Levels Of Galactose Research Articles

Innovative modified pectins enriched with galactose: Preparation, characterization, and gel-based properties of curcumin-delivered hydrogel beads

Gelling and thickening are the most important properties of pectin, and these properties are dependent on the molecule's structure. In this study, new types of pectin with varying degrees of modification were prepared by reducing the methyl ester groups on the galacturonic acid residues in pectin to hydroxyl groups using NaBH4, thereby converting methylated galacturonic acid to galactose. The physicochemical properties, rheological behavior, and gelling properties of the modified pectins were investigated. Galacturonic acid levels gradually decreased from 68.98% ± 0.40%–7.49% ± 0.09%, whereas galactose levels increased from 10.29% ± 0.48%–72.26% ± 0.62% with increasing degree of modification. The molecular weight exceeded 500 kDa, and solubility was high. The novel modified pectins underwent gelation without requiring sucrose addition and were calcium and pH sensitive. A higher degree of modification promoted gelation at higher temperatures. Addition of Ca2+ at 60 mg/g caused R-55 to gel at 37 °C and pH 2 but disintegrate at pH 6. Three types of R-55 hydrogel beads encapsulating curcumin were prepared. These beads effectively protected curcumin in the stomach; two allowed for its rapid and slow release in the small intestine, whereas one allowed for its release in the colon. Thus, R-55 can be used as a carrier for the targeted delivery of drugs to the small intestine and colon. The modified pectins are suitable for innovative applications in the food and pharmaceutical fields.

Determining Carbohydrates for Increasing Safety: GC-FID Quantification of Lactose, Galactose, Glucose, Tagatose and Myo-Inositol in ‘Maturo’ PDO Pecorino Sardo Cheese

Although PDO Pecorino Sardo is one of the oldest traditional cheeses of Sardinia, Italy, data on its nutritional properties and food safety are lacking. In particular, significant amounts of lactose and galactose may be a health concern for consumers. The primary objective of this study is to quantify, using a validated GC-FID method, the residual lactose and galactose content in “maturo” (i.e., ripened for at least two months) Protected Denomination of Origin (PDO) Pecorino Sardo cheese. A statistically representative sampling from seven dairies distributed throughout Sardinia has been selected for this aim. In addition to lactose and galactose, two of their metabolites (i.e., glucose and tagatose, respectively) and a bioactive polyol like myo-inositol were quantified. The concentration of lactose (mean 26 mg kg−1, range 4–90 mg kg−1) was below the strictest limit set in the European Union (i.e., 100 mg kg−1), while the galactose content was found to be in an amount (mean: 76 mg kg−1, range: 10–200 mg kg−1) that even patients afflicted with severe galactosemia, albeit with some circumspection, could consume this cheese. Ripening (two to four months) had no significant effect on the amount of all analytes, while a slight decrease in galactose levels was observed during the manufacturing season. Finally, the amounts of glucose, tagatose, and myo-inositol are constant in the range of a few tens of mg kg−1.

Hydrogen sulfide alleviates pericarp browning in lichi fruit by modulating energy and sugar metabolisms.

Postharvest litchi is susceptible to browning that limits the development of litchi industry. Hydrogen sulfide (H2S) is an important bioactive molecule that can regulate many physiological processes. This study examined the effects of exogenous H2S on pericarp browning and related physiological mechanisms in postharvest litchi. The results exhibited that exogenous H2S treatment delayed the browning of litchi pericarp and reduced the damage to cell membrane integrity during storage. This treatment inhibited the energy losses of litchi fruit by increasing the activities of H+-ATPase, Ca2+- ATPase, cytochrome C oxidase (CCO) and succinate dehydrogenase (SDH) and regulating the expression of energy metabolism-related genes, including LcAtpB, LcSnRK2, LcAAC1, LcAOX1 and LcUCP1. In addition, H2S treatment increased the levels of fructose, glucose, sucrose, inositol, galactose and sorbose in litchi fruit, and promoted sucrose synthesis by regulating the activities of sucrose phosphate synthase (SPS), sucrose synthase (SS), acid invertase (AI) and neutral invertase (NI). Based on the current findings, we suggest that exogenous H2S enhances the energy supply and antioxidant activity of litchi by modulating energy and sugar metabolism, thereby inhibiting fruit browning and senescence. These results indicated that H2S treatment is an effective approach to maintaining the quality of litchi fruit and extending its shelf life.

