ABSTRACT
INTRODUCTION: Spinocerebellar ataxia type 3 (SCA3) is a neurodegenerative disease characterized by increasingly worsening ataxia and non-ataxia features, negatively impacting patients' quality of life. This study was designed to test formally evaluate whether oral trehalose was effective in SCA3 patients. METHODS: In this double-blind, randomized controlled trial, SCA3 patients received either 100 g oral trehalose or 30 g maltose to improve ataxia severity over six months. We also measured other clinical (non-ataxia), patient-reported (quality of life, motivations), and safety endpoints. An unscheduled interim analysis was conducted using two-way ANOVAs to analyze the interaction between time (baseline, 3-months, 6-months) and intervention (Trehalose vs. Placebo). RESULTS: Fifteen participants (Trehalose = 7 vs. Placebo = 8) completed the study at the time of interim analysis. There was no interaction effect on the ataxia severity, and available data suggested an estimated sample size of 132 (66 per arm) SCA3 patients required to demonstrate changes in a 6-month trial. There were significant interaction effects for executive function (Æ2 = 0.28-0.43). Safety data indicated that 100 g oral trehalose was well-tolerated. CONCLUSION: We performed an unplanned interim analysis due to a slow recruitment rate. The new estimated sample size was deemed unfeasible, leading to premature termination of the clinical trial. In this small, current sample of SCA3 patients, 100 g oral trehalose did not differentially impact on ataxia severity compared to placebo. Interestingly, our findings may suggest an improvement in executive function. Future efforts will require a large multi-country, multi-center study to investigate the potential effect of trehalose.
Subject(s)
Machado-Joseph Disease , Trehalose , Humans , Trehalose/administration & dosage , Trehalose/pharmacology , Double-Blind Method , Male , Female , Middle Aged , Machado-Joseph Disease/drug therapy , Adult , Administration, Oral , Aged , Severity of Illness Index , Quality of Life , Outcome Assessment, Health CareABSTRACT
Carbohydrate (CHO) supplementation during endurance exercise can improve performance. However, it is unclear whether low glycemic index (GI) CHO leads to differential ergogenic and metabolic effects compared with a standard high GI CHO. This study investigated the ergogenic and metabolic effects of CHO supplementation with distinct GIs, namely, (a) trehalose (30 g/hr), (b) isomaltulose (30 g/hr), (c) maltodextrin (60 g/hr), and (d) placebo (water). In this double-blind, crossover, counterbalanced, placebo-controlled study, 13 male cyclists cycled a total of 100 min at varied exercise intensity (i.e., 10-min stages at 1.5, 2.0, and 2.5 W/kg; repeated three times plus two 5-min stages at 1.0 W/kg before and after the protocol), followed by a 20-min time trial on four separated occasions. Blood glucose and lactate (every 20 min), heart rate, and ratings of perceived exertion were collected throughout, and muscle biopsies were taken before and immediately after exercise. The results showed that trehalose improved time-trial performance compared with placebo (total work done 302 ± 39 vs. 287 ± 48 kJ; p = .01), with no other differences between sessions (all p ≥ .07). Throughout the 100-min protocol, blood glucose was higher with maltodextrin compared with the other supplements at all time points (all p < .05). Heart rate, ratings of perceived exertion, muscle glycogen content, blood glucose, and lactate were not different between conditions when considering the 20-min time trial (all p > .05). Trehalose supplementation throughout endurance exercise improved cycling performance and appears to be an appropriate CHO source for exercise tasks up to 2 hr. No ergogenic superiority between the different types of CHO was established.
