scholarly journals Changes in porcine nutrient transport physiology in response to Ascaris suum infection

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Sarina Koehler ◽  
Andrea Springer ◽  
Nicole Issel ◽  
Stefanie Klinger ◽  
Christina Strube ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Ascaris Suum ◽  
Short Circuit ◽  
Ussing Chamber ◽  
Nutrient Transport ◽  
Transport Processes ◽  
Infected Group ◽  
Peptide Transporter ◽  
Control Group ◽  
Expression Levels

Abstract Background The roundworm Ascaris suum is one of the parasites with the greatest economic impact on pig farming. In this context, lower weight gain is hypothesized to be due to decreased nutrient absorption. This study aims at characterizing the effects of A. suum infection on intestinal nutrient transport processes and potential molecular mechanisms. Methods Three groups of six piglets each were infected orally (10,000 embryonated A. suum eggs) in a single dose (“single infection”). Another three groups were infected orally (1000 embryonated eggs) for 10 consecutive days (“trickle infection”). Animals were necropsied 21, 35 and 49 days post-infection (dpi). Three groups served as respective controls. The Ussing chamber technique was applied for the functional characterization of small intestinal tissues [short-circuit currents (Isc) as induced by glucose, alanine and peptides; 3H-glucose net flux rates; tissue conductance (Gt)]. Transcription and expression levels of relevant cytokines and nutrient transporters were evaluated (qPCR/western blot). Results Peptide- and alanine-induced changes in Isc were significantly decreased in the jejunum and ileum of the trickle-infected group at 49 dpi and in the ileum of the single-infected group at 49 dpi. No significant differences regarding glucose transport were observed between the Ascaris-infected groups and the control group in Ussing chamber experiments. Transcription levels of the glucose and peptide transporters as well as of selected transcription factors (transcription of signal transducer and activator of transcription 6 [STAT6] and hypoxia-inducible factor 1-alpha [Hif-1α]) were significantly increased in response to both infection types after some periods. The transcription of interleukins 4 and 13 varied between decrease and increase regarding the respective time points, as did the protein expression of glucose transporters. The expression of the peptide transporter PepT1 was significantly decreased in the ileal single-infected group at 35 dpi. Hif-1α was significantly increased in the ileal tissue from the single-infected group at 21 dpi and in the trickle-infected group at 35 dpi. The expression levels of Na+/K+-ATPase and ASCT1 remained unaffected. Conclusions In contrast to the current hypothesis, these results indicate that the nutrient deprivation induced by A. suum cannot be explained by transcriptional or expression changes alone and requires further studies. Graphical abstract

Basolateral Cl− uptake mechanisms in Xenopus laevis lung epithelium

2010 ◽  
Vol 299 (1) ◽  
pp. R92-R100 ◽  
Author(s):  
Jens Berger ◽  
Martin Hardt ◽  
Wolfgang G. Clauss ◽  
Martin Fronius
Keyword(s):  
Xenopus Laevis ◽  
Ion Transport ◽  
Anion Exchanger ◽  
Short Circuit ◽  
Ussing Chamber ◽  
Short Circuit Current ◽  
Transport Processes ◽  
Lung Epithelium ◽  
Active Ion Transport ◽  
Uptake Mechanisms

A thin liquid layer covers the lungs of air-breathing vertebrates. Active ion transport processes via the pulmonary epithelial cells regulate the maintenance of this layer. This study focuses on basolateral Cl− uptake mechanisms in native lungs of Xenopus laevis and the involvement of the Na+/K+/2 Cl− cotransporter (NKCC) and HCO3−/Cl− anion exchanger (AE), in particular. Western blot analysis and immunofluorescence staining revealed the expression of the NKCC protein in the Xenopus lung. Ussing chamber experiments demonstrated that the NKCC inhibitors (bumetanide and furosemide) were ineffective at blocking the cotransporter under basal conditions, as well as under pharmacologically stimulated Cl−-secreting conditions (forskolin and chlorzoxazone application). However, functional evidence for the NKCC was detected by generating a transepithelial Cl− gradient. Further, we were interested in the involvement of the HCO3−/Cl− anion exchanger to transepithelial ion transport processes. Basolateral application of DIDS, an inhibitor of the AE, resulted in a significantly decreased the short-circuit current (ISC). The effect of DIDS was diminished by acetazolamide and reduced by increased external HCO3− concentrations. Cl− secretion induced by forskolin was decreased by DIDS, but this effect was abolished in the presence of HCO3−. These experiments indicate that the AE at least partially contributes to Cl− secretion. Taken together, our data show that in Xenopus lung epithelia, the AE, rather than the NKCC, is involved in basolateral Cl− uptake, which contrasts with the common model for Cl− secretion in pulmonary epithelia.

