scholarly journals Change in Proteolytic Profile in Heifers After Oligofructose Overload

2020 ◽  
Vol 7 ◽  
Author(s):  
Shuaichen Li ◽  
Xiaoyan Zheng ◽  
Mengyuan Ding ◽  
Ze Tao ◽  
Jiantao Zhang ◽  
...  
Keyword(s):  
Body Weight ◽  
Basement Membrane ◽  
Western Blot ◽  
Neutrophil Elastase ◽  
Proteolytic Enzymes ◽  
Tap Water ◽  
Plasma Samples ◽  
Tissue Inhibitors Of Metalloproteinase ◽  
Blood Neutrophils ◽  
And Control

Laminitis in cattle is an important underlying cause of lameness, which leads to a significant reduction in economic and animal welfare. Nevertheless, the disordered pathological processes of laminitis remain unclear. Several proteinases are probably involved in the disorder of basement membrane (BM) metabolism in laminitis, for instance, matrix metalloproteinases (MMPs), neutrophil elastase (NE), and myeloperoxidase (MPO). This study aimed to investigate the change in proteolytic profile in circulating and lamellar tissues using an oligofructose (OF) overload-induced laminitis model in heifers. Twelve clinically healthy and nonlame Chinese Holstein heifers were recruited and randomly divided into two groups: OF-induced and control (CON). The OF-induced heifers group (n = 6) was administered 17 g/kg of body weight (BW) of OF dissolved in 2 L/100 kg of BW of tap water via the oral-rumen tube. The CON group (n = 6) was given an equal volume of tap water. The plasma samples were collected 0, 6, 12, 18, 24, 36, 48, 60, and 72 h after administration, and the lamellar samples were collected 72 h after euthanasia. The plasma samples were analyzed by zymography and reverse zymography. Histological examination, zymography, reverse zymography, and Western blot of lamellar samples were conducted. In the plasma of the OF-induced group, the pro-MMP9 activity increased from 36 h (P < 0.001) to 60 h (P < 0.05). Moreover, the plasma tissue inhibitors of metalloproteinase 1 (TIMP1) activity decreased after 18 h (P < 0.05), while the ratio of pro-MMP9 to TIMP1 and TIMP2 increased after 18 h (P < 0.001) and 48 h (P < 0.05), respectively. The act-MMP2, pro-MMP9, and act-MMP9 activities increased in the lamellar tissue of the OF-induced group compared with the CON group (P < 0.05). In addition, the expression of lamellar NE protein was higher in the OF-induced group (P < 0.01), while no change was found in lamellar MPO protein compared with the CON group. In conclusion, increased pro-MMP9 combined with decreased TIMP1 activity in the circulation might have caused the activation of blood neutrophils, while the activation of proteolytic enzymes in lamellar tissue probably led to the dysfunction of BM in the OF-induced group.

Electron microscopic radioautographic studies on the incorporation of 3H proline in the glomerular basement membrane (GBM) in antistreptococcal-cell-membrane (SCM) injected mice

1984 ◽  
Vol 42 ◽  
pp. 188-189
Author(s):  
R.P. Nayyar ◽  
C.F. Lange ◽  
J. L. Borke
Keyword(s):  
Body Weight ◽  
Cell Membrane ◽  
Basement Membrane ◽  
Glomerular Basement Membrane ◽  
Mesangial Matrix ◽  
Electron Microscopic ◽  
Group A ◽  
Injection Protocol

Streptococcal cell membrane (SCM) antiserum injected mice show a significant thickening of glomerular basement membrane (GBM) and an increase in mesangial matrix within 4 to 24 hours of antiserum administration (1,2,3). This study was undertaken to evaluate the incorporation of 3H proline into glomerular cells and GBM under normal and anti-SCM induced conditions. Mice were administered, intraperitoneally, 0.1 ml of normal or anti-SCM serum followed by a 10 µC/g body weight injection of 3H proline. Details of the preparation of anti-SCM (Group A type 12 streptococcal pyogenes) and other sera and injection protocol have been described elsewhere (2). After 15 minutes of isotope injection a chase of cold proline was given and animal sacrificed at 20 minutes, 1,2,4,8,24 and 48 hours. One of the removed kidneys was processed for immunofluorescence, light and electron microscopic radioautographic studies; second kidney was used for GBM isolation and aminoacid analysis.