Subject(s)
Athletic Performance , Bicycling , Blood Glucose , Cross-Over Studies , Heart Rate , Isomaltose , Lactic Acid , Polysaccharides , Trehalose , Humans , Male , Bicycling/physiology , Double-Blind Method , Trehalose/administration & dosage , Trehalose/pharmacology , Athletic Performance/physiology , Adult , Blood Glucose/metabolism , Blood Glucose/drug effects , Heart Rate/drug effects , Lactic Acid/blood , Polysaccharides/administration & dosage , Polysaccharides/pharmacology , Isomaltose/analogs & derivatives , Isomaltose/administration & dosage , Isomaltose/pharmacology , Dietary Supplements , Glycemic Index , Physical Endurance/drug effects , Physical Endurance/physiology , Muscle, Skeletal/drug effects , Muscle, Skeletal/metabolism , Sports Nutritional Physiological Phenomena , Performance-Enhancing Substances/administration & dosage , Performance-Enhancing Substances/pharmacology , Dietary Carbohydrates/administration & dosage , Young Adult , Physical Exertion/physiology , Physical Exertion/drug effects , Glycogen/metabolismABSTRACT
Trehalose has been widely studied as a treatment for a variety of human disorders due to its ability to stimulate autophagy. Trehalose, however, is poorly adsorbed and is hydrolyzed in the intestinal mucosa, and oral delivery requires relatively high doses to induce autophagy. The parenteral injection of trehalose-releasing nanogels proposed in this study offers an alternative mode of delivery. This study aimed to develop stable colloidal dispersions of trehalose-rich nanogels that could sustainably release trehalose under physiologically relevant conditions. The nanogel design was based on the covalent incorporation of 6-O-acryloyl-trehalose within a polymer network. A series of nine trehalose-rich nanogels with highly conjugated trehalose (up to 59 % w/w) were synthesized and shown to sustainably release trehalose at a rate that is not dose dependent. The nanogels were optimized to keep colloidal stability in serum-enriched cell culture media. The stable nanogels were not cytotoxic to primary HUVECs. Two selected nanogels with opposite surface charges were subjected to extended in vitro characterization that included a cellular uptake study and a hemocompatibility assay. Both nanogels were efficiently taken up by HUVECs during a short incubation. They also proved not to be hemolytic to human RBCs in concentrations up to 2.0 mg/mL. Finally, an in vivo autophagy stimulation study employing transgenic zebrafish and Drosophila larvae demonstrated that prolonged exposure to a cationic trehalose-releasing nanogel can induce autophagic activity in in vivo systems without any detectable toxicity.
Subject(s)
Excipients , Trehalose , Animals , Autophagy , Drosophila , Humans , Nanogels , Polymers , Trehalose/administration & dosage , ZebrafishABSTRACT
Trehalose is added in drug formulations to act as fillers or improve aerosolization performance. Its characteristics as a carrier molecule have been explored; however, the fate of trehalose in human airway tissues has not been thoroughly investigated. Here, we investigated the fate of nebulized trehalose using in vitro human air-liquid bronchial epithelial cultures. First, a tracing experiment was conducted using 13C12-trehalose; we measured trehalose distribution in different culture compartments (apical surface liquid, epithelial culture, and basal side medium) at various time points following acute exposure to 13C12-labeled trehalose. We found that 13C12-trehalose was metabolized into 13C6-glucose. The data was then used to model the kinetics of trehalose disappearance from the apical surface of bronchial cultures. Secondly, we evaluated the potential adverse effects of nebulized trehalose on the bronchial cultures after they were acutely exposed to nebulized trehalose up to a level just below its solubility limit (50 g/100 g water). We assessed the ciliary beating frequency and histological characteristics. We found that nebulized trehalose did not lead to marked alteration in ciliary beating frequency and morphology of the epithelial cultures. The in vitro testing approach used here may enable the early selection of excipients for future development of inhalation products.
Subject(s)
Bronchi/drug effects , Respiratory Mucosa/drug effects , Trehalose/pharmacology , Aerosols/administration & dosage , Aerosols/pharmacokinetics , Aerosols/pharmacology , Bronchi/metabolism , Cells, Cultured , Humans , Nebulizers and Vaporizers , Respiratory Mucosa/metabolism , Trehalose/administration & dosage , Trehalose/pharmacokineticsABSTRACT
BACKGROUND: Oral administration of bioactive peptides has potential clinical advantages, but its applicability is limited due to gastric and pancreatic enzyme proteolysis. OBJECTIVE: To examine whether the co-packaging of bovine colostrum (BC), a rich source of IgG, immune and growth factors, with the food additives trehalose (carbohydrate), stearine (fat), casein (protein present in BC) or soy flour (plant based with high protease inhibitory activity) enhances the stability of BC against digestion. DESIGN: Samples alone and in combination (BC+ 10% wt/wt trehalose, stearine, casein or soy) were exposed to HCl/pepsin, followed by trypsin and chymotrypsin ("CT"). Assessment of proliferation used gastric AGS cells (Alamar blue), IgG function measured bovine IgG anti-E.coli binding and ELISAs quantified growth factor constituents. In vivo bioassay assessed ability of BC alone or with soy to reduce injury caused by dextran sodium sulphate (DSS, 4% in drinking water, 7 days, test products started 2 days prior to DSS). RESULTS: Proliferative activity of BC reduced 61% following HCl/pepsin and CT exposure. This was truncated 50% if soy was co-present, and also protected against loss of total IgG, IgG E.coli binding, TGFß, lactoferrin and EGF (all P<0.01 vs BC alone). Co-packaging with trehalose was ineffective in preventing digestion whereas casein or stearine provided some intermediate protective effects. Rats given BC alone showed beneficial effects on weight gain, disease activity index, tissue histology and colonic MPO. Soy alone was ineffective. BC+ soy combination showed the greatest benefit with a dose of 7 mg/kg (6.4 BC + 0.6 soy flour) having the same degree of benefit as using 20 mg/kg BC alone. CONCLUSION: Soy, and to a lesser extent casein, enhanced the biostability of BC against digestive enzymes. Co-packaging of BC with other food products such as soy flour could result in a decreased dose being required, improving cost-effectiveness and patient compliance.