scholarly journals Expression of ADAMTS13 in human endometrium and its potential role in recurrent pregnancy loss

2021 ◽  
Author(s):  
Yun Feng ◽  
Xueyin Li ◽  
Xi Chen ◽  
Mengling Zhao ◽  
Qian Wang ◽  
...  
Keyword(s):  
Pregnancy Loss ◽  
Molecular Mechanisms ◽  
Recurrent Pregnancy Loss ◽  
Gene Knockout ◽  
Embryo Implantation ◽  
Human Endometrium ◽  
Control Group ◽  
Expression Levels ◽  
Knockout Animal ◽  
Mrna And Protein Expression

IntroductionThe mechanisms underlying the pathogenesis of recurrent pregnancy loss (RPL) and the effective approaches to treat this disease still remain vague and absent. Proteinases of ADAMTS family play important roles in embryonic growth and development. Our previous study suggest a role of ADAMTS13 during pregnancy. Current Study was to determine the expression of ADAMTS13 in human endometrium and its association with RPL.Material and methodsThe spatiotemporal expression of ADAMTS13 in human endometrium was examined by immunohistochemistry. real-time PCR sand western blot were then employed to determine the mRNA and protein expression levels of ADAMTS13 in human endometrium. Proteolytic cleavage of FRETS-VWF73 were performed to determine the activity of ADAMTS13 in plasma and that secreted by human endometrium. ELISA was carried out to measure plasma VWF antigen.ResultsWe show that proteolytically active ADAMTS13 is expressed in human endometrium throughout the menstrual cycle and pregnancy. The decidual expression levels of mRNA and protein in women with RPL were significantly lower compared with women with uncomplicated pregnancies (P<0.01, P<0.05, respectively). Furthermore, significantly reduced plasma ADAMTS13 activity (median [range] 69.09 [65.2–93.7]% versus 93.62 [88.1–115.6]%, P<0.001) and elevated plasma VWF antigen levels (median [range] of 125.5 [54.2–262.8]% versus 91.9[80.4–138.7]%, P < 0.01) were detected in RPL patients compared with the control group.ConclusionsThese findings suggest that ADAMTS13 may play a role in embryo implantation and the pathogenesis of recurrent pregnancy loss. Further investigation on ADAMTS13 gene knockout animal models is necessary for understanding the molecular mechanisms of the biological roles of ADAMTS13 during gestation.

scholarly journals Ascaris suum Nutrient Uptake and Metabolic Release, and Modulation of Host Intestinal Nutrient Transport by Excretory-Secretory and Cuticle Antigens In Vitro

Pathogens ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1419
Author(s):  
Sarina Koehler ◽  
Andrea Springer ◽  
Nicole Issel ◽  
Stefanie Klinger ◽  
Michael Wendt ◽  
...  
Keyword(s):  
Nutrient Uptake ◽  
Molecular Mechanisms ◽  
Ascaris Suum ◽  
Nutrient Transport ◽  
Ammonia Excretion ◽  
Electrogenic Transport ◽  
Ussing Chambers ◽  
Net Flux

Ascaris suum, the most important pig parasite, also infects humans as a zoonotic pathogen. Malabsorption upon infection probably results from impaired nutrient transport, presumably mediated by the parasite´s excretory-secretory (ES) or cuticle somatic (CSO) antigens. The present study investigated the electrogenic transport (∆Isc) of glucose, alanine and the dipeptide glycyl-l-glutamine (glygln), as well as glucose net flux rates in pig jejunal tissue after in vitro exposure to adult A. suum total ES or CSO antigens in Ussing chambers. ∆Isc of glucose, alanine and glucose net flux rate were significantly decreased after one hour of exposure to total ES antigen. In contrast, CSO antigens increased the transport of glygln. Additionally, nutrient uptake and ES antigen pattern were compared in culture medium from untreated adult worms and those with sealed mouth and anal openings. Untreated worms completely absorbed glucose, while cuticular absorption in sealed worms led to 90% reduction. Amino acid absorption was 30% less effective in sealed worms, and ammonia excretion decreased by 20%. Overall, the results show that A. suum total ES antigen rapidly impairs nutrient transport in vitro. Future studies confirming the results in vivo, narrowing down the ES components responsible and investigating underlying molecular mechanisms are needed.