Safety, Pharmacokinetics, and Pharmacodynamics of the New Aldose Reductase Inhibitor Govorestat (AT-007) After a Single and Multiple Doses in Participants in a Phase 1/2 Study.

In classic galactosemia (CG) patients, aldose reductase (AR) converts galactose to galactitol. In a phase 1/2, placebo-controlled study (NCT04117711), safety, pharmacokinetics (PK), and pharmacodynamics (PD) of govorestat were evaluated after single and multiple ascending doses (0.5-40 mg/kg) in healthy adults (n = 81) and CG patients (n = 14). Levels of govorestat in plasma and cerebrospinal fluid (CSF) and blood levels of galactitol, galactose, and galactose-1-phosphate (Gal-1p) were measured for population PK and PK/PD analyses. Govorestat was well tolerated. Adverse event frequency was comparable between placebo and govorestat. Govorestat PK displayed a 2-compartment model with sequential zero- and first-order absorption, and no effect of demographic factors. Multiple-dose PK of govorestat was linear in the 0.5-40 mg/kg range, and CSF levels increased dose dependently. Elimination half-life was ∼10 h. PK/PD modeling supported once-daily dosing. Change from baseline in galactitol was -15% ± 9% with placebo and -19% ± 10%, -46% ± 4%, and -51% ± 5% with govorestat 5, 20, and 40 mg/kg, respectively, thus was similar for 20 and 40 mg/kg. Govorestat did not affect galactose or Gal-1p levels. In conclusion, govorestat displayed a favorable safety, PK, and PD profile in humans, and reduced galactitol levels in the same magnitude (∼50%) as in a rat model of CG that demonstrated an efficacy benefit on neurological, behavioral, and ocular outcomes.

Age-specific composition of milk microbiota in Tibetan sheep and goats

This study investigates the dynamic changes in milk nutritional composition and microbial communities in Tibetan sheep and goats during the first 56 days of lactation. Milk samples were systematically collected at five time points (D0, D7, D14, D28, D56) post-delivery. In Tibetan sheep, milk fat, protein, and casein contents were highest on D0, gradually decreased, and stabilized after D14, while lactose and galactose levels showed the opposite trend. Goat milk exhibited similar initial peaks, with significant changes particularly between D0, D7, D14, and D56. 16S rRNA gene sequencing revealed increasing microbial diversity in both species over the lactation period. Principal coordinates analysis identified distinct microbial clusters corresponding to early (D0–D7), transitional (D14–D28), and mature (D56) stages. Core phyla, including Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria, dominated the milk microbiota, with significant temporal shifts. Core microbes like Lactobacillus, Leuconostoc, and Streptococcus were common in both species, with species-specific taxa observed (e.g., Pediococcus in sheep, Shewanella in goats). Furthermore, we observed a highly shared core microbiota in sheep and goat milk, including Lactobacillus, Leuconostoc, and Streptococcus. Spearman correlation analysis highlighted significant relationships between specific microbial genera and milk nutrients. For instance, Lactobacillus positively correlated with total solids, non-fat milk solids, protein, and casein, while Mannheimia negatively correlated with protein content. This study underscores the complex interplay between milk composition and microbial dynamics in Tibetan sheep and goats, informing strategies for livestock management and nutritional enhancement.Key points• The milk can be classified into three types based on the microbiota composition• The changes of milk microbiota are closely related to the variations in nutrition• Filter out microbiota with species specificity and age specificity in the milk

Suaeda maritima polysaccharides attenuate LPS-induced inflammation of RAW264.7 cells and antioxidative activity

