RESUMO
We recently established a method for the isolation of serum-free oligosaccharides, and characterized various features of their structures. However, the precise mechanism for how these glycans are formed still remains unclarified. To further investigate the mechanism responsible for these serum glycans, here, we utilized rat primary hepatocytes to examine whether they are able to secrete free glycans. Our findings indicated that a diverse array of free oligosaccharides such as sialyl/neutral free N-glycans (FNGs), as well as sialyl lactose/LacNAc-type glycans, were secreted into the culture medium by primary hepatocytes. The structural features of these free glycans in the medium were similar to those isolated from the sera of the same rat. Further evidence suggested that an oligosaccharyltransferase is involved in the release of the serum-free N-glycans. Our results indicate that the liver is indeed secreting various types of free glycans directly into the serum.
Assuntos
Hepatócitos , Oligossacarídeos , Animais , Ratos , Hepatócitos/metabolismo , Oligossacarídeos/sangue , Oligossacarídeos/química , Oligossacarídeos/metabolismo , Células Hep G2 , Humanos , Masculino , Ratos WistarRESUMO
(1) Background: Human milk oligosaccharides (HMOs) are present in maternal serum during pregnancy and their composition is altered in gestational diabetes (GDM). HMOs are also in fetal cord blood and in contact with the feto-placental endothelium, potentially affecting its functions, such as angiogenesis. We hypothesized that cord blood HMOs are changed in GDM and contribute to increased feto-placental angiogenesis, hallmark of GDM. (2) Methods: Using HPLC, we quantified HMOs in cord blood of women with normal glucose tolerance (NGT, n = 25) or GDM (n = 26). We investigated in vitro angiogenesis using primary feto-placental endothelial cells (fpECs) from term placentas after healthy pregnancy (n = 10), in presence or absence of HMOs (100 µg/mL) isolated from human milk, 3'-sialyllactose (3'SL, 30 µg/mL) and lactose (glycan control) and determined network formation (Matrigel assay), proliferation (MTT assays), actin organization (F-actin staining), tube formation (fibrin tube formation assay) and sprouting (spheroid sprouting assay). (3) Results: 3'SL was higher in GDM cord blood. HMOs increased network formation, HMOs and 3'SL increased proliferation and F-actin staining. In fibrin assays, HMOs and 3'SL increased total tube length by 24% and 25% (p < 0.05), in spheroid assays, by 32% (p < 0.05) and 21% (p = 0.056), respectively. Lactose had no effect. (4) Conclusions: Our study suggests a novel role of HMOs in feto-placental angiogenesis and indicates a contribution of HMO composition to altered feto-placental vascularization in GDM.
Assuntos
Indutores da Angiogênese/sangue , Diabetes Gestacional/sangue , Sangue Fetal/química , Oligossacarídeos/sangue , Circulação Placentária/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Células Endoteliais/química , Feminino , Humanos , Lactose/sangue , Leite Humano/química , Placenta/irrigação sanguínea , Placenta/citologia , GravidezRESUMO
Sialyllactose (SL), an acidic oligosaccharide, has immune-protective effects against pathogens and helps with the development of the immune system and intestinal microorganisms. To elucidate the pharmacokinetic characterization after oral administration to rats, the simultaneous quantification method for 3'-SL and 6'-SL in rat plasma was validated, using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in an electrospray ionization (ESI) mode. Several types of columns [C18, amide, and hydrophilic interaction liquid chromatography (HILIC) phase] were used to separate the peaks of 3'-SL and 6'-SL, which improved chromatographic selectivity. Ultimately, the HILIC phase column had a good peak shape and quick resolution, with a mobile phase comprising ammonium acetate buffer and acetonitrile obtained by gradient elution. In addition, the simultaneous quantification of 3'-SL and 6'-SL in rat plasma samples were adequately applied to pharmacokinetic study.
Assuntos
Lactose/análogos & derivados , Oligossacarídeos/sangue , Oligossacarídeos/farmacocinética , Administração Oral , Animais , Configuração de Carboidratos , Cromatografia Líquida , Relação Dose-Resposta a Droga , Lactose/administração & dosagem , Lactose/sangue , Lactose/farmacocinética , Masculino , Oligossacarídeos/administração & dosagem , Ratos , Espectrometria de Massas em TandemRESUMO
OBJECTIVES: The study compared the diagnostic efficiency of serum oligosaccharide chain (G-test) and alpha-fetoprotein (AFP) for hepatitis B-related hepatocellular carcinoma (HCC). METHODS: Serum samples from 100 patients (divided into five groups of 20 each, namely the hepatitis, liver cirrhosis, liver cancer, health, and interference groups) who were admitted to the Second Affiliated Hospital of Nanchang University from October 2019 to January 2020 were collected, and the levels of G-test and AFP were determined. The sensitivity and specificity of the two indicators were compared, and the receiver operating characteristic curve of the subjects was drawn to evaluate the diagnostic values of G-test and AFP for HCC. RESULTS: The diagnostic ability of G-test (area under the curve [AUC]: 0.88 ± 0.05) was better than that of AFP (AUC: 0.76 ± 0.05). When G-test and AFP were combined for detection, the AUC was larger than that of either indicator. The G-test was superior to AFP in the differential diagnosis of early HCC and cirrhosis. A combination of the two indicators (AUC: 0.769 ± 0.05) significantly improved the diagnostic rate for early HCC, indicating that G-test and AFP complemented each other. CONCLUSION: G-test was better than AFP for screening HCC in patients with chronic hepatitis B and cirrhosis. The combination of the two further improved the diagnostic rate of hepatitis B-related liver cancer. The G-test improves the screening rate of early HCC in patients with cirrhosis. Therefore, these markers are of great clinical significance and can improve the sensitivity of HCC detection and reduce missed diagnosis rates.
