Markers Of Tissue Damage Research Articles

Paracetamol and Hydrogen peroxide-induced tissue alterations are reduced by Costus afer extract

Aim/Objective: To evaluate the effect of the methanolic extract of Costus afer stem on paracetamol-induced tissue injury in rats (in-vivo) and its ability to reduce hydrogen peroxide-induced erythrocyte damage (in-vitro). Methods: Twenty-five rats were acclimatized for two weeks and randomized into five groups. Group I were untreated controls, while each rat in groups III - V was given a single intraperitoneal-administration of paracetamol (800 mg/kg b.w.). Prior to the administration of paracetamol, rats in groups II and IV were orally given Costus afer extract at 200 mg/kg b.w while rats in group V were given 400 mg/kg b.w. extract once daily for four days. In the other experiment, hydrogen peroxide was used to induce erythrocyte damage (after isolating erythrocytes from whole blood) and subsequently incubated with the extract at different concentrations. Markers for tissue and erythrocyte damage were assessed in serum, tissue homogenates and erythrocytes. Results: Paracetamol caused marked elevations (p < 0.05) in the levels of alanine aminotransaminase (ALT), aspartate aminotransaminase (AST), alkaline phosphatase (ALP), urea and creatinine in serum. The levels of thiobarbituric acid reactive species (TBARS) in both liver and kidney were also raised over control (p < 0.05) while the activities of superoxide dismutase (SOD) and catalase were reduced over untreated controls (p < 0.05). Pre-treatment of rats with Costus afer extract (p.o.) caused significant reduction in serum levels of ALT, AST, ALP, urea, creatinine and tissue TBARS closer to control values (p < 0.05). Pre-treatment of the rats with the plant extract also increased the activities of SOD and catalase in both liver and kidney. Apart from the levels of TBARS in liver and catalase in kidney, rats pre-treated with 400 mg/kg b.w. extract showed greater response than pre-treatment with 200 mg/kg b.w. The extract also reduced hydrogen peroxide-induced erythrocyte damage assessed as haemolysis and lipid peroxidation which was concentration-dependent (p < 0.05). Conclusion: This indicated that Costus afer possesses the ability to protect the various tissues against chemical-induced injury.

Cerebral damage pathogeny peculiarities in local and globalcerebral perfusion impairment models in rats

A complex study of clinical and laboratory pathogenic features of cerebrovascular disease has been performed in rats using two experimental models: local ischemia with cerebral infarction and global ischemia caused by acute or chronic impairment of cerebral circulation without verified site of brain damage. A pronounced difference has been demonstrated between the two studied models of cerebral impairment. The development of brain damage site constituting 20-30% of the total cortex of the impaired hemisphere was demonstrated to be accompanied by a considerable increase of basic nervous tissue damage markers (neuron-specific enolase, S100β-protein), development of endothelial dysfunction and hemostasiological disturbances. Irreparable cerebral circulation impairment by ligation of carotid arteries was followed by considerably lower lethality in the experimental animals, neurospecific markers’ moderate dynamics but however side by side with pronounced and inadequately compensated manifestations in the hemostasis system and vascular endothelium caused by brain hypoxia.

Prediction of bleeding and thrombosis by standard biochemical coagulation variables in haematological intensive care patients.

We assessed the value of standard biochemical coagulation parameters in predicting bleeding, thrombosis and mortality in adult Intensive Care Unit (ICU) patients with haematological malignancies. We screened all patients with acute leukaemia and myelodysplastic syndrome admitted to a university hospital ICU during 2008-2012. Data were obtained from the clinical chemistry laboratory database and patient files. We graded bleeding according to the World Health Organisation (WHO)-system within 24-h, within 5-days and during the whole ICU stay. We analysed the predictive values of laboratory parameters using multiple logistic regression and receiver operator characteristics (ROC) curves. As we previously have established that platelet count at admission was associated with bleeding, we focused on International Normalised Ratio (INR), activated pro-thrombin time (APTT), anti-thrombin, D-dimer and fibrinogen, and markers of infection (C-reactive protein, pro-calcitonin), kidney function (creatinine) and tissue damage (lactate dehydrogenase (LDH)). We included 116 patients; 66 (57%) had at least one bleeding episode and 11 (9%) patients had at least one thrombotic event. The differences in coagulation values when bleeding compared to baseline values were minor. INR was the only variable we found associated with subsequent bleeding within 24 h from admission to ICU (odds ratio 2.91, 95% CI: 1.01-8.43, P = 0.048). ROC analyses did not show predictive value of any of the other variables with regards to bleeding and none of the variables were associated with thrombosis in adjusted analyses. Increased levels of LDH at admission were associated with increased 7-day and 30-day mortality. Increased INR at admission was associated with a higher rate of bleeding in ICU patients with haematological malignancies. No other biochemical coagulation or other parameter had any association with bleeding, thrombosis or mortality except increased LDH, which at ICU admission was associated with increased 30-day mortality.

