scholarly journals Differential Hippocampal Protein Expression Betweennormal Mice and Mice With the Perioperative Neurocognitive Disorder: a Proteomic Analysis

2021 ◽  
Author(s):  
Chuan Li ◽  
Jingzhu Li ◽  
He Tao ◽  
Jinghua Shan ◽  
Fanghao Liu ◽  
...  
Keyword(s):  
Protein Expression ◽  
High Performance ◽  
Neuronal Cell ◽  
Pure Oxygen ◽  
Control Group ◽  
Neurocognitive Disorder ◽  
Mitochondrial Oxidative Stress ◽  
Erbb Signaling ◽  
Differential Expression Protein ◽  
And Control

Abstract ObjectivesTo compare differential expression protein in hippocampal tissues from mice of perioperative neurocognitive disorder (PND) and normal control mice and to explore the possible mechanism of PND. MethodsMice were randomly divided into a PND group(n=9) and a control group(n=9).The mice in the PND group were treated with open tibial fracture with intramedullary fixation under isoflurane anesthesia, while the mice in the control group received pure oxygen without surgery. The cognitive functions of the two groups were examined using Morris water maze experiment,Open field test and Fear conditioning test. The protein expression of the hippocampus of mice were analyzed by High-Performance Liquid-Chromatography Mass Spectrometry (HPLC-MS/MS) .Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed to explore the principal functions of dysregulated proteins. ResultsA total of 21 proteins were differentially expressed between PND and control mice on days 1, 3, and 7 after the operation. These proteins were involved in many pathological processes, such as neuroinflammatory responses, mitochondrial oxidative stress, impaired synaptic plasticity, and neuronal cell apoptosis. Also, the dysregulated proteins were involved in MAPK, AMPK, and ErbB signaling pathways. ConclusionThe occurrence of PND could be attributed to multiple mechanisms.

scholarly journals SUZ12 Participates in PNH Cloning Proliferation By Regulating Histone H3K27me3 Methylation Level

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 1106-1106
Author(s):  
Rong Fu ◽  
Yingying Chen ◽  
Zonghong Shao ◽  
Hui Liu ◽  
Lijie Zeng ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Protein Expression ◽  
Peripheral Blood ◽  
Methylation Level ◽  
Cell Model ◽  
Gene Mutations ◽  
Control Group ◽  
Mrna And Protein Expression ◽  
And Control

Abstract Paroxysmal nocturnal hemoglobinuria (PNH) is a disease of hematopoietic stem cell membrane defects due to acquired PIG-Amutation. Our previous study found some secondary gene mutations in PNH patients by WES. However, it is not clear exactly which mutations are associated with the disease. So, 97 target genes were selected as a target gene panel and tested in 23 PNH patients by DNA sequencing of specific target regions. We found that all PNH patients had other gene mutations except PIG-Amutations, including TTN, NCOR2, CPS1, MUC4, SUZ12, LFNG, CELSR2, JAK2, SETBP1 and KMT2D (Figure1A). Through harmful analysis, KEGG enrichment, GO enrichment analysis and protein interaction analysis, we screened out the secondary mutant gene SUZ12 that may be involved in the cloning proliferation of PNH. We detected the mRNA and protein expression levels of SUZ12 and H3K27me3 methylation in PNH patients and health volunteers, the results showed that the mRNA and protein expression levels of SUZ12 and H3K27me3 methylation in peripheral blood CD59 -cells of PNH patients were higher than those in CD59 + cells of PNH patients and healthy controls (Figure1B). The relative expression level of SUZ12 in peripheral blood CD59 -cells of PNH patients was correlated with (r=0.4162, p=0.0385), CD59 -erythrocyte ratio (r=0.4636, p=0.0196), CD59 -monocyte ratio (r=0.4052, p=0.0495), Flaer -monocyte ratio (r=0.6769, p=0.0004) and Flaer -granulocytic ratio (r=0.6146, p=0.0018), indicating that SUZ12 may be involved in abnormal PNH cloning and proliferation by regulating H3K27me3. To verify the role of SUZ12 in the proliferation of PNH cloning, we used CRISPR/Cas9 to knockdown PIG-A expression in THP-1 cells to construct A PNH cell model, the expression level of PIG-A protein in the cell model was significantly decreased, and the proportion of CD59 - cells accounted was stable at 95%. Then lentivirus transfection was used to knockdown the expression of SUZ12 in PNH cell model. The results showed when the SUZ12 expression was knockdown, the methylation level of histone H3K27me3 was decreased, the cell proliferation activity was decreased, apoptosis was increased, and the cell cycle was arrested at G0/G1 phase. The proportion of CD59 + cells increased gradually from 3 weeks after transfection, and significantly increased at 4 weeks after transfection, while no changes were observed in the empty virus group and control group (Figure1C). Four weeks after lentivirus transfection, the expression of PIG-A protein recovered in SUZ12 knockdown group compared with empty virus group and control group (Figure1D). In conclusion, SUZ12 mutation leads to the overexpression of SUZ12, which can affect cell proliferation, apoptosis and cell cycle by regulating the methylation level of histone H3K27me3, thereby promoting the proliferation of PNH abnormal cloning and participating in the pathogenesis of PNH. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

