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1.
MEHMET ÇI˙FTÇI˙ YAVUZ DEMI˙R I˙SMAIL ÖZMEN ÖKKEŞ ATICI 《Journal of enzyme inhibition and medicinal chemistry》2013,28(1):71-76
The present study was undertaken to determine in vivo and in vitro effects of some plant growth regulators on rat erythrocyte carbonic anhydrase (CA) and glucose-6-phosphate dehydrogenase (G6PD) activities. Both in vivo and in vitro, spermidine and kinetin did not affect enzymatic activities of CA and G6PD, whereas putrescine decreased these activities, and abscisic acid increased them. Since plants use such growth regulators, their effects should be considered on mammals consuming them since they may possess important biological effects. 相似文献
2.
Carl PC Chen Chih-Chin Hsu Wen-Lin Yeh Hsiu-Chu Lin Sen-Yung Hsieh Shih-Cherng Lin Tai-Tzung Chen Max JL Chen Simon FT Tang 《Proteome science》2011,9(1):1-10
Background
Prenatal screening for Down Syndrome (DS) would benefit from an increased number of biomarkers to improve sensitivity and specificity. Improving sensitivity and specificity would decrease the need for potentially risky invasive diagnostic procedures.Results
We have performed an in depth two-dimensional difference gel electrophoresis (2D DIGE) study to identify potential biomarkers. We have used maternal plasma samples obtained from first and second trimesters from mothers carrying DS affected fetuses compared with mothers carrying normal fetuses. Plasma samples were albumin/IgG depleted and expanded pH ranges of pH 4.5 - 5.5, pH 5.3 - 6.5 and pH 6 - 9 were used for two-dimensional gel electrophoresis (2DE). We found no differentially expressed proteins in the first trimester between the two groups. Significant up-regulation of ceruloplasmin, inter-alpha-trypsin inhibitor heavy chain H4, complement proteins C1s subcomponent, C4-A, C5, and C9 and kininogen 1 were detected in the second trimester in maternal plasma samples where a DS affected fetus was being carried. However, ceruloplasmin could not be confirmed as being consistently up-regulated in DS affected pregnancies by Western blotting.Conclusions
Despite the in depth 2DE approach used in this study the results underline the deficiencies of gel-based proteomics for detection of plasma biomarkers. Gel-free approaches may be more productive to increase the number of plasma biomarkers for DS for non-invasive prenatal screening and diagnosis. 相似文献3.
Neurodegenerative diseases are caused by proteinaceous aggregates, usually consisting of misfolded proteins which are often typified by a high proportion of β-sheets that accumulate in the central nervous system. These diseases, including Morbus Alzheimer, Parkinson disease and Transmissible Spongiform Encephalopathies (TSEs)—also termed prion disorders—afflict a substantial proportion of the human population and, as such, the etiology and pathogenesis of these diseases has been the focus of mounting research. Although many of these diseases arise from genetic mutations or are sporadic in nature, the possible horizontal transmissibility of neurodegenerative diseases poses a great threat to population health. In this article we discuss recent studies that suggest that the “non-transmissible” status bestowed upon Alzheimer and Parkinson diseases may need to be revised as these diseases have been successfully induced through tissue transplants. Furthermore, we highlight the importance of investigating the “natural” mechanism of prion transmission including peroral and perenteral transmission, proposed routes of gastrointestinal uptake and neuroinvasion of ingested infectious prion proteins. We examine the multitude of factors which may influence oral transmissibility and discuss the zoonotic threats that Chronic Wasting disease (CWD), Bovine Spongiform Encephalopathy (BSE) and Scrapie may pose resulting in vCJD or related disorders. In addition, we suggest that the 37 kDa/67 kDa laminin receptor on the cell surface of enterocytes, a major cell population in the intestine, may play an important role in the intestinal pathophysiology of alimentary prion infections.Key words: prion, 37 kDa/67 kDa laminin receptor, CJD, BSE, CWD, scrapie, Alzheimer disease, Parkinson disease, intestine, enterocytesMany different mechanisms exist which underlie the etiology of the numerous neurodegenerative diseases affecting the human population. Amongst the most prominent are Morbus Alzheimer, prion disorders, Parkinson disease, Chorea Huntington, frontotemporal dementia and amylotrophic lateral sclerosis. The molecular mechanisms underlying these diseases vary; however, all neurodegenerative diseases share a common feature: they are caused by protein aggregation. The only neurodegenerative diseases proven to be transmissible are prion disorders. In contrast to frontotemporal dementia, recent evidence suggests that Alzheimer and Parkinson diseases may also be transmissible. Pre-symptomatic