Blockade of Inflammation and Apoptosis Pathways by siRNA Prolongs Cold Preservation Time and Protects Donor Hearts in a Porcine Model.

In donor hearts from mini pigs, overtime cold preservation and ischemia-reperfusion injury cause poor graft quality and impaired heart function. Blockage of complement, apoptosis, and inflammation is considered a strategy for attenuating ischemia-reperfusion injury and protecting cardiac function. Minipig donor hearts were perfused and preserved in Celsior solution or transfection reagent containing Celsior solution with scramble siRNA or siRNAs targeting complement 3, caspase-8, caspase-3, and nuclear factor κB-p65 genes at 4°C and subsequently hemo-reperfused ex vivo (38°C) or transplanted into recipients. The protective effect of the siRNA solution was evaluated by measuring cell apoptosis, structural alteration, protein markers for tissue damage and oxidative stress, and cardiac function. We found a reduction in cell apoptosis, myocardial damage, and tissue inflammation by reduced biochemistry and markers and protein expression of proinflammatory cytokines and improvement in cardiac function, as shown by the improved hemodynamic indices in 12-hr-preserved siRNA-treated hearts of both ex vivo and orthotopic transplantation models. These findings demonstrate that blockade of inflammation and apoptosis pathways using siRNA can prolong cold preservation time and better protect donor heart function in cardiac transplantation of large animals, which may be beneficial for human heart preservation.

Earliest Phase of Systemic Sclerosis Typified by Increased Levels of Inflammatory Proteins in the Serum.

Patients with definite systemic sclerosis (SSc) who lack fibrotic features can be stratified into an intermediate stage of disease severity between preclinical/early SSc (EaSSc) and fibrotic subsets (limited cutaneous SSc [lcSSc] and diffuse cutaneous SSc [dcSSc]). The aim of the present study was to molecularly characterize nonfibrotic SSc and EaSSc on the basis of a broad panel of serum markers of inflammation and tissue damage, in order to increase the knowledge of the pathophysiologic mechanisms underlying SSc progression before the development of fibrosis. An 88-plex immunoassay was performed in serum samples from a discovery cohort composed of 21 patients with EaSSc (meeting the LeRoy and Medsger criteria), 15 with nonfibrotic SSc (meeting the American College of Rheumatology/European League Against Rheumatism 2013 classification criteria, without skin or lung fibrosis), and 11 healthy controls. Analyte concentrations that were consistently significantly different at the exploratory P value threshold of 0.1 were selected for replication analysis in a larger group composed of 47 patients with EaSSc, 48 with nonfibrotic SSc, and 43 healthy controls, as well as 51 patients with lcSSc and 35 with dcSSc. The value of the replicated molecules in predicting SSc progression (at a family-wise error rate of 0.05) was tested. Based on the results of the explorative analysis, 16 molecules were selected for testing in the replication set. The results showed that CXCL10, CXCL11, tumor necrosis factor receptor type II (TNFRII), and chitinase 3-like protein 1 levels were significantly increased in patients with EaSSc and those with nonfibrotic SSc as compared to healthy controls. The disease in patients with high concentrations of CXCL10 and TNFRII was also characterized by a faster rate of progression from EaSSc and from nonfibrotic SSc to worse disease stages. SSc patients with preclinical/early SSc and those with established, yet nonfibrotic, disease exhibit clear molecular alterations that are associated with faster rates of disease evolution. These data open novel avenues for disease interception in SSc.

Activation of planarian TRPA1 by reactive oxygen species reveals a conserved mechanism for animal nociception.

