scholarly journals Impact of Tigecycline Versus Other Antibiotics on the Fecal Metabolome and on Colonization Resistance to Clostridium difficile in Mice

2017 ◽  
Vol 2 (1) ◽  
pp. 1 ◽  
Author(s):  
Robin L.P. Jump ◽  
David Kraft ◽  
Kelly Hurless ◽  
Alex Polinkovsky ◽  
Curtis J. Donskey
Keyword(s):  
Clostridium Difficile ◽  
Intestinal Microbiota ◽  
High Performance ◽  
The Other ◽  
Colonization Resistance ◽  
Bacterial Metabolites ◽  
Metabolic Functions ◽  
Short Period ◽  
Lipid Metabolites

Background: The glycylcycline antibiotic tigecycline may have a relatively low propensity to promote Clostridium difficile infection in part because it causes less disruption of the indigenous intestinal microbiota than other broad-spectrum antibiotics.  We used a mouse model to compare the compare the effects of tigecycline versus other commonly used antibiotics on colonization resistance to C. difficile and on metabolic functions of the intestinal microbiota.   Methods: To assess in vivo colonization resistance to C. difficile, mice were challenged with oral C. difficile spores 1, 7, or 12 days after completion of 3 days of treatment with subcutaneous saline, tigecycline, ceftriaxone, piperacillin-tazobactam, or linezolid.  Levels of bacterial metabolites in fecal specimens of mice treated with the same antibiotics were analyzed using non-targeted metabolic profiling by gas chromatograph (GC)/mass spectrometry (MS) and ultra-high performance liquid chromatography-tandem MS (UPLC-MS/MS).  Results:  All of the antibiotics disrupted colonization resistance to C. difficile when challenge occurred 2 days after treatment.  Only piperacillin/tazobactam and ceftriaxone-treated mice had disturbed colonization resistance at 7 days after treatment.  All of the antibiotics altered fecal metabolites in comparison to controls, but tigecycline caused significantly less alteration than the other antibiotics, including less suppression of multiple amino acids, bile acids, and lipid metabolites.   Conclusions:  Tigecycline and linezolid caused transient disruption of colonization resistance to C. difficile, whereas ceftriaxone and piperacillin/tazobactam caused disruption that persisted for 7 days post-treatment.  Tigecycline caused less profound alteration of fecal bacterial metabolites than the other antibiotics, suggesting that the relatively short period of disruption of colonization resistance might be related in part to reduced alteration of the metabolic functions of the microbiota

scholarly journals Targeted Metabolomics Analysis Identifies Intestinal Microbiota-Derived Urinary Biomarkers of Colonization Resistance in Antibiotic-Treated Mice

2017 ◽  
Vol 61 (8) ◽  
Author(s):  
Mark E. Obrenovich ◽  
MaryAnn Tima ◽  
Alex Polinkovsky ◽  
Renliang Zhang ◽  
Steven N. Emancipator ◽  
...  
Keyword(s):  
Intestinal Tract ◽  
Fold Increase ◽  
Urinary Metabolites ◽  
Complete Recovery ◽  
Colonization Resistance ◽  
Bacterial Metabolites ◽  
Content Type ◽  
Metabolic Functions ◽  
Liquid Chromatography Tandem Mass

ABSTRACT Antibiotics excreted into the intestinal tract may disrupt the microbiota that provide colonization resistance against enteric pathogens and alter normal metabolic functions of the microbiota. Many of the bacterial metabolites produced in the intestinal tract are absorbed systemically and excreted in urine. Here, we used a mouse model to test the hypothesis that alterations in levels of targeted bacterial metabolites in urine specimens could provide useful biomarkers indicating disrupted or intact colonization resistance. To assess in vivo colonization resistance, mice were challenged with Clostridium difficile spores orally 3, 6, and 11 days after the completion of 2 days of treatment with piperacillin-tazobactam, aztreonam, or saline. For concurrent groups of antibiotic-treated mice, urine samples were analyzed by using liquid chromatography-tandem mass spectrometry (LC-MS/MS) to quantify the concentrations of 11 compounds targeted as potential biomarkers of colonization resistance. Aztreonam did not affect colonization resistance, whereas piperacillin-tazobactam disrupted colonization resistance 3 days after piperacillin-tazobactam treatment, with complete recovery by 11 days after treatment. Three of the 11 compounds exhibited a statistically significant and >10-fold increase (the tryptophan metabolite N-acetyltryptophan) or decrease (the plant polyphenyl derivatives cinnamoylglycine and enterodiol) in concentrations in urine 3 days after piperacillin-tazobactam treatment, followed by recovery to baseline that coincided with the restoration of in vivo colonization resistance. These urinary metabolites could provide useful and easily accessible biomarkers indicating intact or disrupted colonization resistance during and after antibiotic treatment.