Alzheimer disease (APP23) mice exhibited an increase in the Alzheimer phenotype when brain homogenate of autopsied human Alzheimer disease patients and older, amyloid beta- (Aβ-) laden APP23 mice was injected into their hippocampi.1 These findings suggest that the Aβ-abundant brain homogenate of Alzheimer disease patients may possess the ability to induce or supplement the overproduction of Aβ, possibly leading to the onset of Alzheimer disease.The pathological feature associated with Parkinson disease is the formation of Lewy bodies in cell bodies and neuronal processes in the brain.2 The main component of these protein aggregates is α-synuclein (reviewed in ref. 2). Autopsies of Parkinson disease patients revealed that Lewy bodies had formed on healthy embryonic neurons that had been grafted onto the brain tissue of the patients several years before (prior to said examination).3–5 It may thus be proposed that α-synuclein transmission is possible from diseased to healthy neurons, suggesting that Parkinson disease may be transmissible from a Parkinson disease patient to a healthy individual. These findings imply that Alzheimer and Parkinson diseases may be transmissible through tissue transplants and the use of contaminated surgical tools.6Prion disorders, also termed Transmissible Spongiform Encephalopathies (TSEs), are fatal neurodegenerative diseases that affect the central nervous system (CNS) of multiple animal species. In lieu of the social, economic and political ramifications of such infections, as well as the possible intra- and interspecies transmissibility of such disorders, various routes of experimental transmission have been investigated including intracerebral, intraperitoneal, intraventricular, intraocular, intraspinal and subcutaneous injections (reviewed in ref. 7–9). However, such routes of transmission are not representative of the “natural” mechanism as the majority of prion disorders are contracted through ingestion of infectious prion (PrPSc) containing material. Thus, the peroral and perenteral prion transmission is of greatest consequence with respect to TSE disease establishment. Moreover, the presence of PrPSc in the buccal cavity of scrapie-infected sheep10 (reviewed in ref. 11) and the possible horizontal transfer as a result hereof, as may be similarly proposed for animals suffering from other TSEs, may further contribute to the oral transmissibility of TSEs.A number of model systems have been employed to study TSE transmissibility. Owing to ethical constraints, TSE transmissibility to humans via the oral route may not be directly investigated and as a result hereof, alternative model systems are needed. These may include the use of transgenic mice, cell lines which are permissive to infection12 and experimental animals such as sheep, calves, goats, minks, ferrets and non-human primates (reviewed in ref. 9).Intestinal entry of PrPSc has been proposed to occur via two pathways, the membranous (M) cell-dependent and M cell-independent pathways (Fig. 1).13,14 The former involves endocytic M (microfold)-cells, which cover the intestinal lymphoid follicles (Peyer''s patches)14 and may take up prions and thereby facilitate the translocation of these proteins across the intestinal epithelium into the lymphoid tissues (reviewed in ref. 9) as has been demonstrated in a cellular model.13 Following such uptake by the M cells, the prions may subsequently pass to the dendritic cells and follicular dendritic cells (FDCs) (Fig. 1), which allow for prion transport to the mesenteric lymph nodes and replication, respectively.15 The prion proteins may subsequently gain access to the enteric nervous system (ENS) and ultimately the central nervous system (CNS).15Open in a separate windowFigure 1Proposed routes of gastrointestinal entry of ingested infectious prions (PrPSc) as well as possible pathways of amplification and transport to the central nervous system.However, prion intestinal translocation has been observed in the absence of M cells and has been demonstrated to be as a result of the action of polar, 37 kDa/67 kDa LRP/LR (non-integrin laminin receptor; reviewed in ref. 16–18) expressing enterocytes. Enterocytes are the major cell population of the intestinal epithelium and due to their ability to endocytose pathogens, nutrients and macromolecules,19 it has been proposed that these cells may represent a major entry site for alimentary prions (Fig. 1).Since enterocyte prion uptake has been demonstrated to be dependent on the presence of LRP/LR on the apical brush border of the cells,14,20 the interaction between varying prion protein strains and the receptor21–23 may be employed as a model system to study possible oral transmissibility of prion disorders across species as well as the intestinal pathophysiology of alimentary prion infections.24 Moreover, the blockage of such interactions through the use of anti-LRP/LR