Release Of Intracellular Contents Research Articles

Partners in Crime: The Interplay of Proteins and Membranes in Regulated Necrosis.

Pyroptosis, necroptosis, and ferroptosis are well-characterized forms of regulated necrosis that have been associated with human diseases. During regulated necrosis, plasma membrane damage facilitates the movement of ions and molecules across the bilayer, which finally leads to cell lysis and release of intracellular content. Therefore, these types of cell death have an inflammatory phenotype. Each type of regulated necrosis is mediated by a defined machinery comprising protein and lipid molecules. Here, we discuss how the interaction and reshaping of these cellular components are essential and distinctive processes during pyroptosis, necroptosis, and ferroptosis. We point out that although the plasma membrane is the common target in regulated necrosis, different mechanisms of permeabilization have emerged depending on the cell death form. Pore formation by gasdermins (GSDMs) is a hallmark of pyroptosis, while mixed lineage kinase domain-like (MLKL) protein facilitates membrane permeabilization in necroptosis, and phospholipid peroxidation leads to membrane damage in ferroptosis. This diverse repertoire of mechanisms leading to membrane permeabilization contributes to define the specific inflammatory and immunological outcome of each type of regulated necrosis. Current efforts are focused on new therapies that target critical protein and lipid molecules on these pathways to fight human pathologies associated with inflammation.

Acute Kidney Injury and Rhabdomyolysis

Background and objectives: Rhabdomyolysis is clinical syndrome characterized by breakdown of skeletal muscle and the release of intracellular contents into the circulation. Acute kidney injury (AKI) is one of the common systemic complication of rhabdomyolysis. The aim of the study is to report the varied aetiologies of rhabdomyolysis and to study their presentation, clinical profile and outcome in patients with AKI. Methods: Sixty-four patients are identified with rhabdomyolysis over a period of one year and out of them AKI was seen in 24 patients (37.5%%). The association of rhabdomyolysis and AKI was studied retrospectively in eight patients based on the data in our charts. One patient from different aetiologies of rhabdomyolysis leading to AKI was taken to report the varied aetiologies and presentation of rhabdomyolysis. Hence, eight patients included in the study. Data was expressed as median. Results: Our results show that AKI associated with rhabdomyolysis was related with comorbidity in all cases (bacterial sepsis, statin induced, chemotherapy induced, herbicide poisoning, cocaine abuse, road traffic accident, strenuous exercise, metabolic myopathy). Most common presenting symptom was myalgia followed by oliguria. Maximum median serum creatinine was 4.7 mg/dl before treatment and 1.7 mg/dl after treatment. Maximum median creatine phosphokinase levels was 2451 IU/L before treatment and 262 IU/L after treatment. Treatment given for rhabdomyolysis and acute kidney injury include either conservative treatment with adequate hydration and alkaline diuresis or invasive treatment such as renal replacement therapy or both. All the patients were treated with hydration and six patients required haemodialysis during hospital stay. Serum creatinine fell simultaneously with the decrease in creatine phosphokinase levels in all cases. Conclusion: In conclusion, rhabdomyolysis is an uncommon and preventable cause of AKI. It is caused by varied aetiologies, hence high index of suspicion regarding rhabdomyolysis is required in patients with AKI. The presence of rhabdomyolysis associated with AKI is indicative of the presence of comorbidity, as observed in all eight patients studied here. There was an improvement of renal function in all cases as creatine phosphokinase levels decreased. Hence, timely intervention is required to prevent adverse outcomes in rhabdomyolysis.

Necroptosis in pancreatic cancer promotes cancer cell migration and invasion by release of CXCL5.