All animals must detect noxious stimuli to initiate protective behavior, but the evolutionary origin of nociceptive systems is not well understood. Here, we show that noxious heat and irritant chemicals elicit robust escape behaviors in the planarian Schmidtea mediterranea, and that the conserved ion channel TRPA1 is required for these responses. TRPA1 mutant flies (Drosophila) are also defective in noxious heat responses. Unexpectedly, we find that either planarian or human TRPA1 can restore noxious heat avoidance to TRPA1 mutant Drosophila, even though neither is directly activated by heat. Instead, our data suggest that TRPA1 activation is mediated by H2O2/Reactive Oxygen Species, early markers of tissue damage rapidly produced as a result of heat exposure. Together, our data reveal a core function for TRPA1 in noxious heat transduction, demonstrate its conservation from planarians to humans, and imply that animal nociceptive systems may share a common ancestry, tracing back to a progenitor that lived more than 500 million years ago.

Protective Effect of Glyceollins in a Mouse Model of Dextran Sulfate Sodium-Induced Colitis.

Glyceollins, which are derived from daidzein in soybean in response to various stimuli or stresses, have been reported to activate antioxidant/detoxifying enzymes in a nuclear factor (erythroid-derived 2)-like 2 (Nrf2)-dependent manner, in addition to exerting anti-inflammatory effects in murine macrophages. As the Nrf2 signaling pathway is known to antagonize nuclear factor (NF)-κB signaling, glyceollins likely have the potential to prevent or treat inflammatory bowel disease. Thus, this study was conducted to examine whether glyceollins could inhibit dextran sulfate sodium (DSS)-induced colitis in a mouse model. Ulcerative colitis (UC) was induced in male BALB/c mice by administering drinking water with 4% DSS for 5 days. Glyceollins (4 or 10 mg/kg of body weight) were orally administered 48 h before and after DSS treatment. We found that glyceollins alleviated histological colon damage and inflammation induced by DSS treatment. More specifically, glyceollins reduced plasma levels of inflammatory cytokines, such as tumor necrosis factor-α and interleukin-6, which were otherwise markedly increased by DSS treatment. Markers of tissue damage, including malondialdehyde and 8-hydroxy-2-guanosine, were significantly increased by DSS treatment; however, this effect was mitigated through concomitant treatment with glyceollins. Furthermore, nuclear accumulation of NF-κB p65 and the expression of inducible nitric oxide synthase were upregulated by glyceollins, consistent with the observed modulation of inflammatory markers. In conclusion, glyceollins have therapeutic potential for UC and merit further clinical study.

In Situ Imaging of Tissue Remodeling with Collagen Hybridizing Peptides.

Collagen, the major structural component of nearly all mammalian tissues, undergoes extensive proteolytic remodeling during developmental states and a variety of life-threatening diseases such as cancer, myocardial infarction, and fibrosis. While degraded collagen could be an important marker of tissue damage, it is difficult to detect and target using conventional tools. Here, we show that a designed peptide (collagen hybridizing peptide: CHP), which specifically hybridizes to the degraded, unfolded collagen chains, can be used to image degraded collagen and inform tissue remodeling activity in various tissues: labeled with 5-carboxyfluorescein and biotin, CHPs enabled direct localization and quantification of collagen degradation in isolated tissues within pathologic states ranging from osteoarthritis and myocardial infarction to glomerulonephritis and pulmonary fibrosis, as well as in normal tissues during developmental programs associated with embryonic bone formation and skin aging. The results indicate the general correlation between the level of collagen remodeling and the amount of denatured collagen in tissue and show that the CHP probes can be used across species and collagen types, providing a versatile tool for not only pathology and developmental biology research but also histology-based disease diagnosis, staging, and therapeutic screening. This study lays the foundation for further testing CHP as a targeting moiety for theranostic delivery in various animal models.

Seizure activity per se does not induce tissue damage markers in human neocortical focal epilepsy.

The contribution of recurring seizures to the progression of epileptogenesis is debated. Seizure-induced brain damage is not conclusively demonstrated either in humans or in animal models of epilepsy. We evaluated the expression of brain injury biomarkers on postsurgical brain tissue obtained from 20 patients with frequent seizures and a long history of drug-resistant focal epilepsy. The expression patterns of specific glial, neuronal, and inflammatory molecules were evaluated by immunohistochemistry in the core of type II focal cortical dysplasias (FCD-II), at the FCD boundary (perilesion), and in the adjacent normal-appearing area included in the epileptogenic region. We also analyzed surgical specimens from cryptogenic patients not presenting structural alterations at imaging. Astroglial and microglial activation, reduced neuronal density, perivascular CD3-positive T-lymphocyte clustering, and fibrinogen extravasation were demonstrated in the core of FCD-II lesions. No pathological immunoreactivity was observed outside the FCD-II or in cryptogenetic specimens, where the occurrence of interictal and ictal epileptiform activity was confirmed by either stereo-electroencephalography or intraoperative electrocorticography. Recurrent seizures do not induce the expression of brain damage markers in nonlesional epileptogenic cortex studied in postsurgical tissue from cryptogenic and FCD patients. This evidence argues against the hypothesis that epileptiform activity per se contributes to focal brain injury, at least in the neocortical epilepsies considered here. Ann Neurol 2017;82:331-341.

