Browsing by Subject "METABOLITES"

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  • Noman, Omar M.; Herqash, Rashed N.; Shahat, Abdelaaty A.; Ahamad, Syed Rizwan; Mechchate, Hamza; Almoqbil, Abdulaziz N.; Alqahtani, Ali S. (2022)
    Centaurea is one of the most important genera within the family Asteraceae. An investigation of the phytochemical composition of Centaurea bruguieriana using Gas-Chromatography coupled to Mass spectrometry (GC-MS) was performed. Antimicrobial activity was evaluated using the minimum inhibitory concentration method (MIC) and validated by molecular docking for the major compounds of the most active fraction (1,10-di-epi-cubenol and methyl 8-oxooctanoate) of C. bruguieriana against three bacterial receptors (TyrRS, DNA gyrase, and dihydrofolate reductase (DHFR)). Evaluation of antioxidant activity was conducted using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2 '-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. High-performance liquid chromatography (HPLC) was used to identify and quantify the contents of major compounds from ethyl acetate fraction (luteolin 7-O-glucoside, chlorogenic acid, kaempferol and isorhamnetin). The antimicrobial activity test showed that the chloroform fraction was more active against all microbial strains. The results of the molecular docking of two major compounds from chloroform fraction showed that good affinities were made between 1,10-di-epi-cubenol and the three selected receptors (TyrRs: -6.0 Kcal/mol against -8.2 Kcal/mol obtained with clorobiocin (standard); DNA gyrase: -6.6 Kcal/mol against -9.1 Kcal/mole obtained with clorobiocin; DHFR: -7.4 Kcal/mol against -6.3 Kcal/mol obtained with SCHEMBL2181345 Standard). Antioxidant evaluation showed that the ethyl acetate fraction was the most active fraction in DPPH (IC50 49.4 mu g/mL) and ABTS (IC50 52.8 mu g/mL) models. HPLC results showed the contents of luteolin 7-O-glucoside (7.4 mu g/mg), and chlorogenic acid (3.2 mu g/mg). Our study demonstrated that C. bruguierana is a promising source of bioactive compounds.
  • Limonte, Christine P.; Valo, Erkka; Montemayor, Daniel; Afshinnia, Farsad; Ahluwalia, Tarunveer S.; Costacou, Tina; Darshi, Manjula; Forsblom, Carol; Hoofnagle, Andrew N.; Groop, Per-Henrik; Miller, Rachel G.; Orchard, Trevor J.; Pennathur, Subramaniam; Rossing, Peter; Sandholm, Niina; Snell-Bergeon, Janet K.; Ye, Hongping; Zhang, Jing; Natarajan, Loki; de Boer, Ian H.; Sharma, Kumar (2020)
    Background: Individuals with type 1 diabetes (T1D) demonstrate varied trajectories of estimated glomerular filtration rate (eGFR) decline. The molecular pathways underlying rapid eGFR decline in T1D are poorly understood, and individual-level risk of rapid eGFR decline is difficult to predict. Methods: We designed a case-control study with multiple exposure measurements nested within 4 well-characterized T1D cohorts (FinnDiane, Steno, EDC, and CACTI) to identify biomarkers associated with rapid eGFR decline. Here, we report the rationale for and design of these studies as well as results of models testing associations of clinical characteristics with rapid eGFR decline in the study population, upon which "omics" studies will be built. Cases (n = 535) and controls (n = 895) were defined as having an annual eGFR decline of >= 3 and
  • van der Lugt, Benthe; van Beek, Adriaan A.; Aalvink, Steven; Meijer, Ben; Sovran, Bruno; Vermeij, Wilbert P.; Brandt, Renata M. C.; de Vos, Willem M.; Savelkoul, Huub F. J.; Steegenga, Wilma T.; Belzer, Clara (2019)
    BackgroundThe use of Akkermansia muciniphila as potential therapeutic intervention is receiving increasing attention. Health benefits attributed to this bacterium include an improvement of metabolic disorders and exerting anti-inflammatory effects. The abundance of A. muciniphila is associated with a healthy gut in early mid- and later life. However, the effects of A. muciniphila on a decline in intestinal health during the aging process are not investigated yet. We supplemented accelerated aging Ercc1(-/7) mice with A. muciniphila for 10weeks and investigated histological, transcriptional and immunological aspects of intestinal health.ResultsThe thickness of the colonic mucus layer increased about 3-fold after long-term A. muciniphila supplementation and was even significantly thicker compared to mice supplemented with Lactobacillus plantarum WCFS1. Colonic gene expression profiles pointed towards a decreased expression of genes and pathways related to inflammation and immune function, and suggested a decreased presence of B cells in colon. Total B cell frequencies in spleen and mesenteric lymph