scholarly journals A Pilot Study on Characteristics of Metabolomics and Lipidomics according to Sasang Constitution

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Min Jung Kim ◽  
Da-Hye Lee ◽  
Jiyun Ahn ◽  
Tae-Youl Ha ◽  
Young Jin Jang ◽  
...  
Keyword(s):  
Pilot Study ◽  
High Performance ◽  
The Other ◽  
Metabolomics Data ◽  
Flight Mass Spectrometry ◽  
Sasang Constitution ◽  
Canonical Pathways ◽  
Phenylalanine Metabolism ◽  
Lipid Metabolites

Although classification of an individual’s Sasang constitution is a key step in the prescription of traditional Korean medicine, the classifying process is complex and not objective. Identification of metabolic-based biomarkers could allow the development of a reliable and sensitive classification technique and even therapeutic management. Our pilot study investigated whether metabolites in plasma are characteristic of Sasang constitutions. Ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry-based metabolic analysis was conducted against 15 Soyangin (SY), 15 Taeeumin (TE), and 18 Soeumin (SE) individuals, as classified according to the Questionnaire for Sasang Constitution Classification II (QSCC II) and specialist diagnosis. Metabolomics data showed that the TE group was significantly separated from the SY and SE groups. Nine canonical pathways related to constitution; phenylalanine metabolism, aminoacyl-tRNA, tyrosine, and tryptophan biosynthesis were activated in the TE group as compared with the other groups. Similar to the results of the metabolomics analysis, the TE group was also significantly separated from the other two groups by lipidomic analysis. On the other hand, the intensity of lipid metabolites was higher in the SY group than in the other groups. Our findings suggest that the combined analysis of metabolomics and lipidomics can provide useful information for characteristics of Sasang constitutions.

Selection against sprouting damage in wheat. III.* Dormancy, germinative alpha-amylase, grain redness and flavanols

1979 ◽  
Vol 30 (3) ◽  
pp. 387 ◽  
Author(s):  
IL Gordon
Keyword(s):  
The Other ◽  
Grain Development ◽  
Germination Inhibitors ◽  
Development Patterns ◽  
White Wheat ◽  
Long Period ◽  
Short Period ◽  
Embryo Dormancy ◽  
Different Levels

Grain development of embryo dormancy, germinative α-amylase, pigmentation and flavanols was examined in the wheats Timgalen and Gamut (white-grained, non-dormant), and Pembina and Sonora (red-grained with different levels of dormancy). It was found that each trait had distinctive patterns of development. The net result at harvest ripeness depended on the synchronizations amongst the traits. Dormancy, as judged by embryo response (i.e. embryo dormancy), was restricted to the red wheats. Three ways of expressing it were noted: (1) in terms of development patterns, (2) as levels at harvest ripeness or at harvest, and (3) by the length of the period of embryo dormancy after harvest ripeness. Two hypotheses linking embryo dormancy and grain redness appeared plausible from the results. One was that the polyphenol oxidase complex, which polymerizes flavanols to the putative pigment phlobaphene, contributes towards embryo dormancy, probably through enhancement of hypo-oxia. The other was that the pigment itself and its tanning complexes cause the hypo-oxia. Flavanols did not appear to be in vivo germination inhibitors. Dormancy, as judged by α-amylase response (i.e. amylase dormancy), was not always present together with embryo dormancy. A long period of amylase dormancy was found in the more embryo-dormant red wheat, but not in the other. Conversely, a short period of amylase dormancy was found in one white wheat, but it was not embryo-dormant. Possible relationships between these physiological traits and the classical genes for red grain-coat were discussed. Implications concerning selection against sprouting damage were considered. _________________ *Part II, Aust. J. Agric. Res 30: 1-17 (1979).