Our research focused on extracting polysaccharides from Suaeda maritima (SMP) to obtain crude polysaccharides (SMP-C), which were subsequently purified into SMP-F1 and SMP-F2. SMPs were evaluated for anti-inflammatory effects and SMP-F1 showed the highest inhibitory effects on nitric oxide (NO) production. The monosaccharide composition analysis of SMP-F1 (molecular weight of 112.2 × 103 g/mol) revealed predominant levels of glucose (45.4 %), arabinose (20.5 %), mannose (14.2 %), and galactose (12.7 %). The primary backbone of SMP-F1 consisted of (1 → 4)-D-glucopyranoside, (1 → 4,6)-D-glucopyranoside, (1 → 3)-D-mannopyranoside, (1 → 3,6)-D-mannopyranoside, and (1 → 5)-L-arabifuranoside. In addition, we hydrolysed SMP-F1 to SMP-H1, SMP-H2, and SMP-H3 and investigated their anti-inflammatory effects on RAW264.7 macrophages. Following SMP-F1 hydrolysis, SMP-H3 (molecular weight of 25.8 × 103 g/mol) exhibited superior anti-inflammatory properties compared to SMP-H1 and SMP-H2, demonstrating a significant decrease in NO production. SMP-H3 also demonstrated a remarkable reduction in the secretion of inflammatory mediators including NO, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2), and pro-inflammatory cytokines including tumour necrosis factor-alpha (TNF-α), interleukin (IL-1β and IL-6), while increasing IL-10 expression. Furthermore, SMP-H3 significantly inhibited LPS-stimulated cluster of differentiation (CD) 11b and CD40 expression. Our subsequent investigation unveiled the involvement of SMP-H3-activated macrophages in the nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) pathways. Additionally, SMP-H3 exhibited antioxidant activity by scavenging 2,2-diphenyl-1-picrylhydrazyl (DPPH), superoxide, and 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) free radicals. These findings suggest the potential of SMP-H3 as an ingredient in the development of alternative drugs or functional foods.

Pollinator Diversity and Phenological Interplay: Exploring Mineral, Hormonal, Sugar, and Vitamin Contents in Vitis vinifera L. cv Bozcaada Çavuşu.

Unraveling the intricate physiological and biochemical intricacies associated with female dominance in grape berries across diverse developmental stages is imperative for optimizing grape production and ensuring the attainment of high-quality yields. This study conducted a thorough analysis of grape berries across phenological stages (BBCH-79, BBCH-81, BBCH-89) and cultivars. At BBCH-89, Bozcaada Çavuşu*Vasilâki demonstrated the highest berry weight and total soluble solids (TSS) levels, emphasizing its enological potential. Acidity peaked at BBCH-79 (28.16) and declined at BBCH-89 (6.11), signaling a shift towards lower acidity in later stages. Bozcaada Çavuşu*Vasilâki consistently showed the highest maturity index (MI). Mineral content variations were observed across nitrogen (N), calcium (Ca), potassium (K), phosphorus (P), magnesium (Mg), sulfur (S), iron (Fe), manganese (Mn), boron (B), zinc (Zn), and copper (Cu), with Bozcaada Çavuşu*Vasilâki often having the highest concentrations, particularly in potassium, calcium, and boron. Hormonal analysis revealed a significant surge in concentrations at BBCH-89, with Bozcaada Çavuşu*Vasilâki standing out. Notably, Indole-3-acetic acid (IAA) concentrations increased by 106%, and abscisic acid (ABA) levels peaked at BBCH-79 with a 38% increase in Bozcaada Çavuşu*Kuntra. Sugar content analysis showed variations in fructose, glucose, sucrose, rhamnose, xylose, galactose, and arabinose levels across sampling times and cultivars. Bozcaada Çavuşu*Vasilâki consistently exhibited higher sugar levels, especially at BBCH-81 and BBCH-89. Vitamin concentrations varied temporally and among cultivars, with BBCH-89 displaying the highest vitamin A concentration (6.24 mg/100 g FW), and Bozcaada Çavuşu*Vasilâki often exhibiting maximum values for vitamin B1, B2, B6, and C. Further research and targeted cultivation practices focusing on the unique attributes of Bozcaada Çavuşu*Vasilâki could enhance grape production efficiency, emphasizing its potential contribution to achieving consistently high-quality yields across various phenological stages.