nodes were not altered after A. muciniphila supplementation. Mature and immature B cell frequencies in bone marrow were increased, whereas B cell precursors were unaffected. These findings implicate that B cell migration rather than production was affected by A. muciniphila supplementation. Gene expression profiles in ileum pointed toward a decrease in metabolic- and immune-related processes and antimicrobial peptide production after A. muciniphila supplementation. Besides, A. muciniphila decreased the frequency of activated CD80(+)CD273(-) B cells in Peyer's patches. Additionally, the increased numbers of peritoneal resident macrophages and a decrease in Ly6C(int) monocyte frequencies in spleen and mesenteric lymph nodes add evidence for the potentially anti-inflammatory properties of A. muciniphila.ConclusionsAltogether, we show that supplementation with A. muciniphila prevented the age-related decline in thickness of the colonic mucus layer and attenuated inflammation and immune-related processes at old age. This study implies that A. muciniphila supplementation can contribute to a promotion of healthy aging.
  • Pöhö, Paivi; Scholz, Karen; Kärkkäinen, Niina; Haapala, Markus; Räikkönen, Heikki; Kostiainen, Risto; Vaikkinen, Anu (2019)
    A new heated capillary photoionization (CPI) ion source design was developed to photoionize analytes inside a transfer capillary between a gas chromatograph (GC) and a mass spectrometer (MS). The CPI setup included a wide, oval-shaped vacuum-ultraviolet (VUV) transparent magnesium fluoride (MgF2) window to maximize photoionization efficiency and thus sensitivity. The source contained a nitrogen housing around the ionization chamber inlet to avoid undesirable hydrolysis and oxidation reactions with ambient air and to maximize the proportion of formed molecular radical cations of analytes. The feasibility of the ion source was studied by analyzing 18 endogenous steroids in urine as their trimethylsilyl (TMS) derivatives with gas chromatography-tandem mass spectrometry (GC-MS/MS). The method was validated and applied to human urine samples. To our best knowledge, this is the first time that a capillary photoionization ion source has been applied for quantitative analysis of biological samples. The GC-CPI-MS/MS method showed good chromatographic resolution (peak half-widths between 3.1 to 5.3 s), acceptable linearity (coefficient of determination between 0.981 to 0.996), and repeatability (relative standard deviation (RSD%) between 5 to 18%). Limits of detection (LOD) were between 2 to 100 pg mL(-1) and limits of quantitation (LOQ) were between 0.05 to 2 ng mL(-1). In total, 15 steroids were quantified either as a free steroid or glucuronide conjugate from the urine of volunteers. The new CPI source design showed excellent sensitivity for analysis of steroids in complex biological samples. (C) 2019 Elsevier B.V. All rights reserved.
  • Alitalo, Olga-Sofia; Rantalainen, Anna-Lea; Pellinen, Jukka (2022)
    The occurrence of three anticancer drugs (gemcitabine, letrozole, tamoxifen) was studied in wastewater samples from two local wastewater treatment plants (WWTPs) in Finland. Studied pharmaceuticals were selected, as anticancer drugs are potential to cause adverse effects on organisms even at low concentrations, but they are seldom included in the analysis of emerging contaminants. The concentration of anticancer drugs was determined by liquid chromatography-triple quadrupole mass spectrometer (LC-MS/MS). Tamoxifen and letrozole were detected from influent samples ranging from 0.5 to 5.0 ng/L, respectively. Letrozole was detected from effluent samples at a concentration up to 2.4 ng/L. Letrozole has been detected in wastewater effluent only once before, at a lower concentration of 0.28 ng/L. Gemcitabine was not detected in any of the samples. UV irradiation is used in many wastewater treatment plants to disinfect the effluent. Such tertiary treatment might degrade also these potentially harmful drugs and, therefore, photodegradation of the chosen pharmaceuticals was studied in laboratory-scale experiments. Tamoxifen showed high degradation rates, 94% in spiked wastewater with UV fluence 4830 mJ/cm(2) and 98% in pure water with UV fluence 2520 mJ/cm(2), respectively. Letrozole showed the lowest degradation rates of 24% in wastewater and 34% in pure water, respectively. The degradation rate at the fluence level typical for UV disinfection stage of wastewater treatment plants was 37% for tamoxifen but only 5% for letrozole. To the best of the authors' knowledge, this is the first report to show the effectiveness of UV irradiation to degrade letrozole.
