyprogesterone acetate, a drug that contains polyethylene glycol as an excipient. Three of five participants given the highest dose of rAvPAL-PEG (0·100 mg/kg) developed a generalised skin rash. By the end of the study, all participants had developed antibodies against polyethylene glycol, and some against phenylalanine ammonia lyase as well. Drug concentrations peaked about 89-106 h after administration of the highest dose. Treatment seemed to be effective at reducing blood phenylalanine in all five participants who received the highest dose (mean reduction of 54·2% from baseline), with a nadir about 6 days after injection and an inverse correlation between drug and phenylalanine concentrations in plasma. Phenylalanine returned to near-baseline concentrations about 21 days after the injection. INTERPRETATION: Subcutaneous administration of rAvPAL-PEG in a single dose of up to 0·100 mg/kg was fairly safe and well tolerated in adult patients with phenylketonuria. At the highest dose tested, rAvPAL-PEG reduced blood phenylalanine concentrations. In view of the development of antibodies against polyethylene glycol (and in some cases against phenylalanine ammonia lyase), future studies are needed to assess the effect of repeat dosing. FUNDING: BioMarin Pharmaceutical.

Silicon waveguide filter based on cladding modulated anti-symmetric long-period grating.

In this paper, we demonstrate an optical filter using cladding modulated anti-symmetric long-period grating in a two-mode silicon waveguide. The filter consists of a two-mode waveguide connected with an input and output single-mode waveguide through two linear tapers. The anti-symmetric grating is formed by placing two periodic arrays of silicon squares offset by half of a grating pitch along the two-mode waveguide. Light coupling occurs between two co-propagating modes at the coupling wavelength through the grating and results in a rejection band at the output. The grating pitch, coupling coefficient, transmission spectrum and 3-dB bandwidth of the grating are investigated with the coupled-mode theory. By using a cladding modulated grating, the grating coupling strength can be controlled over a wide range by the two-mode waveguide width or separation distance between the grating and waveguide. Band-rejection filters are experimentally demonstrated in 1-µm, 0.8-µm and 0.7-µm wide two-mode silicon waveguides and rejection bands with different bandwidths and maximal attenuation contrasts larger than 15 dB (~97% coupling efficiency) have been achieved.

Overexpression of glucosylceramide synthase in associated with multidrug resistance of leukemia cells.

Ceramide, as a second messenger, initiates one of the major signal transduction pathways in tumor apoptosis. Glucosylceramide synthase (GCS) catalyzes glycosylation of ceramide and produces glucosylceramide. Through GCS, ceramide glycosylation allows cellular escape from ceramide-induced programmed cell death. Here we investigated the expression of GCS in human leukemia cells and an association between GCS and multidrug resistance of leukemia cells. Using RT-PCR technique the level of GCS gene was detected in 65 clinical multidrug resistance/non-resistance cases with leukemia, and in K562 and K562/A02 cell lines. AlamarBlue Assay was applied to confirm the multidrug resistant of K562/A02 cells. PPMP, which is a chemical inhibitor for GCS, was used to determine the relationship between GCS and drug-resistance in K562/A02 cells. In addition, multidrug resistance gene (mdr1), Bcl-2 and Bax mRNA was also analyzed by RT-PCR. The expression of GCS and mdr1 mRNA in clinic multidrug resistance samples exhibited significantly increased compared with clinic drug sensitive group (P<0.05). There was the positive correlation both the expression of GCS and mdr1 genes in leukemia samples (P<0.01, gamma=0.7). AlamarBlue Assay showed that the K562/A02 cell line was 115-fold more resistant to adriamycin and 36-fold more resistant to vincristine compared with drug-sensitive K562 cell line. There also was significant expression difference of GCS and mdr1 genes between K562 and K562/A02 cells. Bcl-2 gene exhibited higher expressions whatever in clinic drug-resistance samples or K562/A02 cells, whereas the expressions of Bax gene were higher in drug-sensitive samples and K562 cells. PPMP increased sensitivity to adriamycin toxicity by inhibiting GCS in K562/A02 cells. Therefore, it is suggested that a high level of GCS in leukemia is possible contributed to multidrug resistance of leukemia cells. Abnormally expressions of the genes in associated with cell apoptosis might be one of the main molecular pathology mechanisms of multidrug resistance caused by GCS gene.

[Relation between glucosylceramide synthase and multidrug resistance in leukemia cells].

This study was purposed to explore the expression of glucosylceramide synthase (GCS) in human leukemia cells and its relationship with multidrug resistance. RT-PCR was used to analyze peripheral blood samples from 53 leukemia patients with multidrug resistance/non-resistance, and to detect the expression level of GCS gene in HL-60 cells and HL-60/ADR cells, the expression level was compared with the level of mdr-1. The expressions of GCS protein and P-gp protein in HL-60 cells and HL-60/ADR cells were assayed by Western blot analysis. The results showed that the relative optical density ratio of GCS gene amplified bands in samples of leukemia patients with drug-resistance was significantly higher than that in samples of leukemia patients with drug non-resistance group (P < 0.05), meanwhile the significant enhancement of optical density value of GCS gene amplified bands accompanied by high expression of mdr-1 gene. Their correlation showed positive (P < 0.01, r = 0.6). The GCS mRNA and protein were overexpressed in HL-60/ADR cells, and their expression levels were obviously higher than that in HL-60 cells, meanwhile the expression of mdr-1 mRNA and P-gp also significantly increased in HL-60/ADR cells. It is concluded that the high level of GCS in leukemia patients possibly is associated with multidrug resistance of leukemia cells.