Melatonin and resveratrol alleviate molecular and metabolic toxicity induced by Bisphenol A in endometrial organoids

Bisphenol A (BPA), a widespread environmental contaminant, poses concerns due to its disruptive effects on physiological functions of the uterine endometrium. In contrast, melatonin (MT) and Resveratrol (RSV) are under scrutiny for their potential protective roles against BPA-induced damage. For the efficacy and ethical concerns in the animal test, endometrial organoids, three-dimensional models mimicking endometrium, serve as crucial tools for unraveling the impact of environmental factors on reproductive health. This study aimed to comprehensively characterize the morphological, molecular and metabolic responses of porcine endometrial organoids to BPA and assess the potential protective effects of MT and RSV. Porcine uteri were prepared, digested with collagenase, mixed with Matrigel, and incubated at 38°C with 5 % CO2. Passaging involved dissociation through trypsin-EDTA treatment and subculturing. The culture medium was refreshed every 2–3 days. To investigate the environmental impact on reproductive health, endometrial organoids were treated with BPA (0.5 µM), MT (with/without BPA at 0.1 µM), and/or RSV (10 µM). Various molecular screening using gene expression, western blotting, immunofluorescence staining, and metabolites profiling were assessed the effects of BPA, MT, and RSV in terms of cell viability, morphology, reproductivity, and metabolism alteration in the endometrial organoids. As expected, BPA induced structural and molecular disruptions in organoids, affecting cytoskeletal proteins, Wnt/β-catenin signaling, and epithelial/mesenchymal markers. It triggered oxidative stress and apoptotic pathways, altered miRNA expression, and disrupted the endocannabinoid system. The level of glucose, galactose, and essential amino acids were increased or decreased by approximately 1.5–3 times in BPA-treated groups compared to the control groups (p-value < 0.05), indicating metabolic changes. Moreover, MT and RSV treated groups exhibited protective effects, mitigating BPA-induced disruptions across multiple pathways. For the first time, our study models endometrial organoids, advancing understanding of environmental impacts on reproductive health.

Dynamic patterns of carbohydrate metabolism genes in bacterioplankton during marine algal blooms

Carbohydrates play a pivotal role in nutrient recycling and regulation of algal–bacterial interactions. Despite their ecological significance, the intricate molecular mechanisms governing regulation of phycosphere carbohydrates by bacterial taxa linked with natural algal bloom have yet to be fully elucidated. Here, a comprehensive temporal metagenomic analysis was conducted to explore the carbohydrate-active enzyme (CAZyme) genes in two discrete algal bloom microorganisms (Gymnodinium catenatum and Phaeocystis globosa) across three distinct bloom stages: pre-bloom, peak bloom, and post-bloom. Elevated levels of extracellular carbohydrates, primarily rhamnose, galactose, glucose, and arabinose, were observed during the initial and post-peak stages. The prominent CAZyme families identified—glycoside hydrolases (GH) and carbohydrate-binding modules (CBMs)—were present in both algal bloom occurrences. In the G. catenatum bloom, GH23/24 and CBM13/14 were prevalent during the pre-bloom and peak bloom stages, whereas GH2/3/30 and CBM12/24 exhibited increased prevalence during the post-bloom phase. In contrast, the P. globosa bloom had a dominance of GH13/23 and CBM19 in the initial phase, and this was succeeded by GH3/19/24/30 and CBM54 in the later stages. This gene pool variation—observed distinctly in specific genera—highlighted the dynamic structural shifts in functional resources driven by temporal alterations in available substrates. Additionally, ecological linkage analysis underscored a correlation between carbohydrates (or their related genes) and phycospheric bacteria, hinting at a pattern of bottom-up control. These findings contribute to understanding of the dynamic nature of CAZymes, emphasizing the substantial influence of substrate availability on the metabolic capabilities of algal symbiotic bacteria, especially in terms of carbohydrates.

