Epithelial Necrosis Research Articles

Novel Alginate-Chitosan Composite Microspheres for Implant Delivery of Vancomycin and In Vivo Evaluation.

In this study, vancomycin loaded alginate-chitosan composite microspheres were developed by emulsion cross-linking method. The in vitro and vivo characterizations were done to evaluate the feasibility of application. Our experimental results showed that the emulsification cross-linking technique appeared to be a feasible method for the preparation of alginate-chitosan composite microspheres. The microspheres were spherical in shape and the mean particle size and drug loading were 25.3 ± 5.4 μm and 18.5 ± 2.3% respectively. A sustained vancomycin release was realized i.e. the amount of cumulative release increased in a time frame of 24 h to reach an amount i.e. ~68%. The model that fit best for vancomycin released from the microspheres was the Higuchi kinetic model with a correlation coefficient r = 0.9996. In vivo results showed that the application of microspheres not only reduced the toxicity, but also maintained effective drug concentration. In addition, no severe signs of epithelial necrosis and sloughing of epithelial cells were detected in histological studies.

An Early Passage Human Isolate of Kyasanur Forest Disease Virus Shows Acute Neuropathology in Experimentally Infected CD-1 Mice

Kyasanur Forest disease virus (KFDV) is a tick-borne Flavivirus that causes a severe illness in humans. Disease spectrum can vary from subclinical infection to fatal cases with hemorrhagic complications. The pathology of KFDV remains incompletely understood. This study describes the histopathologic and immunohistochemical findings in experimentally infected infant CD-1 mice with an early passage human KFDV isolate. Acute histological changes were primarily seen in the brain. The spectrum of changes included gliosis, inflammatory response, necrosis, neural loss, and syncytium formation in mid and hind brain structures. Microscopic lesions observed in the liver were mainly necrosis and vacuolation of hepatocytes and in small intestine, prominent epithelial cell necrosis. KFDV antigens could be stained by a sensitive immunohistochemical labeling in the same organs. Findings from this study are suggestive of neuropathology as the main manifestation of an early passaged human KFDV isolate. Importantly, this suggests that KFDV may be causing primarily a neurologic disease and secondary organ damage could be because of disease pathology per se. The use of primary low passage human isolates and neuropathology profile could also be more apt in developing a challenge model for testing potential antivirals and therapeutic agents.

Rat Urinary Osteopontin and Neutrophil Gelatinase-Associated Lipocalin Improve Certainty of Detecting Drug-Induced Kidney Injury

Traditional kidney biomarkers are insensitive indicators of acute kidney injury, with meaningful changes occurring late in the course of injury. The aim of this work was to demonstrate the diagnostic potential of urinary osteopontin (OPN) and neutrophil gelatinase-associated lipocalin (NGAL) for drug-induced kidney injury (DIKI) in rats using data from a recent regulatory qualification submission of translational DIKI biomarkers and to compare performance of NGAL and OPN to five previously qualified DIKI urinary biomarkers. Data were compiled from 15 studies of 11 different pharmaceuticals contributed by Critical Path Institute's Predictive Safety Testing Consortium (PSTC) Nephrotoxicity Working Group (NWG). Rats were given doses known to cause DIKI or other target organ toxicity, and urinary levels of the candidate biomarkers were assessed relative to kidney histopathology and serum creatinine (sCr) and blood urea nitrogen (BUN).OPN and NGAL outperformed sCr and BUN in identifying DIKI manifested as renal tubular epithelial degeneration or necrosis. In addition, urinary OPN and NGAL, when used with sCr and BUN, increased the ability to detect renal tubular epithelial degeneration or necrosis. NGAL and OPN had comparable or improved performance relative to Kim-1, clusterin, albumin, total protein, and beta-2 microglobulin. Given these data, both urinary OPN and NGAL are appropriate for use with current methods for assessing nephrotoxicity to identify and monitor DIKI in regulatory toxicology studies in rats. These data also support exploratory use of urinary OPN and NGAL in safety monitoring strategies of early clinical trials to aid in the assurance of patient safety.