Subject(s)
Caseins/administration & dosage , Cell Proliferation/drug effects , Colostrum/chemistry , Stomach/drug effects , Trehalose/administration & dosage , Animals , Cattle , Cell Line , Cell Line, Tumor , Humans , Stomach/cytologyABSTRACT
Trehalose, a nonreducing disaccharide consisting of d-glucose with α,α-1,1 linkage, was evaluated as a functional material to improve the gut environment in preweaned calves. In experiment 1, 173 calves were divided into two groups; the trehalose group was fed trehalose at 30 g/animal/d with milk replacer during the suckling period, and the control group was fed nonsupplemented milk replacer. Medication frequency was lower in the trehalose group (P < 0.05). In experiment 2, calves (n = 20) were divided into two groups (control group [n = 10] and trehalose group [n = 10]) based on their body weight and reared under the same feeding regimens as in experiment 1. Fresh feces were collected from individual animals at the beginning of the trial (average age 11 d), 3 wk after trehalose feeding (experimental day 22), and 1 d before weaning, and the fecal score was recorded daily. Fecal samples were analyzed for fermentation parameters and microbiota. The fecal score was significantly lower in the trehalose group than in the control group in the early stage (at an age of 14 to 18 d; P < 0.05) of the suckling period. Calves fed trehalose tended to have a higher proportion of fecal butyrate on day 22 than calves in the control group (P = 0.08). Population sizes of Clostridium spp. were significantly lower (P = 0.036), whereas those of Dialister spp. and Eubacterium spp. tended to be higher in the feces of calves in the trehalose group on day 22 (P = 0.060 and P = 0.083). These observations indicate that trehalose feeding modulated the gut environment and partially contributed to the reduction in medication frequency observed in experiment 1.
Subject(s)
Cattle Diseases/epidemiology , Diarrhea/veterinary , Feces/microbiology , Microbiota , Milk , Trehalose/administration & dosage , Animal Feed/analysis , Animals , Body Weight , Cattle , Diarrhea/epidemiology , Diet/veterinary , Dietary Supplements , Incidence , WeaningABSTRACT
Purpose: To compare the efficacy and safety of HU00701 (0.01% cyclosporin A + 3% trehalose), HU007 (0.02% cyclosporin A + 3% trehalose) (all w/v), and placebo in patients with moderate to severe dry eye disease (DED). Methods: This was a multicenter, randomized, double-masked, parallel, placebo-controlled phase II study. In total, 114 patients were randomly assigned to the HU00701, HU007, placebo, or reference group. There was a 2-week run-in period before the 12-week intervention. Efficacy and safety were evaluated every 4 weeks. Results: The primary endpoint, change in corneal staining score from baseline to week 12, did not differ significantly among the control, HU00701, and HU007 groups in the full analysis. Of the secondary endpoints, only the tear film breakup time differed significantly at week 12 between the placebo and HU00701 groups. Twenty adverse events were reported by 15 patients, but the rate did not differ significantly among the 4 groups. The laboratory test, vital signs, and physical examination data showed no significant changes during the study. Conclusions: HU00701 and HU007 are safe, and HU007 effectively reduces the corneal staining score in patients with moderate-to-severe DED (NCT02917512).