Phenotypic and genetic spectra of galactose mutarotase deficiency: A nationwide survey conducted in Japan

PurposeGalactose mutarotase (GALM) deficiency was first reported in 2019 as the fourth type of galactosemia. This study aimed to investigate the clinical and genotypic spectra of GALM deficiency. MethodsThis was a questionnaire-based retrospective survey conducted in Japan between February 2022 and March 2023. ResultsWe identified 40 patients with GALM deficiency in Japan (estimated prevalence: 1:181,835). Four of 38 patients (10.5%) developed cataracts, which resolved with lactose restriction in 3 out of 4 patients. Transient transaminitis was the most common symptom (23.1%). All of the patients followed lactose restriction; discontinuation of the restriction after infancy did not cause any complications. Moreover, none of the participants experienced long-term complications. Two variants, GALM NM_138801.3: c.294del and c.424G>A, accounted for 72.5% of the identified pathogenic variants. The patients showed moderately elevated blood galactose levels with lactose intake; however, the elevation was lower than that observed in galactokinase deficiency. ConclusionGALM deficiency is characterized by a similar but milder phenotype and lower blood galactose elevation than in galactokinase deficiency. Diagnosis and initiation of lactose restriction in early infancy should be essential for prevention of cataracts, especially in cases of irreversible opacity.

GALDAR: A genetically encoded galactose sensor for visualizing sugar metabolism in vivo.

Sugar metabolism plays a pivotal role in sustaining life. Its dynamics within organisms is less understood compared to its intracellular metabolism. Galactose, a hexose stereoisomer of glucose, is a monosaccharide transported via the same transporters with glucose. Galactose feeds into glycolysis and regulates protein glycosylation. Defects in galactose metabolism are lethal for animals. Here, by transgenically implementing the yeast galactose sensing system into Drosophila, we developed a genetically encoded sensor, GALDAR, which detects galactose in vivo. Using this heterologous system, we revealed dynamics of galactose metabolism in various tissues. Notably, we discovered that intestinal stem cells do not uptake detectable levels of galactose or glucose. GALDAR elucidates the role for galactokinase in metabolism of galactose and a transition of galactose metabolism during the larval period. This work provides a new system that enables analyses of in vivo sugar metabolism.

Development of a New Amperometric Biosensor for Measurement of Plasma Galactose Levels.

Galactosemia is an inherited disease that occurs as a result of insufficient or no synthesis of some enzymes (GALT, GALK, and GALE) in galactose metabolism. Failure to make an early diagnosis, especially in newborns, can lead to severe clinical and even fatal consequences. The aim of this study is to develop a biosensor for measuring free galactose in plasma. The immobilization components of the developed free galactose biosensor are screen printed carbon electrode (SCPE), Prussian blue (PB), chitosan (CHIT), Nafion (NAF), gold nanoparticle (GNP), and galactose oxidase (GaOX). The CHIT/GaOX/NAF-GNP/GaOX/CHIT-GNP/SCPE-PB electrode showed a sensitive amperometric response to detect galactose. While the surface characterization of the biosensor was performed with cyclic voltammetry and scanning electron microscopy, the optimization and performance characterizations were made by applying an amperometry technique. The amperometric operating potential for the free galactose biosensor was determined as -0.05 V. The linear detection range for the free galactose biosensor is between 0.025 and 10 mM. This range includes galactose levels in plasma of both healthy and patients. The percent coefficient of variation values calculated for intraday and interday repeatability of the developed biosensor are below 10%. The practical use of the biosensor, for which optimization and characterization studies were carried out, was tested in 10 healthy 11 patients with galactosemia, and the results were compared with the colorimetric method. In conclusion, the unique analytical properties and effortless preparation of the new galactose biosensor developed in this study make them serious candidates for point-of-care diagnostic testing.