Histopathology biomarker responses in Asian sea bass, Lates calcarifer (Bloch) exposed to copper

Copper is a trace element necessary for the normal growth and metabolism of living organisms. If exceeds its limit it becomes pollutant and causes pathological effects. Asian sea bass (Lates calcarifer) was exposed to sublethal concentrations of copper for 28days and histopathological alterations were noticed in the gill, liver, muscle and intestine. Epithelial necrosis, hypertrophy, rupture of gill epithelium and haemorrhage at primary lamellae were observed after 7days of exposure. Lifting of epithelium, oedema and fusion of adjacent secondary lamellae were conspicuous at 28days of exposure. The experimental liver tissue showed reduction in the size of hepatocytes, vacuolisation, and hypertrophy. The intestine displayed fused microvilli, sloughing of mucus membrane and swollen cells. Muscle bundles with severe oedema and their thickening and separation were more pronounced in sublethal treatment of copper in the muscle. Several histopathological changes observed in various fish organs would serve a useful purpose in evaluating the toxic effects of copper. The present study clearly demonstrated that all the treated body organs exhibited significant damage with response; amongst the body organs the liver is an important target organ for copper toxicity in L. calcarifer and this species could be possibly used as a model organism for toxicity studies.

Amikacin induced renal damage and the role of the antioxidants on neonatal rats

Amikacin (AK) is frequently used on the treatment of Gram-negative infections on neonates, but its usage is restricted because of nephrotoxicity. In this study, on neonatal rats, we aimed to investigate the effects of erythropoietin and vitamin E on AK induced nephrotoxicity. A total of 35 newborn Wistar Albino rats were divided into four groups: (1) injected with saline (serum physiological was administered to placebo controls), (2) injected with AK (1200 mg/kg), (3) injected with AK + vitamin E (150 mg/kg), (4) injected with AK + erythropoietin (EPO) (300 IU/kg/day). In renal tissue, AK levels were significantly high in all groups except the control. Tissue malondialdehyde (MDA) and nitric oxide (NO) levels were statistically higher in AK -treated group than the control. MDA and NO levels were significantly decreased with the administration of vitamin E and EPO. Glutathione peroxidase (GPX) levels were statistically low in AK group compared with the controls. The levels of GPX, in vitamin E group, were increased significantly. However, superoxide dismutase and catalase levels were not significantly different in none of the groups. Insulin-like growth factor-1 values in AK, EPO and vitamin E groups were significantly higher than the control group. Histomorphological changes such as tubular epithelial necrosis were seen in AK treated group. Histopathological improvements observed with EPO and vitamin E administration. AK nephrotoxicity is related to oxidative stress and is supported with biochemical and histopathological findings. Vitamin E and EPO, as antioxidants, can be useful renoprotective agents for ameliorating AK induced nephrotoxicity in neonates.

Ozone-Induced Type 2 Immunity in Nasal Airways. Development and Lymphoid Cell Dependence in Mice.

Inhalation exposures to ozone commonly encountered in photochemical smog cause airway injury and inflammation. Elevated ambient ozone concentrations have been epidemiologically associated with nasal airway activation of neutrophils and eosinophils. In the present study, we elucidated the temporal onset and lymphoid cell dependency of eosinophilic rhinitis and associated epithelial changes in mice repeatedly exposed to ozone. Lymphoid cell-sufficient C57BL/6 mice were exposed to 0 or 0.5 parts per million (ppm) ozone for 1, 2, 4, or 9 consecutive weekdays (4 h/d). Lymphoid cell-deficient, Rag2(-/-)Il2rg(-/-) mice were similarly exposed for 9 weekdays. Nasal tissues were taken at 2 or 24 hours after exposure for morphometric and gene expression analyses. C57BL/6 mice exposed to ozone for 1 day had acute neutrophilic rhinitis, with airway epithelial necrosis and overexpression of mucosal Ccl2 (MCP-1), Ccl11 (eotaxin), Cxcl1 (KC), Cxcl2 (MIP-2), Hmox1, Il1b, Il5, Il6, Il13, and Tnf mRNA. In contrast, 9-day ozone exposure elicited type 2 immune responses in C57BL/6 mice, with mucosal mRNA overexpression of Arg1, Ccl8 (MCP-2), Ccl11, Chil4 (Ym2), Clca1 (Gob5), Il5, Il10, and Il13; increased density of mucosal eosinophils; and nasal epithelial remodeling (e.g., hyperplasia/hypertrophy, mucous cell metaplasia, hyalinosis, and increased YM1/YM2 proteins). Rag2(-/-)Il2rg(-/-) mice exposed to ozone for 9 days, however, had no nasal pathology or overexpression of transcripts related to type 2 immunity. These results provide a plausible paradigm for the activation of eosinophilic inflammation and type 2 immunity found in the nasal airways of nonatopic individuals subjected to episodic exposures to high ambient ozone.