Necroptosis is a form of programmed cell death that is accompanied by release of intracellular contents, and reportedly contributes to various diseases. Here, we investigate the significance of necroptosis in pancreatic cancer. We used immunohistochemistry and western blot analysis to evaluate expression of the key mediators of necroptosis-receptor-interacting serine/threonine protein kinase 3 (RIP3) and mixed lineage kinase domain-like (MLKL)-in human pancreatic cancer. We also tested the effects of conditioned media (CM) from necroptotic cells on pancreatic cancer cells in Transwell migration and Matrigel invasion assays. Protein array analysis was used to investigate possible mediators derived from necroptotic cells. RIP3 and MLKL are highly expressed in human pancreatic cancer tissues compared with normal pancreas. MLKL expression was particularly intense at the tumor invasion front. CM derived from necroptotic cells promoted cancer cell migration and invasion, but not CM derived from apoptotic cells. C-X-C motif chemokine 5 (CXCL5) was upregulated in CM derived from necroptotic cells compared with CM derived from control or apoptotic cells. Moreover, expression of the receptor for CXCL5, C-X-C-motif chemokine receptor-2 (CXCR2), was upregulated in pancreatic cancer cells. Inhibition of CXCR2 suppressed cancer cell migratory and invasive behavior enhanced by necroptosis. These findings indicate that necroptosis at the pancreatic cancer invasion front can promote cancer cell migration and invasion via the CXCL5-CXCR2 axis.

Inhibition of biofilm- and hyphal- development, two virulent features of Candida albicans by secondary metabolites of an endophytic fungus Alternaria tenuissima having broad spectrum antifungal potential

Fungal resistance against frequently used antifungal medicines used for invasive candidiasis and other fungal infections is directing scientist for searching and developing novel antifungal drugs. An endophytic fungal strain Alternaria tenuissima OE7 has been isolated from leaves of Ocimum tenuiflorum L. which showed antifungal activity against numbers of human pathogenic fungi including Trichophyton rubrum, Microsporum gypseum, Aspergillus parasiticus, A. flavus, A. fumigates, Candida albicans and C. tropicalis. Thermostable, non-proteinacious antifungal metabolites produced zones of inhibition against all pathogenic fungi tested. The ethyl acetate extract of the cell free supernatant was found inhibitory to the radial growth and conidial germination of T. rubrum and M. gypseum. It also showed cidal mode of action against C. albicans at a concentration of 1.0 mg/ml. Most interestingly, inhibition of biofilm formation and hyphal development of C. albicans were observed upon treatment with EA fraction at comparatively lower concentrations (100–500 μg/ml). Release of intracellular contents from treated cells of Candida and scanning electron microscopic observation suggested cellular disruptions by antifungal metabolites. Checkerboard study revealed synergy between EA fraction of OE7 (150 μg/ml) and fluconazole (30 μg/ml) with ƩFIC of 0.45. Two active fractions viz. band ‘C’ and band ‘G’ derived after thin layer chromatographic analysis showed inhibitory activity against C. albicans with MIC values of 80 μg/ml and 130 μg/ml respectively. GCMS analysis suggested presence of numbers of compounds in each active fraction. Overall observations attest the prospective role of the isolate OE7 as a potent candidate for the production of antifungal metabolites against human pathogenic fungi.

Membrane Disruption by Very Long Chain Fatty Acids during Necroptosis.

Necroptosis is a form of regulated cell death which results in loss of plasma membrane integrity, release of intracellular contents, and an associated inflammatory response. We previously found that saturated very long chain fatty acids (VLCFAs), which contain ≥20 carbons, accumulate during necroptosis. Here, we show that genetic knockdown of Fatty Acid (FA) Elongase 7 (ELOVL7) reduces accumulation of specific very long chain FAs during necroptosis, resulting in reduced necroptotic cell death and membrane permeabilization. Conversely, increasing the expression of ELOVL7 increases very long chain fatty acids and membrane permeabilization. In vitro, introduction of the VLCFA C24 FA disrupts bilayer integrity in liposomes to a greater extent than a conventional C16 FA. To investigate the microscopic origin of these observations, atomistic Molecular Dynamics (MD) simulations were performed. MD simulations suggest that fatty acids cause clear differences in bilayers based on length and that it is the interdigitation of C24 FA between the individual leaflets that results in disorder in the region and, consequently, membrane disruption. We synthesized clickable VLCFA analogs and observed that many proteins were acylated by VLCFAs during necroptosis. Taken together, these results confirm the active role of VLCFAs during necroptosis and point to multiple potential mechanisms of membrane disruption including direct permeabilization via bilayer disruption and permeabilization by targeting of proteins to cellular membranes by fatty acylation.