Metabolomics and Physiological Changes Underlying Increased Tolerance to Salt Stress Induced by Applied Nitric Oxide in Fatsia japonica Seedlings

Fatsia japonica (Thunb.) Decne. et Planch. is an important woody landscape plant, and its distribution is commonly limited by salt stress. Although the application of exogenous nitric oxide (NO) has been known to be effective in alleviating abiotic stress in plants, the underlying mechanism by which NO induces salt resistance in F. japonica remains unknown. In this study, the physiological and metabolic characteristics of F. japonica seedlings with the application of NO under salt stress conditions were investigated. We demonstrated that exogenous NO (0.1 mM sodium nitroprusside, SNP) mitigated the growth inhibition caused by 0.4% NaCl. This alleviation could be attributed to NO-induced enhancement in photosynthesis, osmotic adjustment, antioxidant enzyme activities, and a reduction in oxidative damage when exposed to salt stress. Furthermore, the metabolomic analysis revealed that salt stress significantly disrupts the growth of F. japonica by downregulating sugars, sugar alcohols, amino acids, and organic acids. However, the application of exogenous NO improves sugar metabolism, enhancing the levels of fructose, glucose, mannose, galactose, xylose, ribose, inositol, and sorbitol, as well as the metabolism of amino acids and organic acids. These findings provide new insights into the physiological and metabolic homeostasis adjustments induced by NO that promote salt stress tolerance in F. japonica, enhancing our understanding of plant resilience mechanisms.

Identification of a novel biomarker for sarcopenia diagnosis using serum metabolomic analysis: a pilot study.

This pilot study compared serum metabolites in participants with and without sarcopenia. Metabolomic techniques were applied to identify serum metabolites and novel biomarkers specific to patients with sarcopenia. In accordance with AWGS2019 criteria, sarcopenia was defined as low muscle mass plus either low muscle strength/low physical function, and severe sarcopenia was defined as low muscle mass, low muscle strength, and low physical function all together. The sarcopenia group had higher hypoxanthine, galactose, and mannose levels but lower triethanolamine and homogentisic acid levels than the non-sarcopenia group. The severe sarcopenia group had lower levels of alpha-tocopherol than the mild and moderate sarcopenia groups. This study is the first to identify hypoxanthine as a potential biomarker for sarcopenia in humans and provides new insights into the pathophysiology of sarcopenia. Furthermore, the identified metabolites may be useful for the early detection of sarcopenia.

Twelve-year review of galactosemia newborn screening in Taiwan: Evolving methods and insights

BackgroundGalactosemia was introduced into Taiwan's routine newborn screening (NBS) program in 1985. This study presents a 12-year experience, emphasizing disease diagnosis and screening performance. MethodNBS for galactosemia utilized dried blood spot samples taken 48–72 h post-delivery, with total galactose (TGal) level as the primary marker. Newborns with critical TGal levels were referred immediately, while those with borderline TGal underwent a recall test. GALT activity measurement was applied simultaneously as the second-tier marker. Further confirmatory tests, such as whole exome sequencing (WES), were conducted upon referral. ResultsFrom January 1st, 2011, to December 31st, 2022, 51 cases were identified from 817,906 newborns. Of these, nine individuals had persistently elevated TGal. Diagnoses included one case of GALT deficiency, one of GALM deficiency, and seven of GALE deficiencies. Notably, the classic galactosemia patient (GALT deficiency) presented with extreme high TGal and was referred to the hospital for diet management immediately. All affected patients were instructed to adopt a galactose-restricted diet. By the median age of 2.5 years, all exhibited normal development and liver function. ConclusionThe incidence of classical galactosemia and its variants is extremely low in Taiwan. Incorporating WES into NBS has improved our ability to detect various galactosemia forms, enriching our understanding of the genetic underpinnings. While these newly discovered forms often present with milder initial elevations in TGal, specific biochemical investigations and regular monitoring are essential to understanding the long-term implications and outcomes.