scholarly journals Polyomic Analyses of Dopaminergic Neurons Isolated From Human Substantia Nigra in Parkinson’s Disease: An Exploratory Study.

2021 ◽  
Author(s):  
Affif ZACCARIA ◽  
Paola Antinori Malaspina ◽  
Virginie Licker ◽  
Enikö Kovari ◽  
Johannes A Lobrinus ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Substantia Nigra ◽  
Protein Expression ◽  
Differential Expression ◽  
Exploratory Study ◽  
Substantia Nigra Pars Compacta ◽  
Control Group ◽  
Gene And Protein Expression ◽  
And Control

Abstract Background Dopaminergic (DA) neurons of the substantia nigra pars compacta (SNpc) selectively and progressively degenerate in Parkinson’s disease (PD). Until now, molecular analyses of DA neurons in PD have been limited to genomic and transcriptomic approaches, whereas, to the best of our knowledge, no proteomic or combined polyomic study examining the protein profile of these neurons, is currently available. Methods In this exploratory study, we used laser microdissection to extract DA neurons from 10 human SNpc samples obtained at autopsy in PD patients and control subjects. Extracted RNA and proteins were identified by RNA sequencing and nano-LC-MS/MS, respectively, and the differential expression between the PD and control group was assessed. Results Qualitative analyses confirmed that the microdissection protocol preserves the integrity of our samples and offers access to specific molecular pathways. This polyomic analysis highlighted differential expression of 52 genes and 33 proteins, including molecules of interest already known to be dysregulated in PD, such as LRP2, PNMT, CXCR4, MAOA and CBLN1 genes, or the Aldehyde dehydrogenase 1 protein. On the other hand, despite the same samples were used for both analyses, correlation between RNA and protein expression was low, as exemplified by the CST3 gene encoding for the cystatin C protein. Conclusion This is the first exploratory study analyzing both gene and protein expression of LMD-dissected DA neurons from SNpc in PD. Although correlation between RNA and protein expressions was limited, this polyomic study provides an extensive and integrated overview of molecular changes identified in the PD SNpc and may offer novel insights into specific pathological processes at work in PD degeneration.

scholarly journals Mitigating Effects of Liriope platyphylla on Nicotine-Induced Behavioral Sensitization and Quality Control of Compounds

2020 ◽  
Vol 10 (9) ◽  
pp. 654
Author(s):  
Dahye Yoon ◽  
In Soo Ryu ◽  
Woo Cheol Shin ◽  
Minhan Ka ◽  
Hyoung-Geun Kim ◽  
...  
Keyword(s):  
Quality Control ◽  
Protein Expression ◽  
High Performance ◽  
Behavioral Sensitization ◽  
Dorsal Striatum ◽  
Control Group ◽  
Tandem Mass ◽  
Nicotine Administration ◽  
Liriope Platyphylla ◽  
Spicatoside A