Assuntos
Biomarcadores Tumorais/sangue , Carcinoma Hepatocelular/diagnóstico , Vírus da Hepatite B/isolamento & purificação , Hepatite B/complicações , Neoplasias Hepáticas/diagnóstico , Oligossacarídeos/sangue , alfa-Fetoproteínas/análise , Carcinoma Hepatocelular/sangue , Carcinoma Hepatocelular/epidemiologia , Carcinoma Hepatocelular/virologia , Feminino , Hepatite B/virologia , Humanos , Neoplasias Hepáticas/sangue , Neoplasias Hepáticas/epidemiologia , Neoplasias Hepáticas/virologia , Masculino , Pessoa de Meia-Idade , Curva ROC , Estudos RetrospectivosRESUMO
Red raspberries (RRB) are high in anthocyanin- and ellagitannin- type (poly)phenols. This study aimed to investigate the effect of 4-week RRB supplementation on (poly)phenolic metabolism in adults with prediabetes and insulin-resistance (PreDM-IR); and whether adding fructo-oligosaccharides (FOS), prebiotics, would augment the microbial metabolites of RRB (poly)phenols. In a randomized crossover clinical trial, subjects (n = 35: PreDM-IR, n = 25; healthy Reference group, n = 10) consumed 1 cup RRB (fresh weight equivalence) per day and RRB with 8 g FOS per day each for 4 weeks in random order separated by 4-week washout. Plasma and urinary (poly)phenolic metabolites were characterized after (0-24h) consuming a RRB-based test drink (2 cups RRB) at baseline/week 0 and again after 4-week supplementations. A total of 123 (poly)phenolic metabolites were quantified. After 4-week RRB supplementation, several metabolite groups were significantly increased (p < 0.05), including urolithins, phenyl-γ-valerolactones, and phenolic acids. Supplementing FOS with RRB for 4 weeks enhanced benzoic acid derivatives compared to the baseline (p < 0.05). Specific effects of supplementation by metabolic status indicated 4-week RRB supplementation significantly increased microbial metabolites that were lower in PreDM-IR group. Our results suggest alterations in the capacity of PreDM-IR group to metabolize and render bioavailable raspberry-derived (poly)phenols when consumed regularly.
Assuntos
Oligossacarídeos/administração & dosagem , Polifenóis/sangue , Estado Pré-Diabético/dietoterapia , Rubus/química , Adulto , Suplementos Nutricionais/efeitos adversos , Ingestão de Alimentos/efeitos dos fármacos , Ingestão de Alimentos/genética , Feminino , Frutas/química , Humanos , Resistência à Insulina/genética , Masculino , Pessoa de Meia-Idade , Oligossacarídeos/sangue , Fenóis/sangue , Prebióticos/administração & dosagem , Estado Pré-Diabético/sangue , Estado Pré-Diabético/genética , Estado Pré-Diabético/metabolismoRESUMO
This study assessed the effects of galacto-oligosaccharides (Oligomate) on hematocrit, serum enzymes, total bilirubin levels, and serum electrolytes in controls and severely malnourished infants, with emphasis on gastrointestinal symptoms. Oligomate doses and phases did not affect stools frequency per day, indicating that prebiotic effect on stool may be due to the prebiotic type. The number of vomits per day during phases 2 and 3 were significantly reduced (p<0.05) in response to prebiotics, despite the prebiotic dose effect was not significant (p>0.05). Moreover, prebiotics administration during phases 2 and 3 markedly improved hemoglobin levels (p<0.05), but not the dose. Similarly, hematocrit levels and white blood cells were significantly improved during the last 2 phases, but dose have no effects on blood hematocrit levels. Erythrocyte sedimentation rate significantly decreased (p<0.05) in phases 2 and 3 compared to phase 1. No dose-related effect was stated on erythrocytes sedimentation rate. Regarding the serum enzymes, SGPT significantly decreased (p<0.05) in phases 2 and 3 compared to phase 1, whereas SGOT significantly decreased only in phase 3. Total bilirubin levels increased significantly (p<0.05) in phase 3 when compared to phases 1 or 2. Prebiotics significantly decreased (p<0.05) sodium levels in the treated group, while potassium levels did not change in all groups, excepting during phase 2, where it increased significantly. Thus, our results confirm the hypothesis that prebiotic supplementation improves blood parameters and health status, consequently decreasing the infection risk and number of vomit per day in infants.