Genotoxicity and Cell Proliferative Activity of 3-Chloro-4-(Dichloromethyl)-5-Hydroxy-2(5H)-Furanone [MX] in Rat Glandular Stomach

1992 ◽  
Vol 25 (11) ◽  
pp. 341-345 ◽  
Author(s):  
C. Furihata ◽  
M. Yamashita ◽  
N. Kinae ◽  
T. Matsushima
Keyword(s):  
Cell Proliferation ◽  
Body Weight ◽  
Dna Synthesis ◽  
Ornithine Decarboxylase ◽  
Tap Water ◽  
Fold Increase ◽  
Single Strand ◽  
Glandular Stomach ◽  
Decarboxylase Activity ◽  
Pyloric Mucosa

MX is a strong direct acting mutagen on Salmonella typhimurium TA100 and is present in chlorinated tap water which contains organic compounds. MX was administered orally to 7-week-old male F344 rats, and its geno-toxicity in the pyloric mucosa of stomach was examined by analysis of DNA single strand scissions by the alkaline elution method. The effect of MX on cell proliferation was examined by assays of the inductions of replicative DNA synthesis and ornithine decarboxylase. MX at closes of 20-48 mg/kg body weight induced DNA single strand scissions dose-dependently (p<0.02) in the pyloric mucosa of the stomach 2 h after its administration. Moreover at doses of 10-60 mg/kg body weight, it induced up to 21-fold increase in replicative DNA synthesis (p<0.01) 16 h after its administration. At doses of 10-60 mg/kg body weight, it induced up to 100-fold increase in ornithine decarboxylase activity with a maximum 16 h after its administration. These results suggest that MX is genotoxic and induces cell proliferation in the glandular stomach of rats.

UPLC-Q-TOF/MS based Untargeted Metabolite and Lipid Analysis on Premature Ovarian Insufficiency Plasma Samples.

2020 ◽  
Vol 16 ◽  
Author(s):  
Yasemin Taşcı ◽  
Rahime Bedir Fındık ◽  
Meryem Kuru Pekcan ◽  
Ozan Kaplan ◽  
Mustafa Çelebier
Keyword(s):  
Lipid Analysis ◽  
Principal Component ◽  
Premature Ovarian Insufficiency ◽  
Plasma Samples ◽  
Ovarian Insufficiency ◽  
Analytical Methodology ◽  
Drug Molecules ◽  
New Treatment ◽  
And Control ◽  
Phenyl Alanine

Background: Metabolomics is one of the main areas to understand cellular process at molecular level by analyzing metabolites. In recent years metabolomics has been emerged as key tool to understand molecular basis of disease, find diagnostic and prognostic biomarkers, and develop new treatment opportunities and drug molecules. Objective: In this study, an untargeted metabolite and lipid analysis were performed to identify potential biomarkers on premature ovarian insufficiency plasma samples. 43 POI subject plasma samples were compared with 32 healthy subject plasma samples. Methods: Plasma samples were pooled and extracted using chloroform:methanol:water (3:3:1 v/v/v) mixture. Agilent 6530 LC/MS Q-TOF instrument equipped with ESI source was used for analysis. A C18 column (Agilent Zorbax 1.8 μM, 50 x 2.1 mm) was used for separation of metabolites and lipids. XCMS, an “R software” based freeware program, was used for peak picking, grouping and comparing the findings. Isotopologue Parameter Optimization (IPO) software was used in order to optimize XCMS parameters. The analytical methodology and data mining process were validated according to the literature. Results: 83 metabolite peaks and 213 lipid peaks were found to be in semi-quantitatively and statistically different (fold change >1.5, p <0.05) between the POI plasma samples and control subjects. Conclusion: According to the results, two groups were successfully separated through principal component analysis. Among the peaks, phenyl alanine, decanoyl-L-carnitine, 1-palmitoyllysophosphatidylcholine and PC(O-16:0/2:0) were identified through auto MS/MS and matched with human metabolome database and proposed as plasma biomarker for POI and monitoring the patients in treatment period.