  • Welsh, Paul; Rankin, Naomi; Li, Qiang; Mark, Patrick B.; Würtz, Peter; Ala-Korpela, Mika; Marre, Michel; Poulter, Neil; Hamet, Pavel; Chalmers, John; Woodward, Mark; Sattar, Naveed (2018)
    Aims/hypotheses We aimed to quantify the association of individual circulating amino acids with macrovascular disease, microvascular disease and all-cause mortality in individuals with type 2 diabetes. Methods We performed a case-cohort study (N = 3587), including 655 macrovascular events, 342 microvascular events (new or worsening nephropathy or retinopathy) and 632 all-cause mortality events during follow-up, in a secondary analysis of the Action in Diabetes and Vascular Disease: Preterax and Diamicron Modified Release Controlled Evaluation (ADVANCE) study. For this study, phenylalanine, isoleucine, glutamine, leucine, alanine, tyrosine, histidine and valine were measured in stored plasma samples by proton NMR metabolomics. Hazard ratios were modelled per SD increase in each amino acid. Results In models investigating associations and potential mechanisms, after adjusting for age, sex and randomised treatment, phenylalanine was positively, and histidine inversely, associated with macrovascular disease risk. These associations were attenuated to the null on further adjustment for extended classical risk factors (including eGFR and urinary albumin/creatinine ratio). After adjustment for extended classical risk factors, higher tyrosine and alanine levels were associated with decreased risk of microvascular disease (HR 0.78; 95% CI 0.67, 0.91 and HR 0.86; 95% CI 0.76, 0.98, respectively). Higher leucine (HR 0.79; 95% CI 0.69, 0.90), histidine (HR 0.89; 95% CI 0.81, 0.99) and valine (HR 0.79; 95% CI 0.70, 0.88) levels were associated with lower risk of mortality. Investigating the predictive ability of amino acids, addition of all amino acids to a risk score modestly improved classification of participants for macrovascular (continuous net reclassification index [NRI] +35.5%, p <0.001) and microvascular events (continuous NRI +14.4%, p = 0.012). Conclusions/interpretation We report distinct associations between circulating amino acids and risk of different major complications of diabetes. Low tyrosine appears to be a marker of microvascular risk in individuals with type 2 diabetes independently of fundamental markers of kidney function.
  • Becker, Anna; Schalin-Jäntti, Camilla; Itkonen, Outi (2021)
    Context: Patients with serotonin-secreting neuroendocrine neoplasms (NENs) have increased serum 5-hydroxyindoleacetic acid (5HIAA) concentrations. Serum 5HIAA thus serves as a biomarker in NEN. Objective: To evaluate an improved tandem mass spectrometric serum 5HIAA assay for diagnosis and follow-up of NEN in a clinical cohort. Design: A retrospective study during 2016-2018 at the Diagnostic Center and Department of Endocrinology at Helsinki University Hospital, Finland. Methods: Detailed patient data was obtained from 116 patients. Serum 5HIAA was analyzed by 2 different liquid chromatography with tandem mass spectrometry (LC-MS/MS) assays with samples prepared either by protein precipitation or solid phase extraction. Twenty-four-hour urine 5HIAA samples (n = 33) were analyzed by amperometric LC, and the results were compared. Specificity and sensitivity were calculated by receiver operating characteristic (ROC) analysis. Results: We achieved 5 to 10 000 nmol/L linearity and Conclusion: Serum 5HIAA by LC-MS/MS after protein precipitation performs equally well for the diagnosis of NEN as urinary 5HIAA LC assay. The outcome and sensitivity for serum and 24-h urine assays are convergent. Due to much more reliable and convenient sampling, we recommend serum instead of 24-h urine 5HIAA for diagnosis and follow-up of NEN patients.