Small conductance potassium channels drive ATP-activated chloride secretion in the oviduct

2012 ◽  
Vol 302 (1) ◽  
pp. C100-C109 ◽  
Author(s):  
N. Keating ◽  
L. R. Quinlan
Keyword(s):  
Potassium Channels ◽  
Intracellular Calcium ◽  
Molecular Mechanisms ◽  
Short Circuit ◽  
Ussing Chamber ◽  
Chloride Secretion ◽  
Sodium Absorption ◽  
Oviduct Epithelium ◽  
Small Conductance ◽  
High Conductance

The molecular mechanisms controlling fluid secretion within the oviduct have yet to be determined. As in other epithelia, both secretory and absorptive pathways are likely to work in tandem to drive appropriate ionic movement to support fluid movement across the oviduct epithelium. This study explored the role of potassium channels in basolateral extracellular ATP (ATPe)-stimulated ion transport in bovine oviduct epithelium using the Ussing chamber short-circuit current ( ISC) technique. Basal ISC in bovine oviduct epithelium comprises both chloride secretion and sodium absorption and was inhibited by treatment with basolateral K+ channel inhibitors tetrapentlyammonium chloride (TPeA) or BaCl2. Similarly, ATP-stimulated chloride secretion was significantly attenuated by pretreatment with BaCl2, tetraethylammonium (TEA), tolbutamide, and TPeA. Basolateral K+ current, isolated using nystatin-perforation technique, was rapidly activated by ATPe, and pretreatment of monolayers with thapsigargin or TPeA abolished this ATP-stimulated K+ current. To further investigate the type of K+ channel involved in the ATP response in the bovine oviduct, a number of specific Ca2+-activated K+ channel inhibitors were tested on the ATP-induced Δ ISC in intact monolayers. Charbydotoxin, (high conductance and intermediate conductance inhibitor), or paxilline, (high conductance inhibitor) did not significantly alter the ATPe response. However, pretreatment with the small conductance inhibitor apamin resulted in a 60% reduction in the response to ATPe. The presence of small conductance family member KCNN3 was confirmed by RT-PCR and immunohistochemistry. Measurements of intracellular calcium using Fura-2 spectrofluorescence imaging revealed the ability of ATPe to increase intracellular calcium in a phospholipase C-inositol 1,4,5-trisphosphate pathway-sensitive manner. In conclusion, these results provide strong evidence that purinergic activation of a calcium-dependent, apamin-sensitive potassium conductance is essential to promote chloride secretion and thus fluid formation in the oviduct.

Computational studies of ion-water flux coupling in the airway epithelium. I. Construction of model

1996 ◽  
Vol 270 (6) ◽  
pp. C1751-C1763 ◽  
Author(s):  
J. A. Novotny ◽  
E. Jakobsson
Keyword(s):  
Water Transport ◽  
Airway Epithelium ◽  
Short Circuit ◽  
Ussing Chamber ◽  
Ionic Composition ◽  
Membrane Potentials ◽  
Transport Processes ◽  
Model Parameters ◽  
Constant Field ◽  
Sodium Potassium

A mathematical model of ion and water transport across the airway epithelium is presented. The model consists of 12 state variables representing ion concentrations, volumes, and membrane potentials. All osmotically significant membrane transport processes for which there is conclusive experimental evidence are included: passive apical sodium and chloride movement, basolateral sodium-potassium pumping, basolateral sodium-potassium-chloride cotransport, passive basolateral potassium movement, nonselective passive paracellular ion motion, and water transport across all membranes. Ion movements are described by Michaelis-Menten kinetics or by the constant field flux equation. Model parameters are established with Ussing chamber data. Model behavior is validated by comparing in vitro simulations with experimental results. The model accurately reproduces short-circuit chloride and sodium fluxes, short-circuit current, and open-circuit membrane potentials from Ussing chamber data in the secreting and nonsecreting states. The model is then used to describe the behavior of the airway epithelium in vivo, in which case the apical electrolyte compartment is small and of variable size and ionic composition.

