Haptoglobin is a plasma protein of mammals that plays a crucial role in vascular homeostasis by binding free haemoglobin released from ruptured red blood cells. Trypanosoma brucei can exploit this by internalising haptoglobin-haemoglobin complex to acquire host haem. Here, we investigated the impact of haptoglobin deficiency (Hp-/-) on T. brucei brucei infection and the parasite ́s capacity to internalise haemoglobin in a Hp-/- mouse model. The infected Hp-/- mice exhibited normal disease progression, with minimal weight loss and no apparent organ pathology, similarly to control mice. While the proteomic profile of mouse sera significantly changed in response to T. b. brucei, no differences in the infection response markers of blood plasma between Hp-/- and control Black mice were observed. Similarly, very few quantitative differences were observed between the proteomes of parasites harvested from Hp-/- and Black mice, including both endogenous proteins and internalised host proteins. While haptoglobin was indeed absent from parasites isolated from Hp-/-mice, haemoglobin peptides were unexpectedly detected in parasites from both Hp-/- and Black mice. Combined, the data support the dispensability of haptoglobin for haemoglobin internalisation by T. b. brucei during infection in mice. Since the trypanosomes knock-outs for their haptoglobin-haemoglobin receptor (HpHbR) internalised significantly less haemoglobin from Hp-/- mice compared to those isolated from Black mice, it suggests that T. b. brucei employs also an HpHbR-independent haptoglobin-mediated mode for haemoglobin internalisation. Our study reveals a so-far hidden flexibility of haemoglobin acquisition by T. b. brucei and offers novel insights into alternative haemoglobin uptake pathways.

• MeSH
• haptoglobiny * genetika   metabolismus   MeSH
• hemoglobiny * metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední C57BL MeSH
• myši knockoutované * MeSH
• myši MeSH
• proteomika metody   MeSH
• Trypanosoma brucei brucei * metabolismus   MeSH
• trypanozomóza africká * parazitologie   imunologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Methods of structural mass spectrometry have become more popular to study protein structure and dynamics. Among them, fast photochemical oxidation of proteins (FPOP) has several advantages such as irreversibility of modifications and more facile determination of the site of modification with single residue resolution. In the present study, FPOP analysis was applied to study the hemoglobin (Hb) - haptoglobin (Hp) complex allowing identification of respective regions altered upon the complex formation. FPOP footprinting using a timsTOF Pro mass spectrometer revealed structural information for 84 and 76 residues in Hp and Hb, respectively, including statistically significant differences in the modification extent below 0.3%. The most affected residues upon complex formation were Met76 and Tyr140 in Hbα, and Tyr280 and Trp284 in Hpβ. The data allowed determination of amino acids directly involved in Hb - Hp interactions and those located outside of the interaction interface yet affected by the complex formation. Also, previously modeled interaction between Hb βTrp37 and Hp βPhe292 was not confirmed by our data. Data are available via ProteomeXchange with identifier PXD021621.

• MeSH
• aminokyseliny chemie   metabolismus   MeSH
• footprinting proteinů metody   MeSH
• haptoglobiny chemie   metabolismus   MeSH
• hemoglobiny chemie   metabolismus   MeSH
• hmotnostní spektrometrie metody   MeSH
• hydroxylový radikál chemie   MeSH
• lidé MeSH
• molekulární modely MeSH
• molekulární struktura MeSH
• oxidace-redukce MeSH
• vazba proteinů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Unconjugated hyperbilirubinemia, a feature of neonatal jaundice or Crigler-Najjar syndrome, can lead to neurotoxicity and even death. We previously demonstrated that unconjugated bilirubin (UCB) can be eliminated via transintestinal excretion in Gunn rats, a model of unconjugated hyperbilirubinemia, and that this is stimulated by enhancing fecal fatty acid excretion. Since transintestinal excretion also occurs for cholesterol (TICE), we hypothesized that increasing fecal cholesterol excretion and/or TICE could also enhance fecal UCB disposal and subsequently lower plasma UCB concentrations. METHODS: To determine whether increasing fecal cholesterol excretion could ameliorate unconjugated hyperbilirubinemia, we treated hyperbilirubinemic Gunn rats with ezetimibe (EZE), an intestinal cholesterol absorption inhibitor, and/or a liver X receptor (LXR) and farnesoid X receptor (FXR) agonist (T0901317 (T09) and obeticholic acid (OCA), respectively), known to stimulate TICE. RESULTS: We found that EZE treatment alone or in combination with T09 or OCA increased fecal cholesterol disposal but did not lower plasma UCB levels. CONCLUSIONS: These findings do not support a link between the regulation of transintestinal excretion of cholesterol and bilirubin. Furthermore, induction of fecal cholesterol excretion is not a potential therapy for unconjugated hyperbilirubinemia. IMPACT: Increasing fecal cholesterol excretion is not effective to treat unconjugated hyperbilirubinemia. This is the first time a potential relation between transintestinal excretion of cholesterol and unconjugated bilirubin is investigated. Transintestinal excretion of cholesterol and unconjugated bilirubin do not seem to be quantitatively linked. Unlike intestinal fatty acids, cholesterol cannot "capture" unconjugated bilirubin to increase its excretion. These results add to our understanding of ways to improve and factors regulating unconjugated bilirubin disposal in hyperbilirubinemic conditions.