In this study we investigated the mitigating effects of Liriope platyphylla Wang et Tang extract on behavioral sensitization and the quantification of its major compounds. The extract of L. platyphylla reduces the expression of tyrosine hydroxylase (TH) protein, which is increased by nicotine, back to normal levels, and increases the expression of dopamine transporter (DAT) protein, which is reduced by nicotine, back to normal levels in PC12 cells. In this study, rats received nicotine (0.4 mg/kg, subcutaneously) only for seven days and then received extract of L. platyphylla (200 or 400 mg/kg, oral) 1 h prior to nicotine administration for an additional seven days. The extract of L. platyphylla reduced locomotor activity compared to the nicotine control group in rats. The extract of L. platyphylla significantly attenuated the repeated nicotine-induced DAT protein expression in the nucleus accumbens (NAc), but there was no effect on increased TH protein expression in the dorsal striatum. These findings suggest that L. platyphylla extract has a mitigating effect on nicotine-induced behavioral sensitization by modulating DAT protein expression in the NAc. For quality control of L. plathyphylla, spicatoside A and D, which are saponin compounds, were quantified in the L. platyphylla extract. The amounts of spicatoside A and D in L. platyphylla extract obtained from ultra-high-performance liquid chromatography with tandem mass spectrometry were 0.148 and 0.272 mg/g, respectively. The identification of these compounds in L. platyphylla, which can be used for quality control, provides important information for the development of drugs to treat nicotine dependence.

scholarly journals An exploratory study of neopterin and kynurenine pathway in pterygium

Pteridines ◽  
2019 ◽  
Vol 30 (1) ◽  
pp. 153-157
Author(s):  
Bilge Kilicarslan ◽  
Aziz Cardak ◽  
Gozde Girgin ◽  
Ozlem Evren Kemer ◽  
Terken Baydar
Keyword(s):  
High Performance ◽  
Degradation Process ◽  
Serum Levels ◽  
Kynurenine Pathway ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Cell Immunity ◽  
Degenerative Disorder ◽  
And Control ◽  
Clinical Potential

Abstract Pterygium is an inflammatory, vascular and degenerative disorder with unknown aetiology. The aim of this study was to evaluate the changes in neopterin levels, reflecting T-cell immunity, and the kynurenine pathway, the main degradation process of tryptophan, in pterygium. For this purpose, neopterin concentrations were measured in serum and tear samples by enzyme-linked immunosorbent assay (ELISA) in pterygium patients (n=31) and control group (n=32). Kynurenine (KYN) and tryptophan (TRP) serum levels were simultaneously determined by high-performance liquid chromatography (HPLC) for evaluation of the kynurenine pathway. Serum neopterin concentrations and kynurenine to tryptophan ratio (KYN/TRP) as an index of tryptophan breakdown were found increased in pterygium compared to controls (p<0.05). Although there was a 3-fold difference observed between serum and tear neopterin levels, no significant relationship was found. It can be concluded that neopterin may be used as a nonspecific biomarker that reflects immunological activity in pterygium and has clinical potential for evaluation of pterygium pathogenesis. These immune- or inflammatory-mediated changes were also supported by an increased KYN/TRP ratio in pterygium patients.

scholarly journals P6 Electroacupuncture Improved QTc Interval Prolongation by Upregulation of Connexin43 in Droperidol Treated Rats

2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
Feng Zhao ◽  
Suyang Cui ◽  
Libing Huang
Keyword(s):  
Protein Expression ◽  
Control Group ◽  
Qtc Interval ◽  
Interval Prolongation ◽  
Cx43 Expression ◽  
Qtc Interval Prolongation ◽  
Mrna And Protein Expression ◽  
Cx43 Mrna ◽  
And Control ◽  
Group D