scholarly journals Clostridium difficile Colonizes Alternative Nutrient Niches during Infection across Distinct Murine Gut Microbiomes

mSystems ◽  
2017 ◽  
Vol 2 (4) ◽  
Author(s):  
Matthew L. Jenior ◽  
Jhansi L. Leslie ◽  
Vincent B. Young ◽  
Patrick D. Schloss
Keyword(s):  
Clostridium Difficile ◽  
Nosocomial Infection ◽  
Diarrheal Disease ◽  
Nutrient Requirements ◽  
Colonization Resistance ◽  
Nutrient Sources ◽  
Infection Susceptibility ◽  
Life Threatening ◽  
Multiple Groups

ABSTRACT Infection by the bacterium Clostridium difficile causes an inflammatory diarrheal disease which can become life threatening and has grown to be the most prevalent nosocomial infection. Susceptibility to C. difficile infection is strongly associated with previous antibiotic treatment, which disrupts the gut microbiota and reduces its ability to prevent colonization. In this study, we demonstrated that C. difficile altered pathogenesis between hosts pretreated with antibiotics from separate classes and exploited different nutrient sources across these environments. Our metabolite score calculation also provides a platform to study nutrient requirements of pathogens during an infection. Our results suggest that C. difficile colonization resistance is mediated by multiple groups of bacteria competing for several subsets of nutrients and could explain why total reintroduction of competitors through fecal microbial transplant currently is the most effective treatment for recurrent CDI. This work could ultimately contribute to the identification of targeted, context-dependent measures that prevent or reduce C. difficile colonization, including pre- and probiotic therapies. Clostridium difficile is the largest single cause of hospital-acquired infection in the United States. A major risk factor for Clostridium difficile infection (CDI) is prior exposure to antibiotics, as they disrupt the gut bacterial community which protects from C. difficile colonization. Multiple antibiotic classes have been associated with CDI susceptibility, many leading to distinct community structures stemming from variation in bacterial targets of action. These community structures present separate metabolic challenges to C. difficile. Therefore, we hypothesized that the pathogen adapts its physiology to the nutrients within different gut environments. Utilizing an in vivo CDI model, we demonstrated that C. difficile highly colonized ceca of mice pretreated with any of three antibiotics from distinct classes. Levels of C. difficile spore formation and toxin activity varied between animals based on the antibiotic pretreatment. These physiologic processes in C. difficile are partially regulated by environmental nutrient concentrations. To investigate metabolic responses of the bacterium in vivo, we performed transcriptomic analysis of C. difficile from ceca of infected mice across pretreatments. This revealed heterogeneous expression in numerous catabolic pathways for diverse growth substrates. To assess which resources C. difficile exploited, we developed a genome-scale metabolic model with a transcriptome-enabled metabolite scoring algorithm integrating network architecture. This platform identified nutrients that C. difficile used preferentially between pretreatments, which were validated through untargeted mass spectrometry of each microbiome. Our results supported the hypothesis that C. difficile inhabits alternative nutrient niches across cecal microbiomes with increased preference for nitrogen-containing carbon sources, particularly Stickland fermentation substrates and host-derived glycans. IMPORTANCE Infection by the bacterium Clostridium difficile causes an inflammatory diarrheal disease which can become life threatening and has grown to be the most prevalent nosocomial infection. Susceptibility to C. difficile infection is strongly associated with previous antibiotic treatment, which disrupts the gut microbiota and reduces its ability to prevent colonization. In this study, we demonstrated that C. difficile altered pathogenesis between hosts pretreated with antibiotics from separate classes and exploited different nutrient sources across these environments. Our metabolite score calculation also provides a platform to study nutrient requirements of pathogens during an infection. Our results suggest that C. difficile colonization resistance is mediated by multiple groups of bacteria competing for several subsets of nutrients and could explain why total reintroduction of competitors through fecal microbial transplant currently is the most effective treatment for recurrent CDI. This work could ultimately contribute to the identification of targeted, context-dependent measures that prevent or reduce C. difficile colonization, including pre- and probiotic therapies.

scholarly journals The cbiS Gene of the Archaeon Methanopyrus kandleri AV19 Encodes a Bifunctional Enzyme with Adenosylcobinamide Amidohydrolase and α-Ribazole-Phosphate Phosphatase Activities