Subject(s)
Cyclosporine/therapeutic use , Dry Eye Syndromes/drug therapy , Ophthalmic Solutions/therapeutic use , Trehalose/therapeutic use , Cyclosporine/administration & dosage , Cyclosporine/adverse effects , Double-Blind Method , Female , Humans , Male , Middle Aged , Ophthalmic Solutions/administration & dosage , Ophthalmic Solutions/adverse effects , Prospective Studies , Trehalose/administration & dosageABSTRACT
Autophagy is a degradation pathway for long-lived cytoplasmic proteins or damaged organelles and also for many aggregate-prone and disease-causing proteins. Endoplasmic reticulum (ER) stress and oxidative stress are associated with the pathophysiology of various liver diseases. These stresses induce the accumulation of abnormal proteins, Mallory-Denk body (MDB) formation and apoptosis in hepatocytes. A disaccharide trehalose had been reported to induce autophagy and decrease aggregate-prone proteins and cytotoxicity in neurodegenerative disease models. But the effects of trehalose in hepatocytes have not been fully understood. We examined the effect of trehalose on autophagy, ER stress and oxidative stress-mediated cytotoxicity and MDB formation in hepatocytes using mice model with 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) treatment for 3 months. We administered trehalose by intraperitoneal injection of water containing 10% trehalose (0.02 mg/g body weight) every other day for 3 months. Our results demonstrated that trehalose induced autophagy and reduced ER stress, oxidative stress, MDB formation and apoptosis in hepatocytes of DDC-fed mice by Western blotting and immunostaining analyses. Electron microscopy revealed that trehalose induced autolysosome formation, which located is close to the MDBs. Thus, our findings suggest that trehalose can become a therapeutic agent for oxidative stress-related liver diseases via activating autophagy.
Subject(s)
Autophagy/drug effects , Liver Diseases/prevention & control , Liver/pathology , Mallory Bodies/drug effects , Trehalose/administration & dosage , Administration, Oral , Animals , Apoptosis/drug effects , Cell Line, Tumor , Disease Models, Animal , Endoplasmic Reticulum Stress/drug effects , Hepatocytes/cytology , Hepatocytes/drug effects , Hepatocytes/pathology , Humans , Liver/cytology , Liver/drug effects , Liver Diseases/pathology , Mice , Oxidative Stress/drug effects , Pyridines/administration & dosage , Pyridines/toxicityABSTRACT
AIM: To evaluate the short-term effect of 0.15% sodium hyaluronate (SH), 0.20% SH, and Trehalose + 0.15% SH on anterior corneal aberrations in dry eye patients. METHODS: 76 eyes of 76 dry eye patients were divided into three groups. Non-preserved 1.5 mg/mL SH was administered in group 1, non-preserved 2.0 mg/mL SH was administered in group 2, and non-preserved trehalose 30 mg/mL and 1.5 mg/mL SH was administered in group 3. Aberrometry measurements were performed before and 10 min after application of the artificial tear drop. Using the Pentacam Scheimpflug imaging system, total root mean square (RMS), lower-order aberration (LOA), higher-order aberrations (HOAs), spherical aberration (SA), trefoil, and coma aberrations were investigated. RESULTS: In each group; the RMS of total, LOA, HOAs, and spherical aberration were significantly decreased after the artificial drop instillation, compared with those of them at baseline; and in groups 1 and 2, vertical trefoil term was also significantly increased, compared with those of them at baseline. According to intergroup analyzes, there was no significant outcome. CONCLUSIONS: It was observed that three artificial tears reduced the anterior corneal aberrations, in a 10-min period. The short-term effect of three artificial tears on the anterior corneal aberration was similar.
Subject(s)
Dry Eye Syndromes/drug therapy , Hyaluronic Acid/administration & dosage , Lubricant Eye Drops/administration & dosage , Trehalose/administration & dosage , Aberrometry , Adult , Aged , Cornea/diagnostic imaging , Cornea/drug effects , Dry Eye Syndromes/diagnosis , Female , Humans , Male , Middle Aged , Prospective Studies , Treatment Outcome , Young AdultABSTRACT
Insects, due to their small size, have limited energy storage space, but they also have high metabolic rate, so their hemolymph sugars are incredibly dynamic and play a number of important physiological functional roles in maintaining energetic homeostasis. In contrast to vertebrates, trehalose is generally the primary sugar found in insect hemolymph, which is followed by glucose and fructose. Many analytical chemistry methods exist to measure sugars, yet a direct comparison of methods that can measure all three simultaneously, and trehalose in particular, from low sample volumes, are sparse. Using the honey bee as a model, we directly compare the leading current methods of using High Performance Liquid Chromatography (HPLC) with an evaporative light-scattering detector and Gas Chromatography coupled with Mass Spectrometry (GC-MS) to determine which method would be better for measuring trehalose, glucose, and fructose in terms of reproducibility, accuracy, and sensitivity. Furthermore, we injected the enzyme inhibitors trehalozin (a trehalase inhibitor) and sorbose (a trehalase p-synthase inhibitor) to manipulate the trehalose levels in honey bee foragers as a proof of concept that this sugar can be altered independently of hemolymph glucose and fructose levels. Overall the HPLC method was less reproducible for measuring fructose and glucose, and it also had lower sensitivity for measuring trehalose. Consequently, significant differences in trehalose levels within the forager class were only detected with the GC-MS and not the HPLC method. Lastly, using the GC-MS method in the follow up study we found that trehalozin and sorbose causes a significant increase and decrease of trehalose levels respectively, in forager honey bees, independent of the glucose and fructose levels, ten minutes after injection. Taken together, these methods will provide useful tools for future studies exploring the many different physiological functional roles that trehalose can play in maintaining insect energetic homeostasis.