Relation between postoperative ileus and anastomotic leakage after colorectal resection: a post hoc analysis of a prospective randomized controlled trial.

Anastomotic leakage (AL) following abdominal surgery is a critical determinant of postoperative recovery, of which the aetiology is largely unknown. Interestingly, interventions aimed at reducing the inflammatory response and postoperative ileus (POI) have an unexpected effect on AL. The aim of this study was to investigate the relation of POI with inflammation and AL after colorectal resection. A post hoc analysis of a prospective randomized controlled trial in which patients underwent a colorectal resection was performed. Patients undergoing a colorectal resection were stratified into having or not having POI. The incidence of AL and other clinical parameters was registered prospectively. Intestinal fatty acid binding protein (I-FABP, a marker for tissue damage) and the inflammatory response in plasma and colon tissue were determined. AL was present in nine of 43 patients in the POI group, and in one of 65 in the group without POI (P<0.001). There was a significant association between POI and AL (OR 12.57, 95% CI: 2.73-120.65; P=0.0005). Patients with POI had significantly higher plasma levels of soluble tumour necrosis factor receptor 1 (TNFRSF1A) at 4h postoperatively (0.89ng/l, interquartile range 0.56) than patients without POI (0.80ng/l, interquartile range 0.37; P=0.04) and higher plasma levels of C-reactive protein on the second day postoperatively (234±77 vs 163±86mg/l; P=0.001). Patients who developed AL had significantly higher plasma levels of I-FABP compared with patients without AL at 24h after onset of surgery. POI is associated with a higher prevalence of AL and an increased inflammatory response.

Cytokine IL1α and lactate as markers for tissue damage in spineboard immobilisation. A prospective, randomised open-label crossover trial

BackgroundSpinal immobilisation using a rigid long spineboard is a well-established procedure in trauma care. During immobilisation, the body is exposed to high tissue-interface pressures. This may lead to a localised inflammatory response of the skin, which may be used to monitor the body's response to different types of immobilisation device. AimIn this study we compared the standard rigid spineboard with a new soft-layered spineboard regarding tissue-interface pressures, skin redness as an indicator of reactive hyperaemia and cutaneous IL1α and lactate release. MethodsTwelve healthy male participants were asked to lie supine on both a rigid and a soft-layered spineboard, loading the sacrum for one hour, followed by one hour in unloaded position. Tissue-interface pressures on the buttocks during loading were measured continuously using a pressure mapping mat. Cutaneous IL1α and lactate concentrations were assessed using Sebutapes, during 20-min periods. After each 20-min period, a photo of the buttocks was taken, which was later assessed for redness by two observers. ResultsSignificant differences in tissue-interface pressure and reactive hyperaemia were found between the two types of spineboard. Release of IL1α and lactate were found to increase with prolonged exposure to pressure, and to decrease in the unloaded prone position. A significant relationship was found between tissue-interface pressure and reactive hyperaemia, but not with IL1α nor lactate release. Time course of IL1α and lactate release was similar for both types of spineboard. ConclusionsIL1α and lactate both have a strong relationship with pressure exposure time, but not with pressure magnitude. Furthermore, IL1α was measured even in the absence of visible redness of the skin. The study offers the potention of biomarkers, reflecting inflammation and/or tissue metabolism, for use in assessing the effects of prolonged spineboard support.