Assuntos
Desnutrição/sangue , Oligossacarídeos/sangue , Prebióticos , Índice de Gravidade de Doença , Doença Aguda , Alanina Transaminase/sangue , Fosfatase Alcalina/sangue , Aspartato Aminotransferases/sangue , Sedimentação Sanguínea , Criança , Pré-Escolar , Eletrólitos/sangue , Fezes , Humanos , Lactente , Leucócitos/metabolismo , Potássio/sangue , Albumina Sérica/metabolismo , Sódio/sangueRESUMO
BACKGROUND: The use of biomarkers of food intake (BFIs) in blood and urine has shown great promise for assessing dietary intake and complementing traditional dietary assessment tools whose use is prone to misreporting. OBJECTIVE: Untargeted LC-MS metabolomics was applied to identify candidate BFIs for assessing the intake of milk and cheese and to explore the metabolic response to the ingestion of these foods. METHODS: A randomized controlled crossover study was conducted in healthy adults [5 women, 6 men; age: 23.6 ± 5.0 y; BMI (kg/m2): 22.1 ± 1.7]. After a single isocaloric intake of milk (600 mL), cheese (100 g), or soy-based drink (600 mL), serum and urine samples were collected postprandially up to 6 h and after fasting after 24 h. Untargeted metabolomics was conducted using LC-MS. Discriminant metabolites were selected in serum by multivariate statistical analysis, and their mass distribution and postprandial kinetics were compared. RESULTS: Serum metabolites discriminant for cheese intake had a significantly lower mass distribution than metabolites characterizing milk intake (P = 4.1 × 10-4). Candidate BFIs for milk or cheese included saccharides, a hydroxy acid, amino acids, amino acid derivatives, and dipeptides. Two serum oligosaccharides, blood group H disaccharide (BGH) and Lewis A trisaccharide (LeA), specifically reflected milk intake but with high interindividual variability. The 2 oligosaccharides showed related but opposing trends: subjects showing an increase in either oligosaccharide did not show any increase in the other oligosaccharide. This result was confirmed in urine. CONCLUSIONS: New candidate BFIs for milk or cheese could be identified in healthy adults, most of which were related to protein metabolism. The increase in serum of LeA and BGH after cow-milk intake in adults calls for further investigations considering the beneficial health effects on newborns of such oligosaccharides in maternal milk. The trial is registered at clinicaltrials.gov as NCT02705560.
Assuntos
Queijo , Dieta , Leite , Oligossacarídeos/sangue , Oligossacarídeos/metabolismo , Adolescente , Adulto , Animais , Biomarcadores/sangue , Estudos Cross-Over , Feminino , Humanos , Masculino , Oligossacarídeos/química , Adulto JovemRESUMO
: Low molecular weight heparins are used during haemodialysis for thromboprophylaxis of the dialysis circuit, with plasma antifactor-Xa (anti-Xa) activity used as a surrogate measure for effective anticoagulation. However, this pharmacokinetic parameter does not always correlate with pharmacodynamic effects in patients. The aim of this study was to investigate the relationship between actual plasma levels of the low molecular weight heparins enoxaparin, anti-Xa activity, and global coagulation measurement of thrombin generation during haemodialysis. Blood was analysed from 16 adult patients with end-stage kidney disease at 0, 2, 4âh, and at completion of 31 dialysis sessions where single fixed doses of 20 (nâ=â3), 40 (nâ=â16), 60 (nâ=â6), or 80 (nâ=â6)âmg of enoxaparin (equating to 0.23-1.07âmg/kg) were used as thromboprophylaxis. Plasma enoxaparin oligosaccharides [degree of polymerization (dp)6-dp16] were measured by high-performance size exclusion chromatography, anti-Xa activity by colourimetric assay, and thrombin generation by calibrated automated thrombogram. Plasma enoxaparin fragments were undetectable at the beginning of each dialysis, peaked at 2âh to levels that correlated with dose (râ=â0.68, Pâ<â0.001) then remained relatively stable. In contrast, therapeutic anti-Xa levels achieved at 2âh in 18 cases (58%) quickly dropped to only six cases (19%) at the end of dialysis, by which time thrombin generation had also recovered in 81% of patients. Statistical modelling revealed a threshold value of anti-Xa at 0.53âIU/ml that supressed thrombin generation to 15.28% of baseline (Pâ<â0.001). Despite loss of anticoagulant activity in the majority of patients, plasma levels of enoxaparin oligosaccharides remained detectable and relatively unchanged throughout dialysis.