• MeSH
• bilirubin chemie   MeSH
• cholesterol metabolismus   MeSH
• Criglerův-Najjarův syndrom metabolismus   terapie   MeSH
• dietní tuky farmakokinetika   MeSH
• ezetimib farmakologie   terapeutické užití   MeSH
• feces chemie   MeSH
• fluorované uhlovodíky farmakologie   terapeutické užití   MeSH
• haptoglobiny analýza   MeSH
• hyperbilirubinemie terapie   MeSH
• jaterní receptor X metabolismus   MeSH
• krysa rodu rattus MeSH
• kyselina chenodeoxycholová analogy a deriváty   farmakologie   terapeutické užití   MeSH
• lipidy krev   MeSH
• náhodné rozdělení MeSH
• potkani Gunn MeSH
• PPAR delta metabolismus   MeSH
• receptory cytoplazmatické a nukleární metabolismus   MeSH
• střeva účinky léků   metabolismus   MeSH
• sulfonamidy farmakologie   terapeutické užití   MeSH
• žluč chemie   MeSH
• žlučové kyseliny a soli metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The aim of our study was to assess the presence and degree of intestinal leakage in subjects suffering from short bowel syndrome (SBS) and its modification by parenteral nutrition. To this end we assessed circulating levels of selected makers of intestinal permeability including zonulin, fatty acid binding protein 2 (FABP-2), citrulline and glucagon-like peptide 2 (GLP-2). We also measured lipopolysaccharide binding protein (LBP) as a marker of circulating levels of lipopolysaccharide acting through the CD14 molecule. Eleven SBS and 10 age- and BMI-matched control subjects were included into the study. The effect of parenteral nutrition was assessed after 14 days, 6 and 12 months from its initiation, respectively. At baseline, SBS patients had increased gut permeability as measured by zonulin (47.24+/-2.14 vs. 39.48+/-1.20 ng/ml, p=0.006) and LBP (30.32+/-13.25 vs. 9.77+/-0.71 microg/ml, p<0.001) compared to healthy controls. Furthermore, SBS subjects had reduced FABP-2, unchanged citrulline and increased sCD14 and GLP-2 relative to control group. Throughout the whole study period the administered parenteral nutrition had no significant effect on any of the studied parameters. Taken together, our data show that patients with short bowel syndrome have increased intestinal permeability that is not affected by parenteral nutrition.

• MeSH
• biologické markery krev   MeSH
• citrulin krev   MeSH
• glukagonu podobný peptid 2 krev   MeSH
• haptoglobiny MeSH
• intestinální absorpce * MeSH
• lidé středního věku MeSH
• lidé MeSH
• membránové glykoproteiny krev   MeSH
• parenterální výživa * MeSH
• permeabilita MeSH
• proteinové prekurzory krev   MeSH
• proteiny akutní fáze MeSH
• proteiny vázající mastné kyseliny krev   MeSH
• senioři MeSH
• studie případů a kontrol MeSH
• syndrom krátkého střeva diagnóza   krev   patofyziologie   terapie   MeSH
• tenké střevo metabolismus   patofyziologie   MeSH
• transportní proteiny krev   MeSH
• výsledek terapie MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Recent studies show that the haptoglobin phenotype in individuals with diabetes mellitus is an important factor for predicting the risk of myocardial infarction, cardiovascular death, and stroke. Current methods for haptoglobin phenotyping include PCR and gel electrophoresis. A need exists for a reliable method for high-throughput clinical applications. Mass spectrometry (MS) can in principle provide fast phenotyping because haptoglobin α 1 and α 2, which define the phenotype, have different molecular masses. Because of the complexity of the serum matrix, an efficient and fast enrichment technique is necessary for an MS-based assay. METHODS: MALDI plates were functionalized by ambient ion landing of electrosprayed antihaptoglobin antibody. The array was deposited on standard indium tin oxide slides. Fast immunoaffinity enrichment was performed in situ on the plate, which was further analyzed by MALDI-TOF MS. The haptoglobin phenotype was determined from the spectra by embedded software script. RESULTS: The MALDI mass spectra showed ion signals of haptoglobin α subunits at m/z 9192 and at m/z 15 945. A cohort of 116 sera was analyzed and the reliability of the method was confirmed by analyzing the identical samples by Western blot. One hundred percent overlap of results between the direct immunoaffinity desorption/ionization MS and Western Blot analysis was found. CONCLUSIONS: MALDI plates modified by antihaptoglobin antibody using ambient ion landing achieve low nonspecific interactions and efficient MALDI ionization and are usable for quick haptoglobin phenotyping.