Effect of cypermethrin on the histoarchitecture of gills and liver of a freshwater catfish, Pangasianodon hypophthalmus

Histoarchitecture effect of cypermethrin was investigated in the gills and liver of freshwater catfish, Pangasianodon hypophthalmus (weight 60-70g) over an exposure period of 96 h as an endpoint of toxicity. The fish were exposed to five acute concentrations (0.00ml/L, 0.025ml/L, 0.050ml/L, 0.075ml/L and 0.10ml/L) and the 96 h LC50 was 0.075 ml/L. The physical reactions observed in the treated fish were erratic swimming, discolorations of the skin, loss of reflex, hyperactivities, surfacing, increasing opercula ventilation and these effects increased with increasing concentration of the toxicants and duration of exposure. The most common changes in gills histoarchitecture at all doses of cypermethrin were curl shape gill lamellae, bulged with the fusion of secondary gill lamellae, severe epithelial necrosis in gill lamellae and hypertrophy. The changes observed in the liver tissues were cloudy swelling, focal necrosis and hypertrophy of hepatocytes, degeneration of hepatocytes and cytoplasmic, extensive vacuolation of hepatocytes and pyknotic nuclei. The results of this histoarchitecture analysis of gills and liver tissues indicate a direct correlation between cypermethrin exposure and the histoarchitecture disorders.Asian J. Med. Biol. Res. December 2015, 1(3): 641-647

Comparison between cryoablation and irreversible electroporation of rabbit livers at a location close to the gallbladder.

BackgroundThe ablation of liver tumors located close to the gallbladder is likely to lead to complications. The aim of this article is to compare the safety and efficacy of irreversible electroporation (IRE) and cryoablation in rabbit livers at a location close to the gallbladder.Materials and methodsWe performed cryoablation (n = 12) and IRE (n = 12) of the area of the liver close to the gallbladder in 24 New Zealand white rabbits in order to ensure gallbladder damage. Serum aminotransferase and serum bilirubin levels were measured before and after the ablation. Histopathological examination of the ablation zones in the liver and gallbladder was performed on the 7th day after the ablation.ResultSeven days after the ablation, all 24 animals were alive. Gallbladder perforation did not occur in the IRE group; only mucosal epithelial necrosis and serous layer edema were found in this group. Gallbladder perforation occurred in four rabbits in the cryoablation group. Serum aminotransferase and serum bilirubin levels obviously increased in both groups by Day 3 and decreased gradually thereafter. The elevation in aminotransferase and bilirubin levels was greater in the cryoablation group than the IRE group. Pathological examination revealed complete necrosis of the liver parenchyma from the ablation center to the gallbladder in both groups, but bile duct and granulation tissue hyperplasia were observed in only the IRE group. Full-thickness gallbladder-wall necrosis was seen in the cryoablation group.ConclusionsFor ablation of the liver area near the gallbladder, IRE is superior to cryoablation, both in terms of safety (no gallbladder perforation in the IRE group) and efficacy (complete necrosis and rapid recovery in the IRE group).

Histopathological and Molecular Diagnosis of Infectious Laryngotracheitis in Tunisia - First Report

Infectious laryngotracheitis (ILT), an acute infection of the upper respiratory tract of chickens, was absent before the end of 2013, in Tunisia. Severe respiratory signs were observed in 48 layer and broiler flocks (3 weeks to 80 weeks old) between June 2013 and June 2016, and characterized with dyspnea, bloody mucus excretion and decrease in egg production. Clinical signs were dominated by congestive and hemorrhagic inflammation of the upper respiratory tract. Histopathological analyses and PCR tests were performed on respiratory tissues of suspected birds. Suggestive lesions of ILT, such as epithelial necrosis, sero-fibrinous and sero-hemorrhagic exudates, syncytia and intranuclear inclusion bodies were observed in layers of the 25 flocks visited. ILTV was detected in 12 out of 31 flocks by conventional PCR, using specific primers for the gB gene of ILTV. Eight of these were positive in histopathology. Our findings confirmed the introduction for the first time of the ILT virus in commercial poultry farms, in Tunisia.