Subject(s)
Chromatography, High Pressure Liquid/methods , Disaccharides/administration & dosage , Gas Chromatography-Mass Spectrometry/methods , Hemolymph/chemistry , Sorbose/metabolism , Trehalose/metabolism , Age Factors , Animals , Bees , Disaccharides/pharmacology , Food Deprivation/physiology , Hemolymph/metabolism , Sorbose/administration & dosage , Sugars/metabolism , Trehalose/administration & dosage , Trehalose/antagonists & inhibitorsABSTRACT
The inhibitory effects of trehalose liposomes (TL) comprising l-α-dimyristoylphosphatidylcholine (DMPC) and α-D-glucopyranosyl-α-D-glucopyranoside monomyritate (TreC14) were investigated on breast cancer MDA-MB-453 cells in vitro and in vivo. The IC50 values of TL for MDA-MB-453 cells were remarkably lower than those of DMPC liposomes. The inhibitory effects of TL on the proliferation of MDA-MB-453 cells mediated via apoptosis induction were observed following their accumulation on MDA-MB-453 cell membranes. The membrane fluidity of MDA-MB-453 cells increased after TL treatment, as evident from a fluorescence depolarization assay. TL induced the apoptosis of MDA-MB-453 cells through caspase activation and mitochondrial membrane potential reduction, and suppressed the nuclear factor kappa B activity. A remarkable reduction in tumor volume was observed in a human breast cancer mouse model topically treated with TL. Induction of apoptosis was evident in TL-treated breast cancer tumors of mice using the TUNEL assay.
Subject(s)
Antineoplastic Agents/therapeutic use , Breast Neoplasms/drug therapy , Trehalose/therapeutic use , Animals , Antineoplastic Agents/administration & dosage , Antineoplastic Agents/metabolism , Apoptosis/drug effects , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Cell Line, Tumor , Cell Membrane/metabolism , Female , Humans , Liposomes , Mice, Inbred BALB C , Signal Transduction/drug effects , Trehalose/administration & dosage , Trehalose/metabolismABSTRACT
Purpose: Autophagy plays an important role in balancing the inflammatory response to restore homeostasis. The aim of this study was to explore the mechanism by which trehalose suppresses inflammatory cytokines via autophagy activation in primary human corneal epithelial cells (HCECs) exposed to hyperosmotic stress. Methods: An in vitro dry eye model was used in which HCECs were cultured in hyperosmolar medium with the addition of sodium chloride (NaCl). Trehalose was applied in different concentrations. The levels of TNF-α, IL-1ß, IL-6, and IL-8 were detected using RT-qPCR and ELISA. Cell viability assays, immunofluorescent staining of LC3B, and western blots of Beclin1, Atg5, Atg7, LC3B, and P62 were conducted. The key factors in upstream signaling pathways of autophagy activation were measured: P-Akt, Akt, and transcription factor EB (TFEB). Results: Trehalose reduced the proinflammatory mediators TNF-α, IL-1ß, IL-6, and IL-8 in primary HCECs at 450 mOsM. This effect was osmolarity dependent, and a level of 1.0% trehalose showed the most suppression. Trehalose promoted autophagosome formation and autophagic flux, as evidenced by increased production of Beclin1, Atg5, and Atg7, as well as higher LC3B I protein turnover to LC3B II, with decreased protein levels of P62/SQSTM1. The addition of 3-methyladenine blocked autophagy activation and increased the release of proinflammatory cytokines. Trehalose further activated TFEB, with translocation from cytoplasm to the nucleus, but diminished Akt activity. Conclusions: Our findings demonstrate that trehalose, functioning as an autophagy enhancer, suppresses the inflammatory response by promoting autophagic flux via TFEB activation in primary HCECs exposed to hyperosmotic stress, a process that is beneficial to dry eye.