MON-237 A Series of Eight Cases with Pigment Nephropathy Due to Different Aetiologies – Clinical Profile and Outcome

Rhabdomyolysis is clinical syndrome characterised by breakdown of skeletal muscle and the release of intracellular contents into the circulation.It is type of pigment nephropathy. Details about the global incidence of rhabdomyolysis is limited. There are eight commonly reported causes of rhabdomyolysis. Systemic complications of rhabdomyolysis are acute kidney injury, compartment syndrome, cardiac arrest and others . Here, we report the incidence of acute kidney injury(AKI), clinical profile and outcome of eight cases with different aetiologies of rhabdomyolysis.

Antibacterial activity of lysozyme-chitosan oligosaccharide conjugates (LYZOX) against Pseudomonas aeruginosa, Acinetobacter baumannii and Methicillin-resistant Staphylococcus aureus.

The recent emergence of antibiotic-resistant bacteria requires the development of new antibiotics or new agents capable of enhancing antibiotic activity. This study evaluated the antibacterial activity of lysozyme-chitosan oligosaccharide conjugates (LYZOX) against Pseudomonas aeruginosa, Acinetobacter baumannii and methicillin-resistant Staphylococcus aureus (MRSA), which should resolve the problem of antibiotic-resistant bacteria. Bactericidal tests showed that LYZOX killed 50% more P. aeruginosa (NBRC 13275), A. baumannii and MRSA than the control treatment after 60 min. In addition, LYZOX was shown to inhibit the growth of P. aeruginosa (NBRC 13275 and PAO1), A. baumannii and MRSA better than its components. To elucidate the antibacterial mechanism of LYZOX, we performed cell membrane integrity assays, N-phenyl-1-naphthylamine assays, 2-nitrophenyl β-D-galactopyranoside assays and confocal laser scanning microscopy. These results showed that LYZOX affected bacterial cell walls and increased the permeability of the outer membrane and the plasma membrane. Furthermore, each type of bacteria treated with LYZOX was observed by electron microscopy. Electron micrographs revealed that these bacteria had the morphological features of both lysozyme-treated and chitosan oligosaccharide-treated bacteria and that LYZOX destroyed bacterial cell walls, which caused the release of intracellular contents from cells. An acquired drug resistance test revealed that these bacteria were not able to acquire resistance to LYZOX. The hemolytic toxicity test demonstrated the low hemolytic activity of LYZOX. In conclusion, LYZOX exhibited antibacterial activity and low drug resistance in the presence of P. aeruginosa, A. baumannii and MRSA and showed low hemolytic toxicity. LYZOX affected bacterial membranes, leading to membrane disruption and the release of intracellular contents and consequent bacterial cell death. LYZOX may serve as a novel candidate drug that could be used for the control of refractory infections.

Design, Synthesis and Biological Evaluation of Substituted (1‐(4‐chlorobenzyl)‐1H‐indol‐3‐yl) 1H‐(1,2,3‐triazol‐4‐yl)methanones as Antifungal Agents

AbstractIn the perusal of our endeavour towards the synthesis of new bioactive agents, a new series of substituted (1‐(4‐chlorobenzyl)‐1H‐indol‐3‐yl) 1H‐(1,2,3‐triazol‐4‐yl) methanones (8a‐u) were designed, synthesized and investigated for their antimicrobial activity with special emphasis on their anti‐Candida potential and Candida biofilm inhibition. Among them, the hybrids 8e, 8f, and 8o demonstrated significant antifungal activity on most of the tested fungal strains with MIC values ranging between 1.9–7.8 μg/mL. In addition, the most promising compounds were found to be effective biofilm inhibitors. We have studied the Field emission Scanning Electron Microscope (FE‐SEM) micrographs for 8f to know the damage of the cells with altered membrane integrity and release of intracellular contents thereby validating the in vitro biofilm inhibition assay. Further, in enzymatic assay, active compounds inhibited ergosterol biosynthesis. Furthermore, docking studies revealed that they effectively bind to 14α‐demethylase (CYP51), thereby inhibiting ergosterol biosynthesis. Interestingly, they exhibited lower cytotoxicity in the normal cell line (MRC5).