Assuntos
Enoxaparina/sangue , Inibidores do Fator Xa/sangue , Diálise Renal , Trombina/biossíntese , Adulto , Anticoagulantes/sangue , Anticoagulantes/uso terapêutico , Enoxaparina/uso terapêutico , Inibidores do Fator Xa/uso terapêutico , Humanos , Falência Renal Crônica/sangue , Falência Renal Crônica/terapia , Cinética , Pessoa de Meia-Idade , Modelos Estatísticos , Oligossacarídeos/sangueRESUMO
BACKGROUND: Pancreatic ductal adenocarcinoma is a devastating malignancy with an extremely poor prognosis. Although the most widely used biomarker for pancreatic cancer is carbohydrate antigen CA19-9, it is elevated mainly in the late stage of pancreatic cancer. Some serum natural antibodies against carbohydrates have been shown to be possible diagnostic markers for cancer. OBJECTIVE: This study was conducted to determine whether the level of natural antibodies against carbohydrates fluctuates in pancreatic ductal adenocarcinoma. METHODS: Serum from pancreatic cancer subjects (n= 55) and 43 subjects free of malignant disease were studied. The contents of natural antibodies against sialyl glycans and CA19-9 in serum were determined by enzyme-linked immunosorbent assay. RESULTS: The level of serum anti-3'-sialyllactose antibodies in pancreatic cancer subjects was significantly lower than that in healthy controls. In contrast, the amounts of serum antibodies against other sialyl glycans were comparable between the two groups. Concentration of serum anti-3'-sialyllactose IgG provided excellent AUC of 0.86, with sensitivity 82%, specificity 81%, and accuracy 82%. The combination of serum anti-3'-sialyllactose IgG with CA19-9 improved the sensitivity of pancreatic cancer detection at an early stage. CONCLUSIONS: Natural antibodies against 3'-sialyllactose constitute a promising biomarker for pancreatic cancer detection. The measurement of serum anti-3'-sialyllactose antibodies could play a supportive role in diagnostics and complement the performance of CA19-9 for the early detection of pancreatic ductal adenocarcinoma.
Assuntos
Adenocarcinoma/sangue , Antígeno CA-19-9/sangue , Carcinoma Ductal Pancreático/sangue , Detecção Precoce de Câncer , Adenocarcinoma/imunologia , Adulto , Idoso , Anticorpos/sangue , Anticorpos/imunologia , Biomarcadores Tumorais/sangue , Carcinoma Ductal Pancreático/imunologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Oligossacarídeos/sangue , Oligossacarídeos/imunologia , Pâncreas/patologia , PrognósticoRESUMO
Human milk oligosaccharides (HMOs) are present in maternal serum in early gestation, raising the question of whether HMOs can cross the placental barrier and reach fetal circulation. Here, we aimed to detect HMOs in cord blood, and assess HMO composition and concentration in relation to maternal HMOs. In an ex-vivo placental perfusion model, we asked whether HMOs can pass over the placenta. Using HPLC, we measured HMOs in maternal serum and matching venous cord blood samples collected at delivery from normal pregnancies (n = 22). To investigate maternal-to-fetal transport, we perfused isolated placental cotyledons from term pregnancies (n = 3) with 2'-fucosyllactose (2'FL) in a double closed setting. We found up to 18 oligosaccharides typically present in maternal serum in all cord serum samples investigated. Median total cord blood HMO concentration did not differ from the concentration in maternal serum. HMO composition resembled the composition in maternal serum, with the strongest correlations for 2'FL and LDFT. After 180 min perfusion, we found 22% of maternally offered 2'FL in the fetal circuit without reaching equilibrium. Our results provide direct evidence of HMOs in cord blood, and suggest that the placenta transfers HMOs from the maternal to fetal circuit. Future studies will investigate potential differences in the transfer of specific HMOs, or in pregnancy disorders.
Assuntos
Sangue Fetal/química , Troca Materno-Fetal , Leite Humano/química , Oligossacarídeos/sangue , Oligossacarídeos/química , Feminino , Humanos , Recém-Nascido , Placenta , GravidezRESUMO
BACKGROUND: Human milk oligosaccharides (HMOs) were recently found in serum of normal-weight pregnant women, with concentrations increasing from early to mid- and late pregnancy. Whether HMOs have effects on maternal metabolism is unknown. OBJECTIVES: We aimed to study the presence and changes in HMOs throughout pregnancy and assess associations with maternal glucose metabolism throughout pregnancy. METHODS: The study was a prospective longitudinal cohort study including 87 overweight or obese women. Blood samples were taken at 15, 24, and 32 wk of pregnancy. In serum, 4 HMOs [2'-fucosyllactose (2'FL), lactodifucotetraose (LDFT), 3'-sialyllactose (3'SL), and 3'-sialyllactosamine (3'SLN)] were measured. In linear regression models, the associations between HMOs and (changes in) maternal metabolic parameters were assessed. RESULTS: All 4 HMOs showed a significant increase from 15 to 32 weeks of gestation. 3'SL and 3'SLN, but not 2'FL or LDFT, at 15 wk were positively associated with (changes in) fasting glucose at 24 and 32 wk. LDFT was positively associated with (changes in) insulin and HOMA-index at 24 but not 32 wk. A model to predict the development of gestational diabetes mellitus (GDM) that included fasting glucose, prepregnancy BMI, gestational weight gain, age, parity, smoking, and history of macrosomia resulted in an area under the curve (AUC) of 0.81 (95% CI: 0.70, 0.92). Adding 3'SL to this model increased the AUC to 0.91 (95% CI: 0.84, 0.97). CONCLUSIONS: The sialylated HMOs 3'SL and 3'SLN were associated with fasting glucose; LDFT was associated with fasting insulin and HOMA-index. Furthermore, 3'SL was more predictive of future GDM diagnoses than was fasting glucose in early pregnancy. Causal relations are unclear and need further investigation.