Subject(s)
Autophagy/drug effects , Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/metabolism , Epithelium, Corneal/drug effects , Signal Transduction/drug effects , Trehalose/pharmacology , Adolescent , Adult , Aged , Cells, Cultured , Dose-Response Relationship, Drug , Enzyme-Linked Immunosorbent Assay , Epithelium, Corneal/cytology , Epithelium, Corneal/metabolism , Humans , Inflammation/etiology , Inflammation/metabolism , Inflammation/prevention & control , Interleukin-1beta/metabolism , Interleukin-6/metabolism , Interleukin-8/metabolism , Middle Aged , Osmotic Pressure , Real-Time Polymerase Chain Reaction , Trehalose/administration & dosage , Tumor Necrosis Factor-alpha/metabolism , Young AdultABSTRACT
Glutamine is a major dietary amino acid that is both a fuel and nitrogen donor for healing tissues damaged by chemotherapy and radiation. Evidence supports the benefit of oral (enteral) glutamine to reduce symptoms and improve and/or maintain quality of life of cancer patients. Benefits include not only better nutrition, but also decreased mucosal damage (mucositis, stomatitis, pharyngitis, esophagitis, and enteritis). Glutamine supplementation in a high protein diet (10 grams/day) + disaccharides, such as sucrose and/or trehalose, is a combination that increases glutamine uptake by mucosal cells. This increased topical effect can reduce painful mucosal symptoms and ulceration associated with chemotherapy and radiation in the head and neck region, esophagus, stomach and small intestine. Topical and oral glutamine seem to be the preferred routes for this amino acid to promote mucosal healing during and after cancer treatment.
Subject(s)
Antineoplastic Agents/adverse effects , Dietary Supplements , Glutamine/administration & dosage , Mucositis/etiology , Mucositis/therapy , Neoplasms/drug therapy , Neoplasms/radiotherapy , Radiotherapy/adverse effects , Diet, High-Protein , Dietary Proteins/administration & dosage , Dietary Sucrose/administration & dosage , Glutamine/metabolism , Glutamine/pharmacology , Humans , Malnutrition/etiology , Mucositis/physiopathology , Mucositis/prevention & control , Mucous Membrane/metabolism , Trehalose/administration & dosage , Wound Healing/drug effectsABSTRACT
Autophagy is a critical process in early mammalian embryogenesis. Mammalian target of rapamycin (mTOR) inhibitors are major regulators of autophagy. However, mTOR plays a vital role in major signaling pathways controlling cell growth and metabolism; thus, more secure autophagy activation methods should be considered. The present study investigated the effects of supplementary trehalose, a novel mTOR-independent autophagy enhancer, on oocyte maturation and embryonic development after parthenogenetic activation (PA). Trehalose treatment during in vitro maturation (IVM) did not affect the nuclear maturation rates of oocytes. Oocytes treated with 25â¯mM trehalose during IVM had a significantly higher (Pâ¯<â¯0.05) blastocyst formation rate (64.2%) after PA compared to that in control oocytes (52.0%). Blastocyst quality was also improved in the trehalose-treated group. The total cell numbers for blastocyst formation and expanded blastocyst formation were significantly increased in the trehalose-treated group (52.2% and 27.7%, respectively) compared to those in the control group (36.9% and 11.0%, respectively). Trehalose treatment led to the increased expression of LC3, an autophagy marker, in metaphase II oocytes and 4-cell stage embryos. Gene expression analysis revealed that the expression of several autophagy related genes (LAMP2, pATG5, and LC3) increased, while the Bax/Bcl2 ratio and pro-apoptotic Bak transcript levels were decreased in the trehalose-treated group. In conclusion, these results indicate that treatment with trehalose during IVM improved the developmental potential of porcine embryos by down-regulation of pro-apoptotic genes and up-regulation of autophagy-related genes and marker. Trehalose may be useful for the large-scale production of high-quality porcine blastocysts in vitro.