Toxic effects of glyphosate-based herbicide, Excel Mera 71 on gill, liver, and kidney of Heteropneustes fossilis under laboratory and field conditions

The effects of glyphosate-based herbicide Excel Mera 71 under field and laboratory conditions were investigated to evaluate the pathological symptoms through light and electron microscopic study in the gill, liver, and kidney of Heteropneustes fossilis (Bloch) for a period of 30 days. Histological alterations like hypertrophy and fusion in secondary lamellae, damage in chloride cells were more prominent in laboratory conditions under light microscopy. Topological changes such as complete loss of microridges, swelling, and irregular arrangement of microridges in the gills were prominent under scanning electron microscopic study under laboratory conditions. Transmission electron microscopy (TEM) study depicted vacuolation and degeneration in chloride cells, dilation in rough endoplasmic reticulum (RER), and mitochondria in gill epithelium. The liver showed enlarged and pyknotic hepatocytes, vacuolation, excess fat deposition, and necrosis under laboratory conditions, while enlarged acentric nuclei, increased sinusoidal space, and less vacuolation in cytoplasm were observed under field conditions. TEM displayed cytoplasmic vacuolation and a reduced number of endoplasmic reticulum and glycogen droplets in the laboratory, but this was less pronounced under field conditions. In the kidneys, loss of hematopoietic tissue, degenerative changes in glomeruli, proximal and distal convoluted tubule, and epithelial cell lining of the renal tubules were comparatively less prominent under field conditions. Under TEM, epithelial cell necrosis, endoplasmic reticulum fragmentation, and mitochondrial degeneration were more prominent under laboratory conditions. The present study evaluated the comparative toxicity under field and laboratory conditions under long-term exposure to glyphosate herbicide and identified pathological responses as indicators in monitoring the herbicidal contamination in aquatic ecosystems.

Protective Effect of Spirodela polyrhiza on Various Organs of Arsenicinduced Wistar Albino Rats

Our study was undertaken to assess the protective effect of Spirodela polyrhiza against sodium arseniteinduced toxicity in experimental model. Animals were divided into six groups. The first group was used as control group while the other groups were treated with S. polyrhiza, sodium arsenite (10 ppm), 10 ppm As plus S. polyrhiza, sodium arsenite (30 ppm), and 30 ppm As plus S. polyrhiza respectively. Organs (kidney, spleen and heart) were collected at the end of the experiment for conducting histopathological analysis and measuring the amount of deposited arsenic. Tubular epithelium necrosis was seen affecting less than 50% of all renal specimens, on the other hand slightly enlarged white pulp region in splenic tissue were found in the groups of Wistar rat where S. polyrhiza and arsenic was administered concomitantly. Moreover, the observed moderate tissue changes (edema and muscle necrosis) in cardiac specimens of arsenic treated groups were found to be mild in the S. polyrhiza +arsenic treated groups. It is noteworthy to mention that S. polyrhiza not only reversed the arsenic-induced toxicity but also reduced the tissue arsenic load in Wistar rats to a considerable extent. Overall, the results indicated that S. polyrhiza might become useful in mitigating the toxic effects of arsenic.

Kinetics of Respiratory Syncytial Virus (RSV) Memphis Strain 37 (M37) Infection in the Respiratory Tract of Newborn Lambs as an RSV Infection Model for Human Infants.