2006 ◽  
Vol 188 (12) ◽  
pp. 4227-4235 ◽  
Author(s):  
Jesse D. Woodson ◽  
Jorge C. Escalante-Semerena
Keyword(s):  
High Performance ◽  
Biochemical Characterization ◽  
The Other ◽  
Bifunctional Enzyme ◽  
Visible Spectroscopy ◽  
Uv Visible ◽  
Cobyric Acid ◽  
Methanopyrus Kandleri

ABSTRACT Here we report the initial biochemical characterization of the bifunctional α-ribazole-P (α-RP) phosphatase, adenosylcobinamide (AdoCbi) amidohydrolase CbiS enzyme from the hyperthermophilic methanogenic archaeon Methanopyrus kandleri AV19. The cbiS gene encodes a 39-kDa protein with two distinct segments, one of which is homologous to the AdoCbi amidohydrolase (CbiZ, EC 3.5.1.90) enzyme and the other of which is homologous to the recently discovered archaeal α-RP phosphatase (CobZ, EC 3.1.3.73) enzyme. CbiS function restored AdoCbi salvaging and α-RP phosphatase activity in strains of the bacterium Salmonella enterica where either step was blocked. The two halves of the cbiS genes retained their function in vivo when they were cloned separately. The CbiS enzyme was overproduced in Escherichia coli and was isolated to >95% homogeneity. High-performance liquid chromatography, UV-visible spectroscopy, and mass spectroscopy established α-ribazole and cobyric acid as the products of the phosphatase and amidohydrolase reactions, respectively. Reasons why the CbiZ and CobZ enzymes are fused in some archaea are discussed.

Immunologie Studies in von Willebrand’s Disease

1976 ◽  
Vol 35 (01) ◽  
pp. 110-119 ◽  
Author(s):  
Y Sultan ◽  
J Simeon ◽  
P Maisonneuve ◽  
J. P Caen
Keyword(s):  
Factor Viii ◽  
Bleeding Time ◽  
The Other ◽  
Von Willebrand's Disease ◽  
Factor Viii Related Antigen ◽  
Von Willebrand’S Disease ◽  
Short Period ◽  
Von Willebrand ◽  
Immunologic Properties

SummaryTwo patients with a severe von Willebrand’s disease characterized by no detectable factor VIII related antigen in their plasma received transfusions of cryoprecipitate. The bleeding time was corrected for a short period of time and returned to its pretransfusional value although the other parameters of the disease were still corrected. Electrophoretic and immunologic properties of factor VIII related antigen infused were determined serially after transfusion. Modifications of these properties occurred progressively after transfusion. The half disappearance time of F. VIIIR. A. was determined and found to be considerably shorter than in hemophilic recipients. This study suggests an alteration in vivo of F. VIIIR. A. infused into von Willebrand recipients.

scholarly journals Biodistribution and Anticancer Characteristics of Les-3833, A Novel 4-thiazolidinone-Based Lead Compound

2020 ◽  
Vol 88 (2) ◽  
pp. 18 ◽  
Author(s):  
Lesya Kobylinska ◽  
Andrii Lozynskii ◽  
Roman Lesyk ◽  
Rostyslav Stoika ◽  
Sandor G. Vari
Keyword(s):  
Maximum Concentration ◽  
High Performance ◽  
Maximum Level ◽  
Lead Compound ◽  
Short Period ◽  
Liquid Chromatography Tandem Mass ◽  
Liver And Kidney ◽  
The Brain

Recently, we identified the promising anticancer potential of the synthetic 4-thiazolidinone-based anticancer lead compound Les-3833 which demonstrated tumor-suppressing action in vitro and in vivo. Based on the results of previous studies, the aim of this research was to investigate the cytotoxicity in vitro and the biodistribution in laboratory mice to support the biotherapeutic drug development of Les-3833. Les-3833 (2.5 mg/kg) was intravenously injected into male Balb/c mice. Measurements were performed at 5 min, 15 min, 1 h, 4 h, and 24 h time points in blood plasma, brain, liver, and kidney using high-performance liquid chromatography/tandem mass spectrometry. After the administration of Les-3833, the maximum level of this compound was observed in plasma at 2.08 min. In the brain, the mean maximum concentration of Les-3833 was 7.17 ng/mL at 5 min, while after 15 min, it was not found. In the liver, at 5 min, the maximum concentration was 1190 ng/g. At 15 min, concentration of Les-3833 in the liver decreased by 14.3%; at 6 h by 22.8%; and after 24 h by 64.7%. Its maximum concentration in kidney was 404 ng/g within 5–15 min, at 1 h it decreased by 36.1%, and after 24 h by 49.3%. Thus, Les-3833 was rapidly taken up by different organs from the bloodstream, partially metabolized in the liver, and excreted mainly through the kidneys, while in the brain, a very low concentration could be observed for only a short period of time.