Subject(s)
In Vitro Oocyte Maturation Techniques/veterinary , Oocytes/drug effects , Parthenogenesis , Swine , Trehalose/pharmacology , Animals , Blastocyst/drug effects , Blastocyst/metabolism , Cumulus Cells/drug effects , Cumulus Cells/metabolism , Dose-Response Relationship, Drug , Embryo Culture Techniques/veterinary , Embryonic Development/drug effects , Embryonic Development/physiology , Gene Expression Regulation, Developmental/drug effects , Oocytes/physiology , Trehalose/administration & dosageABSTRACT
This study evaluated the postprandial glycemic response and physical properties of the high-amylose rice, Koshinokaori (KK), cooked under different conditions. Twelve healthy subjects (Japanese, 6 males, 6 females) were given cooked, white KK rice or tomato chicken rice (TCR) using KK rice. The Japanese standard rice, Koshihikari (KH), was used as reference. All meals contained the same amount (50 g) of available carbohydrate. Blood glucose levels were measured at 0 (fasting), 15, 30, 45, 60, 90, and 120 min after each meal. The results from the cooked, white KK rice showed a significant difference in blood glucose variation at 60, 90, and 120 min and the incremental area under the curve (IAUC) of blood glucose concentration for KK cooked at optimal water to rice ratio was observed. Blood glucose variation and IAUC after intake of TCR-KK rice was lower than that after TCR-KH rice intake. Addition of 5% trehalose to KK rice resulted in a smaller decrease in adhesiveness and stickiness of cooked rice after 180 min at 20ºC. The addition of 5% trehalose to KK rice also produced favorable results in the sensory evaluation. KK rice produces favourable postprandial glycemic responses and physical properties under varied cooking condition and thus, may be beneficial in the prevention of lifestyle-related diseases such as type 2 diabetes.
Subject(s)
Amylose/pharmacology , Cooking/methods , Glycemic Index/drug effects , Oryza/chemistry , Postprandial Period/drug effects , Adult , Area Under Curve , Blood Glucose/drug effects , Diabetes Mellitus, Type 2/prevention & control , Female , Healthy Volunteers , Humans , Male , Trehalose/administration & dosageABSTRACT
Freeze-dried immunoglobulin G (IgG) incorporating trehalose and human serum albumin (HSA) was statistically evaluated regarding the existence of synergism between additives on the stability profile. The levels of HSA (X1) and trehalose (X2) were independent variables. Aggregation following the process (Y1), after 2 and 3 months at 40°C (Y2) and (Y3), respectively, along with the rate constant of aggregation (Y4) were dependent variables. Aggregation and beta-sheet conformation were quantified through size-exclusion chromatography (SEC-HPLC) and Fourier transform infrared spectroscopy (FTIR). Central composite design (CCD) suggested the best formulation. The integrity and thermodynamic stability of optimized formulation were investigated through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and differential scanning calorimetry (DSC). The calculated responses were Y1, 0-0.90%; Y2, 0.4-4.3%; Y3, 2.10-13.46%; and Y4, 0.16-0.69 1/month. The optimized formulation had 10 mg IgG, 86 mg trehalose, and 1 mg HSA with observed responses of Y1, 0.01%; Y2, 0.51%; Y3, 3.08%; and Y4, 0.33 1/month. The models were statistically well-fitted. The optimized formulation was amorphous during freeze-drying (FD), and no fragmentation was observed. Trehalose and HSA demonstrated statistical synergism. CCD was successfully employed to recommend the best ratio of stabilizers and achieve the maximum stabilization of IgG as a model freeze-dried antibody.
Subject(s)
Drug Design , Immunoglobulin G/chemistry , Serum Albumin, Human/chemical synthesis , Trehalose/chemical synthesis , Calorimetry, Differential Scanning/methods , Drug Combinations , Drug Stability , Freeze Drying/methods , Humans , Immunoglobulin G/administration & dosage , Serum Albumin, Human/administration & dosage , Spectroscopy, Fourier Transform Infrared/methods , Trehalose/administration & dosageABSTRACT
This work demonstrates that an HSV-2 candidate vaccine can be thermostabilized by spray drying to reduce cold chain demands. This work is also to optimize the process responses by varying spray dry parameters for pre-screened suitable excipients; and to determine the validity of current prescreening techniques. Vaccine activity losses were measured by in vitro plaque forming assay with Vero cell line. An accelerated storage condition of 45⯰C for 10â¯days was used to determine spray dried sample stability. Prescreening studies demonstrated that trehalose and sucrose were superior to other tested excipients spray dry thermal stabilization of HSV-2. Subsequent optimization by design of experiments (DOE) of activity responses to spray dry parameter changes demonstrated significant differences between trehalose and sucrose for stability of the viral vaccine. Model parameters included the drying conditions inlet temperature, spray gas flow rate, and solids concentration for the model responses of vaccine stabilization. Trehalose was an effective and robust stabilizing excipient for spray drying HSV-2 vaccine. In contrast, stabilization by sucrose was greatly dependent on the spray dry process parameters. These DOE differences indicated inadequate excipient selection by prescreening methods and the variability demonstrated current prescreening techniques may not be adequate for determining optimal excipients.