scholarly journals Impact of Campylobacter spp. on the Integrity of the Porcine Gut

Animals ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 2742
Author(s):  
Alexandra Rath ◽  
Silke Rautenschlein ◽  
Janina Rzeznitzeck ◽  
Gerhard Breves ◽  
Marion Hewicker-Trautwein ◽  
...  
Keyword(s):  
Short Circuit ◽  
Ussing Chamber ◽  
Clinical Signs ◽  
Short Circuit Current ◽  
Transport Processes ◽  
Transepithelial Transport ◽  
Infection Model ◽  
Food Borne ◽  
The Impact ◽  
Campylobacter Spp

Campylobacter (C.) is the most common food-borne zoonosis in humans, which mainly manifests with watery to bloody diarrhoea. While C. jejuni is responsible for most cases of infection, C. coli is less frequently encountered. The object of the study was to prove the clinical impact of mono- and co-colonisation of C. coli and C. jejuni on weaned piglets in an infection model and to investigate the impact on transepithelial transport processes in the jejunum and caecum. At an age of eight weeks, eight pigs were infected with C. coli (ST-5777), 10 pigs with C. jejuni (ST‑122), eight pigs with both strains, and 11 piglets served as control. During the four-week observation period, no clinical signs were observed. During dissection, both strains could be isolated from the jejunum and the caecum, but no alteration of the tissue could be determined histopathologically. Mono-infection with C. jejuni showed an impact on transepithelial ion transport processes of the caecum. An increase in the short circuit current (Isc) was observed in the Ussing chamber resulting from carbachol- and forskolin-mediated Cl− secretion. Therefore, we speculate that caecal colonisation of C. jejuni might affect the transport mechanisms of the intestinal mucosa without detectable inflammatory reaction.

scholarly journals lncRNA-GPAND Promotes Gastric Cancer Progression via RUNX2 and MMP13

2021 ◽  
Author(s):  
Ying Wang ◽  
Xiaojuan Pei ◽  
Chunbing Wang ◽  
Zhibo Tan
Keyword(s):  
Gastric Cancer ◽  
Cell Proliferation ◽  
Cell Line ◽  
Cell Lines ◽  
Molecular Mechanisms ◽  
Tissue Array ◽  
Control Group ◽  
Cancer Tissue ◽  
Expression Levels ◽  
Ags Cell Line

Abstract Background: Gastric cancer (GC) is still one major reason for cancer-related death worldwide and in China, which ranks the second highest common cancer death rate. It is of great importance to study the molecular mechanisms by which gastric cancer develops. Methods: In this study, in situ hybridization histochemistry (ISHH) was used to examine the lncRNA-GPAND expression levels using gastric cancer tissue array. The real-time live-cell imaging system was used to investigate the effect of GPAND on cell proliferation and apoptosis of GC cell lines. Cell cycle of AGS cell line was examined after GPAND was suppressed using the flow cytometry (FCM). Transwell method was used to study the effect of GPAND on the invasion characteristics of GC cell line. Then the next generation sequencing (NGS) was used to study the potential molecular mechanism and the pathway, and the RT-qPCR was performed to verify the potential targets found by NGS method. Results: It was shown that GPAND was significantly over-expressed in the gastric cancer (GC) tissues (n=215) compared with the paired non-cancerous tissues (n=215), the expression levels of GPAND of GC tissues of TNM stage I-II (n=45) were significantly higher than that of stage III-IV (n=147). It has shown that knockdown of GPAND inhibited the AGS and N87 cell proliferation and promoted the cell apoptosis of AGS and N87 cell lines significantly, and the G1 phase percentage was remarkably increased in GPAND knockdown group of AGS cell line compared with control group. Moreover, suppression of GPAND inhibited the AGS cell invasion significantly. It was found via the NGS method that RUNX2 and MMP13 were significantly up-regulated when the GPAND was over-expressed. Conclusions: These observations suggest the lncRNA-GPAND/RUNX2/MMP13 axis to be a viable therapeutic target for gastric cancer.