  • Lindström, Mikael; Tohmola, Niina; Renkonen, Risto; Hämäläinen, Esa; Schalin-Jäntti, Camilla; Itkonen, Outi (2018)
    Background: Serotonin (5-hydroxytyramine) is a mediator of gastrointestinal smooth muscle contraction, and is secreted by neuroendocrine neoplasms (NENs). We developed a liquid chromatography tandem mass spectrometry (LC-MS/MS) assay for serum serotonin to be used in NEN diagnostics and follow-up. Methods: We used serum samples from healthy volunteers (n = 31) and patients suspected or monitored for NEN (n = 98). Serotonin-D-4 internal standard was added to samples before solid phase extraction (SPE) and quantification by LC-MS/MS. The effects of sample handling and preparation on serotonin stability were studied. Finally, we established a provisional reference range for serum serotonin and compared our assay with serum 5hydroxyindoleacetic acid (5-HIAA) for detection of NENs. Results: Our assay is sensitive and has a wide linear range (10-10,000 nmo1/1). Serum serotonin is stable for 7 days at room temperature and for 3 months at -20 degrees C. Sampling temperature is not critical. Normal range for serum serotonin was 270-1490 nmo1/1. We found that serum serotonin and 5-HIAA performed equally well as diagnostic tests for NENs. Conclusions: Our LC-MS/MS assay for serum serotonin is well suited for clinical research and patient diagnostics. Our results confirm that it can complement 5-HIAA in diagnosis of NENs.
  • Kauhanen, Dimple; Sysi-Aho, Marko; Koistinen, Kaisa M.; Laaksonen, Reijo; Sinisalo, Juha; Ekroos, Kim (2016)
    Monitoring the levels of the ceramides (Cer) d18:1/16:0, Cer d18:1/18:0, Cer d18:1/24:0, and Cer d18:1/24:1 and ratios thereof in human plasma empowers the prediction of fatal outcome of coronary artery disease (CAD). We describe a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) methodology for clinical-scaled measurement of the four distinct ceramides. Rapid plasma precipitation was accomplished in 96-well format. Excellent extraction recoveries in the range of 98-109 % were achieved for each ceramide. Addition of corresponding D-7-labeled ceramide standards facilitated precise quantification of each plasma ceramide species utilizing a novel short 5-min LC-MS/MS method. Neither matrix interference nor carryover was observed. Robust intra- and inter-assay accuracy and precision <15 % at five different concentrations were obtained. Linear calibration lines with regressions, R (2) > 0.99, were achieved for all analytes. Short-term bench top, long-term plasma, and extract stability demonstrated that the distinct ceramides were stable in the conditions evaluated. The validity of the methodology was demonstrated by determining the precise ceramide concentrations in a small CAD case-control study. Thus, our LC-MS/MS methodology features simple sample preparation and short analysis time for accurate quantification of Cer d18:1/16:0, Cer d18:1/18:0, Cer d18:1/24:0, and Cer d18:1/24:1, designed for routine analysis.
  • Manca, Maria Laura; Solini, Anna; Haukka, Jani K.; Sandholm, Niina; Forsblom, Carol; Groop, Per Henrik; Ferrannini, Ele (2021)
    Background: Chronic kidney disease (CKD) shows different clinical features in Types1 (T1D) and 2 diabetes (T2D). Metabolomics have recently provided useful contribution to the identification of biomarkers of CKD progression in either form of the disease. However, no studies have so far compared plasma metabolomics between T1D and T2D in order to identify differential signatures of progression of estimated glomerular filtration rate (eGFR) decline. Methods: We used two large cohorts of T1D (from Finland) and T2D (from Italy) patients followed up to 7 and 3 years, respectively. In both groups, progression was defined as the top quartile of yearly decline in eGFR. Pooled data from the two groups were analysed by univariate and bivariate random forest (RF), and confirmed by bivariate partial least squares (PLS) analysis, the response variables being type of diabetes and eGFR progression. Results: In progressors, yearly eGFR loss was significantly larger in T2D [-5.3 (3.0), median (interquartile range)mL/min/1.73 m2/year] than T1D [-3.7 (3.1) mL/min/1.73 m2/year; P = 0.018]. Out of several hundreds, bivariate RF extracted 22 metabolites associated with diabetes type (all higher in T1D than T2D except for 5-methylthioadenosine, pyruvate and β-hydroxypyruvate) and 13 molecules associated with eGFR progression (all higher in progressors than non-progressors except for sphyngomyelin). Three of the selected metabolites (histidylphenylalanine, leucylphenylalanine, tryptophylasparagine) showed a significant interaction between disease type and progression. Only eight metabolites were common to both bivariate RF and PLS. Conclusions: Identification of metabolomic signatures of CKD progression is partially dependent on the statistical model. Dual analysis identified molecules specifically associated with progressive renal impairment in both T1D and T2D.