Impact of an Infant Formula Containing a Novel Fat Blend (Cow’s Milk Fat, Fish and Vegetable Oil) and Prebiotics on Stool Fatty Acid Soaps and Erythrocyte Fatty Acid Profiles in Full-Term Healthy Newborns

2021 ◽  
pp. 1-8
Author(s):  
Maroula Lambidou ◽  
Birgit Alteheld ◽  
Rolf Fimmers ◽  
Frank Jochum ◽  
Antonia Nomayo ◽  
...  
Keyword(s):  
Body Weight ◽  
Fatty Acid ◽  
Infant Formula ◽  
Milk Fat ◽  
Control Group ◽  
Human Breast Milk ◽  
Infant Formulas ◽  
Fa Composition ◽  
And Control ◽  
Breast Fed

<b><i>Introduction:</i></b> Recently, new commercial infant formulas have been composed considering novel fat blends and oligosaccharides to better resemble the fatty acid (FA) composition and stereospecific distribution (e.g., increased amount of ß-palmitate) as well as probiotics content of human breast milk. We hypothesized that these newly composed infant formulas may decrease fecal FA soap excretion and may positively affect erythrocyte FA profiles compared with regular formulas. <b><i>Methods:</i></b> Healthy infants were randomly assigned to receive a high-sn-2-palmitate formula (&#x3e;25% of the PA is esterified to the sn-2 position of the glycerol backbone, verum: <i>n</i> = 30) or a “standard” formula containing &#x3c;10% of PA in sn-2 position and no oligosaccharides (control: <i>n</i> = 27); a non-randomized group of breast-fed infants served as control. Anthropometric data of the infants (body weight, recumbent length, and head circumference) were recorded at inclusion (visit 1) and 6 and 12 weeks after onset of intervention (visits 2 and 3). Blood samples for erythrocyte FA analysis (gas chromatography) were taken at visits 1 and 2; stool samples were collected at visit 2. <b><i>Results:</i></b> Quantitative formula intake (mL/kg body weight × day) at visit 2 (verum: 155 ± 30, control: 164 ± 30) and visit 3 (verum: 134 ± 26, control: 134 ± 21) was comparable. Six weeks after onset of intervention, stool total FA soaps, palmitate soaps, and total FAs were similar in both formula-fed groups but significantly higher than in breast-fed infants. During the 6-week intervention, erythrocyte palmitate decreased significantly from baseline in all 3 groups with no group differences (verum: 29.20 ± 1.17 to 27.12 ± 0.66, control: 29.88 ± 2.00 to 27.01 ± 0.94, breast-fed: 30.20 ± 0.86 to 26.84 ± 0.98). For selected FAs, significant changes over time in verum and control group were obvious but without formula effects. Some variations in the FA profile of breast-fed infants compared to both verum and control groups were observed. <b><i>Conclusions:</i></b> In contrast to our hypothesis, feeding a newly composed infant formula based on a fat blend with 25% of PA in the sn-2 position of triacylglycerols and supplemented with a prebiotic could not decrease insoluble FA soap excretion compared with a standard product; in this respect, breastfeeding is obviously the best choice. Surprisingly, erythrocyte FA profiles were comparable in formula-fed and breast-fed infants; obvious alterations in FA composition of the respective fat sources and structure did not affect FA incorporation into membranes. Caution should be, however, exercised in drawing robust conclusions in the absence of larger, adequately powered intervention studies.

scholarly journals Sirtuin 2 in Endometrial Cancer: A Potential Regulator for Cell Proliferation, Apoptosis and RAS/ERK Pathway

2020 ◽  
Vol 19 ◽  
pp. 153303382098078
Author(s):  
Yanjuan Guo ◽  
Nannan Zhao ◽  
Jianli Zhou ◽  
Jianxin Dong ◽  
Xing Wang
Keyword(s):  
Cell Proliferation ◽  
Endometrial Cancer ◽  
Western Blot ◽  
Cell Line ◽  
Cell Lines ◽  
Erk Pathway ◽  
Uterine Epithelial Cell ◽  
Knock Down ◽  
Ishikawa Cells ◽  
And Control