Assuntos
Glucose/metabolismo , Leite Humano/química , Obesidade/sangue , Oligossacarídeos/sangue , Sobrepeso/sangue , Gravidez/sangue , Adulto , Feminino , Humanos , Estudos Longitudinais , Leite Humano/metabolismo , Obesidade/metabolismo , Oligossacarídeos/metabolismo , Sobrepeso/metabolismo , Gravidez/metabolismo , Estudos ProspectivosRESUMO
Melioidosis and glanders, respectively caused by the Gram-negative bacteria Burkholderia pseudomallei (Bp) and Burkholderia mallei (Bm), are considered as urgent public health issues in developing countries and potential bioterrorism agents. Bp and Bm lipopolysaccharides (LPS) have been identified as attractive vaccine candidates for the development of prophylactic measures against melioidosis and glanders. Bp and Bm express structurally similar LPSs wherein the O-antigen (OAg) portion consists of a heteropolymer whose repeating unit is a disaccharide composed of d-glucose and 6-deoxy-l-talose residues, the latter being diversely acetylated and methylated. Herein we report the synthesis of two tetrasaccharides mimicking the main substitution epitopes of Bp and Bm LPS OAgs. The assembly of the tetrasaccharides was achieved using a sequential glycosylation strategy while relying on the late-stage epimerization of the inner rhamnose into a 6-deoxy-l-talose residue. We show that these synthetic compounds strongly react with culture-confirmed Thai melioidosis patient serum and closely mimic the antigenicity of native Bp OAg. Our results suggest that these tetrasaccharides could be suitable candidates for the development of vaccines and/or diagnostic tools against melioidosis and glanders.
Assuntos
Burkholderia mallei/imunologia , Burkholderia pseudomallei/imunologia , Epitopos/química , Melioidose/sangue , Melioidose/imunologia , Antígenos O/imunologia , Oligossacarídeos/química , Oligossacarídeos/imunologia , Burkholderia mallei/química , Burkholderia pseudomallei/química , Epitopos/sangue , Epitopos/imunologia , Humanos , Antígenos O/química , Oligossacarídeos/sangue , TailândiaRESUMO
Sanfilippo syndrome or mucopolysaccharidosis type III (MPS III) is a childhood metabolic disorder marked by neuropathology arising due to impaired heparan sulphate (HS) catabolism. Consequently, partially degraded HS accumulates in the lysosomes of affected cells and is excreted in the urine. The measurement of HS in urine has long been considered a biomarker of Sanfilippo syndrome although it is largely non-specific. Using blood, urine and CSF collected from a cohort of Sanfilippo patients we investigated the utility of primary and secondary biomarkers to inform on disease activity. These included enzyme activity, specific oligosaccharides with non-reducing end residues reflective of the enzyme deficiency, and gangliosides. The diagnostic oligosaccharides - a HS disaccharide and tetrasaccharide - were elevated in the urine, plasma and CSF of all MPS IIIA and IIIB patients, respectively. There was no correlation between the concentrations in any of the matrices suggesting they reflect specific tissues and not overall disease burden. Enzyme activity did not inform on disease severity, with no measurable activity in CSF and activity approaching normal in MPS IIIA plasma. The concentration of gangliosides, GM2 and GM3, were significantly higher in the CSF of all MPS III subjects when compared to controls and correlated with the age of onset of first symptoms. Given that these gangliosides reflect delayed brain development they may be useful measures of disease burden, within the limitations of the clinical surrogates. Observation of these biochemical measurements in MPS III patients enrolled in clinical trials may determine whether they represent true pharmacodynamics biomarkers.