  • Kriikku, Pirkko; Rasanen, Ilpo; Ojanperä, Ilkka; Thelander, Gunilla; Kronstrand, Robert; Vikingsson, Svante (2020)
    Flualprazolam is a novel designer benzodiazepine, structurally related to alprazolam, flubromazolam and triazolam. In the last couple of years, it has been frequently detected in seizures and in forensic cases in Sweden and Finland. However, there is a lack of published blood concentrations for the drug, which presents difficulties when assessing its relevance for the cause of death. A quantitative method for the determination of flualprazolam in post-mortem blood was developed and validated, and subsequently used to analyse samples from 33 deaths previously screened as testing positive for flualprazolam in Sweden and Finland. Most of the cases in the study were accidental deaths (61 %) or suicides (18 %). The median (range) flualprazolam concentration was 18.0 (3.0-68) ng/g. The majority of the deceased were male (82 %) and the median age was 30 years. The median age in the Swedish cases was significantly higher (35 years) than in the Finnish cases (23 years) (p <0.05). Poly-drug use and particularly the concomitant use of flualprazolam and opioids were very common in the study population. Most of the cases that were positive for flualprazolam were fatal poisonings by a drug (N = 23), and in 13 cases, flualprazolam was implicated in the cause of death. Combining the resources of two countries in which all post-mortem toxicology is centralised provided a more comprehensive insight into the toxicology of flualprazolam. Research on novel psychoactive substances, such as flualprazolam, is required in order to be able to provide scientific evidence on the risks of these new substances for drug administration and potential users. (C) 2019 Elsevier B.V. All rights reserved.
  • Jonsson, Martina; Jestoi, Marika; Anthoni, Minna; Welling, Annikki; Loivamaa, Iida; Hallikainen, Ville; Kankainen, Matti; Lysoe, Erik; Koivisto, Pertti; Peltonen, Kimmo (2016)
    The mycotoxin enniatin B, a cyclic hexadepsipeptide produced by the plant pathogen Fusarium, is prevalent in grains and grain-based products in different geographical areas. Although enniatins have not been associated with toxic outbreaks, they have caused toxicity in vitro in several cell lines. In this study, the cytotoxic effects of enniatin B were assessed in relation to cellular energy metabolism, cell proliferation, and the induction of apoptosis in Balb 3T3 and HepG2 cells. The mechanism of toxicity was examined by means of whole genome expression profiling of exposed rat primary hepatocytes. Enniatin B altered cellular energy metabolism and reduced cell proliferation in Balb 3T3 and HepG2 cell lines. Furthermore, the proportion of apoptotic cell populations of Balb 3T3 cells slightly increased. On the other hand, enniatin B caused necrotic cell death in primary hepatocytes. Gene expression studies revealed the alteration of energy metabolism due to effects on mitochondrial organization and function and the assembly of complex I of the electron transport chain. (C) 2016 Elsevier Ltd. All rights reserved.