Objective: The present study aimed to explore the function of sirtuin 2 (SIRT2) on cell proliferation, apoptosis, rat sarcoma virus (RAS)/ extracellular signal-regulated kinase (ERK) pathway in endometrial cancer (EC). Methods: SIRT2 expression in human EC cell lines and human endometrial (uterine) epithelial cell (HEEC) line was assessed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot. SIRT2 knock-down and control knock-down plasmids were transfected into HEC1A cells, respectively; SIRT2 overexpression and control overexpression plasmids were transfected into Ishikawa cells, respectively. After transfection, SIRT2, HRas proto-oncogene, GTPase (HRAS) expressions were evaluated by RT-qPCR and western blot. ERK and phosphorylated ERK (pERK) expressions were evaluated by western blot. Meanwhile, cell proliferation and cell apoptosis were measured. Results: Compared to normal HEEC cell line, SIRT2 mRNA and protein expressions were increased in most human EC cell lines (including HEC1A, RL952 and AN3CA), while were similar in Ishikawa cell line. In HEC1A cells, SIRT2 knock-down decreased cell proliferation but increased apoptosis. In Ishikawa cells, SIRT2 overexpression induced cell proliferation but inhibited apoptosis. For RAS/ERK pathway, SIRT2 knock-down reduced HRAS and inactivated pERK in HEC1A cells, whereas SIRT2 overexpression increased HRAS and activated pERK in Ishikawa cells, suggesting that SIRT2 was implicated in the regulation of RAS/ERK pathway in EC cells. Conclusion: SIRT2 contributes to the EC tumorigenesis, which appears as a potential therapeutic target.

scholarly journals Clinical and in Vivo Response Following Surgery or Surgery Plus Adjuvant Chemotherapy or Immunotherapy for Colorectal Carcinoma in a Rat Model

1983 ◽  
Vol 76 (10) ◽  
pp. 833-840 ◽  
Author(s):  
A K House ◽  
M A L Maley
Keyword(s):  
Body Weight ◽  
Adjuvant Chemotherapy ◽  
Surgical Excision ◽  
The Body ◽  
Recurrent Tumour ◽  
Mesenteric Node ◽  
And Control ◽  
Tumour Weight

Two cohorts of rats, 240 with colon cancer and 150 controls, were assessed clinically and immunologically for their response to tumour and its management which was either by surgical excision alone or by surgical excision combined with either adjuvant chemotherapy or immunotherapy. The histology and invasion characteristics were observed for similarity with those of human lesions. Metastases were found in liver, lymph nodes, the peritoneum or lungs in 27% of animals during follow up. Significantly fewer adjuvant-treated rats had metastases than those receiving surgery alone ( P < 0.05), and less total tumour weight was found in the adjuvant-treated rats at four ( P < 0.03) and six ( P < 0.001) weeks postoperatively. Animals in the adjuvant immunotherapy group survived longer than in either other group ( P < 0.001). The crude parameters of host response to tumour, body, spleen and mesenteric lymph node weight were recorded and the latter two indexed to body weight. The body weight of tumour and control rats increased significantly with time ( P < 0.04). The spleen and mesenteric node indices were significantly ( P < 0.04) greater in tumour than control rats and were varied by recurrent tumour growth and by the adjuvant treatment administered postoperatively.