RationaleRespiratory syncytial virus (RSV) infection in preterm and newborn infants can result in severe bronchiolitis and hospitalization. The lamb lung has several key features conducive to modeling RSV infection in human infants, including susceptibility to human strains of RSV such as the A2, Long, and Memphis Strain 37 (M37). In this study, the kinetics of M37 infection was investigated in newborn lambs in order to better define clinical, viral, physiological, and immunological parameters as well as the pathology and lesions.MethodsNewborn lambs were nebulized with M37 hRSV (6 mL of 1.27 x 107 FFU/mL), monitored daily for clinical responses, and respiratory tissues were collected from groups of lambs at days 1, 3, 4, 6, and 8 post-inoculation for the assessment of viral replication parameters, lesions and also cellular, immunologic and inflammatory responses.ResultsLambs had increased expiratory effort (forced expiration) at days 4, 6, and 8 post-inoculation. Nasal wash lacked RSV titers at day 1, but titers were present at low levels at days 3 (peak), 4, and 8. Viral titers in bronchoalveolar lavage fluid (BALF) reached a plateau at day 3 (4.6 Log10 FFU/mL), which was maintained until day 6 (4.83 Log10 FFU/mL), and were markedly reduced or absent at day 8. Viral RNA levels (detected by RT-qPCR) in BALF were indistinguishable at days 3 (6.22 ± 0.08 Log10 M37 RNA copies/mL; mean ± se) and 4 (6.20 ± 0.16 Log10 M37 RNA copies/mL; mean ± se) and increased slightly on day 6 (7.15 ± 0.2 Log10 M37 RNA copies/mL; mean ± se). Viral antigen in lung tissue as detected by immunohistochemistry was not seen at day 1, was present at days 3 and 4 before reaching a peak by day 6, and was markedly reduced by day 8. Viral antigen was mainly present in airways (bronchi, bronchioles) at day 3 and was increasingly present in alveolar cells at days 4 and 6, with reduction at day 8. Histopathologic lesions such as bronchitis/bronchiolitis, epithelial necrosis and hyperplasia, peribronchial lymphocyte infiltration, and syncytial cells, were consistent with those described previously for lambs and infants.ConclusionThis work demonstrates that M37 hRSV replication in the lower airways of newborn lambs is robust with peak replication on day 3 and sustained until day 6. These findings, along with the similarities of lamb lung to those of infants in terms of alveolar development, airway branching and epithelium, susceptibility to human RSV strains, lesion characteristics (bronchiolitis), lung size, clinical parameters, and immunity, further establish the neonatal lamb as a model with key features that mimic RSV infection in infants.

Protective effect of allicin against gentamicin-induced nephrotoxicity in rats

In this study, the modulator effect of allicin on the oxidative nephrotoxicity of gentamicin in the kidneys of rats was investigated by determining indices of lipid peroxidation and the activities of antioxidant enzymes, as well as by histological analyses. Furthermore, the effect of allicin on gentamicin induced hypersensitivity of urinary bladder rings to ACh was estimated. Twenty-four male Wistar albino rats were randomly divided into three groups, control, gentamicin (100mg/kg, i.p.) and gentamicin+allicin (50mg/kg, orally). At the end of the study, all rats were sacrificed and then urine, blood samples and kidneys were taken. Gentamicin administration caused a severe nephrotoxicity as evidenced by an elevated kidney/body weight ratio, serum creatinine, blood urea nitrogen (BUN), serum lactate dehydrogenase (LDH) and proteinuria with a reduction in serum albumin and creatinine clearance as compared with control group. In addition, a significant increase in renal contents of malondialdehyde (MDA), myeloperoxidase (MPO), nitric oxide (NOx) and tumor necrosis factor-alpha (TNF-α) concomitantly with a significant decrease in renal reduced glutathione (GSH) and superoxide dismutase (SOD) activities was detected upon gentamicin injection. Exposure to gentamicin increased the sensitivity of isolated urinary bladder rings to ACh and induced acute renal tubular epithelial cells necrosis. Administration of allicin significantly decreased kidney/body weight ratio, serum creatinine, LDH, renal MDA, MPO, NOx and TNF-α while it significantly increased creatinine clearance, renal GSH content and renal SOD activity when compared to gentamicin-treated group. Additionally, allicin significantly reduced the responses of isolated bladder rings to ACh and ameliorated tissue morphology as evidenced by histological evaluation. Our study indicates that allicin exerted protection against structural and functional damage induced by gentamicin possibly due to its antioxidant, anti-inflammatory and immunomodulatory properties in addition to its ability to retaining nitric oxide level.

Histopathological effects of silver and copper nanoparticles on the epidermis, gills, and liver of Siberian sturgeon.

The influence of nanoparticles (NPs) on aquatic environments is still poorly documented. The aim of the study was to determine the effects of silver (AgNPs) and copper (CuNPs) nanoparticles on larval Siberian sturgeon (Acipenser baerii) after 21 days of exposure. Acute toxicity of AgNPs on Siberian sturgeon was investigated in a 96-h static renewal study and compared with the toxicity of CuNPs. The AgNPs and CuNPs 96 h mean lethal concentrations (96 h LC50) were 15.03 ± 2.91 and 1.41 ± 0.24 mg L−1, respectively. Toxicity tests were done in triplicates for each concentration of AgNPs 0.1, 0.5, 1.5 mg L−1 and CuNPs 0.01, 0.05, 0.15 mg L−1. The control group was exposed in freshwater. The results indicate that AgNPs and CuNPs exposure negatively influenced survival; body length and mass; and morphology and physiology of the epidermis, gills, and liver of Siberian sturgeon larvae. Fish exposed to AgNPs and CuNPs showed similar pathological changes: irregular structure and pyknotic nuclei of epidermis, aplasia and/or fusion of lamellae, telangiectasis, epithelial necrosis and lifting of the gills, dilation of sinusoidal space, overfilled blood vessels, and pyknotic nuclei of the liver. Fish exposed to CuNPs only demonstrated hyaline degeneration in the gills epithelium and liver. The study shows that CuNPs were more toxic to Siberian sturgeon larvae than AgNPs.