  • Haggman, J.; Christensen, J. M.; Mantysaari, E. A.; Juga, J. (2019)
    High-yielding cows may suffer from negative energy balance during early lactation, which can lead to ketosis and delayed ability of returning to cyclicity after calving. Fast recovery after calving is essential when breeding for improved fertility. Traditionally used fertility traits, such as the interval from calving to first insemination (CFI), have low heritabilities and are highly influenced by management decisions. Herd Navigator (TM) management program samples and analyses milk progesterone and beta-hydroxybutyrate (BHB) automatically during milking. In this study, the genetic parameters of endocrine fertility traits (measured from milk progesterone) and hyperketonemia (measured from milk BHB) in early lactation were evaluated and compared with traditional fertility traits (CFI, interval from calving to the last insemination and interval from first to last insemination) and the milk yield in red dairy cattle herds in Finland. Data included observations from 14 farms from 2014 to 2017. Data were analyzed with linear animal models using DMU software and analyses were done for first parity cows. Heritability estimates for traditional fertility traits were low and varied between 0.03 and 0.07. Estimated heritabilities for endocrine fertility traits (interval from calving to the first heat (CFH) and commencement of luteal activity (C-LA)) were higher than for traditional fertility traits (0.19 to 0.33). Five slightly different hyperketonemia traits divided into two or three classes were studied. Linear model heritability estimates for hyperketonemia traits were low, however, when the threshold model was used for binary traits the estimates became slightly higher (0.07 to 0.15). Genetic correlation between CFH and C-LA for first parity cows was high (0.97) as expected since traits are quite similar. Moderate genetic correlations (0.47 to 0.52) were found between the endocrine fertility traits and early lactation milk yield. Results suggest that the data on endocrine fertility traits measured by automatic systems is a promising tool for improving fertility, specifically when more data is available. For hyperketonemia traits, dividing values into three classes instead of two seemed to work better. Based on the current study and previous studies, where higher heritabilities have been found for milk BHB traits than for clinical ketosis, milk BHB traits are a promising indicator trait for resistance to ketosis and should be studied more. It is important that this kind of data from automatic devices is made available to recording and breeding organizations in the future.
  • Ware, Jennifer J.; Chen, Xiangning; Vink, Jacqueline; Loukola, Anu; Minica, Camelia; Pool, Rene; Milaneschi, Yuri; Mangino, Massimo; Menni, Cristina; Chen, Jingchun; Peterson, Roseann E.; Auro, Kirsi; Lyytikainen, Leo-Pekka; Wedenoja, Juho; Stiby, Alexander I.; Hemani, Gibran; Willemsen, Gonneke; Hottenga, Jouke Jan; Korhonen, Tellervo; Heliovaara, Markku; Perola, Markus; Rose, Richard J.; Paternoster, Lavinia; Timpson, Nic; Wassenaar, Catherine A.; Zhu, Andy Z. X.; Smith, George Davey; Raitakari, Olli T.; Lehtimaki, Terho; Kahonen, Mika; Koskinen, Seppo; Spector, Timothy; Penninx, Brenda W. J. H.; Salomaa, Veikko; Boomsma, Dorret I.; Tyndale, Rachel F.; Kaprio, Jaakko; Munafo, Marcus R. (2016)
    Genome-wide association studies (GWAS) of complex behavioural phenotypes such as cigarette smoking typically employ self-report phenotypes. However, precise biomarker phenotypes may afford greater statistical power and identify novel variants. Here we report the results of a GWAS meta-analysis of levels of cotinine, the primary metabolite of nicotine, in 4,548 daily smokers of European ancestry. We identified a locus close to UGT2B10 at 4q13.2 (minimum p = 5.89 x 10(-10) for rs114612145), which was consequently replicated. This variant is in high linkage disequilibrium with a known functional variant in the UGT2B10 gene which is associated with reduced nicotine and cotinine glucuronidation activity, but intriguingly is not associated with nicotine intake. Additionally, we observed association between multiple variants within the 15q25.1 region and cotinine levels, all located within the CHRNA5-A3-B4 gene cluster or adjacent genes, consistent with previous much larger GWAS using self-report measures of smoking quantity. These results clearly illustrate the increase in power afforded by using precise biomarker measures in GWAS. Perhaps more importantly however, they also highlight that biomarkers do not always mark the phenotype of interest. The use of metabolite data as a proxy for environmental exposures should be carefully considered in the context of individual differences in metabolic pathways.