specific antibodies has been reported to reduce PrPSc endocytosis19 and thus these antibodies may serve as potential therapeutics to prevent infectious prion internalization and thereby prevent prion infections. It must be emphasized that the adhesion of prion proteins to cells is not solely dependent on the LRP/LR-PrPSc interactions;24 however, this interaction is of importance with regards to internalization and subsequent pathogenesis.We applied the aforementioned cell model to study the possible oral transmission of PrPBSE, PrPCWD and ovine PrPSc to cervids, cattle, swine and humans.24 The direct transmission of the aforementioned animal prion disorders to humans as a result of dietary exposure and the possible establishment of zoonotic diseases is of great public concern. It must however be emphasized that the study investigated the co-localization of LRP/LR and various prion strains and not the actual internalization process.PrPBSE was shown to co-localize with LRP/LR on human enterocytes24, thereby suggesting that PrPBSE is transmissible to humans via the oral route which is widely accepted as the manner by which variant CJD originated. This suspicion was previously investigated using a macaque model, which was successfully perorally infected by BSE-contaminated material and subsequently lead to the development of a prion disorder that resembles vCJD.25 These results, due to the evolutionary relatedness between macaques and humans, allowed researchers to confirm the oral transmissibility of PrPBSE to humans. PrPBSE may also potentially lead to prion disorder establishment in swine,24 livestock of great economic and social importance.The prion disorder affecting elk, mule deer and white-tailed deer is termed CWD. Cases of the disease are most prevalent in the US but are also evident in Canada and South Korea.26,27 As the infectious prion isoform is reported to be present in the blood28 and skeletal muscle,29 hunting, consumption of wild venison and contact with other animal products derived from CWD-infected elk and deer may thereby pose a public health risk. Our studies demonstrate that PrPCWD co-localizes with LRP/LR on human enterocytes24 thereby suggesting a possible oral transmissibilty of this TSE to humans. This is, however, inconsistent with results obtained during intra-cerebral inoculation of the brains and spinal cords of transgenic mice overexpressing the human cellular prion protein (PrPc),26,27 which is essential for TSE disease establishment and progression. Further, discrepancies have also been reported with respect to non-human primates, as squirrel monkeys have been successfully intracerebrally inoculated with mule-deer prion homogenates,30 while cynolmolgus macaques were resistant to infection.31 CWD has been transmitted to ferrets, minks and goats32 and as these animals may serve as domestic animals or livestock, secondary transmission from such animals to humans, through direct contact or ingestion of infected material, may be an additional risk factor that merits further scientific investigation.Ovine PrPSc co-localization with LRP/LR on human and bovine enterocytes may be indicative of the infectious agents'' ability to effect cross-species infections. The oral transmissibility of Scrapie has been confirmed in hamsters fed with sheep-scrapie-infected material.33The discrepancies with regards to the transmissibility of certain infectious prion proteins when assessed by different model systems may be due to the experimental transmission route employed. Oral exposure often results in significantly prolonged incubation times when compared to intracerebral inoculation techniques and thus failure of transgenic mice and normal experimental animals to develop disease phenotypes after being fed TSE-contaminated material may not necessarily indicate that the infection process failed.14 Apart from the route of infection, numerous other factors may influence transmission between species, including dose, PrP polymorphisms and genetic factors, the prion strain employed as well as the efficacy of prion transport to the CNS.34 The degree of homology between the PrPc protein in the animals serving as the infectious prion source and recipient has also been described as a feature limiting cross-species transmission.34 The negative results, as referred to above, obtained upon prion-protein inoculation of animal models may have resulted due to the slow rate at which the infectious prion induces conformational conversion of the endogenous PrPc in the animal cells and this in turn results in low levels of infectious prion replication and symptom development.27Furthermore, even in the event that certain prion disorders are not directly transmissible to humans, most are transmissible to at least a single species of domestic animal or livestock. The infectious agents properties may be altered in the secondary host such that it becomes