Unilateral Renal Ischemia as a Model of Acute Kidney Injury and Renal Fibrosis in Cats.

The objectives of this study were to define the acute and chronic effects of 1-hour unilateral in vivo renal ischemia on renal function and histology in cats. Twenty-one adult purpose-bred research cats were anesthetized, and 1 kidney underwent renal artery and vein occlusion for 1 hour. Serum creatinine and urea concentrations, urine protein:creatinine ratio, urine-specific gravity, glomerular filtration rate, hematocrit, platelet concentration and function, and white blood cell count were measured at baseline and variable time points after ischemia. Renal histopathology was evaluated on days 3, 6, 12, 21, 42, and 70 postischemia; changes in smooth muscle actin and interstitial collagen were examined. Following ischemia, whole animal glomerular filtration rate was significantly reduced (57% of baseline on day 6; P < .05). At the early time points, the ischemic kidneys exhibited severe acute epithelial necrosis accompanied by evidence of regeneration of tubules predominantly within the corticomedullary junction. At later periods, postischemic kidneys had evidence of tubular atrophy and interstitial inflammation with significantly more smooth muscle actin and interstitial collagen staining and interstitial fibrosis when compared with the contralateral control kidneys. This study characterizes the course of ischemic acute kidney injury in cats and demonstrates that ischemic acute kidney injury triggers chronic fibrosis, interstitial inflammation, and tubular atrophy in feline kidneys. These late changes are typical of those observed in cats with naturally occurring chronic kidney disease.

Thearubigins protect against acetaminophen-induced hepatic and renal injury in mice: biochemical, histopathological, immunohistochemical, and flow cytometry study

Context: Acetaminophen toxicity is used as a model for studying chemical toxicity. N-acetylcysteine (NAC) is used for the treatment of hepatotoxicity; however, there is no specific therapy for nephrotoxicity. Objective: This study was designed to investigate the potential protective effect of black tea extract (BTE) and its main phenolic pigment, thearubigins (TRs), against acetaminophen (APAP)-induced hepatic and renal injury in mice. Materials and methods: Besides control groups, six groups (n = 8) were given intraperitoneally APAP (300 mg/kg) and then after 1.5 hours were treated intraperitoneally as follows: NAC (318 mg/kg), BTE (3%, 4.5%), and TRs (50, 60, and 70 mg/kg). Six hours post-APAP injection, blood was collected for biochemical measurements. Later, liver and kidneys were removed for histopathological, immunohistochemical, and flow cytometry studies. Results: APAP increased alanine aminotransferase and malondialdehyde and decreased glutathione levels in blood. Treatments significantly reversed these changes mostly with NAC and TRs70. TRs showed dose-dependent significant differences. The APAP-induced central lobular hepatic necrosis and increased TUNEL positivity were mild with co-administration of NAC and TRs (60, 70) while moderate with co-administration of BTE (3, 4.5) and TRs50. The APAP-increased serum creatinine level was significantly reversed by treatments (mostly TRs60, 70). The APAP-induced renal tubular epithelial degeneration and necrosis were mild with co-administration of TRs (60, 70) while moderate with co-administration of NAC, BTE (3, 4.5), and TRs50. The APAP-accumulated apoptotic cells in sub-G1 phase were significantly decreased by treatments, mostly by NAC and TRs70 in the liver and TRs (60, 70) in kidneys. Conclusion: Thearubigins protected against acetaminophen-induced hepatotoxicity and nephrotoxicity in mice possibly through their antioxidant activity.