  • Humaloja, Jaana; Vento, Maximo; Kuligowski, Julia; Andersson, Sture; Pineiro-Ramos, Jose David; Sanchez-Illana, Angel; Litonius, Erik; Jakkula, Pekka; Hästbacka, Johanna; Bendel, Stepani; Tiainen, Marjaana; Reinikainen, Matti; Skrifvars, Markus B. (2021)
    The products of polyunsaturated fatty acid peroxidation are considered reliable biomarkers of oxidative injury in vivo. We investigated ischemia-reperfusion-related oxidative injury by determining the levels of lipid peroxidation biomarkers (isoprostane, isofuran, neuroprostane, and neurofuran) after cardiac arrest and tested the associations between the biomarkers and different arterial oxygen tensions (PaO2). We utilized blood samples collected during the COMACARE trial (NCT02698917). In the trial, 123 patients resuscitated from out-of-hospital cardiac arrest were treated with a 10-15 kPa or 20-25 kPa PaO2 target during the initial 36 h in the intensive care unit. We measured the biomarker levels at admission, and 24, 48, and 72 h thereafter. We compared biomarker levels in the intervention groups and in groups that differed in oxygen exposure prior to randomization. Blood samples for biomarker determination were available for 112 patients. All four biomarker levels peaked at 24 h; the increase appeared greater in younger patients and in patients without bystander-initiated life support. No association between the lipid peroxidation biomarkers and oxygen exposure either before or after randomization was found. Increases in the biomarker levels during the first 24 h in intensive care suggest continuing oxidative stress, but the clinical relevance of this remains unresolved.
  • Kuitunen, Marja-Leena; Dutoit, Jean-Claude; Siegenthaler, Peter; Rapinoja, Marja-Leena; Vanninen, Paula Sinikka (2022)
    Sensitive and reliable analysis of alkylphosphonic acids (APAs) and 2-(N,N-dialkylamino)ethanesulfonic acids (SAs), the degradation products of chemical warfare agents (CWAs), is one of the most important tasks for verification of the Chemical Weapons Convention (CWC). Unambiguous identification of these chemicals is required in a variety of environmental matrices, including soil and water. These acids with low volatility are very polar, and efficient and reliable methylation methods for their derivatization are needed for analysis with gas chromatography-mass spectrometry (GC-MS). In this study, the derivatization conditions for trimethylsilyldiazomethane (TMSDAM) methylation were optimized for rapid GC-MS screening. Optimized methylation of APAs and SAs with TMSDAM was compared with methylation with diazomethane. The TMSDAM methylation of SAs and benzilic acid was further compared with silylation with N-methyl-N-(tert-butyldimethylsilyl)trifluoroacetamide. The significance and necessity of cation exchange prior to derivatization and analysis were tested on samples with a high inorganic background. A recommendation to use the method for methylation of water samples and aqueous extracts using TMSDAM is given. The robustness of the method was illustrated by the successful identification of APAs and SAs in aqueous samples from proficiency tests organized by the Organisation for the Prohibition of Chemical Weapons.
  • Omidi, Azam; Esterhuizen-Londt, Maranda; Pflugmacher, Stephan (2019)
    In lakes, cyanobacterial blooms are frequently associated with green algae and dominate the phytoplankton community in successive waves. In the present study, the interactions between Microcystis aeruginosa PCC 7806 and Desmodesmus subspicatus were studied to clarify the probable ecological significance of algal secondary metabolites; focusing on the role of cyanotoxin ‘microcystin-LR’(MC-LR). A dialysis co-cultivation technique was applied where M. aeruginosa was grown inside and D. subspicatus was cultured outside of the dialysis tubing. The concentration of the intra- and extracellular MC-LR and the growth of two species were measured at different time points over a period of one month. Additionally, the growth of the two species in the culture filtrate of one another and the effect of the purified MC-LR on the growth of the green alga were studied. The results indicated that the co-existing species could affect each other depending on the growth phases. Despite the early dominance of D. subspicatus during the logarithmic phase,M. aeruginosa suppressed the growth of the green alga at the stationary phase, which coincided with increased MC production and release. However, the inhibitory effects of Microcystis might be related to its other extracellular metabolites rather than, or possibly in addition to, MC
  • Jalanka, Jonna; Mattila, Eero; Jouhten, Hanne; Hartman, Jorn; de Vos, Willem M.; Arkkila, Perttu; Satokari, Reetta (2016)
    Background: Faecal microbiota transplantation (FMT) is an effective treatment for recurrent Clostridium difficile infection (rCDI). It restores the disrupted intestinal microbiota and subsequently suppresses C. difficile. The long-term stability of the intestinal microbiota and the recovery of mucosal microbiota, both of which have not been previously studied, are assessed herein. Further, the specific bacteria behind the treatment efficacy are also investigated. Methods: We performed a high-throughput microbiota profiling using a phylogenetic microarray analysis of 131 faecal and mucosal samples from 14 rCDI patients pre- and post-FMT during a 1-year follow-up and 23 samples from the three universal donors over the same period. Results: The FMT treatment was successful in all patients. FMT reverted the patients' bacterial community to become dominated by Clostridium clusters IV and XIVa, the major anaerobic bacterial groups of the healthy gut. In the mucosa, the amount of facultative anaerobes decreased, whereas Bacteroidetes increased. Post-FMT, the patients' microbiota profiles were more similar to their own donors than what is generally observed for unrelated subjects and this striking similarity was retained throughout the 1-year follow-up. Furthermore, the universal donor approach allowed us to identify bacteria commonly established in all CDI patients and revealed a commonly acquired core microbiota consisting of 24 bacterial taxa. Conclusions: FMT induces profound microbiota changes, therefore explaining the high clinical efficacy for rCDI. The identification of commonly acquired bacteria could lead to effective bacteriotherapeutic formulations. FMT can affect microbiota in the long-term and offers a means to modify it relatively permanently for the treatment of microbiota-associated diseases.