• MeSH
• chromatografie afinitní MeSH
• fenotyp MeSH
• haptoglobiny analýza   imunologie   MeSH
• lidé MeSH
• povrchové vlastnosti MeSH
• protilátky imunologie   MeSH
• software MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice metody   MeSH
• western blotting MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• Klíčová slova
• D-dimery,
• MeSH
• antikoagulancia terapeutické užití   MeSH
• Aspirin * farmakologie   MeSH
• biologické markery MeSH
• haptoglobiny účinky léků   MeSH
• inhibitory agregace trombocytů MeSH
• kardiostimulace umělá * škodlivé účinky   MeSH
• králíci MeSH
• počet leukocytů MeSH
• tromboembolie * diagnóza   krev   prevence a kontrola   MeSH
• vyšetření krevní srážlivosti klasifikace   metody   MeSH
• zánět krev   MeSH
• žilní tromboembolie diagnóza   prevence a kontrola   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH
• srovnávací studie MeSH

• MeSH
• adenokarcinom diagnóza   MeSH
• C-reaktivní protein diagnostické užití   MeSH
• časná detekce nádoru MeSH
• haptoglobiny diagnostické užití   MeSH
• hemoglobiny diagnostické užití   MeSH
• imunologické testy trendy   MeSH
• infekce dýchací soustavy diagnóza   MeSH
• kolorektální nádory diagnóza   MeSH
• okultní krev MeSH
• Publikační typ
• informační letáky pro pacienty MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• diagnostika

AIM: To assess vitamin D status and health correlates in a sample of apparently healthy Caucasian participants residing in an urban area, Athens, with latitude 370 58' 0" N and longitude 230 43' 0" E, after taking into consideration a broad range of purported biological, behavioural and environmental factors. METHOD: Men and women 35+ years from a selected population (n = 490) were studied. Participants completed a detailed questionnaire regarding socio-demographic, lifestyle, clinical and dietary characteristics. Biomarkers were measured after 12 h fasting. Linear and multinomial regression models were used to evaluate the association between 25(OH)D and determinants of vitamin D status. RESULTS: Results revealed that one hour increase of sunlight exposure decreased the odds of having D deficiency (i.e., < 20 ng/mL) by 70% (OR = 0.30, 95% Cl: 0.20-0.45), adjusted for age, sex, family status, physical activity, smoking habits, BMI, triglycerides, parathyroid hormone, uric acid, haptoglobin, folate acid and haemoglobin, as compared to sufficient levels (i.e., >30 ng/mL). Regarding biomarkers, parathyroid hormone and haptoglobin were found to be related with the odds of having vitamin D deficiency (OR = 1.11, 95% CI: 1.05-1.16; OR = 1.02, 95% CI: 1.00-1.03, respectively) as compared to the sufficient levels. CONCLUSIONS: Sufficient serum vitamin D levels were observed among participants with characteristics associated with reduced cardiovascular risk, such as normal BMI, increased physical activity, decreased parathyroid hormone and decreased inflammatory markers. Even a slight increase in sunlight exposure could have beneficial effects on serum vitamin D concentrations and eventually on haemoglobin and inflammatory markers levels, thus providing a simple and inexpensive lifestyle intervention that promotes public health.