transmissible to humans (reviewed in ref. 35). Thus, interspecies transmission between animals may indirectly influence human health.It is noteworthy to add that although the oral route of PrPSc transmission may result in prolonged incubation times, it may broaden the range of susceptible hosts. A common constituent of food is ferritin, a protein that is resistant to digestive enzyme hydrolysis and, due to its homology across species, it may serve as co-transporter of PrPSc and facilitate enterocyte internalization of the infectious prion.36 It may thus be proposed that prion internalization may occur via a ferritin-PrPSc complex even in the absence of co-localization between the infectious agent and LRP/LR such that many more cross-species infections (provided that the other infection factors are favorable) may be probable.37 In addition, digestive enzymes in the gastrointestinal tract facilitate PrPSc binding to the intestinal epithelium and subsequent intestinal uptake36 and thus depending on the individuals'' digestive processes, the susceptibility to infection and the rate of disease development may vary accordingly. As a result hereof, though laboratory experiments in cell-culture and animal models may render a particular prion disorder non-infectious to humans, this may not be true for all individuals.In lieu of the above statements, with particular reference to inconsistencies in reported results and the multiple factors influencing oral transmissibility of TSEs, further transmission studies are required to evaluate the zoonotic threat which CWD, BSE and Scrapie may pose through ingestion. 相似文献
4.
Differential interference contrast micrographs from stretched animals, serially sectioned semi-thin and ultrathin sections revealed that the cerebral ganglia (supraoesophageal mass) of the eulardigrade Milnesium tardigradum lie above the buccal tube and adjacent tissue like a saddle. It has an anterior indentation which is penetrated by two muscles that arise from the cuticle of the forehead. The cerebral ganglia consist of lateral outer lobes bearing an eye on each side, and two inner lobes which extend caudally. Between the inner lobes a cone-like projection tapers into a nerve bundle. Each outer lobe is joined with the first ventral ganglion. From the outer lobe near the eye the ganglion for a posterolateral sensory field extends to the epidermis. Anterior to the supraoesophageal mass are three dorsal ganglia for the upper three peribuccal papillae. Two additional ganglia attached to the cerebral mass supply the lateral cephalic papillae. The cerebral ganglia are covered by a thin neural lamella. The pericarya which surround the neuropil have large nuclei. Near the axons in the centre of the supraoesophageal mass the cytoplasm is crowded with vesicles of different size and appearance. Some of them resemble synaptic vesicles while others resemble dense core bodies. Structurally different types of synapses and axons can be distinguished within the neuropil. 相似文献
5.
Catimel B; Scott AM; Lee FT; Hanai N; Ritter G; Welt S; Old LJ; Burgess AW; Nice EC 《Glycobiology》1998,8(9):927-938
We describe a novel immobilization technique to investigate interactions
between immobilized gangliosides (GD3, GM1, and GM2) and their respective
antibodies, antibody fragments, or binding partners using an optical
biosensor. Immobilization was performed by direct injection onto a
carboxymethyldextran sensor chip and did not require derivatization of the
sensor surface or the ganglioside. The ganglioside appeared to bind to the
sensor surface by hydrophobic interaction, leaving the carbohydrate epitope
available for antibody or, in the case of GM1, cholera toxin binding. The
carboxyl group of the dextran chains on the sensor surface did not appear
to be involved in the immobilization as evidenced by equivalent levels of
immobilization following conversion of the carboxyl groups into acyl amino
esters, but rather the dextran layer provided a hydrophilic coverage of the
sensor chip which was essential to prevent nonspecific binding. This
technique gave better reactivity and specificity for anti- ganglioside
monoclonal antibodies (anti-GD3: KM871, KM641, R24; and anti-GM2: KM966)
than immobilization by hydrophobic interaction onto a gold sensor surface
or photoactivated cross-linking onto carboxymethydextran. This rapid
immobilization procedure has facilitated detailed kinetic analysis of
ganglioside/antibody interactions, with the surface remaining viable for a
large number of cycles (>125). Kinetic constants were determined from
the biosensor data using linear regression, nonlinear least squares and
equilibrium analysis. The values of kd, ka, and KAobtained by nonlinear
analysis (KAKM871 = 1.05, KM641 = 1.66, R24 = 0.14, and KM966 = 0.65 x
10(7) M- 1) were essentially independent of concentration and showed good
agreement with data obtained by other analytical methods.