Protein appetite is increased after central leptin-induced fat depletion

2007 ◽  
Vol 293 (4) ◽  
pp. R1468-R1473 ◽  
Author(s):  
Michael F. Wiater ◽  
Bryan D. Hudson ◽  
Yvette Virgin ◽  
Sue Ritter
Keyword(s):  
Body Weight ◽  
Food Intake ◽  
Body Fat ◽  
Caloric Intake ◽  
Sprague Dawley ◽  
Body Protein ◽  
Macronutrient Selection ◽  
Daily Diet ◽  
Nadir Point ◽  
And Control

Leptin reduces body fat selectively, sparing body protein. Accordingly, during chronic leptin administration, food intake is suppressed, and body weight is reduced until body fat is depleted. Body weight then stabilizes at this fat-depleted nadir, while food intake returns to normal caloric levels, presumably in defense of energy and nutritional homeostasis. This model of leptin treatment offers the opportunity to examine controls of food intake that are independent of leptin's actions, and provides a window for examining the nature of feeding controls in a “fatless” animal. Here we evaluate macronutrient selection during this fat-depleted phase of leptin treatment. Adult, male Sprague-Dawley rats were maintained on standard pelleted rodent chow and given daily lateral ventricular injections of leptin or vehicle solution until body weight reached the nadir point and food intake returned to normal levels. Injections were then continued for 8 days, during which rats self-selected their daily diet from separate sources of carbohydrate, protein, and fat. Macronutrient choice differed profoundly in leptin and control rats. Leptin rats exhibited a dramatic increase in protein intake, whereas controls exhibited a strong carbohydrate preference. Fat intake did not differ between groups at any time during the 8-day test. Despite these dramatic differences in macronutrient selection, total daily caloric intake did not differ between groups except on day 2. Thus controls of food intake related to ongoing metabolic and nutritional requirements may supersede the negative feedback signals related to body fat stores.

A novel coagulation inhibitor fromSchistosoma japonicum

Parasitology ◽  
2015 ◽  
Vol 142 (14) ◽  
pp. 1663-1672 ◽  
Author(s):  
SHIWANTHI L. RANASINGHE ◽  
KATJA FISCHER ◽  
GEOFFREY N. GOBERT ◽  
DONALD P. MCMANUS
Keyword(s):  
Human Blood ◽  
Neutrophil Elastase ◽  
Molecular Mechanisms ◽  
Genomic Sequence ◽  
Proteolytic Enzymes ◽  
Venous Blood ◽  
Coagulation Factors ◽  
Intestinal Wall ◽  
Hemostatic Disorders

SUMMARYLittle is known about the molecular mechanisms whereby the human blood flukeSchistosoma japonicumis able to survive in the host venous blood system. Protease inhibitors are likely released by the parasite enabling it to avoid attack by host proteolytic enzymes and coagulation factors. Interrogation of theS. japonicumgenomic sequence identified a gene,SjKI-1, homologous to that encoding a single domain Kunitz protein (Sjp_0020270) which we expressed in recombinant form inEscherichia coliand purified.SjKI-1is highly transcribed in adult worms and eggs but its expression was very low in cercariae and schistosomula.In situimmunolocalization with anti-SjKI-1 rabbit antibodies showed the protein was present in eggs trapped in the infected mouse intestinal wall. In functional assays, SjKI-1 inhibited trypsin in the picomolar range and chymotrypsin, neutrophil elastase, FXa and plasma kallikrein in the nanomolar range. Furthermore, SjKI-1, at a concentration of 7·5µm, prolonged 2-fold activated partial thromboplastin time of human blood coagulation. We also demonstrate that SjKI-1 has the ability to bind Ca++. We present, therefore, characterization of the first Kunitz protein fromS. japonicumwhich we show has an anti-coagulant properties. In addition, its inhibition of neutrophil elastase indicates SjKI-1 have an anti-inflammatory role. Having anti-thrombotic properties, SjKI-1 may point the way towards novel treatment for hemostatic disorders.