Assuntos
Biomarcadores/análise , Gangliosídeos/análise , Mucopolissacaridose III/diagnóstico , Oligossacarídeos/análise , Pré-Escolar , Gangliosídeos/sangue , Gangliosídeos/líquido cefalorraquidiano , Gangliosídeos/urina , Heparitina Sulfato/metabolismo , Humanos , Lactente , Mucopolissacaridose III/sangue , Mucopolissacaridose III/líquido cefalorraquidiano , Mucopolissacaridose III/urina , Oligossacarídeos/sangue , Oligossacarídeos/líquido cefalorraquidiano , Oligossacarídeos/urinaRESUMO
Sepsis induces heparanase-mediated degradation of the endothelial glycocalyx, a heparan sulfate-enriched endovascular layer critical to vascular homeostasis, releasing highly sulfated domains of heparan sulfate into the circulation. These domains are oligosaccharides rich in heparin-like trisulfated disaccharide repeating units. Using a chemoenzymatic approach, an undecasaccharide containing a uniformly 13C-labeled internal 2-sulfoiduronic acid residue was synthesized on a p-nitrophenylglucuronide acceptor. Selective periodate cleavage afforded a heparin nonasaccharide having a natural structure. This 13C-labeled nonasaccharide was intravenously administered to septic (induced by cecal ligation and puncture, a model of polymicrobial peritonitis-induced sepsis) and nonseptic (sham) mice. Selected tissues and biological fluids from the mice were harvested at various time points over 4 hours, and the 13C-labeled nonasaccharide was recovered and digested with heparin lyases. The resulting 13C-labeled trisulfated disaccharide was quantified, without interference from endogenous mouse heparan sulfate/heparin, using liquid chromatography-mass spectrometry with sensitive and selective multiple reaction monitoring. The 13C-labeled heparin nonasaccharide appeared immediately in the blood and was rapidly cleared through the urine. Plasma nonasaccharide clearance was only slightly prolonged in septic mice (t1/2 â¼ 90 minutes). In septic mice, the nonasaccharide penetrated into the hippocampus but not the cortex of the brain; no hippocampal or cortical brain penetration occurred in sham mice. The results of this study suggest that circulating heparan sulfates are rapidly cleared from the plasma during sepsis and selectively penetrate the hippocampus, where they may have functional consequences.
Assuntos
Heparina/sangue , Hipocampo/fisiologia , Oligossacarídeos/sangue , Sepse/sangue , Sepse/psicologia , Animais , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Cognição , Heparitina Sulfato/metabolismo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Sepse/metabolismoRESUMO
Human milk oligosaccharides (HMOs) are bioactive glycans linked with health benefits to both the breast-fed infant and lactating mother. We hypothesized that HMOs are present before lactation, already during pregnancy, and are influenced by maternal body composition. In a pilot study, we investigated individual and temporal variations in HMO composition and concentration in maternal serum at gestational weeks 10-14 ( visit 1), 20-24 ( visit 2), and 30-35 (visit 3) (V1, V2, and V3, respectively) and associations with maternal body composition. HMOs were quantified by HPLC and confirmed by enzymatic digest and mass spectrometry. Associations of maternal prepregnancy body mass index (BMI), subcutaneous adipose tissue (SAT) thickness, and adipokines with absolute and relative HMO concentrations were analyzed by Spearman correlation. We identified 16 HMOs and 2 oligosaccharides not common to human milk. HMO concentration and composition varied with gestational age and secretor status. HMO concentration increased with gestational age and changed from a predominantly sialylated profile at V1 to a more balanced fucosylated-to-sialylated ratio at V3, mostly due to a profound increase in 2'-fucosyllactose (2'-FL), reflecting secretor phenotype. In secretor-positive women, BMI was negatively correlated with 2'-FL at V2. SAT at V1 and V2 were strongly negatively correlated with 2'-FL concentrations. This pilot study shows that prenatal HMOs are present in maternal serum, suggesting roles for HMOs already during pregnancy. Our result that maternal body composition is associated with prenatal HMOs might indicate that maternal metabolism modulates HMO composition with unknown implications for maternal and fetal health already during pregnancy.