Synergistic effects of tea polyphenol epigallocatechin 3-O-gallate and azole drugs against oral Candida isolates

BackgroundThe antifungal drug resistance has become an emerging problem in the management of candida infections worldwide. The objective of this study was to examine the efficacy of epigallocatechin 3-O-gallate (EGCG) alone and in combination with fluconazole/ketoconazole drugs against oral Candida isolates. MethodsMinimum inhibitory concentration (MIC) and minimum fungicidal concentrations (MFC) of EGCG against 60 oral Candida isolates and 4 ATCC strains were determined. Synergism of EGCG with azole drugs was evaluated by checkerboard micro-dilution method and calculated fractional inhibitory concentration index (FICI). Candida cells’ ultrastructure was studied by electron microscopy. ResultsMIC and MFC values of EGCG were in the range of 3.91–15.63 and 15.63–31.25μg/mL, respectively. Minimum biofilm inhibitory concentration (MBIC) range of EGCG (62.5–125μg/mL), was less than the ketoconazole (64–256μg/mL) and fluconazole (128–512μg/mL). The combination of EGCG with fluconazole/ketoconazole exhibited synergistic effects (ΣFICI≤0.50). EGCG with azole drugs showed high sensitivity against the tested isolates in growth curve assays. Against the biofilm, the susceptibility of fluconazole/ketoconazole significantly increased (3 to 5 fold), after combination with EGCG (MBIC/4) (P≤0.001). Electron microscopy of EGCG treated cells showed deformation of cell structure, ruptured cell wall and release of intracellular content. In molecular docking experiments, a strong interaction was observed between EGCG and fungal cell membrane molecule ergosterol. ConclusionWe conclude that EGCG synergistically enhanced the antifungal potential of azole drugs. The synergistic potential of EGCG might be helpful in preventing the development of drug resistance, in lowering the drug dosage, and thus minimizing adverse effects.

Pulsed electric fields processing of apple tissue: Spatial distribution of electroporation by means of magnetic resonance imaging and computer vision system

Abstract The optimal application of Pulsed Electric Fields (PEF) technology, used by food industry to assist mass transfer processes, depends on the effectiveness of the induced electroporation. The present work aimed at exploring the application of Magnetic Resonance Imaging (MRI) combined with Computer Vision System (CVS) analysis to assess the spatial distribution of electroporation in apple tissue. PEF-treated apple samples were compared with Dipping (Dip) and Vacuum Impregnation (VI) to gain insight into the spatial distribution of mechanisms that lead to microstructural modifications over time. CVS showed that electroporation modified heterogeneously apple microstructure, causing enzymatic browning unevenly across the samples. MRI transverse relaxation times (T2) maps and longitudinal relaxation times (T1)-weighted images throughout apple tissue confirmed the inhomogeneous distribution and extent of the cell disruption, along with the release of intracellular content toward the external solution. Industrial relevance The novel applications of pulsed electric fields require fast and reliable methods to detect and estimate the breakage of the membranes integrity in order to boost their industrial adoption and optimization. The present study provided analytical tools able to monitor the spatial distribution of electroporation in plant tissue samples within minutes and consequently to speed up and improve the assessment of different PEF treatments.