scholarly journals Bioinformatics Prediction and Experimental Verification Identify MAD2L1 and CCNB2 as Diagnostic Biomarkers of Rhabdomyosarcoma

2021 ◽  
Author(s):  
Tian Xia ◽  
Lian Meng ◽  
Zhijuan Zhao ◽  
Yujun Li ◽  
Hao Wen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Molecular Mechanisms ◽  
Clinical Stage ◽  
Gene Set Enrichment Analysis ◽  
Clinical Staging ◽  
Control Group ◽  
Soft Tissue Tumour ◽  
Hub Genes ◽  
Expression Levels ◽  
Differential Genes

Abstract Background: Rhabdomyosarcoma (RMS) is a malignant soft-tissue tumour. In recent years, the tumour microenvironment (TME) has been reported to be associated with the development of tumours. However, the relationship between the occurrence and development of RMS and TME is unclear. The purpose of this study is to identify potential tumor microenvironment-related biomarkers in rhabdomyosarcoma and analyze their molecular mechanisms, diagnostic and prognostic significance.Methods: We first applied bioinformatics method to analyse the tumour samples of 187 patients with rhabdomyosarcoma (RMS) from the Gene Expression Omnibus database (GEO). Then, we used cell function and molecular biology techniques to study the progress of RMS.Results: Bioinformatics results show that the RMS TME key genes were screened, and a TME-related tumour clinical staging model was constructed. The top 10 hub genes were screened through the establishment of a protein-protein interaction network, and then Gene Expression Profiling Interactive Analysis was conducted to measure the overall survival (OS) of the 10 hub genes in the sarcoma cases in The Cancer Genome Atlas (TCGA). Six differential genes of statistical significance were acquired. The correlation between these six differential genes and the clinical stage of RMS was analysed. Our data found that the expression levels of MAD2L1 and CCNB2 negatively correlated with the OS of RMS patients and positively correlated with the clinical stage of RMS patients. Immunohistochemical results also confirmed that the expression levels of MAD2L1 (30/33, 87.5%) and CCNB2 (33/33, 100%) were remarkably higher in RMS group than in normal control group (0/11, 0%). Moreover, the expression of CCNB2 was related to tumour size. Further gene set enrichment analysis revealed that the genes in MAD2L1 and CCNB2 groups with high expression were mainly related to the mechanism of tumour metastasis and recurrence. In the low-expression MAD2L1 and CCNB2 groups, the genes were enriched in the metabolic and immune pathways. Downregulation of MAD2L1 and CCNB2 suppressed the growth, invasion, migration, and cell cycling of RMS cells and promoted their apoptosis. The CIBERSORT immune cell fraction analysis indicated that the expression levels of MAD2L1 and CCNB2 affected the immune status in the TME. Conclusions: The expression levels of MAD2L1 and CCNB2 may help guide the prognosis of patients with RMS and the clinical staging of tumours.

Liver, Kidney Function Tests and Oxidative Damage During and after Treatment of Salmonella typhimurium Infection in Experimental Local Rabbits

2018 ◽  
Vol 9 (4) ◽  
Author(s):  
Mustafa Salah Hasan ◽  
Ayman Barzan Abdulgafor ◽  
Maher Saber Owain ◽  
Mohammed Ali Hussein ◽  
Qusay Mohammed Aboud ◽  
...  
Keyword(s):  
Single Dose ◽  
Oxidative Damage ◽  
Salmonella Typhimurium ◽  
Infected Group ◽  
Post Treatment ◽  
Kidney Damage ◽  
Control Group ◽  
Post Inoculation ◽  
Group 5 ◽  
Group 2