Aim. This study investigated the effect of P6 EA on droperidol-induced QTc interval prolongation and Cx43 expression in ventricular muscle of rats.Methods. Twenty-four rats were randomly divided into control group (C), droperidol group (D), or EA group (E). C group rats were injected with normal saline. D group rats were injected with droperidol 0.13 mg/kg. E group rats were pretreated with EA at left P6 acupoint for 30 min and then injected with droperidol (0.13 mg/kg). QTc intervals were recorded at lead II in ECG within 120 min. Cx43 expression was measured by RT-PCR and western blotting.Result. Droperidol significantly prolonged QTc intervals compared with controls at 5 min, 10 min, 15 min, and 30 min (P<0.05). P6 EA could significantly abbreviate the prolongation of QTc interval compared with droperidol group at 5 min, 10 min, 15 min, and 30 min (P<0.05). Cx43 mRNA and proteins were significantly increased by P6 EA compared with droperidol group at 120 min (P<0.05). There were no significant differences in Cx43 mRNA and protein expression between droperidol and control group at 120 min (P>0.05).Conclusion. P6 EA could improve QTc interval prolongation induced by droperidol, which may relate to upregulation of Cx43 mRNA and protein. Antiemetic dose of droperidol had minor effects on Cx43 mRNA and protein expression at 120 min.

scholarly journals Multiomic Analyses of Dopaminergic Neurons Isolated from Human Substantia Nigra in Parkinson’s Disease: A Descriptive and Exploratory Study

2021 ◽  
Author(s):  
Affif Zaccaria ◽  
Paola Antinori ◽  
Virginie Licker ◽  
Enikö Kövari ◽  
Johannes A. Lobrinus ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Substantia Nigra ◽  
Protein Expression ◽  
Differential Expression ◽  
Dopaminergic Neurons ◽  
Exploratory Study ◽  
Control Group ◽  
Gene And Protein Expression ◽  
And Control

AbstractDopaminergic neurons (DA) of the substantia nigra pars compacta (SNpc) selectively and progressively degenerate in Parkinson’s disease (PD). Until now, molecular analyses of DA in PD have been limited to genomic or transcriptomic approaches, whereas, to the best of our knowledge, no proteomic or combined multiomic study examining the protein profile of these neurons is currently available. In this exploratory study, we used laser capture microdissection to extract regions from DA in 10 human SNpc obtained at autopsy in PD patients and control subjects. Extracted RNA and proteins were identified by RNA sequencing and nanoliquid chromatography–mass spectrometry, respectively, and the differential expression between PD and control group was assessed. Qualitative analyses confirmed that the microdissection protocol preserves the integrity of our samples and offers access to specific molecular pathways. This multiomic analysis highlighted differential expression of 52 genes and 33 proteins, including molecules of interest already known to be dysregulated in PD, such as LRP2, PNMT, CXCR4, MAOA and CBLN1 genes, or the Aldehyde dehydrogenase 1 protein. On the other hand, despite the same samples were used for both analyses, correlation between RNA and protein expression was low, as exemplified by the CST3 gene encoding for the cystatin C protein. This is the first exploratory study analyzing both gene and protein expression of laser-dissected neuronal parts from SNpc in PD. Data are available via ProteomeXchange with identifier PXD024748 and via GEO with identifier GSE 169755.

scholarly journals Studying the pathogenic role of catecholamines in the development of retinopathy of prematurity on an experimental model of the disease

2019 ◽  
Vol 12 (4) ◽  
pp. 64-69
Author(s):  
L. A. Katargina ◽  
N. A. Osipova ◽  
A. J. Panova ◽  
N. S. Bondarenko ◽  
Yu. O. Nikishina ◽  
...  
Keyword(s):  
Retinopathy Of Prematurity ◽  
High Performance ◽  
Amacrine Cells ◽  
Study Groups ◽  
Control Group ◽  
Dopamine Level ◽  
Applied Model ◽  
Rat Pups ◽  
And Control ◽  
Experimental Group