相似文献
6.
Metin BÜLBÜL Olcay HİSAR Şükrü BEYDEMİR Mehmet ÇİFTÇİ Ö. İrfan KÜFREVIOĞLU 《Journal of enzyme inhibition and medicinal chemistry》2013,28(4):371-375
The in vitro and in vivo inhibitory effects of 5-(3α, 12α-dihydroxy-5-β-cholanamido)-1,3,4-thiadiazole-2-sulfonamide (1), 5-(3α, 7α, 12α-trihydroxy-5-β-cholanamido)-1,3,4-thiadiazole-2-sulfonamide (2), 5-(3α, 7α, 12α-triacetoxy-5-β-cholanamido)-1,3,4-thiadiazole-2-sulfonamide (3) and acetazolamide on rainbow trout (Oncorhynchus mykiss) (RT) erythrocyte carbonic anhydrase (CA) were investigated. The RT erythrocyte CA was obtained by affinity chromatography with a yield of 20.9%, a specific activity of 422.5?EU/mg protein and a purification of 222.4-fold. The purity of the enzyme was confirmed by SDS-PAGE. Inhibitory effects of the sulfonamides and acetazolamide on the RT erythrocyte CA were determined using the CO2-Hydratase method in vitro and in vivo studies. From in vitro studies, it was found that all the compounds inhibited CA. The obtained I50 value for the sulfonamides (1), (2) and (3) and acetazolamide were 0.83, 0.049, 0.82 and 0.052?μM, respectively. From in vivo studies, it was observed that CA was inhibited by the sulfonamides (1), (2) and (3) and acetazolamide. 相似文献
7.
Quantitative analysis of sugar constituents of glycoproteins by capillary electrophoresis 总被引:4,自引:0,他引:4
A method for quantitative analysis of monosaccharides including N-
acetylneuraminic acid derived from sialic acid-containing oligosaccharides
and glycoproteins is presented. The analysis is based on the combination of
chemical and enzymatic methods coupled with capillary electrophoretic (CE)
separation and laser-induced fluorescence (LIF) detection. The present
method utilizes a simplified acid hydrolysis procedure consisting of mild
hydrolysis (0.1 M TFA) to release sialic acid and strong acid hydrolysis
(2.0 N TFA) to produce amino and neutral sugars. Amino sugars released from
strong acid hydrolysis of oligosaccharides and glycoproteins were
reacetylated and derivatized with 8-aminopyrene-1,3,6-trisulfonate (APTS)
along with neutral sugars in the presence of sodium cyanoborohydride to
yield quantitatively the highly stable fluorescent APTS adducts. N-
acetylneuraminic acid (Neu5Ac), a major component of most mammalian
glycoproteins, was converted in a fast specific reaction by the action of
neuraminic acid aldolase (N-acylneuraminate pyruvate-lyase EC 4.1.3.3) to
N-acetylmannosamine (ManNAc) and pyruvate. ManNAc was then derivatized with
APTS in the same manner as the other monosaccharides. This method was
demonstrated for the quantitation of pure Neu5Ac and the species derived
from mild acid hydrolysis of 6'-sialyl-N- acetyllactosamine and bovine
fetuin glycan. Quantitative recovery of the N-acetylmannosamine was
obtained from a known amount of Neu5Ac in a mixture of seven other
monosaccharides or from the sialylated oligosaccharides occurring in
glycoproteins. The sequence of procedures consists of acid hydrolysis,
enzymatic conversion and APTS derivatization which produced quantitative
recovery of APTS- monosaccharide adducts. The detection limits for sugars
derivatized with APTS and detected by CE-LIF are 100 pmol for Neu5Ac and 50
pmol for the other sugars.