Cerebrospinal fluid neurofilament light levels mark grey matter volume in clinically isolated syndrome suggestive of multiple sclerosis

Background: Brain atrophy is a known marker of irreversible tissue damage in multiple sclerosis (MS). Cerebrospinal fluid (CSF) osteopontin (OPN) and neurofilament light chain (NF-L) have been proposed as candidate surrogate markers of inflammatory and neurodegenerative processes in MS. Objective: To evaluate the relationship between CSF NF-L and OPN levels and brain grey and white matter volumes in patients with clinically isolated syndrome (CIS) suggestive of MS. Methods: A total of 41 CIS patients and 30 neurological controls (NCs) were included. CSF NF-L and OPN were measured by commercial ELISA. Measures of brain volume (normalized brain volume (NBV), normalized grey matter volume (NGV), peripheral grey matter volume (PGV), normalized white matter volume (WMV), and ventricular volume) were obtained by SIENAX. Corpus callosum index (CCI) was calculated. Brain volumes were categorized into ‘high’ and ‘low’ according to the median value. Results: CSF NF-L and OPN levels were higher in CIS patients in comparison with NCs. CIS patients with ‘low’ TGV, PGV, and TBV showed higher CSF NF-L levels than CIS patients with ‘high’ brain volumes. TGV and PGV correlated inversely with NF-L levels, whereas CCI was inversely related to OPN levels. CSF NF-L was the only independent predictor of TGV and PGV. Conclusion: CSF NF-L tracks mainly grey matter damage in patients with CIS suggestive of MS.

Antioxidant and DNA protective effects of NTX, a proprietary glycyrrhizin/d-mannitol product, in association with alcohol consumption: A randomized, placebo-controlled, double-blind, crossover study

Chronic/heavy alcohol consumption is a risk factor/cause of numerous diseases. Alcohol toxicity involves metabolic activation and production of reactive oxygen species (ROS) with resultant tissue damage. Glycyrrhizin exhibits antioxidant, anti-inflammatory and tissue protective properties. This randomized, double-blind, placebo-controlled, crossover study involving 50 healthy subjects evaluated the ability of a proprietary glycyrrhizin/d-mannitol product to modulate alcohol-induced oxidative stress and tissue damage. One-time acute alcohol consumption at 1.52gm/kg within an hour caused transient elevations in serum ROS and derivatives of reactive oxygen metabolites (dROM), increases in lipid peroxidation and protein carbonyls, and DNA damage in peripheral lymphocytes. Alcohol containing the glycyrrhizin/d-mannitol study product significantly modulated markers of oxidative stress and tissue damage, including a decrease in DNA damage as assessed by three assay techniques, and decreases in lipid and protein oxidative damage. Consumption of the proprietary glycyrrhizin/d-mannitol study product during alcohol consumption may provide protection against the adverse tissue-related effects of alcohol.

Identification of candidate protein markers of Bovine Parainfluenza Virus Type 3 infection using an in vitro model

Bovine Parainfluenza Virus Type 3 (BPI3V) infections are often asymptomatic, causing respiratory tissue damage and immunosuppression, predisposing animals to severe bacterial pneumonia, the leading cause of Bovine Respiratory Disease (BRD) mortality. As with many pathogens, routine BPI3V serology does not indicate the presence of damaged respiratory tissue or active infection. In vitro proteomic marker screening using disease relevant cell models could help identify markers of infection and tissue damage that are also detectable during in vivo infections. This study utilised a proteomic approach to investigate in vitro cellular responses during BPI3V infection to enhance the current understanding of intracellular host-virus interactions and identify putative markers of in vivo infection. Through 2D gel electrophoresis proteomic analysis, BPI3V Phosphoprotein P and host T-complex Protein 1 subunit theta were found to be accumulated at the latter stages of infection within bovine fibroblasts. These proteins were subsequently detected using targeted multiple reaction monitoring (MRM) mass spectrometry in the plasma of animals challenged with BPI3V, with differential protein level profiles observed dependant on animal vaccination status. Potential mechanisms by which BPI3V overcomes host cellular immune response mechanisms allowing for replication and production of viral proteins were also revealed. Assessment of circulating protein marker levels identified through an in vitro approach as described may enable more effective diagnosis of active viral infection and diseased or damaged respiratory tissue in animals and allow for more effective utilisation of preventative therapeutic interventions prior to bacterial disease onset and significantly aid the management and control of BRD.

Detection of Donor-Derived Microparticles in the Peripheral Blood of a Hand Transplant Recipient During Rejection.