Managing Tumor Lysis Syndrome in the Era of Novel Cancer Therapies

Abstract Tumor lysis syndrome (TLS) is a potentially life-threatening emergency that can develop rapidly after the release of intracellular contents from lysed malignant cells. The advent of novel and targeted therapies that have improved tumor-killing efficacy has the potential to increase the risk of TLS when used as part of front-line therapy. A recent review of TLS risk in patients with hematologic malignancies treated with newer targeted agents highlighted the need to revisit TLS risk stratification and to describe the practical challenges of TLS prevention, treatment, and monitoring. Although this era of rapid development of novel cancer therapies provides new hope for patients with hematologic malignancies, it is essential to be prepared for TLS because monitoring and prophylaxis can almost always prevent severe and life-threatening consequences. Heightened awareness of the development of TLS with novel and targeted agents, accompanied by aggressive hydration and rational, risk-appropriate management, are the keys to successful outcomes.

Selective intracellular vaporisation of antibody-conjugated phase-change nano-droplets in vitro

While chemotherapy is a major mode of cancer therapeutics, its efficacy is limited by systemic toxicities and drug resistance. Recent advances in nanomedicine provide the opportunity to reduce systemic toxicities. However, drug resistance remains a major challenge in cancer treatment research. Here we developed a nanomedicine composed of a phase-change nano-droplet (PCND) and an anti-cancer antibody (9E5), proposing the concept of ultrasound cancer therapy with intracellular vaporisation. PCND is a liquid perfluorocarbon nanoparticle with a liquid–gas phase that is transformable upon exposure to ultrasound. 9E5 is a monoclonal antibody targeting epiregulin (EREG). We found that 9E5-conjugated PCNDs are selectively internalised into targeted cancer cells and kill the cells dynamically by ultrasound-induced intracellular vaporisation. In vitro experiments show that 9E5-conjugated PCND targets 97.8% of high-EREG-expressing cancer cells and kills 57% of those targeted upon exposure to ultrasound. Furthermore, direct observation of the intracellular vaporisation process revealed the significant morphological alterations of cells and the release of intracellular contents.

Disinfection effect of pressurized carbon dioxide on Escherichia coli and Enterococcus sp. in seawater

Ship ballast water should be disinfected before being discharged into the ocean to avoid the dispersal of non-native species into the marine environment. This study presents the results of using pressurized carbon dioxide (CO2) at less than 1.0 MPa for inactivating Escherichia coli and Enterococcus sp. in artificial seawater (3.4% salinity). The bactericidal effects of pressurized CO2 were assessed using a liquid-film-forming apparatus under various conditions of pressure, temperature, and working volume ratio (WVR). Additionally, leakage of proteins and nucleic acids from cells was measured. Cell morphology of untreated cells and cells treated with pressurized CO2 was assessed using scanning electron microscopy (SEM). Pressurized CO2 treatment affected both strains; however, Enterococcus sp. exhibited higher resistance to pressurized CO2 treatment than did E. coli. Under identical treatment conditions (0.7 MPa, 20 °C, and 50% WVR), more than 5.0 log reduction in the load of E. coli and Enterococcus sp. was achieved after treatments for 5 min and 20 min, respectively. Release of intracellular contents occurred during the treatment process and SEM images of E. coli and Enterococcus sp. revealed that morphological changes had occurred after the treatment with pressurized CO2. Hence, pressurized CO2 has potential applications for inactivating pathogens in ballast water.

Peach juice processed by the ultrasound technology: Changes in its microstructure improve its physical properties and stability

Ultrasound is a non-conventional processing technology, which can be used not only for food preservation, but also to improve its properties and quality. This study evaluated the physical properties and stability of peach juice processed by the ultrasound technology. The peach juice processed by the ultrasound technology showed changes in its structure, evidenced by the optical microscopy and particle size distribution, involved steps of cell damage and release of intracellular content, particle size reduction, disruption of the whole cells, polysaccharide size reduction and dispersion of constituents. These effects, depending of the processing time, can trigger different mechanisms with a complex behaviour. The interaction among them and the relative importance of each one change during processing, determining the final rheological properties, pulp sedimentation and serum cloudiness (turbidity). The results indicated that the ultrasound technology can be used to improve the physical properties of peach juice, increasing the stability to pulp sedimentation and serum cloudiness, maintaining or increasing the juice consistency, with insignificant colour changes during the storage.