Subject(s)
Herpesvirus 2, Human/immunology , Viral Vaccines/administration & dosage , Animals , Chlorocebus aethiops , Desiccation , Drug Compounding/methods , Drug Stability , Excipients/administration & dosage , Sucrose/administration & dosage , Trehalose/administration & dosage , Vero CellsABSTRACT
L-HBsAg is a third-generation hepatitis vaccine capable of inducing antibodies in non-responders and thus providing potentially therapeutic treatment. In this study, L-HBsAg was administered using microneedles (MN) without an adjuvant to induce intradermal (ID) immunization, and the efficacy of ID immunization was compared with that of intramuscular (IM) immunization that uses a conventional formulation with an adjuvant of aluminum hydroxide (L-HBsAg-AL-IM). The L-HBsAg was dip-coated onto 800-µm-long microneedles made of polylactic acid (PLA). Delivery efficiency and administration time were determined through in vitro experiments using porcine skin. The denaturation of the formulation against sterilization by gamma rays was observed. A storage test and a freeze-thaw cycle test of the microneedles with trehalose as a stabilizer (L-HBsAg-MN-Tre) were observed. An antibody titer of L-HBsAg-MN-Tre was compared with that of the conventional IM immunization of the L-HBsAg solution with aluminum hydroxide (L-HBsAg-AL-IM). The formulation containing L-HBsAg was located on the upper third of the microneedle tips. The formulation on the MN was dissolved and delivered within 30â¯min of insertion into porcine skin in vitro. Trehalose was selected as a stabilizer, and the stabilizing effect increased with the increase of trehalose content in the solidified formulation. L-HBsAg-MN with 15% of trehalose was stable for 7â¯days at 40⯰C and showed increased stability compared to the conventional liquid formulations. L-HBsAg-MN-Tre showed improved stability during the freeze-thaw cycle. The antibody titer of L-HBsAg-MN-Tre at 28â¯days was higher than that of L-HBsAg-AL-IM. ID administration of L-HBsAg-MN-Tre showed better efficacy and improved thermal and freeze thaw stability compared to L-HBsAg-AL-IM. Therefore, L-HBsAg-MN-Tre administration showed the possibility of ID delivery of L-HBsAg without the use of an adjuvant for the efficacy, convenience, and safety of pediatric vaccination.
Subject(s)
Hepatitis B Surface Antigens/administration & dosage , Hepatitis B Surface Antigens/immunology , Hepatitis B Vaccines/administration & dosage , Hepatitis B Vaccines/immunology , Immunization/methods , Skin/immunology , Vaccination/methods , Adjuvants, Immunologic/administration & dosage , Aluminum Hydroxide/administration & dosage , Animals , Female , Hepatitis B Antibodies/immunology , Injections, Intradermal/methods , Injections, Intramuscular/methods , Mice, Inbred BALB C , Needles , Trehalose/administration & dosageABSTRACT
A commercial drinkable yogurt with and without 4% of added trehalose (as cell protectant) was spray-dried obtaining a powder with low water activity (aw). Total bacterial count in the powder was between 8.48-8.90 log cfu/g. The dried yogurt was stored: (i) at 38 °C and aw = 0.33; (ii) at 38 °C in hermetically sealed flasks (aw = 0.21/0.22); (iii) in a cyclic temperature chamber (10-20 °C) in hermetically sealed flasks (aw = 0.21/0.22). Whole milk was then fermented by adding an inoculum of spray-dried yogurt after storage under these different conditions. The kinetics of acidification showed the presence of a lag time which was strongly dependent on storage conditions. The data was fitted with a logistic type equation from which the lag time was calculated. To evaluate structural differences among samples, Fourier Transform Infrared spectra (FTIR) were recorded. Partial Least Squares (PLS) models enabled a good correlation between lag time of fermentation and FTIR spectra. The lag time for yogurt powder stored at aw about 0.21/0.22 and cyclic temperature 10-20 °C remained approximately constant over the 12 weeks of storage, while all the other conditions resulted in a dramatic increase. The addition of trehalose had a small influence on lag time and, therefore, as a protectant of lactobacilli.