Assuntos
Adipocinas/sangue , Leite Humano , Oligossacarídeos/sangue , Gravidez/sangue , Adiponectina/sangue , Adulto , Composição Corporal , Índice de Massa Corporal , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Estudos de Coortes , Feminino , Idade Gestacional , Humanos , Leptina/sangue , Estudos Longitudinais , Tamanho do Órgão , Projetos Piloto , Primeiro Trimestre da Gravidez , Segundo Trimestre da Gravidez , Terceiro Trimestre da Gravidez , Estudos Prospectivos , Gordura Subcutânea/anatomia & histologia , Espectrometria de Massas em TandemRESUMO
BACKGROUND: In 1987, three unrelated English families were reported with a putative blood subgroup called Apae. Swedish researchers later found evidence leading to abolishment of the Apae subgroup and establishment instead of the FORS blood group system (System 31 - ISBT, 2012). It is important to know the prevalence of antibodies in order to make the best decisions in transfusion medicine. Cells expressing the Forssman saccharide, such as sheep erythrocytes, are needed to detect the anti-Forssman antibody. The aim of this study was to define the prevalence of human anti-Forssman antibody. MATERIALS AND METHODS: Plasma samples from 800 individuals were studied. Sheep erythrocytes or Forssman "kodecytes" were mixed with the plasma samples using the tube technique. Plasma from an Apae individual was used as a negative control and monoclonal anti-Forssman antibody (M1/22.25.8HL cell line supernatant) was used as the positive control. RESULTS: Of the 800 individuals tested, one was negative for the presence of anti-Forssman antibody. We compared the anti-Forssman antibody reaction pattern between genders and found that males have weaker reactions than females, both at room temperature (p=0.026) and at 37 °C (p=0.043). We also investigated the reaction pattern of anti-Forssman antibody in relation to ABO and Rh blood group types without finding any significant differences. DISCUSSION: Sheep erythrocytes are suitable for searching for human anti-Forssman antibody. The quantity of anti-Forssman antibodies in plasma is higher in females than in males. In the population (n=800) studied here, we found one individual lacking the anti-Forssman antibody. These results contribute to the data already published, confirming that FORS is a rare blood group.
Assuntos
Antígenos de Grupos Sanguíneos/sangue , Tipagem e Reações Cruzadas Sanguíneas/métodos , Antígeno de Forssman/sangue , Isoanticorpos/sangue , Oligossacarídeos/sangue , Animais , Antígenos de Grupos Sanguíneos/imunologia , Feminino , Antígeno de Forssman/imunologia , Humanos , Isoanticorpos/imunologia , Masculino , Oligossacarídeos/imunologia , Prevalência , OvinosRESUMO
Sepsis and acute kidney injury (AKI) synergistically increase morbidity and mortality in the ICU. How sepsis reduces glomerular filtration rate (GFR) and causes AKI is poorly understood; one proposed mechanism includes tubuloglomerular feedback (TGF). When sodium reabsorption by the proximal tubules is reduced in normal animals, the macula densa senses increased luminal sodium chloride, and then adenosine-1a receptor (A1aR) signaling triggers tubuloglomerular feedback, reducing GFR through afferent arteriole vasoconstriction. We measured GFR and systemic hemodynamics early during cecal ligation and puncture-induced sepsis in wild-type and A1aR-knockout mice. A miniaturized fluorometer was attached to the back of each mouse and recorded the clearance of FITC-sinistrin via transcutaneous fluorescence to monitor GFR. Clinical organ injury markers and cytokines were measured and hemodynamics monitored using implantable transducer telemetry devices. In wild-type mice, GFR was stable within 1 h after surgery, declined by 43% in the next hour, and then fell to less than 10% of baseline after 2 h and 45 min. In contrast, in A1aR-knockout mice GFR was 37% below baseline immediately after surgery and then gradually declined over 4 h. A1aR-knockout mice had similar organ injury and inflammatory responses, albeit with lower heart rate. We conclude that transcutaneous fluorescence can accurately monitor GFR and detect changes rapidly during sepsis. Tubuloglomerular feedback plays a complex role in sepsis; initially, TGF helps maintain GFR in the 1st hour, and over the subsequent 3 h, TGF causes GFR to plummet. By 18 h, TGF has no cumulative effect on renal or extrarenal organ damage.
Assuntos
Injúria Renal Aguda/metabolismo , Taxa de Filtração Glomerular , Rim/metabolismo , Receptor A1 de Adenosina/metabolismo , Sepse/metabolismo , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/genética , Injúria Renal Aguda/fisiopatologia , Animais , Modelos Animais de Doenças , Retroalimentação Fisiológica , Fluoresceínas/administração & dosagem , Corantes Fluorescentes/administração & dosagem , Corantes Fluorescentes/metabolismo , Fluorometria/métodos , Hemodinâmica , Injeções Intravenosas , Rim/fisiopatologia , Camundongos Endogâmicos C57BL , Camundongos Knockout , Oligossacarídeos/administração & dosagem , Oligossacarídeos/sangue , Receptor A1 de Adenosina/deficiência , Receptor A1 de Adenosina/genética , Sepse/complicações , Sepse/genética , Sepse/fisiopatologia , Transdução de Sinais , Fatores de TempoRESUMO
To verify the idea that extracellular free oligosaccharides might be able to reflect the functional status of the endoplasmic reticulum (ER) and lysosomal-endosomal system, HPLC-profiles of serum-derived free oligosaccharides (FOS) in human healthy aging, acute myeloproliferative neoplasms, and cardiovascular pathologies were compared with intracellular glycans. After plasma deproteinization and FOS purification the oligosaccharides were labelled with anthranilic acid, separated into the neutral and charged with QAE Sephadex (Q25-120) chromatography and analysed using high-performance liquid chromatography (HPLC). The charged FOS were digested with a sialidase and compared with free oligosaccharides from transferrin for structural decoding. HPLC-profiles of serum-derived FOS revealed mild delay of the dolichol phosphate cycle in ER, moderate intensification of ER-associated degradation (ERAD) and degradation in endosomal-lysosomal system with aging; an inhibition of the dolichol phosphate cycle, intensification of ERAD and increasing of lysosomal exocytosis in acute myeloproliferative neoplasms; intensification of ERAD and glycocojugate degradation with endosomal-lysosomal system in cardiovascular diseases. As serum free oligosaccharides are able to reflect specifically perturbations in ER and endosomal-lysosomal system under wide range of stressors they can serve as extracellular markers of functionality of these organelles.