BackgroundMicroparticles (MPs) are released from the plasma membrane of activated or dying cells and bear surface molecules from those cells. We examined whether donor-derived MPs in the peripheral blood of the recipient could serve as a marker of tissue damage due to rejection of a transplanted hand.MethodsPlatelet-free plasma from the recipient of the transplanted hand was analyzed for MPs bearing the donor-specific HLA molecule A*02 using flow cytometry. Rejection status of the transplanted hand was monitored by histopathology of skin punch biopsies.ResultsDonor-specific MPs expressing HLA A*02 were quantifiable in the peripheral blood of the recipient. Levels of these MPs increased with worsening rejection of the transplanted hand.ConclusionsThese findings demonstrate the ability to detect donor specific MPs through staining of graft cell-specific HLA and promote further investigation into the potential utility of flow cytometry for donor-derived MPs as a noninvasive tool to assess rejection in solid organ transplantation patients.

Impact of Hot Environment on Fluid and Electrolyte Imbalance, Renal Damage, Hemolysis, and Immune Activation Postmarathon.

Previous studies have demonstrated the physiological changes induced by exercise exposure in hot environments. We investigated the hematological and oxidative changes and tissue damage induced by marathon race in different thermal conditions. Twenty-six male runners completed the São Paulo International Marathon both in hot environment (HE) and in temperate environment (TE). Blood and urine samples were collected 1 day before, immediately after, 1 day after, and 3 days after the marathon to analyze the hematological parameters, electrolytes, markers of tissue damage, and oxidative status. In both environments, the marathon race promotes fluid and electrolyte imbalance, hemolysis, oxidative stress, immune activation, and tissue damage. The marathon runner's performance was approximately 13.5% lower in HE compared to TE; however, in HE, our results demonstrated more pronounced fluid and electrolyte imbalance, renal damage, hemolysis, and immune activation. Moreover, oxidative stress induced by marathon in HE is presumed to be related to protein/purine oxidation instead of other oxidative sources. Fluid and electrolyte imbalance and protein/purine oxidation may be important factors responsible for hemolysis, renal damage, immune activation, and impaired performance after long-term exercise in HE. Nonetheless, we suggested that the impairment on performance in HE was not associated to the muscle damage and lipoperoxidation.

Costus afer extract ameliorates paracetamol-induced tissue injury and H2O2-induced erythrocyte peroxidation

Aim/Objective: To evaluate the effect of the methanolic extract of Costus afer stem on paracetamol-induced tissue injury in rats and its ability to reduce hydrogen peroxide-induced erythrocyte damage. Methods: Twenty rats were acclimatized for two weeks and randomized into five groups. Group I were untreated controls, while each rat in groups III - V was given a single intraperitoneal-administration of paracetamol (800 mg/kg b.w.). Prior to the administration of paracetamol, rats in groups II and IV were orally given Costus afer extract at 200 mg/kg b.w while rats in group V were given 400 mg/kg b.w. extract once daily for four days. In the other experiment, hydrogen peroxide was used to induce erythrocyte damage and subsequently incubated with the extract at different concentrations. Markers for tissue and erythrocyte damage were assessed in serum, tissue homogenates and erythrocytes. Results: Paracetamol caused marked elevations (p < 0.05) in the levels of alanine aminotransaminase, aspartate aminotransaminase, alkaline phosphatas, urea, creatinine in serum. The levels of thiobarbituric acid reactive species (TBARS) in both liver and kidney were also raised over control (p < 0.05) while the activities of superoxide dismutase and catalase were reduced over untreated controls (p < 0.05) Pre-treatment of rats with Costus afer extract (p.o.) caused significant reduction of the markers closer to control values (p < 0.05). Apart from TBARS and catalase in liver kidney respectively, rats pre-treated with 400 mg/kg b.w. extract showed greater alterations in the levels of the markers. The extract also reduced hydrogen peroxide-induced erythrocyte damage assessed as haemolysis and lipid peroxidation which was concentration-dependent (p < 0.05). Conclusion: This indicated that Costus afer possesses the ability to protect the various tissues against injury which is implicated in some diseases. Implications are discussed.

The Piezo Actuator-Driven Pulsed Water Jet System for Minimizing Renal Damage after Off-Clamp Laparoscopic Partial Nephrectomy.