Purpose. To study the involvement of dopamine and noradrenaline in the pathogenesis of retinopathy of prematurity (ROP) on an original rat model of the disease.Material and methods. The study was conducted on 41 newborn Wistar rats (82 eyes), divided into 2 groups: experimental (EROP, rats with experimental ROP, n = 21) and control (n = 20). The rats were taken out of the experiment on the 7th, 14th, 23rd and 28th days of life. All rat pups were given binocular enucleation at the indicated times, whereupon the eyeballs were dissected along the limbus and the cornea, lens, hyaloid system, and vitreous were removed. The retina was isolated from the eye cup. The isolated retinal samples were homogenized in 10 volumes of 0.1 n HClO4 containing 50 pmol/ml (or more) of 3,4-dihydroxybenzylamine (DBA), using an ultrasonic homogenizer (Labsonic M, Sartorius), centrifuged at 2000g for 20 minutes, and the norepinephrine, dopamine and precursor of dopamine — L-3,4 dihydroxyphenylalanine (L-DOPA) were determined in the resulting supernatant. The contents of substances were measured using reverse phase high performance liquid chromatography with electrochemical detection (Amperometric detector LC-4B, Bioanalytical Systems, USA) set at the potential of 850 mV.Results. On the 7th day, on which avascular retinal zones in both groups of animals existed, no significant differences were found in the content of monoamines in the retina of rats with EROP and in the control group. On the 28th day, the content of noradrenaline, dopamine and L-DOPA in the retina of the experimental group was significantly increased compared with the control. On day 23, corresponding to the peak of neovascularization in the EROP model applied, the level of norepinephrine in the retina of experimental group rats was significantly higher, while the level of L-DOPA was significantly lower compared to the control group. The dopamine level was comparable in both study groups and similar to the level of L-DOPA in the control group. On the 28th day, corresponding to the beginning of EROP regression accompanied by vascular activity decrease, the content of dopamine and L-DOPA remained lower than in the control group.Conclusion. During the development of pathological neovascularization of rat pup retina with EROP, the level of noradrenaline is growing, revealing a peak corresponding to the period of pronounced pathological growth of retinal vessels within the applied model, which indicates to the fact of noradrenalin proangiogenic properties and its direct participation in the pathogenesis of ROP. The level of dopamine and its predecessor, L-DOPA, increased towards the 14th day as compared to its level detected on the 7th day, which may be due to the maturation of the amacrine cells producing, and then, on the day 23. i. e. during the period corresponding to the maximum peak of angiogenesis, its relative decrease of L-DOPA was noted. It can be assumed that the lack of this monoamine, and hence insufficient manifestation of its anti-angiogenic properties contributes to the development of uncontrolled neovascularization of the retina.

scholarly journals Immunohistochemical Expression of Gata3 Gene in Patients with Breast Cancer

2021 ◽  
Vol 15 (8) ◽  
pp. 2355-2358
Author(s):  
Zeinab Hemeed Abbas ◽  
Karrar S. Zayed ◽  
Rihab H. Almudhafer ◽  
Hanaa Hemeed Abbas
Keyword(s):  
Breast Cancer ◽  
Protein Expression ◽  
Ductal Carcinoma ◽  
Control Group ◽  
Immunohistochemical Expression ◽  
Tissue Samples ◽  
Proliferation And Differentiation ◽  
System 1 ◽  
And Control ◽  
Gata3 Protein

Breast cancer (BC) is one of the most common cancers in the world. In numerous tissues, including the breast, GATA3 plays an important role in stimulating proliferation and differentiation. The main aims of this study is determining the types of BC (IDC and ILC) then the estimation of the role of GATA3 protein expression by immunohistochemical staining method (IHC) in BC patients and control groups as a biomarker. The present study was done during the period from October 2020 to April 2021. Sixty seven tissue samples block embedded in wax taken from BC female patients and thirty four of normal non-tumoral breast tissue as a control group collected randomly with their data from three private pathological clinics, these blocks have been prepared between (2014 – 2021), three pathologists re-evaluate each pathologic material. Regarding to IHC GATA3 protein expression, after histological re-evaluation of slides, the rate of IDC was 80.6% (54 patients) and of ILC was (19.4%) (13 patients).The scoring system +1 (37.3%) and +2(19.4%) increased significantly in BC patients than control (P=0.001), in addition to, the nuclear positive expression of GATA3 decrease significantly in BC patients than control (Odd ratio 2.55, 95% CI 1.45-2.37, P=0.0001), On the other hand, the positivity of GATA3 protein increased significantly in patients with invasive ductal carcinoma (IDC) (P=0.001). Keyword: GATA3, Breast Cancer, Immunohistochemical Expression.