Optimization of Intracellular Product Release from Neisseria denitrificans Using Microfluidizer

Disruption of Neisseria denitrificans cells by microfluidizer was optimized using a factorial experiments design. The pH, pretreatment time, cell concentration, NaCl, ethylenediamine tetraacetic acid (EDTA) and Triton X-100 concentrations showed significant impact on disruption process and the process was optimized using central composite design and response surface methodology (RSM). Investigation revealed optimum conditions: 90 min pretreatment at pH 9.0 containing 110 g L−1 cells (dry cell weight), 50 mM NaCl, 10 mM EDTA, and 0.2% Triton X-100. At optimized conditions, the disruption rate increased twofold, up to 5.62 ± 0.27 × 10−3 MPa-a; meanwhile, yield of intracellular content was increased by 26%, with 1 g of cells resulting in 113.2 ± 8.2 mg proteins, 12.1 ± 0.7 mg nucleic acids, 21.0 ± 1.2 mg polysaccharides, 0.99 ± 0.08 kU glucose-6-phosphate dehydrogenase (G6PD), and 10,100 ± 110 kU restriction endonuclease NdeI endonuclease. Particle size distribution analysis revealed nearly twofold larger cell lysate particles with diameter of 120 nm. For optimal release of intracellular content, 9200 J/g of energy was needed (95% confidence), yielding 6900 J/g energy savings. Model equations generated from RSM on cell disruption of N. denitrificans were found adequate to determine significant factors and its interaction. The results showed that optimized combination of known pretreatment and disruption methods could considerably improve cell disruption efficiency.

Bacteriocin activity against various pathogens produced by Pediococcus pentosaceus VJ13 isolated from Idly batter

Bacteriocins, an antimicrobial peptide, is known to have wide spectrum antimicrobial activity against various pathogens. Because they are easily digested in the intestine, they are considered as safe and are widely used as food preservatives. Hence their purification and characterization have attracted considerable attraction, especially for those having activity against human pathogens. In this study, the bacteriocin produced by Pediococcus pentosaceus VJ13 was precipitated with cold acetone and purified by gel permeation chromatography and hydrophobic interaction chromatography. The bacteriocin exhibited antimicrobial activity against various pathogens, like Mycobacterium smegmatis, Klebsiella pneumonia, Clostridium perfringens and Staphylococcus epidermidis. The activity of bacteriocin was lost completely after treatment with protease, which revealed its proteinaceous nature. The bacteriocin was stable up to 100°C and exhibited antilisterial property which is a characteristic feature of class IIa bacteriocins. It was active within the pH range of 2-8 and stable against various chemicals and denaturants. Tricine SDS-PAGE revealed its molecular weight to be 4.0 kDa, where the corresponding activity against Listeria monocytogenes was also noted. Treatment of L. monocytogenes with bacteriocin decreased the viable cell count, and scanning electron microscope analysis revealed membrane pore formation that resulted in the release of intracellular content, suggesting its bactericidal effect.

Evaluation of pyroptosis in macrophages using cytosolic delivery of purified flagellin

Pyroptosis is a molecularly controlled form of cell death that exhibits some features of apoptosis as well of necrosis. Pyroptosis is induced by inflammasome-activated caspase-1 or caspase-11 (caspase-4 in humans), as a result of distinct pathogenic or damage stimuli. Although pyroptosis displays some morphological and biochemical features of apoptosis, it has an inflammatory outcome due to the loss of plasma membrane integrity and the consequent release of intracellular contents, reminiscent to necrosis. Here, we use cytosolic delivery of purified flagellin as an experimental tool to trigger pyroptosis and describe potential methods to study this form of cell death. Finally, we discuss the advantages and limitations of these methods.