Molecular characterization of novel and rare DNA variants in patients with galactosemia

Introduction: Galactosemia is an inherited disorder caused by mutations in the three genes that encode enzymes implicated in galactose catabolism. Currently, the only available treatment for galactosemia is life-long dietary restriction of galactose/lactose, and despite treatment, it might result in long-term complications.Methods: Here, we present five cases of newborn patients with elevated galactose levels, identified in the context of the newborn screening program. Genetic analysis concerned a next generation sequencing (NGS) methodology covering the exons and adjacent splice regions of the GALT, GALK1, and GALE genes.Results: Our approach led to the identification of eight rare nonsynonymous DNA variants. Four of these variants, namely, p.Arg204Gln and p.Met298Ile in GALT, p.Arg68Leu in GALK1, and p.Ala180Thr in GALE, were already recorded in relevant databases, yet their clinical significance is uncertain. The other four variants, namely, p.Phe245Leu in GALT, p.Gly193Glu in GALK1, and p.Ile266Leu and p.Ala216Thr in the GALE gene, were novel. In silico analysis of the possible effect of these variants in terms of protein function and stability was performed using a series of bioinformatics tools, followed by visualization of the substituted amino acids within the protein molecule. The analysis revealed a deleterious and/or destabilizing effect for all the variants, supported by multiple tools in each case.Discussion: These results, given the extreme rarity of the variants and the specific phenotype of the respective cases, support a pathogenic effect for each individual variant. Altogether, our study shows that targeted NGS methodologies may offer a time- and cost-effective approach for the genetic investigation of galactosemia and can assist in elucidating the complex genetic background of this disorder.

An oxygen-insensitive amperometric galactose biosensor based on galactose oxidase co-immobilized with an Os-complex modified redox polymer

Monitoring galactose levels in dairy products can help to prevent severe complications with a hereditary metabolic disease such as galactosemia, a life-threatening disease. The current state of the art requires the development of less expensive, more reliable and specific methods to determine galactose levels in food in a practical way. We report the development and optimization of an amperometric biosensor for determination of galactose in dairy products based on galactose oxidase (GaOx) co-immobilized with an osmium-complex modified redox polymer on glassy carbon electrodes. To attain the maximum catalytic currents based on mediated electron transfer, two Os-complex based polymers with different redox potentials and different enzyme:redox polymer ratios were studied. The optimized GaOx-modified electrode gave a maximum electrocatalytic response of galactose oxidation that was not affected by the presence of O2, indicating fast wiring of the enzyme by the Os-complex modified redox polymer. The biosensor that gave the best analytical parameters for galactose detection was further tested for measuring galactose concentration in lactose-containing and lactose-free milk and yogurt samples under aerobic conditions. The results obtained with the amperometric biosensor were validated by high-performance anion-exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD).

Determination of The GALT Gene in Galactosemic Cataract Patients in Azerbaijan

Galactosemia is one of the genetic diseases that can be treated thanks to early detection by genetic screening test. Galactosemia is a hereditary metabolic disease. This disease is heterogeneous and controlled by GALT, GALK, GALE and GALM genes. The presented article is dedicated to galactosemic disease among Azerbaijani patients as a result of GALT (galactose-1-phosphate uridyltransferase) gene mutations. This study reviews discusses the type of cataracts associated with galactosemia. The genetics of galactosemic patients are evaluated and details of galactose metabolism are described. This study describes the results of the GALT gene analysis of two Azerbaijanian patients with Galactosemic cataract. In one of the patients the heterzygous form of the same mutation, and the homozygous form in the other patient were found. In this study, control and experimental groups were determined, and DNA extracted from the peripheral blood of patients belonging to both groups was subjected to PCR amplification according to standard protocols and sequenced by Sanger sequence method. The symptoms of the disease were reviewed in these patients. Patients presented with jaundice, diarrhea, vomiting, malnutrition, seizures, allergies and more cataracts in the neonatal period. Commonly, Cataract disease is idiopathic origin. Also, patients with cataracts may have an underlying genetic abnormality of galactose metabolism. Elevated levels of galactose and galactose-1-phosphate and absence of GALT activity were diagnosed in the studied patients. Early biochemical and molecular-genetic diagnosis and genetic counseling of patients and their family members are extremely important in the treatment of galactosemia. We selected 25 cataract patients aged 0-45 years and 25 age and sex-matched controls for the study. Blood samples taken from the experimental and control groups, were biochemically and then molecular-genetically analyzed. Mutations Q188R, K285N and N314D were investigated in patients. Of these mutations, only the N314D mutation was detected in two patients. Both heterozygous and homozygous forms of N314D mutation were detected.