Assuntos
Doenças Cardiovasculares/sangue , Fosfatos de Dolicol/sangue , Degradação Associada com o Retículo Endoplasmático , Glicoconjugados/sangue , Envelhecimento Saudável/sangue , Transtornos Mieloproliferativos/sangue , Oligossacarídeos/sangue , Biomarcadores/sangue , Biomarcadores/química , Sequência de Carboidratos , Doenças Cardiovasculares/diagnóstico , Estudos de Casos e Controles , Retículo Endoplasmático/metabolismo , Endossomos/metabolismo , Glicosilação , Humanos , Lisossomos/metabolismo , Transtornos Mieloproliferativos/diagnóstico , Oligossacarídeos/química , Coloração e Rotulagem/métodos , ortoaminobenzoatos/químicaRESUMO
BACKGROUND: The aim of this study was to determine the effects of a low fermentable oligo-, di- and monosaccharides and polyols (FODMAP) diet on the nutritional status and body composition, abdominal symptoms, quality of life, anxiety/depression and sleep quality of patients with irritable bowel syndrome (IBS). METHODS: Consecutive patients were given a low FODMAP diet for 8 weeks. At baseline and after 8 weeks, blood tests were taken to evaluate nutritional status and a bioelectrical impedance analysis was performed to assess body composition. Anthropometric data, IBS Symptom Severity Score, results of a bowel habits questionnaire, Bristol Stool Chart classification, SF36, Hamilton Depression Anxiety Scale outcome and Pittsburgh Sleep Quality Index were also recorded. During the 8-week diet period, the patients were phoned periodically by the nutritionist to verify their compliance. RESULTS: Twenty-six IBS patients with a mean age of 46.2 ± 13.8 years were studied. After 8 weeks, there were no abnormalities in anthropometric data, bioelectrical impedance parameters and blood tests. The patients' IBS Symptom Severity Score improved (305.2 ± 84.1 vs 156.3 ± 106.4; p < 0.0001), as did bowel habits, Bristol Stool Chart classification, quality of life and HADS anxiety score, whereas sleeping quality and depression were unchanged. The degree of relief from symptoms and satisfaction with the diet was high. CONCLUSIONS: A low FODMAP diet improved IBS symptoms without effects on nutritional status and body composition.
Assuntos
Composição Corporal/fisiologia , Dieta/métodos , Impedância Elétrica , Síndrome do Intestino Irritável/dietoterapia , Síndrome do Intestino Irritável/fisiopatologia , Adolescente , Adulto , Idoso , Dissacarídeos/efeitos adversos , Dissacarídeos/sangue , Feminino , Fermentação , Humanos , Síndrome do Intestino Irritável/sangue , Masculino , Pessoa de Meia-Idade , Monossacarídeos/efeitos adversos , Monossacarídeos/sangue , Estado Nutricional , Oligossacarídeos/efeitos adversos , Oligossacarídeos/sangue , Projetos Piloto , Qualidade de Vida , Índice de Gravidade de Doença , Resultado do Tratamento , Adulto JovemRESUMO
Human milk oligosaccharides (HMO) are involved in many biological functions influencing infant health. Although HMO act locally at the intestine, recent evidence has demonstrated that HMO are partially incorporated into the systemic circulation of breast-fed infants. In the last few years, a large amount of research has been conducted using preclinical models to uncover new biological functions of HMO. The aim of this study was to evaluate the absorption and urine excretion of HMO in rats. We administered a single oral dose of the following HMO: 2'-fucosyllactose (2'-FL), 6'-sialyllactose and lacto-N-neotetraose at different concentrations to adult rats. The time course of absorption of HMO into the bloodstream and their appearance in urine was studied. Our results showed that rats, similar to human infants, are able to effectively absorb a portion of HMO from the intestine into plasma and to excrete them in urine. On the basis of this, we also conducted a specific kinetic absorption study with 2'-FL, the most predominant HMO in human milk, in 9-11-d-old rat pups. Our results confirmed that a significant amount of 2'-FL was absorbed into the systemic circulation and subsequently excreted in urine during lactation in rats in a dose-depended manner. We also found basal levels of these HMO in plasma and urine of adult rats as well as rat pups as a natural result of nursing. Our data suggest that the rat may be a useful preclinical model that provides new insights into the metabolism and functions of HMO.