In the setting of partial nephrectomy (PN) for renal cell carcinoma, postoperative renal dysfunction might be caused by surgical procedure. The aim of this study was to clarify the technical safety and renal damage after off-clamp laparoscopic PN (LPN) with a piezo actuator-driven pulsed water jet (ADPJ) system. Eight swine underwent off-clamp LPN with this surgical device, while off-clamp open PN was also performed with radio knife or soft coagulation. The length of the removed kidney was 40 mm, and the renal parenchyma was dissected until the renal calyx became clearly visible. The degree of renal degeneration from the resection surface was compared by Hematoxylin-Eosin staining and immunostaining for 1-methyladenosine, a sensitive marker for the ischemic tissue damage. The mRNA levels of neutrophil gelatinase-associated lipocalin (Ngal), a biomarker for acute kidney injury, were measured by quantitative real-time PCR. Off-clamp LPN with ADPJ system was successfully performed while preserving fine blood vessels and the renal calix with little bleeding. In contrast to other devices, the resection surface obtained with the ADPJ system showed only marginal degree of ischemic changes. Indeed, the expression level of Ngal mRNA was lower in the resection surface obtained with the ADPJ system than that with soft coagulation (p = 0.02). Furthermore, using the excised specimens of renal cell carcinoma, we measured the breaking strength at each site of the human kidney, suggesting the applicability of this ADPJ to clinical trials. In conclusion, off-clamp LPN with the ADPJ system could be safely performed with attenuated renal damage.

Microparticle Formation in Peritoneal Dialysis: A Proof of Concept Study.

Background:Injury to the mesothelial layer of the peritoneal membrane during peritoneal dialysis (PD) is implicated in loss of ultrafiltration capacity, but there are no validated biomarkers for mesothelial cell injury. Microparticles (MPs) are 0.1 to 1.0 µm membrane vesicles shed from the cell surface following injury and are sensitive markers of tissue damage. Formation of MPs in the peritoneal cavity during PD has not been reported to date.Methods:We designed a single-center, proof of concept study to assess whether peritoneal solution exposure induces formation of mesothelial MPs suggestive of PD membrane injury. We examined MP levels in PD effluents by electron microscopy, nanoparticle tracking analysis (NTA), flow cytometry, procoagulant activity, and Western blot.Results:NTA identified particles in the size range of 30 to 900 nm, with a mean of 240 (SE: 10 nm). MP levels increased in a progressive manner during a 4-hour PD dwell. Electron microscopy confirmed size and morphology of vesicles consistent with characteristics of MPs as well as the presence of mesothelin on the surface. Western blot analysis of the MP fraction also identified the presence of mesothelin after 4 hours, suggesting that MPs found in PD effluents may arise from mesothelial cells.Conclusions:Our results suggest that MPs are formed and accumulate in the peritoneal cavity during PD, possibly as a stress response. Assessing levels of MPs in PD effluents may be useful as a biomarker for peritoneal membrane damage.

Protective effects of tea polyphenols on exhaustive exercise-induced fatigue, inflammation and tissue damage

ABSTRACTBackground: The beneficial properties of tea polyphenols have been extensively studied; however, less attention has been paid to their effects, especially anti-inflammatory effect during exhaustive exercise.Objective: The present study assessed the potential protective effects of tea polyphenols against the fatigue, inflammation and tissue injury caused by an exhaustive exercise bout in rats.Design: Twenty-four healthy male rats were divided into three groups. Group C was a sedentary control group, Groups E+TP and Group E performed a single exhaustive swimming test; all groups had normal diets, but Group E+TP was supplemented with tea polyphenols. All rats were immediately euthanized after exhaustive exercise, and biochemical and inflammatory parameters, including lactic acid (LA), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-10 (IL-10), lactic dehydrogenase (LDH), and creatine kinase (CK) activity levels, were measured. Reverse transcription (RT) and Real-Time PCR was employed to evaluate the mRNA expression of IL-1β in the liver.Results: The results showed a decrease in serum LA levels (22%, p < 0.05) in rats that consumed dietary tea polyphenols. Interestingly, dietary tea polyphenols decreased the serum levels of pro-inflammatory factors (TNF-α: 13%, p < 0.05; IL-1β: 10%, p < 0.05; and IL-6: 48%, p < 0.05) and shifted the serum IL-10/TNF-α ratio to a predominantly anti-inflammatory milieu (0.52 ± 0.07 vs. 0.67 ± 0.10, p < 0.01). Furthermore, the polyphenols effectively inhibited the release of tissue damage markers (CK: 24%, p < 0.05 and LDH: 28%, p < 0.05) in the serum and decreased IL-1β mRNA expression in the liver.Conclusions: This study indicated that tea polyphenols could significantly protect rats from the fatigue, inflammation and tissue damage induced by acute exhaustive exercise.