THE UPTAKE OF TRITIATED LYSINE VASOPRESSIN BY RAT AND PIG PITUITARY NEURAL LOBES IN VITRO

1971 ◽  
Vol 50 (4) ◽  
pp. 669-677 ◽  
Author(s):  
B. A. EDWARDS
Keyword(s):  
Tap Water ◽  
Exchange Chromatography ◽  
Control Group ◽  
Breakdown Product ◽  
Nacl Solution ◽  
Water Control ◽  
Neural Lobe ◽  
Lysine Vasopressin ◽  
And Control

SUMMARY Uptake of tritiated lysine vasopressin ([3H]LVP) was studied in halved neural lobes of rats (which had been given either tap water (control group) or 2% (w/v) NaCl solution as drinking water for 4 days) as well as in slices of pig neural lobe. Uptake of radioactivity into the neural lobes was shown but analysis of the extracts of incubated lobes of both species by ion exchange chromatography showed that very little of it remained in the tissue as hormone. In addition, some radioactivity was associated with trichloroacetic acid-insoluble proteins. After 90 min of incubation, and after correction for the breakdown, the uptake of unchanged [3H]LVP, expressed as a tissue: medium ratio, was 0·14 ± 0·04 and 0·09 ± 0·03 (mean ± s.e.m.) for the saline-treated and control rats respectively, while the tissue: medium ratios for the breakdown product(s) were 6·47 ± 0·45 and 5·50 ± 0·36. The results suggest uptake of [3H]LVP into the cell with almost complete intracellular breakdown of the hormone.

Abstract 596: Increased Migration of Neutrophils Across Endothelial Cells from Patients with Pulmonary Arterial Hypertension is Related to Reduced Platelet Endothelial Cell Adhesion Molecule -1 and is Prevented by the Neutrophil Elastase Inhibitor Elafin

2016 ◽  
Vol 36 (suppl_1) ◽  
Author(s):  
Shalina Taylor ◽  
Jan-Renier Moonen ◽  
Kazuya Miyagawa ◽  
Mingxia Gu ◽  
Silin Sa ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Neutrophil Elastase ◽  
Cell Adhesion Molecule ◽  
Neutrophil Adhesion ◽  
Elastase Inhibitor ◽  
Platelet Endothelial Cell Adhesion ◽  
Pulmonary Arterial ◽  
And Control ◽  
Cell Adhesion Molecule 1 ◽  
Endothelial Cell Adhesion

Pulmonary arterial hypertension (PAH) is a devastating progressive disease associated with a high mortality despite current vasodilator therapies. Perivascular inflammation and high levels of neutrophil elastase are thought to play a pivotal role in the adverse vascular remodeling of small pulmonary arteries that causes PAH. Despite this, the function of neutrophils and in particular, their interaction with the pulmonary arterial endothelium has not been studied in PAH. We hypothesized that neutrophil functions such as adhesion to a substratum or to endothelial cells and transmigration across a substratum or trans-endothelial migration (TEM) are abnormal in PAH on the basis of dysfunction of both cell types. Using a neutrophil like cell line dHL-60 and isolated human neutrophils from donors and PAH patients, we demonstrated no significant difference in adhesion to PAH vs. control PAECs when stimulated with TNF-α (100μg/mL). However, TEM of both IL-8 (100ng/ml) and fMLP (100nM) stimulated dHL-60 cells and donor neutrophils was enhanced in PAH vs. control PAECs (p<0.01) likely related to reduced expression of Platelet Endothelial Cell Adhesion Molecule -1 (PECAM-1) in PAH PAECs (p <0.001). We therefore further hypothesized that inhibition of neutrophil elastase via recombinant human elafin would be sufficient to reverse TEM in both PAH and control PAEC by reducing neutrophil adhesion, a feature known to be elastase-dependent. Administration of elafin (1μg/mL) attenuated fMLP and IL-8 induced dHL-60 and neutrophil adhesion to fibrinogen and fibronectin (p<0.05 for both) as well as transmigration across both substrates (p<0.05 for both). Elafin reduced adhesion of neutrophil similarly in PAH and control PAEC (p <0.05 for both). Furthermore, in a dose dependent manner, elafin inhibited TEM in PAH PAEC by 40% in control PAECs, by 20% (p<0.001 and p<0.01 respectively) bringing values to within the same range of suppressing but not eliminating TEM. We therefore conclude that despite the reduction in PECAM-1 in PAH PAEC, inhibition of neutrophil adhesion with the elastase inhibitor elafin is sufficient to prevent the enhanced TEM. Elafin may therefore be of benefit in suppressing the deleterious impact of neutrophil in enhanced inflammation seen in PAH.

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