相似文献
8.
Amyloid beta (Aβ) is a major causative agent of Alzheimer disease (AD). This neurotoxic peptide is generated as a result of the cleavage of the Amyloid-Precursor-Protein (APP) by the action of β-secretase and γ-secretase. The neurotoxicity was previously thought to be the result of aggregation. However, recent studies suggest that the interaction of Aβ with numerous cell surface receptors such as N-methyl-D-aspartate (NMDA), receptor for advanced glycosylation end products (RAGE), P75 neurotrophin receptor (P75NTR) as well as cell surface proteins such as the cellular prion protein (PrPc) and heparan sulfate proteoglycans (HSPG) strongly enhances Aβ induced apoptosis and thereby contributes to neurotoxicity. This review focuses on the molecular mechanism resulting in Aβ-shedding as well as Aβ-induced apoptotic processes, genetic risk factors for familial AD and interactions of Aβ with cell surface receptors and proteins, with particular emphasis on the cellular prion protein. Furthermore, comparisons are drawn between AD and prion disorders and the role of laminin, an extracellular matrix protein, glycosaminoglycans and the 37 kDa/67 kDa laminin receptor (LRP/LR) have been highlighted with regards to both neurodegenerative diseases.Key words: Alzheimer disease, amyloid β, apoptosis, 37 kDa/67 kDa laminin receptor, prion proteinsAlzheimer disease (AD), primarily defined by psychiatrist Alois Alzheimer in 1906, is a neurodegenerative disorder and currently exhibits a prevalence that “doubles approximately every five years from 0.5% at the common age of onset-65 years old.”1 This disease is the most common form of dementia afflicting the elderly and at present affects in excess of 37 million people globally2 and it is predicted that 100 million people will be living with the disease by 2050.3AD has received mounting scientific interest and has stimulated tireless research endeavours not only due to the complex mechanism by which it is caused; the multitude of contributing factors and contradictions which have arisen between hypotheses and acquired results, but also due to the rise in life expectancies4 owing to the advent of modern medicine, which has socio-economic implications particularly in terms of strain placed upon national health systems. 相似文献
9.
10.
Richard D. Beger Deborah K. Hansen Laura K. Schnackenberg Brandie M. Cross Javad J. Fatollahi F. Tracy Lagunero Zoltan Sarnyai Laszlo G. Boros 《Metabolomics : Official journal of the Metabolomic Society》2009,5(3):336-345
Previous genetic and proteomic studies identified altered activity of various enzymes such as those of fatty acid metabolism
and glycogen synthesis after a single toxic dose of valproic acid (VPA) in rats. In this study, we demonstrate the effect
of VPA on metabolite synthesis flux rates and the possible use of abnormal 13C labeled glucose-derived metabolites in plasma or urine as early markers of toxicity. Female CD-1 mice were injected subcutaneously
with saline or 600 mg/kg) VPA. Twelve hours later, the mice were injected with an intraperitoneal load of 1 g/kg [U-13C]-d-glucose. 13C isotopomers of glycogen glucose and RNA ribose in liver, kidney and brain tissue, as well as glucose disposal via cholesterol
and glucose in the plasma and urine were determined. The levels of all of the positional 13C isotopomers of glucose were similar in plasma, suggesting that a single VPA dose does not disturb glucose absorption, uptake
or hepatic glucose metabolism. Three-hour urine samples showed an increase in the injected tracer indicating a decreased glucose
re-absorption via kidney tubules. 13C labeled glucose deposited as liver glycogen or as ribose of RNA were decreased by VPA treatment; incorporation of 13C via acetyl-CoA into plasma cholesterol was significantly lower at 60 min. The severe decreases in glucose-derived carbon
flux into plasma and kidney-bound cholesterol, liver glycogen and RNA ribose synthesis, as well as decreased glucose re-absorption
and an increased disposal via urine all serve as early flux markers of VPA-induced adverse metabolic effects in the host. 相似文献