Functions of the osteocyte network in the regulation of bone mass

Osteocytes establish an extensive intracellular and extracellular communication system via gap-junction-coupled cell processes and canaliculi throughout bone and the communication system is extended to osteoblasts on the bone surface. The osteocyte network is an ideal mechanosensory system and suitable for mechanotransduction. However, the overall function of the osteocyte network remains to be clarified, since bone resorption is enhanced by osteocyte apoptosis, which is followed by a process of secondary necrosis attributable to the lack of scavengers. The enhanced bone resorption is caused by the release of intracellular content, including immunostimulatory molecules that activate osteoclastogenesis through the canaliculi. Therefore, a mouse model is required in which the osteocyte network is disrupted but in which no bone resorption is induced, in order to evaluate the overall functions of the osteocyte network. One such model is the BCL2 transgenic mouse, in which the osteocyte network, including both intracellular and extracellular networks, is disrupted. Another model is the osteocyte-specific Gja1 knockout mouse, in which intercellular communication through gap junctions is impaired but the canalicular system is intact. Combining the findings from these mouse models with previous histological observations showing the inverse linkage between osteocyte density and bone formation, we conclude that the osteocyte network enhances bone resorption and inhibits bone formation under physiological conditions. Further, studies with BCL2 transgenic mice show that these osteocyte functions are augmented in the unloaded condition. In this condition, Rankl upregulation in osteoblasts and Sost upregulation in osteocytes are, at least in part, responsible for enhanced bone resorption and suppressed bone formation, respectively.

Chemokines and mitochondrial products activate neutrophils to amplify organ injury during mouse acute liver failure

Acetaminophen (APAP) is a safe analgesic and antipyretic drug. However, APAP overdose leads to massive hepatocyte death. Cell death during APAP toxicity occurs by oncotic necrosis, in which the release of intracellular contents can elicit a reactive inflammatory response. We have previously demonstrated that an intravascular gradient of chemokines and mitochondria-derived formyl peptides collaborate to guide neutrophils to sites of liver necrosis by CXC chemokine receptor 2 (CXCR2) and formyl peptide receptor 1 (FPR1), respectively. Here, we investigated the role of CXCR2 chemokines and mitochondrial products during APAP-induced liver injury and in liver neutrophil influx and hepatotoxicity. During APAP overdose, neutrophils accumulated into the liver, and blockage of neutrophil infiltration by anti-granulocyte receptor 1 depletion or combined CXCR2-FPR1 antagonism significantly prevented hepatotoxicity. In agreement with our in vivo data, isolated human neutrophils were cytotoxic to HepG2 cells when cocultured, and the mechanism of neutrophil killing was dependent on direct contact with HepG2 cells and the CXCR2-FPR1-signaling pathway. Also, in mice and humans, serum levels of both mitochondrial DNA (mitDNA) and CXCR2 chemokines were higher during acute liver injury, suggesting that necrosis products may reach remote organs through the circulation, leading to a systemic inflammatory response. Accordingly, APAP-treated mice exhibited marked systemic inflammation and lung injury, which was prevented by CXCR2-FPR1 blockage and Toll-like receptor 9 (TLR9) absence (TLR9(-/-) mice). Chemokines and mitochondrial products (e.g., formyl peptides and mitDNA) collaborate in neutrophil-mediated injury and systemic inflammation during acute liver failure. Hepatocyte death is amplified by liver neutrophil infiltration, and the release of necrotic products into the circulation may trigger a systemic inflammatory response and remote lung injury.

Meloxicam-Induced Rhabdomyolysis in the Context of an Acute Ross River Viral Infection

Acute rhabdomyolysis is a clinical and laboratory syndrome resulting from the breakdown of skeletal muscle, with the release of intracellular contents into the circulatory system, which can cause potentially lethal complications. Here, we present the case of a patient who developed acute rhabdomyolysis after consumption of meloxicam for jaw pain and experienced generalized myalgias in the context of an acute febrile illness with generalized urticaria. Further investigation indicated elevated muscle enzymes and acute renal failure. Serological analysis revealed that the patient was positive for Ross River virus (RRV) IgM. Genetic studies to detect CYP2C9 polymorphisms were negative. Meloxicam was discontinued. He responded to conservative measures within 2 weeks. Oral aspirin challenge was negative, suggesting a drug-specific effect of meloxicam rather than a class effect. Our case indicates a causative role for meloxicam and/or acute RRV in rhabdomyolysis.