  • Pryazhnikov, Evgeny; Mugantseva, Ekaterina; Casarotto, Plinio; Kolikova, Julia; Fred, Senem Merve; Toptunov, Dmytro; Afzalov, Ramil; Hotulainen, Pirta; Voikar, Vootele; Terry-Lorenzo, Ryan; Engel, Sharon; Kirov, Sergei; Castren, Eero; Khiroug, Leonard (2018)
    Ketamine, a well-known anesthetic, has recently attracted renewed attention as a fast-acting antidepressant. A single dose of ketamine induces rapid synaptogenesis, which may underlie its antidepressant effect. To test whether repeated exposure to ketamine triggers sustained synaptogenesis, we administered a sub-anesthetic dose of ketamine (10 mg/kg i.p.) once-daily for 5 days, and repeatedly imaged dendritic spines of the YFP-expressing pyramidal neurons in somatosensory cortex of awake female mice using in vivo two-photon microscopy. We found that the spine formation rate became significantly higher at 72-132 h after the first ketamine injection (but not at 6-24 h), while the rate of elimination of pre-existing spines remained unchanged. In contrast to the net gain of spines observed in ketamine-treated mice, the vehicle-injected control mice exhibited a net loss typical for young-adult animals undergoing synapse pruning. Ketamine-induced spinogenesis was correlated with increased PSD-95 and phosphorylated actin, consistent with formation of new synapses. Moreover, structural synaptic plasticity caused by ketamine was paralleled by a significant improvement in the nest building behavioral assay. Taken together, our data show that subchronic low-dose ketamine induces a sustained shift towards spine formation.
  • Ruohtula, Terhi; de Goffau, Marcus C.; Nieminen, Janne K.; Honkanen, Jarno; Siljander, Heli; Hämäläinen, Anu-Maaria; Peet, Aleksandr; Tillmann, Vallo; Ilonen, Jorma; Niemelä, Onni; Welling, Gjalt W.; Knip, Mikael; Harmsen, Hermie J.; Vaarala, Outi (2019)
    Recent studies suggest that the cross-talk between the gut microbiota and human immune system during the first year of life is an important regulator of the later development of atopic diseases. We explored the changes in the gut microbiota, blood regulatory T cells, and atopic sensitization in a birth-cohort of Estonian and Finnish children followed from 3 to 36 months of age. We describe here an infant Treg phenotype characterized by high Treg frequency, the maturation of Treg population characterized by a decrease in their frequency accompanied with an increase in the highly activated Treg cells. These changes in Treg population associated first with the relative abundance of Bifidobacterium longum followed by increasing colonization with butyrate producing bacteria. High bifidobacterial abundance in the neonatal microbiota appeared to be protective, while colonization with Bacteroides and E. coli was associated with later risk of allergy. Estonian children with lower risk of IgE mediated allergic diseases than Finnish children showed an earlier maturation of the gut microbiota, detected as earlier switch to an increasing abundance of butyrate-producing bacteria, combined with an earlier maturation of Treg cell phenotype and total IgE production. The children with established allergic diseases by age 3 showed a decreased abundance of butyrate producing Faecalibacterium. These results suggest that as well as the maintenance of a bifidobacterial dominated gut microbiota is important during the first weeks of life, the overtake by butyrate producing bacteria seems to be a beneficial shift, which should not be postponed.