Untargeted metabolomics study of dexamethasone-induced congenital cleft palate in New Zealand rabbits

Objective: To investigate the metabolic disorders in placental tissues of dexamethasone induced cleft palate mode. Methods: Twelve pregnant rabbits were randomly divided into dexamethasone group (experimental group, 8) and saline control group (4), and a certain amount of dexamethasone and saline were administered intramuscularly to the experimental and control groups respectively from embryonic days (ED) 13 to 16, and placental tissue samples were collected on day 21 of gestation. The corresponding profiles of the embryonic placental tissue samples were obtained by liquid chromatography-triple tandem quadrupole(LC-MS), and the metabolites of the embryonic placental tissues were characterized by principal component analysis among the dexamethasone-treated group with cleft palate (D-CP group), the dexamethasone-treated group without cleft palate (D-NCP group) and the control group. Results: There were significant metabolic differences among the D-CP group, D-NCP group and control group, with a total of 133 differential metabolites (VIP>1, P<0.05) involving in important metabolic pathways including vitamin B6 metabolism, lysine metabolism, arginine anabolic metabolism, and galactose metabolism. The four metabolites, vitamin B6, galactose, lysine and urea, differed among the three groups (P<0.05). There were significant differences in vitamin B6 (0.960±0.249, 0.856±0.368, 1.319±0.322), galactose (0.888±0.171, 1.033±0.182, 1.127±0.127), lysine (1.551±0.924, 1.789±1.435, 0.541±0.424) and urea (0.743±0.142, 1.137±0.301, 1.171±0.457, respectively) levels among control group, D-NCP group and D-CP group (F=5.90, P=0.008; F=5.59, P=0.009; F=4.26, P=0.025; F=5.29, P=0.012). Conclusions: The results indicated that dexamethasone induced cleft palate may be highly correlated with metabolic disorders including vitamin B6 metabolism, lysine metabolism, arginine anabolic metabolism and galactose metabolism.

High serum mannose in colorectal cancer: a novel biomarker of lymph node metastasis and poor prognosis.

Lymph node status is an important prognostic indicator and it significantly influences treatment decisions for colorectal cancer (CRC). The objective of this study was to evaluate the ability of serum monosaccharides in predicting lymph node metastasis (LNM) and prognosis. High performance anion exchange chromatography coupled with pulsed amperometric detector (HPAEC-PAD) was used to quantify serum monosaccharides from 252 CRC patients. Receiver operating characteristic (ROC) curves were used to evaluate predictive performance of parameters. Predictors of LNM were evaluated by univariate and multivariate analyses. The prognostic role of the factors was evaluated by survival analysis. The levels of serum mannose (Man) and galactose (Gal) were significantly increased in patients with LNM (p <0.0001, p =0.0017, respectively). The area under the curves (AUCs) of Man was 0.8140, which was higher than carcinoembryonic antigen (CEA) (AUC =0.6523). Univariate and multivariate analyses demonstrated histologic grade (G3) (odds ratio [OR] =2.60, p =0.043), histologic grade (mucin-producing subtype) (odds ratio [OR] =3.38, p =0.032), lymphovascular invasion (LVI) (OR =2.42, p <0.01), CEA (>5ng/ml) (OR =1.85, p =0.042) and high Man (OR =2.65, p =0.006) to be independent risk factors of LNM. The survival analysis showed that the high serum Man was independent risk factor for poor prognosis in CRC patients (HR=1.75, p =0.004). The Man is superior to CEA in prediction of LNM for CRC patients. Man is expected to be a predictor for LNM in CRC. High serum Man is associated with poor prognosis of CRC patients.