Abstract 4077: Adipose-derived stem cells after Paclitaxel treatment demonstrate decreased function and suppression of breast cancer cell viability

Abstract Background: Following neoadjuvant or adjuvant chemotherapy for breast cancer resections, poor soft tissue wound healing can pose a major clinical challenge. Adipose-derived stem cells (ASCs) in clinically applied stromal vascular fractions are considered to play a positive role in wound healing (SVF) as evidenced by their increasing use in breast reconstructive procedures. Our study evaluated the potential confounding effects of Paclitaxel (PTX) on ASC proliferation and differentiation capacities and if PTX treated ASCs may subsequently influence tumor cell viability. Methods: IRB-approved human ASCs were isolated and treated with PTX at different concentrations. Proliferation, cell viability, and cell migration rates were measured by growth curves, MTT assays and scratch assays. ASCs were cultured in derivative-specific differentiation media with or without PTX (1uM) for 3 weeks. Adipogenic, osteogenic and endothelial differentiation levels were measured by quantitative RT-PCR, Oil-Red-O, Alizarin Red staining and cord formation on Matrigel, respectively. MDA-MB-231 (breast cancer cell line) and ASCs were co-cultured at equal cell numbers. ASC conditional media were collected every 3 days from ASC no-PTX treatment and PTX treated - washout groups. Results: PTX decreased proliferation of ASCs in a dose- and time-dependent manner. PTX treatment down-regulated the capacity of ASCs for adipogenic, osteogenic and endothelial differentiation (p&amp;lt;0.05 vs. control). In comparison with controls, PTX treated ASCs had decreased cell migration (relative scratch width, p&amp;lt;0.05 vs. control). Following cessation and washout of PTX after 8 days, treated ASCs exhibited cell growth and differentiating ability, but their capacity was suppressed compared to controls. The cumulative population doubling time was longer for cells co-cultured with PTX treated-washout ASC conditional media. (p&amp;lt;0.01 vs. no-PTX treatment group). Total cellular mRNA expressions of epithelial cell adhesion molecule (EPCAM), synuclein gamma (breast cancer specific protein 1; SNCG) and tumor necrosis factor alpha (TNF-α) were down-regulated in MDA-MB-231 cells when co-cultured directly with PTX treated ASC or indirectly with PTX treated-washout ASC conditional media (p&amp;lt;0.05 vs. control). In cells co-cultured for 3 days, VEGF proteins released in conditioned medium were decreased compared with single cultures (p&amp;lt;0.01 vs. control). Conclusion: Our results indicate that: 1) PTX inhibited ASC proliferation and decreased the multipotency differentiating capacity; and 2) Direct contact or treatment with conditional media from PTX treated ASCs suppressed tumor cell growth. In evaluating autologous ASCs for reconstructive procedures, this study may provide insight into poor soft tissue wound healing immediately following the removal of chemotherapy as well as decreased recurrence in breast cancer patients. Citation Format: William M. Harris, Michael Plastini, Telisha Ortiz, Nikolas Kappy, Jefferson Benites, Shaohua Chang, A. Lelani Fahey, Martha S. Matthews, Alexandre Hageboutros, Jeffrey P. Carpenter, Spencer Brown, Ping Zhang. Adipose-derived stem cells after Paclitaxel treatment demonstrate decreased function and suppression of breast cancer cell viability. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4077. doi:10.1158/1538-7445.AM2015-4077

Morphological and microcirculatory evaluation of the rat testis after detorsion with or without a capsular release with a tunica vaginalis flap.

Testicular torsion may lead to serious ischemia, and the viability depends on the duration of torsion and the effect of ischemia-reperfusion. Testicular decompression and tunica vaginalis flap application technique were introduced in 2008 by Kutikov et al. We aimed to examine the impact of this method on the testicular microcirculation and hemorheological parameters in a rat model. Six adult rats underwent bilateral scrotal exploration. Intravaginal torsion of the testis was created by 720° rotation on both sides for 2 h. After detorsion, the right testes underwent tunica albuginea incision and tunica vaginalis flap application. Testicular microcirculation was monitored and hematological parameters, erythrocyte deformability, and aggregation were determined. Measurements were performed before and after torsion, directly after detorsion, on the 1st–2nd and 8th postoperative day. After the last sampling, testicles were removed to determine their volume for histological examinations. The microcirculatory parameters demonstrated slight differences between testicles. Apical zone of the left (nondecompressed) testicles had elevated compared to the middle zone (P < 0.05). On the 2nd and 8th day, the microcirculation of the testes normalized but not equally. The erythrocyte aggregation and deformability decreased by the 8th day. Both testicles underwent atrophy and epithelial necrosis, but the volume of the decompressed ones was lower (1.07 ± 0.08 vs 1.25 ± 0.31). Histologically, there was no significant difference in epithelial damage score between decompressed and nondecompressed testes. In conclusion, 2-h ischemia led to alteration in testicular microcirculation, reduction in volume, changes in hemorheological parameters and serious epithelial necrosis both in decompressed and nondecompressed testicles without remarkable differences.