THE EFFECT OF LOCAL TEMPERATURE ON THE REACTIVITY TO NORADRENALINE OF DIGITAL VESSELS

1967 ◽  
Vol 45 (1) ◽  
pp. 93-102 ◽  
Author(s):  
Peter Gaskell ◽  
David L. Hoeppner
Keyword(s):  
Intravenous Infusion ◽  
The Other ◽  
Local Temperature ◽  
Continuous Intravenous Infusion ◽  
Opening Pressure ◽  
Simultaneous Measurements ◽  
Critical Opening ◽  
Short Period ◽  
The Mean

The effect of local temperature on the in vivo reactivity of vascular smooth muscle was studied. Reactivity was measured as the increase in critical opening pressure (COP) of digital vessels caused by intravenous infusion of 5 μg of noradrenaline per minute. With one hand cool (22 °C) and the other warm (34 °C) in test experiments or both hands either cool or warm in control experiments, simultaneous measurements were made of the increase in COP of vessels in both middle fingers in response to the noradrenaline. In control experiments the mean increase in COP was similar in right and left fingers, but in test experiments the mean increase was greater in the warm finger than in the cool one. Warm vessels were more reactive to noradrenaline than cool ones (p < 0.01). Because a short period of ischemia is involved in the measurement of COP, other experiments were performed in which the effect of duration of ischemia on the COP, with and without a continuous intravenous infusion of noradrenaline, was ascertained. They suggested that the estimated COP would, in most cases, be about 3 mm Hg less than the COP existing just before the measurement. These results also indicated that although the rate of fall of COP during ischemia was slightly greater for a higher initial COP, the ischemia involved did not invalidate the comparison of the increases in COP caused by noradrenaline in warm and cool fingers, as an index of relative arteriolar reactivity in the test reactivity experiments.

scholarly journals Rapid, Refined, and Robust Method for Expression, Purification, and Characterization of Recombinant Human Amyloid beta 1-42

2019 ◽  
Vol 2 (2) ◽  
pp. 48 ◽  
Author(s):  
Priya Prakash ◽  
Travis C. Lantz ◽  
Krupal P. Jethava ◽  
Gaurav Chopra
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Amyloid Beta ◽  
High Performance ◽  
Force Microscopy ◽  
Atomic Force ◽  
Short Period ◽  
One Step

Amyloid plaques found in the brains of Alzheimer’s disease patients primarily consists of amyloid beta 1-42 (Aβ42). Commercially, Aβ42 is synthesized using high-throughput peptide synthesizers resulting in the presence of impurities and the racemization of amino acids that affects its aggregation properties. Furthermore, the repeated purchase of even a small quantity (~1 mg) of commercial Aβ42 can be expensive for academic researchers. Here, we describe a detailed methodology for robust expression of recombinant human Aβ(M1-42) in Rosetta(DE3)pLysS and BL21(DE3)pLysS competent E. coli using standard molecular biology techniques with refined and rapid one-step analytical purification techniques. The peptide is isolated and purified from transformed cells using an optimized reverse-phase high-performance liquid chromatography (HPLC) protocol with commonly available C18 columns, yielding high amounts of peptide (~15–20 mg per 1 L culture) within a short period of time. The recombinant human Aβ(M1-42) forms characteristic aggregates similar to synthetic Aβ42 aggregates as verified by western blotting and atomic force microscopy to warrant future biological use. Our rapid, refined, and robust technique produces pure recombinant human Aβ(M1-42) that may be used to synthesize chemical probes and in several downstream in vitro and in vivo assays to facilitate Alzheimer’s disease research.

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