Effect of Oral Adsorbent on Blood Metabolites in Hepatic Failure Dogs

1989 ◽  
Vol 12 (2) ◽  
pp. 114-120 ◽  
Author(s):  
M. Hiraishi ◽  
T. Takahama ◽  
Z. Yamazaki ◽  
F. Kanai ◽  
K. Ohnishi ◽  
...  
Keyword(s):  
Body Weight ◽  
Hepatic Failure ◽  
High Performance ◽  
Body Weight Loss ◽  
Blood Metabolites ◽  
Control Group ◽  
Middle Molecules ◽  
Oral Adsorbent ◽  
And Control ◽  
Carbonaceous Adsorbent

AST-120 is a specially synthesized carbonaceous adsorbent for oral use. It mainly adsorbs low to middle molecules in the alimentary tract. In the present study, AST-120 was administered to hepatic failure dogs, and blood metabolites were analyzed by high performance liquid chromatography (HPLC). Thirty adult mongrel dogs underwent posta-cavae (P-C) shunts with 40% and 70% hepatectomies. They were divided into two groups, the AST group (n= 19) and control group (n=11). The AST group received about 0.5 g/kg of the adsorbent intermittently with diet after the operation. The control group was fed the ordinary diet. Body weight, blood ammonia, plasma bile acids were measured, and blood metabolites were analyzed by the multi-column HPLC system. P-C shunt dogs with 70% hepatotectomies died within three months showing about 40–50% body weight loss. HPLC analysis of their plasma showed some specific peaks for middle molecules, about 3000–5000 daltons. After administration of the adsorbent, these peaks were not detected, so it was considered that these substances had been adsorbed.

scholarly journals High-performance inulin and oligofructose prebiotics increase the intestinal absorption of iron in rats with iron deficiency anaemia during the growth phase

2011 ◽  
Vol 108 (6) ◽  
pp. 1008-1016 ◽  
Author(s):  
Karine de Cássia Freitas ◽  
Olga Maria Silvério Amancio ◽  
Mauro Batista de Morais
Keyword(s):  
Intestinal Absorption ◽  
High Frequency ◽  
High Performance ◽  
Fe Deficiency ◽  
Control Group ◽  
Control Groups ◽  
Deficiency Anaemia ◽  
Significant Difference ◽  
Group 2 ◽  
And Control

Considering the high frequency of anaemia due to Fe deficiency, it is important to evaluate the effects of prebiotics on the absorption of Fe. The aim of the present study was to evaluate the effects of high-performance (HP) inulin, oligofructose and synergy1 during recovery from anaemia in rats through the intestinal absorption of Fe, food intake, body growth, caecal pH and weight of the intestine. Wistar rats (n 47) were fed with rations of AIN93-G with no Fe to induce Fe deficiency anaemia. At 36 d of life, anaemic rats were divided into four groups: (1) the HP inulin group; (2) the synergy1 group; and (3) the oligofructose group, all with 100 g of the respective prebiotic per kg of ration; and (4) a control group, in which the prebiotic was replaced by maize starch. Then, 25 mg of elemental Fe/kg of ration was added to all rations to allow recovery from anaemia. The final values of Hb in the HP inulin, synergy1, oligofructose and control groups were, respectively: 98 (94–99); 83 (81–92); 100 (90–114); 77 (72–81) g/l, with a statistically significant difference (P ≤ 0·001) between the oligofructose and control groups and the HP inulin and control groups. The four groups had an increase in weight and body length and had similar consumption of rations. The intestinal weight and caecal pH were significantly different between the groups that consumed prebiotics and the control group. HP inulin and oligofructose increased the intestinal absorption of Fe in rats.

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