IL-1 receptor anatagonist anakinra reduces neutrophilic airway inflammation in mice with cystic fibrosis-like lung disease

Hypoxic degeneration and necrosis of epithelial cells associated with airway mucus obstruction precedes neutrophilic airway inflammation in βENaC-overexpressing (βENaC-Tg) mice with cystic fibrosis (CF)-like lung disease (Mall MA et al., Am J Respir Crit Care Med., 2008). In many organs sterile neutrophilic inflammation is triggered by necrosis via release of Interleukin-1 alpha (IL-1α) and IL-1 receptor (IL-1R) signaling (Chen CJ et al., Nat Med., 2007). Recently we demonstrated that lack of IL-1R improved survival and reduced neutrophilic airway inflammation, structural lung damage and mucus obstruction in βENaC-Tg mice (Zhou-Suckow Z et al., Pediatr Pulmonol., Suppl 2013). In the present day, we hypothesized that treatment with the IL-1R antagonist anakinra may reduce neutrophilic airway inflammation and ameliorate CF-like lung disease in βENaC-Tg mice. To test this hypothesis, we treated 4 to 6 week old βENaC-Tg mice in established lung disease and wild-type littermates with anakinra (200 mg/kg body weight) or vehicle (NaCl 0.9%) alone by subcutaneous injection twice per day for one week. At the endpoint, we performed bronchoalveolar lavage (BAL) to determine differential cell counts and KC levels, and analyzed histology and morphometry for airway epithelial necrosis, airway mucus obstruction and emphysema. As expected from previous studies of βENaC-Tg mice with genetic deletion of IL-1R, treatment with anakinra had no effect on hypoxic epithelial degeneration. However, anakinra treatment significantly reduced neutrophilic inflammation and KC levels in BAL. Furthermore, mean linear intercepts and destructive index were also reduced by anakinra indicating less structural lung damage in βENaC-Tg mice. These results demonstrate that inhibition of IL-1R signalling provides an effective therapy of CF-like lung disease in mice and suggest anakinra as a potential anti-inflammatory therapy to limit neutrophilic inflammation and associated lung damage in patients with CF and potentially other muco-obstructive lung diseases.

Effect of Quyu Chencuo Formula () on Renal Fibrosis in Obstructive Nephropathy Rats.

To observe the effect of Quyu Chencuo Formula (, QCF) on renal fibrosis in rats with obstructive nephropathy. Twenty-four rats were randomly divided into three groups, 4 for sham operation as the control group, 10 for unilateral ureteral obstruction (UUO) model group, and the rest 10 for QCF treating UUO model group. All rats were sacrificed under 3% pentobarbital (50 mg/kg) anesthesia on the 14th day after surgery, then the right kidney samples of rats were harvested for hematoxylin eosin (HE) staining and Masson staining to observe the renal pathological changes. Immunohistochemistry and Western blotting were used to examine the expression of transforming growth factor β1 (TGF-β1), and real-time polymerase chain reaction (RT-PCR) was employed to examine the expressions of TGF-β1, α-smooth muscle actin (α-SMA) and E-cadherin mRNA. HE and Masson staining showed that the renal interstitial of the rats in the control group had no significant fibrotic lesion; in the model group, there were obvious interstitial fibrosis; for the QCF group, there were epithelial cell necrosis, infiltration of lymphocytes and mononuclear cells, aggravated interstitial fibrosis in varied degrees, but the pathological changes were less in the QCF group than in the model group. The immunohistochemistry and Western blotting results showed that the TGF-β1 expression was increased significantly in the model group, while decreased significantly in the QCF group (P<0.05); RT-PCR showed that the mRNA expression of α-SMA and TGF-β1 increased significantly in the model group, while both were significantly decreased in the QCF group compared with the model group (P<0.05). The mRNA expression of E-cadherin was decreased significantly in the model group, and it was significantly increased in the QCF group as compared with the model group (P<0.05). QCF may improve renal fibrosis by regulating the expressions of TGF-β1, α-SMA and E-cadherin, and prevent the progress of kidney fibrosis.