This study aimed to evaluate the liver, kidney damage caused by S. typhimurium and to estimate the oxidative damage in association with this bacteria. A highly virulent isolates of S. typhimurium were obtained from the department of internal and preventive medicine/ College of Veterinary Medicine/ University of Baghdad. A twenty five local rabbits of both genders with age range (2-4 months) weeks old were used for this study, the rabbits were divided randomly into five groups each group contains 5 rabbits :- group 1: drenched orally with 5 ml of normal saline and consider as control group, group 2: were drenched orally with (5 ml) suspension which contain (5��109 CFU) of Salmonella typhimurium and regarded as infected group, group 3 were drenched orally with (5 ml) suspension which have (5��109 CFU) of Salmonella typhimurium then treated with a single dose of gentamicin alone at 0.05ml/kg (5mg/ml) orally after presence of signs (after 24hrs. post inoculation), group 4 were drenched (5 ml) suspension having (5��109 CFU) of Salmonella typhimurium then treated with a single dose of Ca-EDTA alone at 40mg/kg orally after presence of signs (after 24hrs. post inoculation) and group 5 were drenched (5 ml) suspension that contain (5��109 CFU) of Salmonella typhimurium then treated with a single dose of combined gentamicin at 0.05ml/kg (5mg/ml) orally after presence of signs (after 24hrs. post inoculation) and Ca-EDTA 40mg/kg after presence of signs (after 24hrs. post inoculation).The results of biochemical profile showed a significant increase (p less than 0.05) in ALT, creatinine and urea levels in infected group as compared with control group, while, the treated groups especially group 5 showed a significant improvement in ALT, Urea and creatinine levels which returned to relative normal levels as compared with infected group after 96hrs. post treatment. Also, the results of oxidative stress showed a significant increase in the levels of MDA in G2, G3, G4 and G5 after 48 hrs. post treatment, while the level of GSH showed a significant decrease in the level at 48hrs., both were returned to relative normal levels after 96hrs.post treatment especially in group 5.In conclusion, S. typhimurium can causing liver and kidney damage which is manifested by increase ALT, Urea and Creatinine. Also, MDA and GSH is increased due to salmonellosis.

Expression Levels of MicroRNA-125b in Serum Exosomes of Patients with Asthma of Different Severity and its Diagnostic Significance

2019 ◽  
Vol 20 (10) ◽  
pp. 781-784 ◽  
Author(s):  
Meizhen Zhao ◽  
Li Juanjuan ◽  
Fan Weijia ◽  
Xie Jing ◽  
Huang Qiuhua ◽  
...  
Keyword(s):  
Correlation Analysis ◽  
Roc Curve ◽  
Quantitative Polymerase Chain Reaction ◽  
Persistent Asthma ◽  
Asthma Severity ◽  
Control Group ◽  
Spearman Correlation ◽  
Diagnostic Efficacy ◽  
Expression Levels ◽  
Serum Exosomes

Background: This study aimed to investigate the expression levels of microRNA (miRNA)-125b in serum exosomes and its diagnostic efficacy for asthma severity. Methods: The study included 80 patients with untreated asthma and 80 healthy volunteers. The patients were divided into 4 groups according to disease severity: 20 with the intermittent state, 20 with the mildly persistent state, 20 with the moderately persistent state, and 20 with the severely persistent state. The expression levels of miRNA-125b in serum exosomes of each group were detected using a quantitative polymerase chain reaction and compared. The Spearman correlation analysis was used to study the correlation between the expression levels of miRNA-125b in serum exosomes and asthma severity. The diagnostic efficacy of the expression levels of miRNA-125b in exosomes for asthma severity was evaluated using the Receiver Operating Characteristic (ROC) curve. Results: The expression levels of miRNA-125b in serum exosomes of patients with intermittent, mildly persistent, moderately persistent, and severely persistent asthma were all higher than those in the healthy control group, with statistically significant differences. The expression levels of miRNA-125b were also statistically significantly different among patients in each group. The Spearman correlation analysis showed a positive correlation of the relative expression of miRNA-125b in serum exosomes with asthma severity. The area under the ROC curve of the diagnostic efficacy of miRNA-125b in serum exosomes for patients with intermittent, mildly, moderately, and severely persistent asthma was 0.7770, 0.8573, 0.9111, and 0.9995, respectively. Conclusion: The expression levels of miRNA-125b in serum exosomes had a high diagnostic efficacy and might serve as a noninvasive diagnostic marker for asthma severity.

Sign in / Sign up

Export Citation Format

Share Document