• MeSH
• analýza rozptylu MeSH
• biologické markery krev   MeSH
• dieta MeSH
• dospělí MeSH
• haptoglobiny metabolismus   MeSH
• index tělesné hmotnosti MeSH
• lidé středního věku MeSH
• lidé MeSH
• městské obyvatelstvo MeSH
• nedostatek vitaminu D epidemiologie   MeSH
• parathormon krev   MeSH
• podnebí MeSH
• pohybová aktivita MeSH
• průzkumy a dotazníky MeSH
• regresní analýza MeSH
• rizikové faktory MeSH
• rozdělení chí kvadrát MeSH
• sluneční záření MeSH
• životní styl MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Řecko MeSH

Haptoglobin is a glycoprotein involved in the acute phase response. Previously we reported that haptoglobin gene expression was up-regulated during dietary restriction in young female rats. The present study aimed at determining whether chronic dietary restriction affects haptoglobin blood levels through changing levels and/or activities of IL-6-related transcription factors STAT and C/EBP in the liver as is the case during the acute phase response. To this end, we compared a female Wistar rat model of 50% 6-week-long dietary restriction with the standard laboratory model for the acute phase response induced by turpentine administration. During the turpentine-induced acute phase response, the transitory 5.4-fold increase of rat haptoglobin expression was accompanied by a prominent rise of serum IL-6 concentration and the increased binding of STAT3 and 35kD C/EBPbeta/LAP transcription factors to the haptoglobin gene hormone-responsive element. Results obtained after immunoblotting and DNA affinity chromatography (using hormone-responsive element) suggest that the stable 1.7-fold increase of serum haptoglobin level during dietary restriction was the result of increased amounts and activities of constitutive transcription factors C/EBPalpha and STAT5b, and to a smaller extent of STAT3. When dietary restriction rats were administered turpentine, a 8.7-fold increase in haptoglobin expression was followed by a considerable increase in the amount and hormone-responsive element binding activity of STAT3 but not 35kD C/EBPbeta/LAP. We concluded that haptoglobin gene up-regulation during chronic dietary restriction was regulated by different mechanisms than during the acute phase response, and that it depended on the amount(s) and activit(ies) of transcription factor(s) that characterize low-grade inflammatory conditions.

• MeSH
• chromatografie afinitní MeSH
• dráždivé látky farmakologie   MeSH
• financování organizované MeSH
• haptoglobiny genetika   metabolismus   MeSH
• imunoblotting MeSH
• interleukin-6 krev   MeSH
• kalorická restrikce MeSH
• krysa rodu rattus MeSH
• potkani Wistar MeSH
• reakce akutní fáze chemicky indukované   metabolismus   MeSH
• regulace genové exprese MeSH
• terpentýn farmakologie   MeSH
• transkripční faktor STAT3 metabolismus   MeSH
• transkripční faktor STAT5 metabolismus   MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• ženské pohlaví MeSH
• zvířata MeSH

Ovarian cancer is the most lethal of all gynecological cancers. Although serum biomarker CA125 is routinely used, there is a need for sensitive and specific complementary biomarkers. N-glycosylation changes in ovarian cancer serum glycoproteins include a decrease in galactosylation of IgG and an increase in sialyl Lewis X (SLex) on haptoglobin ß-chain, ?1-acid glycoprotein and ?1- antichymotrypsin. These changes are also present in chronic inflammations but not in malignant melanoma with low levels of inflammation. Acute phase proteins carrying increased amounts of SLex have an increased half-life. Sialylation of acute phase proteins decreases apoptosis favouring survival of cancer cells. Cancer cells produce inflammatory cytokines which influence glycosylation in liver parenchymal cells. The decreased galactosylation and sialylation of IgG increases cytotoxicity of natural killer cells and complement activation via mannosebinding lectin. Altered glycosylation of acute phase proteins and IgG suggests that cancer regulates certain pathways favouring survival of cancer cells.

• MeSH
• alfa-1-antichymotrypsin krev   metabolismus   MeSH
• alfa-makroglobuliny metabolismus   MeSH
• galaktosa nedostatek   MeSH
• glykosylace MeSH
• haptoglobiny metabolismus   MeSH
• imunoglobulin G krev   metabolismus   MeSH
• kyseliny sialové nedostatek   MeSH
• lidé MeSH
• metabolismus sacharidů MeSH
• nádorové biomarkery krev   MeSH
• nádory vaječníků imunologie   metabolismus   MeSH
• proteiny akutní fáze metabolismus   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH