Translational Research - Articles in Press

The role of inflammation in health and disease: Translating discovery into novel therapeutic approaches - Uncorrected Proof

Neal L. Weintraub — 2012-05-21

The 2012 Combined Annual Meeting of the Central Society for Clinical Research and the Midwestern Section American Federation for Medical Research was held April 26 to 27 in Chicago, Illinois. The Central Society for Clinical Research (now formally known as the Central Society for Clinical and Translational Research), founded in 1928, is one of the oldest and largest academic medical societies in the Midwest. The primary objectives are to advance medical science, cultivate clinical research by the methods of the natural and behavioral sciences, correlate science with the art of medical practice, encourage scientific investigation, and disseminate this science among the members. The membership is a veritable Who's Who of the academic leadership of American medical schools. Past presidents include Robert Good, the father of modern immunology, Charles Rammelkamp, who identified streptococcus as the etiologic agent for rheumatic fever, John Hickam (after whom the Hickam Lecture is named), who created the prototype for modern departments of medicine as Chairman of Medicine at Indiana, and Donald Seldin, a giant of 20th century medicine who transformed the Department of Medicine at the University of Texas Southwestern into one of the top departments in the nation.

Functional analysis of the novel sequence variants within TBX5 gene promoter in patients with ventricular septal defects - Uncorrected Proof

2012-05-14

Congenital heart disease (CHD) is the most common birth defect that affects 1% to 2% of human live births. Despite advanced surgical treatment, adult patients with CHD are predisposed to late cardiac complications, such as arrhythmias, coronary heart disease, and heart failure. Mutations in cardiac transcription factor genes, including GATA transcription factor 4; NK2 transcription factor, locus 5; and T-box transcription factor 5 (TBX5) genes, have been implicated in a small portion of familiar and sporadic CHD cases. However, genetic causes and underlying molecular mechanisms of CHD remain largely unknown.

Vascular endothelial growth factor and uPA/suPAR system in early and advanced chronic kidney disease patients: a new link between angiogenesis and hyperfibrinolysis? - Uncorrected Proof

Krystyna Pawlak, Blanka Ulazka, Michal Mysliwiec, Dariusz Pawlak — 2012-05-10

Disturbances in hemostasis and abnormal angiogenesis are components in the plaque growth and destabilization. The role of the vascular endothelial growth factor (VEGF) in the perturbation of hemostasis in chronic kidney disease (CKD) is still unknown. In this preliminary study, we investigate the relation between VEGF and the parameters of coagulation: tissue factor (TF), its inhibitor (TFPI), and fibrinolytic system: urokinase-type plasminogen activator (uPA), its soluble receptor (suPAR), tissue-type plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1), plasmin/antiplasmin complexes (PAP) in the patients with mild-to-moderate, and severe CKD and healthy controls. All indices (except TFPI) were raised in CKD patients, particularly in those with severe CKD, compared with controls. The strong positive correlations were between VEGF and some parameters, both coagulation (TF, TFPI, TF/TFPI ratio) and fibrinolytic system (uPA, suPAR, PAP). The relationships were also between the individual hemostatic parameters. In multiple regression analysis, VEGF and kidney dysfunction markers (urea and creatinine levels) were independently associated with uPA, and VEGF was independently associated with suPAR levels. Moreover, PAP was independently associated with age and suPAR. This study represents the first to investigate the relation between VEGF and the activation both coagulation and fibrinolysis in CKD patients. VEGF and the parameters of hemostatic system activation were higher in the CKD group than in the controls with a significant correlation between them. VEGF was independently associated with uPA/suPAR system, whereas suPAR was independently related to PAP levels, suggesting a new link between abnormal angiogenesis and hyperfibrinolysis in this population.

Circulating microRNAs as candidate biomarkers in patients with systemic lupus erythematosus - Uncorrected Proof

Honglei Wang, Wujian Peng, Xin Ouyang, Wuxian Li, Yong Dai — 2012-05-07

Aberrant expression of microRNAs (miRNAs) has been identified in various diseases. Recent studies demonstrated that miRNAs can be detected in the circulation and serve as potential biomarkers of various diseases. Moreover, the detection of circulating miRNAs can provide important novel information concerning diseases. In this study, a miRNA profile was used to determine the aberrantly expressed circulating miRNAs in patients with systemic lupus erythematosus (SLE) compared with patients with rheumatoid arthritis (RA) and healthy controls (HCs). To further confirm the microarray data, we identified 8 miRNAs (miR-126, miR-21, miR-451, miR-223, miR-16, miR-125a-3p, miR-155, and miR-146a) by real-time quantitative PCR (qRT-PCR) in 20 healthy controls and in 55 patients, of whom 30 patients were diagnosed with SLE and 25 were diagnosed with RA. Consistent with the microarray data, miR-126 was specifically enriched only in the blood of the SLE patients, but 4 other miRNAs (miR-21, miR-451, miR-223, and miR-16) were upregulated in the patients with SLE and were also significantly increased in the patients with RA. In contrast, miR-125a-3p, miR-155, and miR-146a showed a trend toward significantly reduced levels in the patients with SLE. In addition, to further estimate the potential roles of these differentially expressed circulating miRNAs in the pathogenesis of SLE, we used a bioinformatics exploratory analysis and identified a number of significantly enriched pathways, which implied that most dysregulated circulating miRNAs might be involved in various signal transduction pathways and cell interactions, particularly the MAPK signaling pathway. Based on these findings, we postulate that aberrantly expressed plasma miRNAs could be attractive as candidates for putative biomarkers of SLE and may help elucidate the possible pathogenesis of SLE.

Emerging view of the human virome - Uncorrected Proof

Kristine M. Wylie, George M. Weinstock, Gregory A. Storch — 2012-04-25

The human virome is the collection of all viruses that are found in or on humans, including both eukaryotic and prokaryotic viruses. Eukaryotic viruses clearly have important effects on human health, ranging from mild, self-limited acute or chronic infections to those with serious or fatal consequences. Prokaryotic viruses can also affect human health by affecting bacterial community structure and function. Therefore, definition of the virome is an important step toward understanding how microbes affect human health and disease. We review progress in virome analysis, which has been driven by advances in high-throughput, deep sequencing technology. Highlights from these studies include the association of viruses with clinical phenotypes and description of novel viruses that may be important pathogens. Together these studies indicate that analysis of the human virome is critical as we aim to understand how microbial communities affect human health and disease. Descriptions of the human virome will stimulate future work to understand how the virome affects long-term human health, immunity, and response to coinfections. Analysis of the virome ultimately may affect the treatment of patients with a variety of clinical syndromes.

Erratum - Uncorrected Proof

2012-04-25

Giuseppina Novo, Francesco Cappello, Manfredi Rizzo, Giovanni Fazio, Sabrina Zambuto, Enza Tortorici, Antonella Marino Gammazza, Simona Corrao, Giovanni Zummo, Everly C. De Macario, Alberto JL Macario,5 Pasquale Assennato, Salvatore Novo, Giovanni L. Volti

Glomerular basement membrane and related glomerular disease - Uncorrected Proof

Ying Maggie Chen, Jeffrey H. Miner — 2012-04-11

The glomerular basement membrane (GBM) is lined by fenestrated endothelium from the capillary-lumen side and by interdigitating foot processes of the podocytes from the urinary- space side. These three layers of the glomerular capillary wall constitute the functional unit of the glomerular filtration barrier. The GBM is assembled through an interweaving of type IV collagen with laminins, nidogen, and sulfated proteoglycans. Mutations in genes encoding LAMB2, COL4A3, COL4A4, and COL4A5 cause glomerular disease in humans as well as in mice. In addition, laminin α5 mutation in podocytes leads to proteinuria and renal failure in mice. Moreover, more neoepitopes in Goodpasture's disease and for the first time alloepitopes in Alport post-transplantation nephritis have been located in the collagen α5(IV) NC1 domain. These discoveries underscore the importance of the GBM in establishing and maintaining the integrity of the glomerular filtration barrier.

The clinical and immunologic features of pulmonary fibrosis in sarcoidosis - Uncorrected Proof

2012-04-11

Sarcoidosis is a multisystem, granulomatous disease that most often affects the lungs. The clinical course is highly variable; many patients undergo spontaneous remission, but up to a third of patients progresses to a chronic disease course. The development of pulmonary fibrosis (PF) in a subset of patients with chronic disease has a negative impact on morbidity and mortality. While sarcoidosis-associated PF can be progressive, it is often referred to as “burnt out” disease, a designation reflecting inactive granulomatous inflammation. The immune mechanisms of sarcoidosis-associated PF are not well understood. It is not clear if fibrotic processes are active from the onset of sarcoidosis in predisposed individuals, or whether a profibrotic state develops as a response to ongoing inflammation. Transforming growth factor β (TGF-β) is an important profibrotic cytokine, and in sarcoidosis, distinct genotypes of TGF-β have been identified in those with PF. The overall cytokine profile in sarcoidosis-associated PF has not been well characterized, although a transition from a T helper 1 to a T helper 2 signature has been proposed. Macrophages have important regulatory interactions with fibroblasts, and the role of alveolar macrophages in sarcoidosis-associated PF is a compelling target for further study. Elucidating the natural history of sarcoidosis-associated PF will inform our understanding of the fundamental derangements, and will enhance prognostication and the development of therapeutic strategies.

Participation of endoplasmic reticulum stress in the pathogenesis of spontaneous glomerulosclerosis–Role of intra-renal angiotensin system - Uncorrected Proof

Mohammad A. Aminzadeh, Tadashi Sato, Nosratola D. Vaziri — 2012-04-09

Endoplasmic reticulum (ER) is the site of synthesis, folding, assembly, and degradation of proteins. Disruption of ER function leads to ER stress, which is marked by accumulation of unfolded proteins in the ER lumen. Detection of unfolded proteins by the ER membrane receptors triggers the “unfolded protein response (UPR)” designed to restore ER function via activation of the adaptive/cytoprotective responses. Failure of UPR or persistent stress triggers activation of ER stress-mediated apoptotic pathway. Several in vivo and in vitro studies have demonstrated the association of ER stress with glomerular diseases. Imai rats develop progressive glomerulosclerosis (GS), which is associated with oxidative stress, inflammation and activation of intra-renal angiotensin system, and can be prevented by AT-1 receptor blockade (ARB). Since persistent oxidative and inflammatory stresses trigger ER stress-induced apoptosis and tissue injury, we hypothesized that kidneys in the Imai rats may exhibit failure of the adaptive and activation of the apoptotic ER stress responses, which could be prevented by ARB. To this end 10-week old Imai rats were randomized to untreated and ARB-treated groups and observed for 24 weeks. At age 34 weeks, untreated rats showed heavy proteinuria, azotemia, advanced GS, impaired ER stress adaptive/cytoprotective responses (depletion of UPR-mediating proteins), and activation of ER stress apoptotic responses. ARB treatment attenuated GS, suppressed intra-renal oxidative stress, restored ER-associated adaptive/cytoprotective system, and prevented the ER stress mediated apoptotic response in this model. Thus, progressive GS in Imai rats is accompanied by activation of ER stress-associated apoptosis, which can be prevented by ARB.

Influence of hepatic and intestinal efflux transporters and their genetic variants on the pharmacokinetics and pharmacodynamics of raloxifene in osteoporosis treatment - Uncorrected Proof

2012-03-29

Raloxifene exhibits a large and unexplained interindividual variability in its pharmacokinetics and pharmacodynamics. The aim of our study was to identify transporters involved in the efflux of raloxifene and its glucuronide metabolites by various in vitro models and by an in vivo study to explore the possible involvement of P-glycoprotein (Pgp), multidrug resistance–associated protein (MRP)1, MRP2, MRP3, and breast cancer resistance protein in the observed high interindividual variability. Experiments with the parallel artificial membrane permeability assay showed the highest passive permeability for raloxifene, followed by raloxifene-6-β-glucuronide (M1), raloxifene-4'-β-glucuronide (M2), and raloxifene-6,4'-diglucuronide (M3). Caco-2 cell monolayer experiments indicated an interaction of raloxifene with Pgp. The ATPase assay confirmed the raloxifene interaction with Pgp and indicated interactions of all raloxifene species with MRP1, MRP2, MRP3, and breast cancer resistance protein, except for M1, which did not show any interactions with MRP2. Furthermore, the vesicular experiments confirmed the interaction of M2 and M3 with MRP2. Although the in vivo study on osteoporotic postmenopausal women on raloxifene could not confirm a significant influence of ABCB1 and ABCC2 genetic polymorphisms on its pharmacokinetics, a clear trend toward higher total raloxifene concentrations was observed in carriers of at least 1 ABCB1 c.3435T allele. Moreover, the same polymorphism effect was also observed as a significant increase in total hip bone mineral density after 1 year of treatment. The results of our study support the involvement of efflux transporters in disposition of raloxifene and its metabolites and may partially explain the observed raloxifene variability by the influence of the ABCB1 c.3435C>T polymorphism.

Miniaturized technology for protein and nucleic acid point-of-care testing - Uncorrected Proof

Felix Olasagasti, Juan Carlos Ruiz de Gordoa — 2012-03-12

The field of point-of-care (POC) testing technology is developing quickly and producing instruments that are increasingly reliable, while their size is being gradually reduced. Proteins are a common target for POC analyses and the detection of protein markers typically involves immunoassays aimed at detecting different groups of proteins such as tumor markers, inflammation proteins, and cardiac markers; but other techniques can also be used to analyze plasma proteins. In the case of nucleic acids, hybridization and amplification strategies can be used to record electromagnetic or electric signals. These techniques allow for the identification of specific viral or bacterial infections as well as specific cancers. In this review, we consider some of the latest advances in the analysis of specific nucleic acid and protein biomarkers, taking into account their trend toward miniaturization and paying special attention to the technology that can be implemented in future applications, such as lab-on-a-chip instruments.

Understanding vaginal microbiome complexity from an ecological perspective - Uncorrected Proof

Roxana J. Hickey, Xia Zhou, Jacob D. Pierson, Jacques Ravel, Larry J. Forney — 2012-03-07

The various microbiota normally associated with the human body have an important influence on human development, physiology, immunity, and nutrition. This is certainly true for the vagina wherein communities of mutualistic bacteria constitute the first line of defense for the host by excluding invasive, nonindigenous organisms that may cause disease. In recent years much has been learned about the bacterial species composition of these communities and how they differ between individuals of different ages and ethnicities. A deeper understanding of their origins and the interrelationships of constituent species is needed to understand how and why they change over time or in response to changes in the host environment. Moreover, there are few unifying theories to explain the ecological dynamics of vaginal ecosystems as they respond to disturbances caused by menses and human activities such as intercourse, douching, and other habits and practices. This fundamental knowledge is needed to diagnose and assess risk to disease. Here we summarize what is known about the species composition, structure, and function of bacterial communities in the human vagina and the applicability of ecological models of community structure and function to understanding the dynamics of this and other ecosystems that comprise the human microbiome.

The Gly460Trp polymorphism of alpha-adducin gene as a predictor of renal function decline over 4 years of follow-up in an apparently healthy Chinese population - Uncorrected Proof

2012-03-05

A large epidemiologic study showed that subjects with reduced renal function, defined as a glomerular filtration rate (GFR) less than 60 mL/min/1.73 m2, are at risk for the development of end-stage renal disease. The reason for this might be partly related to the presence of traditional cardiovascular risk factors, such as hypertension and diabetes. For the elderly without traditional risk factors, it is unknown which factor makes subjects susceptible to reduced renal function.

Pretransplant IFN-γ ELISPOT assay as a potential screening test to select immunosuppression protocols for patients receiving basiliximab induction therapy - Uncorrected Proof

2012-03-05

The use of basiliximab induction therapy has increased in standard immunological risk patients. The objective of this study was to identify whether pretransplant donor-reactive interferon-γ enzyme-linked immunosorbent spot (ELISPOT) assay results were associated with post-transplant clinical outcomes in patients receiving basiliximab induction therapy and whether this could be helpful for choosing an efficacious immunosuppressive regimen. In 154 living donor renal transplant recipients who received basiliximab induction therapy without desensitization, we determined pretransplant ELISPOT frequencies and correlated the results with clinical outcomes based on the use of calcineurin inhibitors (tacrolimus [TAC] or cyclosporine [CSA]). The ELISPOT (+) patients had higher rate of post-transplant biopsy-proven acute rejection (AR) than ELISPOT (−) patients (P = 0.001) regardless of immunosuppressive regimen. In the logistic and multivariate regression analysis, ELISPOT was the only significant correlate of AR (P = 0.002), and the patients with increased ELISPOT results and CSA therapy were associated with AR. Our results suggest that the pretransplant ELISPOT (+) may assess the risk of poor post-transplant outcomes in patients with basiliximab induction.

Therapeutic window of taurine against experimental stroke in rats - Uncorrected Proof

Ming Sun, Yu-Mei Zhao, Yi Gu, Chao Xu — 2012-03-02

The dose-dependent protection of taurine against experimental stroke has been demonstrated previously. The objective of this study was to investigate the therapeutic window of taurine against experimental stroke, and the effects of delayed administration of taurine on inflammatory reaction in a rat model of stroke. Rats received 2-h ischemia by intraluminal filament, and then reperfused. Taurine (50 mg/kg) was administered intravenously 4 h, 8 h, 10 h, or 12 h after ischemia. The neurologic scores and the infarct volumes were evaluated 24 h after ischemia. Then, the effect of administration of taurine at 8 h after ischemia on the neutrophil infiltration in ischemic region was determined. Treatment with taurine 4 h or 8 h after ischemia significantly improved the neurologic function, and decreased the infarct volumes 24 h after ischemia. However, administration of taurine at 10 h or 12 h after ischemia had no significant neuroprotection. Further, taurine administered at 8 h after ischemia markedly reduced myeloperoxidase activity and attenuated neutrophil infiltration in ischemic region. Our data suggest that the therapeutic window of taurine against experimental stroke is of at least 8 h, and suppressing the neutrophil infiltration may be one of the mechanisms of delayed administration of taurine against experimental stroke.

Future directions and treatment strategies for head and neck squamous cell carcinomas - Uncorrected Proof

2012-02-29

Head and neck cancer is a devastating disease that afflicts many individuals worldwide. Conventional therapies are successful in only a limited subgroup and often leave the patient with disfigurement and long lasting adverse effects on normal physiologic functions. The field is in dire need of new therapies. Oncolytic viral as well as targeted therapies have shown some success in other malignancies and are attractive for the treatment of head and neck cancer. Recently, it has been shown that a subset of head and neck cancers is human papillomavirus (HPV) positive and that this subset of cancers is biologically distinct and more sensitive to chemoradiation therapies although the underlying mechanism is unclear. However, chemoresistance remains a general problem. One candidate mediator of therapeutic response, which is of interest for the targeting of both HPV-positive and -negative tumors is the human DEK proto-oncogene. DEK is upregulated in numerous tumors including head and neck cancers regardless of their HPV status. Depletion of DEK in tumor cells in culture results in sensitivity to genotoxic agents, particularly in rapidly proliferating cells. This suggests that tumors with high DEK protein expression may be correlated with poor clinical response to clastogenic therapies. Targeting molecules such as DEK in combination with new and/or conventional therapies, holds promise for novel future therapeutics for head and neck cancer.

Pioglitazone, a PPAR-γ activator, attenuates the severity of cerulein-induced acute pancreatitis by modulating early growth response-1 transcription factor - Uncorrected Proof

Hongyu Wan, Yaozong Yuan, Jiansheng Liu, Guohong Chen — 2012-02-29

The purpose of this study was to test the hypothesis that activation of endogenous peroxisome proliferator-activated receptor (PPARγ) inhibits induction of early growth response factor-1 (Egr-1), which is rapidly induced in the pancreas following cerulein intraperitoneal injection. Acute pancreatitis was induced in mice by hourly intraperitoneal injection of cerulein. Pioglitazone was administered prophylactically and pancreatic inflammation was assessed. AR42J cells were stimulated with caerulein10−8M co-incubated in presence of different concentration of pioglitazone. The expression of PPARγ, Egr-1, and the target genes of Egr-1 were studied by real-time reverse transcriptase polymerase chain reaction (PCR), Western blot, and immunohistochemistry. In vitro, a PPAR-γ activator (pioglitazone) strikingly diminished Egr-1 mRNA and protein expression corresponding to Egr-1. In vivo, treatment with pioglitazone prior to the intraperitoneal injection of cerulein induction of Egr-1 and its target genes such as, monocyte chemotactic protein-1 (MCP-1) and macrophage inflammatory protein-1 (MIP-1). The inhibitory effect of pioglitazone on Egr-1 expression induced by cerulein was almost fully restored by GW9662. Activation of PPAR-γ suppressed the activation of Egr-1 and its inflammatory gene targets and provided potent protection against pancreas injury. These data suggest a new mechanism in which PPAR-γ activation may decrease tissue inflammation in response to a cerulein insult.

Human umbilical cord blood mononuclear cells activate the survival protein Akt in cardiac myocytes and endothelial cells that limits apoptosis and necrosis during hypoxia - Uncorrected Proof

2012-02-29

We have previously reported that human umbilical cord blood mononuclear cells (HUCBC), which contain hematopoietic, mesenchymal, and endothelial stem cells, can significantly reduce acute myocardial infarction size. To determine the mechanism whereby HUCBC increase myocyte and vascular endothelial cell survival, we treated cardiac myocytes and coronary artery endothelial cells in separate experiments with HUCBC plus culture media or culture media alone and subjected the cells to 24 h of hypoxia or normoxia. We then determined in myocytes and endothelial cells activation of the cell survival protein Akt by Western blots. We also determined in these cells apoptosis by annexin V staining and necrosis by propidium iodide staining. Thereafter, we inhibited with API, a specific and sensitive Akt inhibitor, Akt activation in myocytes and endothelial cells cultured with HUCBC during hypoxia and determined cell apoptosis and necrosis. In cells cultured without HUCBC, hypoxia only slightly activated Akt. Moreover, hypoxia increased myocyte apoptosis by ≥226% and necrosis by 58% in comparison with myocytes in normoxia. Hypoxic treatment of endothelial cells without HUCBC increased apoptosis by 94% and necrosis by 59%. In contrast, hypoxia did not significantly affect HUCBC. Moreover, in myocyte + HUCBC cultures in hypoxia, HUCBC induced a ≥135% increase in myocyte phospho-Akt. Akt activation decreased myocyte apoptosis by 76% and necrosis by 35%. In endothelial cells, HUCBC increased phospho-Akt by 116%. HUCBC also decreased endothelial cell apoptosis by 58% and necrosis by 42%. Inhibition of Akt with API in myocytes and endothelial cells cultured with HUCBC during hypoxia nearly totally prevented the HUCBC-induced decrease in apoptosis and necrosis. We conclude that HUCBC can significantly decrease hypoxia-induced myocyte and endothelial cell apoptosis and necrosis by activating Akt in these cells and in this manner HUCBC can limit myocardial ischemia and injury.

The microbiome of the lung - Uncorrected Proof

James M. Beck, Vincent B. Young, Gary B. Huffnagle — 2012-02-29

Investigation of the lung microbiome is a relatively new field. Although the lungs were classically believed to be sterile, recently published investigations have identified microbial communities in the lungs of healthy humans. At the present time, there are significant methodologic and technical hurdles that must be addressed in ongoing investigations, including distinguishing the microbiota of the upper and lower respiratory tracts. However, characterization of the lung microbiome is likely to provide important pathogenic insights into cystic fibrosis, respiratory disease of the newborn, chronic obstructive pulmonary disease, and asthma. In addition to characterization of the lung microbiome, the microbiota of the gastrointestinal tract have profound influence on the development and maintenance of lung immunity and inflammation. Further study of gastrointestinal–respiratory interactions is likely to yield important insights into the pathogenesis of pulmonary diseases, including asthma. As this field advances over the next several years, we anticipate that studies using larger cohorts, multicenter designs, and longitudinal sampling will add to our knowledge and understanding of the lung microbiome.

Cantharidin-induced inflammation in mouse ear model for translational research of novel anti-inflammatories - Uncorrected Proof

2012-02-27

The murine model of cantharidin-induced ear inflammation was profiled in detail for its alignment with the human model and to explore the mechanism of anti-inflammatory activity of the macrolide antibiotics, clarithromycin and azithromycin. Ear swelling in CD1 mice persisted for 7 days, with peak intensity at 16 h after inflammation induction. As in humans, cantharidin (12.5 μg/ear) generated macrophage-inflammatory protein (MIP)-2, monocyte chemoattractant protein (MCP)-1, keratinocyte-derived chemokine (KC), interleukin (IL)-6, IL-1β, and myeloperoxidase (MPO) production, as well as neutrophil accumulation in mouse ear tissue. The tested macrolides, clarithromycin and azithromycin, administered orally (2 × 150 mg/kg) 0.5 h before and 5 h after cantharidin challenge, reduced MIP-2, MCP-1, KC, and MPO concentrations and thereby decreased ear swelling. Our results suggest that cantharidin-induced acute inflammation represents an excellent model for translational research of novel anti-inflammatories.

Accelerated cellular senescence in the kidney: Cause or effect of disease progression? - Uncorrected Proof

Carrie L. Phillips — 2012-02-27

Structure and function are intimately connected in biological systems. When the structural integrity of cells, tissues, and organs is compromised, then physiologic function declines or completely stops. In the human kidney, architectural intactness of nephrons is closely tied to proper renal function. Numbering approximately 1 million per kidney, each nephron begins as a glomerular tuft of capillaries that filters waste products from blood. The resultant urinary filtrate flows through epithelial-lined tubules into a larger, more distal-collecting system. Maintenance of physiologically intact and functional renal tubule epithelial cells (RTECs) requires a careful balance of proliferation, maturation, regeneration, and repair. When these normal processes are disrupted in the kidney, then renal insufficiency or failure ensues. To uncover the underlying source of functional decline, affected patients may undergo a diagnostic biopsy procedure that will serve to guide therapy. Although physiology is static in formalin-fixed kidney biopsy specimens, careful examination of these tissue samples at a microscopic level permits reproducible morphologic assessment of renal tubules and their surrounding interstitium, which can be correlated to a patient’s renal function. In most cases, this correlation seems to hold up regardless of the basic underlying renal disease. In other words, thousands of diverse insults and injuries to the kidney follow along converging pathways that, if untreated, ultimately lead to disruption of the tubulointerstitial compartment. This decline seems to involve cellular senescence.

Erratum - Uncorrected Proof

2012-02-27

Cary Stelloh, Kenneth P. Allen, David L. Mattson, Alexandra Lerch-Gaggl, Sreenivas Reddy, and Ashraf El-Meanawy Prematurity in Mice Leads to Reduction in Nephron Number, Hypertension, and Proteinuria

Biochemical dynamics relevant to the safety of low-dose, intraclot alteplase for deep vein thrombosis - Uncorrected Proof

Jay Nelson Lozier, Ann Cullinane, Khanh Nghiem, Richard Chang, Mcdonald K. Horne — 2012-02-24

Intraclot tissue plasminogen activator (tPA) has been shown to be an effective treatment for deep vein thrombosis (DVT) (Radiology 2008;246:619 and J Vasc Interv Radiol 2011;22:1107). We sought to correlate pharmacokinetics of tPA, fibrinogen, fibrinolytic inhibitors, and D-dimers with the safety and efficacy of intraclot tPA. Thirty subjects received intraclot tPA for lower extremity DVT by infiltrating the thrombus with ≤10 mg doses tPA in an open-label study, using a pulse-spray catheter. We measured various parameters over 8 h following a first dose of tPA. Mean tPA levels of 75 units per mL (95% confidence interval 19–131 units/mL) were seen immediately after administration of a mean tPA dose of 8.0 mg (SD 1.5 mg). tPA levels returned to baseline within 2 h of completion of treatment. Plasminogen activator inhibitor-1 (PAI-1) was consumed following tPA treatment, but rose to levels significantly greater than baseline (P < 0.001). Fibrinogen decreased slightly, but remained >125 mg/dL for all subjects. α2-antiplasmin decreased from a mean of 115 units/mL to 56 units/mL after tPA administration (P < 0.001) and remained decreased for 8 h. Plasminogen at baseline (112 units/mL) decreased to 89 units/mL immediately after tPA administration (P < 0.001) and was unchanged thereafter. D-dimer levels were >20 μg/mL in all but 4 subjects, one of whom was the only one to fail to achieve clot lysis. The safety of low-dose, intraclot tPA is due to its short persistence in the circulation, lack of hypofibrinogenemia, and a reflexive rise PAI-1. Subjects whose D-dimers remain <20 μg/mL are at risk of not achieving thrombolysis.

Blockade of electron transport before ischemia protects mitochondria and decreases myocardial injury during reperfusion in aged rat hearts - Uncorrected Proof

Christine Tanaka-Esposito, Qun Chen, Edward J. Lesnefsky — 2012-02-20

Myocardial injury is increased in the aged heart following ischemia and reperfusion (I-R) in both humans and experimental models. Hearts from aged 24-month-old Fischer 344 rats sustain greater cell death and decreased contractile recovery after I-R compared with 6-month-old adult controls. Cardiac mitochondria incur damage during I-R contributing to cell death. Aged rats have a defect in complex III of the mitochondrial electron transport chain (ETC) localized to the interfibrillar population of cardiac mitochondria (IFM), situated in the interior of the cardiomyocyte among the myofibrils. The defect involves the quinol oxidation site (Qo) and increases the production of reactive oxygen species (ROS) in the baseline state. Ischemia further decreases complex III activity via functional inactivation of the iron-sulfur subunit. We studied the contribution of ischemia-induced defects in complex III with the increased cardiac injury in the aged heart. The reversible blockade of the ETC proximal to complex III during ischemia using amobarbital protects mitochondria against ischemic damage, removing the ischemia component of mitochondrial dysfunction. Reperfusion of the aged heart in the absence of ischemic mitochondrial damage decreases net ROS production from mitochondria and reduces cell death. Thus, even despite the persistence of the age-related defects in electron transport, protection against ischemic damage to mitochondria can reduce injury in the aged heart. The direct therapeutic targeting of mitochondria protects against ischemic damage and decreases cardiac injury during reperfusion in the high risk elderly heart.

Pathogenesis of pre-eclampsia: marinobufagenin and angiogenic imbalance as biomarkers of the syndrome - Uncorrected Proof

2012-02-10

Pre-eclampsia (preE), a pregnancy disorder with the de novo onset of hypertension and proteinuria after 20 weeks of gestation, has multiple triggers that initiate pathophysiologic mechanisms. This review addresses translational aspects of preE by synthesizing information on preE pathogenesis, describing diagnostic biomarkers that predict disease, and suggesting strategies to lessen adverse outcomes. Key to this understanding is the role of cardiotonic bufodienolides, with marinobufagenin (MBG) as the prototype, and angiogenic factors in preE pathogenesis. Data from a rat model believed to mimic human preE show that urinary excretion of MBG increases before the onset of hypertension and proteinuria and that affected animals have an increased vascular leakage and blood brain barrier permeability. Angiogenic imbalance occurs with the onset of the syndrome in this model. Also, we report that MBG levels in preE patients exceed those in normal pregnancy and that angiogenic factors are altered in patients showing signs and symptoms of overt disease. In vitro administration of MBG inhibits cytotrophoblast function and triggers hyperpermeability in endothelial cell monolayers. We advance the hypotheses that MBG precedes preE; MBG causes disruption of tight junction proteins leading to vascular leak via activation of MAPK which triggers apoptotic mechanisms resulting in further endothelial dysfunction leading to edema with the release of angiogenic factors. This review provides new evidence about the role of MBG and vasoactive intermediates in preE pathogenesis including the neurologic sequela and may reveal new therapeutic targets for the prevention of preE complications.

Tanshinone II-A inhibits oxidized LDL-induced LOX-1 expression in macrophages by reducing intracellular superoxide radical generation and NF-κB activation - Uncorrected Proof

2012-02-10

Lectin-like oxidized LDL (oxLDL) receptor-1 (LOX-1), a novel scavenger receptor highly expressed in human and experimental atherosclerotic lesions, is responsible for the uptake of oxLDL in vascular cells. We demonstrated previously that Tanshinone II-A (Tan), a pharmacologically active compound extracted from the rhizome of the Chinese herb Salvia miltiorrhiza Bunge, inhibits atherogenesis in hypercholesterolemic rats, rabbits, and apolipoprotein-E deficient (ApoE−/−) mice. However, the precise mechanism by which Tan protects against atherogenesis remains to be elucidated. Therefore, we hypothesized that Tan can suppress the uptake of oxLDL by diminishing the expression of LOX-1 via suppression of NF-κB signaling pathway, thereby contributing to reduced macrophage foam cell formation. In cultured murine macrophages, oxLDL induced LOX-1 expression at the mRNA and protein levels, was abrogated by addition of Tan or pyrrolidinedithiocarbamic acid ammonium salt (PDTC), a widely used inhibitor of NF-κB, suggesting the involvement of NF-κB. Tan also reduced LOX-1 expression in atherosclerotic lesions of ApoE−/− mice fed a high cholesterol diet. Mechanistically, Tan suppressed the nuclear translocation of NF-κB P65 subunit and phosphorylation of IκB-α induced by oxLDL. Electrophoretic mobility shift assay (EMSA) assay demonstrated that Tan inhibited the nuclear protein binding to NF-κB consensus sequence. Functionally, we observed that Tan inhibited DiI-oxLDL uptake by macrophages in a fashion similar to that produced by LOX-1 neutralizing antibody. Our current findings reveal a novel mechanism by which Tan protects against atherogenesis and shed new light on the potential therapeutic application of Tan to the treatment and prevention of atherosclerotic cardiovascular diseases.

Serum levels of proprotein convertase subtilisin/kexin type 9 in subjects with familial hypercholesterolemia indicate that proprotein convertase subtilisin/kexin type 9 is cleared from plasma by low-density lipoprotein receptor–independent pathways - Uncorrected Proof

2012-02-10

Secreted proprotein convertase subtilisin/kexin type 9 (PCSK9) binds to the low-density lipoprotein receptor (LDLR) at the cell surface and disrupts the normal recycling of the LDLR. When human PCSK9 is injected into LDLR-deficient mice, PCSK9 is still rapidly cleared by the liver. This finding may suggest that PCSK9 is physiologically also cleared by receptors other than the LDLR. An alternative explanation could be that PCSK9 has undergone modifications during purification and is cleared by scavenger receptors on liver endothelial sinusoidal cells when injected into mice. If the only mechanism for clearing PCSK9 in humans is through the LDLR, one would expect that differences in the number of LDLRs would affect the plasma levels of low-density lipoprotein cholesterol (LDLC) and PCSK9 in a similar fashion. In this study, levels of LDLC and PCSK9 were measured in familial hypercholesterolemia (FH) homozygotes, FH heterozygotes, and normocholesterolemic subjects. The ratio between the levels of LDLC and PCSK9 was 1.7-fold higher in FH heterozygotes and 3-fold higher in FH homozygotes than in the normocholesterolemic subjects. Thus, defective LDLRs have a greater impact on the levels of LDLC than on the levels of PCSK9. By assuming that the rate of PCSK9 synthesis is similar in the 3 groups, this finding suggests that in humans, plasma PCSK9 is also cleared by LDLR-independent mechanisms.

An inflammation-based prognostic index predicts survival advantage after transarterial chemoembolization in hepatocellular carcinoma - Uncorrected Proof

David J. Pinato, Rohini Sharma — 2012-02-10

Transarterial chemoembolization (TACE) is the preferred treatment for unresectable, intermediate-stage hepatocellular carcinoma (HCC). However, survival after TACE can be highly variable, suggesting the need for more accurate patient selection to improve therapeutic outcome. We have explored the prognostic ability of the blood neutrophil-to-lymphocyte ratio (NLR), a biomarker of systemic inflammation, as a predictor of survival after TACE. Fifty-four patients with a diagnosis of HCC eligible for TACE were selected. Clinicopathologic variables were collected, including demographics, tumor staging, liver functional reserve, and laboratory variables. Dynamic changes in the NLR before and after TACE were studied as predictors of survival using both a univariate and multivariate Cox regression model. Patients in whom the NLR remained stable or normalized after TACE showed a significant improvement in overall survival of 26 months compared with patients showing a persistently abnormal index (P = .006). Other predictors of survival on univariate analysis were Cancer of the Liver Italian Program score (P = .05), intrahepatic spread (P = .01), tumor diameter > 5 cm (P = .02), > 1 TACE (P = .01), alpha-fetoprotein ≥ 400 (P = .002), and radiologic response to TACE (P < .001). Improved NLR after TACE (P = .03) and radiologic response after TACE (P = .003) remained independent predictors of survival on multivariate analysis. Changes in alpha-fetoprotein after treatment did not predict survival. Patients with a persistently increased NLR have a worse outcome after TACE. NLR is a simple and universally available stratifying biomarker that can help identify patients with a significant survival advantage after TACE.

Relationship between adipokines and coronary artery aneurysm in children with Kawasaki disease - Uncorrected Proof

Ruixi Liu, Bo He, Fang Gao, Qian Liu, Qijian Yi — 2012-02-10

Body fat is an important source of adipokines not only in association with energy balance, but also with inflammatory and immune responses. This study investigated the relationship between serum levels of adipokines and coronary artery aneurysm in patients with Kawasaki disease (KD). Levels of leptin, adiponectin, and resistin were measured in 165 cases, including 4 groups: the control group (n = 85), KD with normal coronary arteries (n = 41), KD with dilatation and/or ectasia (n = 31), and KD with coronary aneurysm (n = 8). White blood cells counts (WBC), red blood cells counts (RBC), hemoglobin (HB), Hematocrit (Hct), platelet count, C reactive protein (CRP), and erythrocyte sedimentation rate (ESR) were tested in children. Levels of adiponectin and resistin levels were significantly elevated; hemoglobin significantly decreased in the group of KD with coronary aneurysm compared with the controls or other KD subgroups. There were markedly positive relationships between levels of resistin and CRP, and negative relationships between levels of resistin and RBC in patients with KD. Levels of adiponectin, resistin, and hemoglobin were associated with the development of coronary aneurysm in children with KD. The up-regulation of resistin secreted from adipose tissue may be closely linked to up-regulation of systemic proinflammatory markers in acute KD.

Serum level of soluble CD26/dieptidyl peptidase-4 (DPP-4) activity correlates with other liver disease biomarkers both in Asian and European patients. Higher doses of DPP-4 inhibitors (more than 50 mg daily sitagliptin) or loss of 4 kg bodyweight with a strict supervised diet are required in those patients with type 2 diabetes mellitus who also present with nonalcoholic fatty liver disease (NAFLD) - Uncorrected Proof

Gabor Firneisz, Aniko Somogyi — 2012-02-10

We read with great interest the online paper titled: “Serum level of soluble CD26/dipeptidyl peptidase-4 (DPP-4) predicts the response to sitagliptin, a DPP-4 inhibitor, in patients with type 2 diabetes controlled inadequately by metformin and/or sulfonylurea” by Aso et al in the Translational Research.

Author reply - Uncorrected Proof

Yoshimasa Aso — 2012-02-10

We appreciate Dr. Firneisz’s interest and comments on our article, as well as pointing out our careless error to us and the readers. As they indicated, our statement that there was a negative correlation between serum sCD26/DPP4 and gGT or ALT in patients with type 2 diabetes is incorrect (Page 30, line 1). In fact, we found a positive correlation between serum sCD26/DPP4 and gGT or ALT in the present study, as shown in Table II.

Identifying and exploiting defects in the Fanconi anemia/BRCA pathway in oncology - Uncorrected Proof

Shane R. Stecklein, Roy A. Jensen — 2012-02-10

Defects in components of DNA repair pathways are responsible for numerous hereditary cancer syndromes and are also common in many sporadic malignancies. Inherited mutations in the breast cancer susceptibility genes BRCA1 and BRCA2 or components of the Fanconi anemia (FA) complex incite genomic instability and predispose to malignancy. The products of the BRCA and FA genes participate in a conserved DNA damage repair pathway that is responsible for repairing interstrand crosslinks and double-strand DNA breaks by homologous recombination. While the genetic instability resulting from FA/BRCA dysfunction contributes to cancer pathogenesis, deficiency of these genes also lends to therapeutic exploitation. Crosslinking agents and ionizing radiation induce damage in cancer cells that requires the FA/BRCA pathway to be resolved; thus cancers that are deficient in BRCA1, BRCA2, or any other component of the FA/BRCA pathway are hypersensitive to these agents. Moreover, emerging synthetic lethal strategies offer opportunities to selectively target cancer cells with defects in homologous recombination. Conversely, enhanced activity of the FA/BRCA pathway is responsible for acquired resistance to specific therapeutic agents, suggesting that both dysfunction and hyperfunction of the FA/BRCA repair machinery are rational targets for cancer therapy. Selection of specific cytotoxic agents based on repair capacity may improve responses and enable personalized cytotoxic chemotherapy. This article reviews the FA/BRCA pathway and current approaches to identify deficiencies within it, discusses synthetic lethality and enhanced repair capacity as causes of therapeutic hypersensitivity and resistance, respectively, and highlights recent studies that have linked FA/BRCA pathway function with therapeutic efficacy.

Translational research in infectious disease: current paradigms and challenges ahead - Uncorrected Proof

Judith M. Fontana, Elizabeth Alexander, Mirella Salvatore — 2012-01-16

In recent years, the biomedical community has witnessed a rapid scientific and technologic evolution after the development and refinement of high-throughput methodologies. Concurrently and consequentially, the scientific perspective has changed from the reductionist approach of meticulously analyzing the fine details of a single component of biology to the “holistic” approach of broadmindedly examining the globally interacting elements of biologic systems. The emergence of this new way of thinking has brought about a scientific revolution in which genomics, proteomics, metabolomics, and other “omics” have become the predominant tools by which large amounts of data are amassed, analyzed, and applied to complex questions of biology that were previously unsolvable. This enormous transformation of basic science research and the ensuing plethora of promising data, especially in the realm of human health and disease, have unfortunately not been followed by a parallel increase in the clinical application of this information. On the contrary, the number of new potential drugs in development has been decreasing steadily, suggesting the existence of roadblocks that prevent the translation of promising research into medically relevant therapeutic or diagnostic application. In this article, we will review, in a noninclusive fashion, several recent scientific advancements in the field of translational research, with a specific focus on how they relate to infectious disease. We will also present a current picture of the limitations and challenges that exist for translational research, as well as ways that have been proposed by the National Institutes of Health to improve the state of this field.

Treating hepatitis C infection by targeting the host - Uncorrected Proof

Shadi Salloum, Andrew W. Tai — 2012-01-11

More than 130 million people worldwide are chronically infected with the hepatitis C virus (HCV), which can lead to cirrhosis, liver failure, and hepatocellular carcinoma. Although recently approved HCV NS3-4A protease inhibitors significantly improve treatment response rates, current HCV treatment is still frequently limited by side effects and by the low genetic barrier to viral resistance against direct-acting antiviral agents. A complementary strategy is to target the host cellular factors that support the HCV life cycle. Several studies, including RNA interference screens, demonstrated that HCV depends on dozens, if not hundreds, of cellular proteins to complete its life cycle. A better understanding of the interactions between HCV proteins and host factors may help to identify host targets for antiviral therapy. In this review, we highlight some of the host factors that are particularly attractive targets for the treatment of HCV.

Effect of oxidative stress and endotoxin on human serum albumin in brain-dead organ donors - Uncorrected Proof

2012-01-09

Albumin, among other molecules, binds and detoxifies endotoxin in healthy people. Oxidative stress leads to protein oxidation and thus to the impaired binding properties of albumin. This property, in combination with increased gut permeability, leads to the appearance of endotoxin in the systemic circulation and to impaired organ function. We hypothesize that these processes occur in the serum of brain-dead organ donors. Endotoxin was determined with an adapted Limulus amoebocyte lysate assay. The albumin fractions and binding capacity were determined by high-performance liquid chromatography (HPLC). FlowCytomix (eBioscience, San Diego, Calif) was used to determine the cytokine levels. Carbonylated proteins (CPs) and myeloperoxidase (MPO) were measured by an enzyme-linked immunosorbent assay (ELISA). Eighty-four brain-dead organ donors were enrolled and categorized by the duration of intensive care unit (ICU) stay. The albumin-binding capacity for dansylsarcosine was reduced in brain-dead patients compared with controls. Endotoxin positivity in 16.7% of donors was associated with decreased binding capacity in donors and worse survival of recipients. The CP and MPO levels of organ donors were significantly higher than in healthy controls. The durations of ICU stay increased albumin oxidation. In addition, interleukin-6 (IL-6), IL-8, IL-10, and IL-1β levels were increased in patients, whereas the interferon-γ (IFN-γ) levels were within the normal range. We conclude that oxidative stress and systemic endotoxemia are present in brain-dead organ donors, which might affect recipient survival. High endotoxin levels might be caused by increased gut permeability and decreased binding capacity of albumin influenced not just by higher albumin oxidation.

Silibinin modulates lipid homeostasis and inhibits nuclear factor kappa B activation in experimental nonalcoholic steatohepatitis - Uncorrected Proof

2012-01-04

Nonalcoholic steatohepatitis (NASH) is associated with increased liver-related mortality. Disturbances in hepatic lipid homeostasis trigger oxidative stress and inflammation (ie, lipotoxicity), leading to the progression of NASH. This study aimed at identifying whether silibinin may influence the molecular events of lipotoxicity in a mouse model of NASH. Eight-week-old db/db mice were fed a methionine-choline deficient (MCD) diet for 4 weeks and treated daily with silibinin (20 mg/kg intraperitoneally) or vehicle. Liver expression and enzyme activity of stearoyl-CoA desaturase-1 and acyl-CoA oxidase, and expression of liver fatty acid-binding protein were assessed. Hepatic levels of reactive oxygen species, thiobarbituric acid-reactive substances (TBARS), 3-nitrotyrosine (3-NT), inducible nitric oxide synthase (iNOS), and nuclear factor kappa B (NFkB) activities were also determined. Silibinin administration decreased serum alanine aminotransferase and improved liver steatosis, hepatocyte ballooning, and lobular inflammation in db/db mice fed an MCD diet. Gene expression and activity of stearoyl-CoA desaturase-1 were reduced in db/db mice fed an MCD diet compared with lean controls and were increased by silibinin; moreover, silibinin treatment induced the expression and activity of acyl-CoA oxidase and the expression of liver fatty acid-binding protein. Vehicle-treated animals displayed increased hepatic levels of reactive oxygen species and TBARS, 3-NT staining, and iNOS expression; silibinin treatment markedly decreased reactive oxygen species and TBARS and restored 3-NT and iNOS to the levels of control mice. db/db mice fed an MCD diet consistently had increased NFkB p65 and p50 binding activity; silibinin administration significantly decreased the activity of both subunits. Silibinin treatment counteracts the progression of liver injury by modulating lipid homeostasis and suppressing oxidative stress–mediated lipotoxicity and NFkB activation in experimental NASH.

Accelerated senescence of renal tubular epithelial cells is associated with disease progression of patients with immunoglobulin A (IgA) nephropathy - Uncorrected Proof

2011-12-28

The aim of this study is to determine the potential correlation between the accelerated senescence of renal tubular epithelial cells (RTECs) and the disease progression of patients with immunoglobulin A nephropathy (IgAN). A total of 108 IgAN patients with different Lee's pathologic grades were enrolled. Additionally, 18 patients with renal resection were recruited as controls. Cellular senescence was evaluated by senescence-associated β-galactosidase (SA-β-gal) staining and an immunohistochemical analysis of p21 and p16 protein expression. The expression of type III collagen (Col III) and fibronectin (FN) in renal interstitium and the levels of serum total and low-density lipoprotein (LDL) cholesterol, serum creatinine concentration (SCr), and 24-h urinary protein excretion were evaluated also. SA-β-gal staining and the expression of p16 and p21 were increased significantly in renal biopsy specimens obtained from grades I–II IgAN patients compared with controls (P < 0.05). The expression of these senescence-associated markers increased gradually with disease progression and correlated with the renal morphologic changes and the expression of Col III and FN in renal interstitium in IgAN patients. A correlation analysis showed that the expressions of p16, p21, and SA-β-gal staining were associated significantly with blood pressure and renal function (P < 0.05), but not with patient age, body mass index (BMI), LDL cholesterol level, or 24-h urinary protein value (P > 0.05). Our results indicated that the RTECs in IgAN patients exhibited features of accelerated senescence, which were unrelated to mechanisms associated with normal aging. Cellular senescence was associated closely with IgAN disease progression, which suggested the accelerated senescence of RTECs may contribute to this progression.

Inflammatory bowel disease and pregnancy: overlapping pathways - Uncorrected Proof

Noha Ahmed Nasef, Lynnette R. Ferguson — 2011-12-26

Several studies have reported on the association between inflammatory bowel disease (IBD) and adverse pregnancy outcomes, such as preterm birth. The exact mechanisms of action are unclear; however, several pathways and processes are involved in both IBD and pregnancy that may help explain this. In this review, we discuss the immune system's T helper cells and human leukocyte antigens, inflammation, its function, and the role of Toll-like receptors (TLRs), NOD-like receptors (NLRs), and prostaglandins in the inflammatory response. For each of these topics, we consider their involvement in IBD and pregnancy, and we speculate as to how they can lead to preterm birth. Finally, we review briefly corticosteroids, biologic therapies, and immunosuppressants for the treatment of IBD, as well as their safety in use during pregnancy, with special focus on preterm birth.

Reply to “Intracardiac T-wave alternans, ischemia, and arrhythmias, in a canine model” - Uncorrected Proof

Michael A. Kwofie, James B. Martins — 2011-12-12

We appreciate the interest in the article on T-wave alternans (TWA) and arrhythmia. The issues of interest that were raised in the initial letter to the editor are addressed in the following paragraphs. As noted in the initial letter, an increase in amplitude of R-waves and T-waves is observed in the hyperacute phase of myocardial ischemia (MI); indeed we noted similar results with R-waves under our experimental conditions as well but in less than 2% of cases. As indicated in Reference 23 in the article of interest, the reduction in electrogram voltage is tightly correlated with blood flow measurements documenting ischemia; this is our routine measurement in all past studies in this model.

Single-nucleotide polymorphisms in inflammatory bowel disease - Uncorrected Proof

Manuela G. Neuman, Radu M. Nanau — 2011-11-24

Strong evidence indicates that inflammatory bowel disease, including Crohn disease and ulcerative colitis, is a result of an inappropriate inflammatory response in which genetic and environmental factors play important roles. This review discusses several single-nucleotide polymorphisms with either susceptibility or protective effects on inflammatory bowel disease.

Genetic origin and interaction of the Filipino β0-thalassemia with Hb E and α-thalassemia in a Thai family - Uncorrected Proof

Supawadee Yamsri, Kanokwan Sanchaisuriya, Goonnapa Fucharoen, Supan Fucharoen — 2011-11-18

We describe hematologic and molecular characteristics of a hitherto undescribed interaction between the Filipino deletional β0-thalassemia with Hb E and α-thalassemia in a Thai family. This study was conducted during the prenatal screening of a pregnant Thai woman and her family members. A prenatal diagnosis was performed at her second pregnancy by amniocentesis. Laboratory investigations identified that the pregnant woman was Hb E heterozygote with α+-thalassemia, whereas her husband was a double heterozygote for the Filipino deletional β0-thalassemia and α+-thalassemia. Their affected son was a patient with a previously undescribed condition of Hb E-β0-thalassemia with α+-thalassemia. Both a combined gap-polymerase chain reaction (PCR) and allele-specific PCR were used successfully in the prenatal diagnosis, which identified an affected fetus with Hb E-β0-thalassemia without α+-thalassemia. Beta globin gene haplotype analysis indicated the same origin of this Filipino β0-thalassemia in Asian populations.

Chronic inflammation and pain in a tumor necrosis factor receptor (TNFR) (p55/p75-/-) dual deficient murine model - Uncorrected Proof

Karin W. Westlund, Liping Zhang, Fei Ma, Helieh S. Oz — 2011-11-10

Many aspects of tissue damage after acute or chronic inflammatory reactions can be attributed directly to the concomitant biosynthesis and release of inducible early proinflammatory cytokine tumor necrosis factor alpha (TNFα). Conversely, systemic inflammation is impacted by the consequences of tissue damage. Dysregulated TNFα contributes to numerous pathophysiologic conditions including inflammatory bowel disease (IBD) and arthritis. Inflammatory stimuli trigger proteolytic cleavage and shedding of extracellular domains of TNFα receptors giving rise to 2 soluble fragments (p55 soluble tumor necrosis factor receptor 1 (sTNFR1) and p75 sTNFR2) that block the additional binding, activity, and synthesis of TNFα. We hypothesized that absence of sTNFR inhibitory feedback control would result in accumulated high levels of TNFα and other inflammatory factors promoting the cardinal signs of chronic inflammation and pain. The current study reports a translational murine model of chronic arthritis precipitated by 2 consecutive inflammatory insults. The “double hit” procedures provoke a chronic inflammatory response and pain-related behaviors in mice that are dually deficient in p55 (TNFR1) and p75 (TNFR2). The inflammation- and pain-related behaviors are transient in similarly treated wild-type (WT) mice. The complete Freund’s adjuvant (CFA) method was used initially to induce knee joint inflammation, tactile mechanical and heat hypersensitivity, and gait disturbance. After these transient effects of the insult were resolved, a recrudescence persisting at least through 23 weeks was promoted by gastrointestinal (GI) insult with dilute intracolonic mustard oil (MO) only in the mutant mice. A serum proteome profiling analysis revealed high levels of serum inflammatory factors TNFα, regulated upon activation normally T-cell expressed and secreted (RANTES), chemokine (C-X-C motif) ligand 9 [CXCL9 (MIG)], chemokine (C-X-C motif) ligand 10 [CXCL10 (IP-10)], and chemokine (C-C motif) ligand 2 [CCL2 (MCP-1)]. These data suggest that impaired signaling of TNFα as a result of the deficit of the 2 protective soluble p55 and p75 sTNFR inhibitory factors plays a pivotal role in the reactivation of the immune response to GI insult that can produce recrudescence of inflammatory injury and a chronic pain state through promotion of high levels of serum inflammatory factors.

Erythromycin ameliorates cigarette-smoke–induced emphysema and inflammation in rats - Uncorrected Proof

Yan Zhou, Xianshu Tan, Wenjuan Kuang, Litao Liu, Lihong Wan — 2011-10-24

The exposure to cigarette smoke (CS) is associated with emphysema. In addition to chronic lung inflammation, emphysema is known mainly for the complex pathogenesis associated with imbalance of proteolytic and antiproteolytic activities, oxidative stress, and apoptosis of lung structural cells. Increasing evidence shows that erythromycin, which is a macrolide antibiotic, ameliorates chronic inflammation via mechanisms independent of its antibacterial activity. We hypothesize that erythromycin protects against CS-induced emphysema and inflammation in rats via its anti-inflammation and antiapoptosis action. Sprague-Dawley (SD) rats were administered lipopolysaccharide (LPS) intratracheally solution twice and exposed to the CS, the control rats were administered saline intratracheally and exposed to ambient air for 3 weeks. Then, all the CS rats were distributed randomly into 3 groups and, respectively, treated orally with saline (LPS + CS + saline), Guilongkechuanning capsule (450 mg/kg) (LPS + CS + GLKCN), or erythromycin (100 mg/kg) (LPS + CS + ERY) 0.5 h before CS exposure for 2 weeks. On day 36, the rats were killed. The cytokines in serum were measured by enzyme-linked immunosorbent assay (ELISA). The middle lobe of the right lung was removed for histology and apoptosis analyses, respectively. Emphysematous lesions and inflammatory cell infiltrations in the CS group were evident by a histologic analysis. Erythromycin protected significantly against the alveolar enlargement levels (P = 0.0017), reduced the pathologic apoptosis (P = 0.0023) related with Bcl-2 (P = 0.0002) and Bax (P = 0.0002), and inhibited the expressions of matrix metalloproteinase (MMP)-9 (P = 0.0019) and TIMP-1 protein (P = 0.04) and the MMP-9/TIMP-1 ratio (P = 0.0002) in the lungs of CS-induced emphysema in rats. The protective effect of erythromycin on CS-induced emphysema and inflammation in rats is associated with a reduction in inflammation, imbalance of MMP-9/TIMP-1, and apoptosis of lung structural cells. However, erythromycin did not recover completely the emphysematous morphologic changes to the levels when compared with control rats. This distinctive pattern implies that erythromycin might have the potential to suppress airway inflammation and maintain the integrity of airway epithelium to some extent.

Inflammatory bowel disease: role of diet, microbiota, life style - Uncorrected Proof

Manuela G. Neuman, Radu M. Nanau — 2011-09-26

Inflammatory bowel disease (IBD) encompassed several chronic inflammatory disorders leading to damage of the gastrointestinal tract (GI). The 2 principal forms of these disorders are ulcerative colitis (UC) and Crohn disease (CD). Bacteria are involved in the etiology of IBD, and the genetic susceptibility, environmental factors, and lifestyle factors can affect the individual's predisposition to IBD. The review discusses the potential role of environmental factors such as diet and microbiota as well as genetics in the etiology of IBD. It is suggested that microbial ecosystem in the human bowel colonizing the gut in many different microhabitats can be influence by diet, leading to formation of metabolic processes that are essential form the bowel metabolism.

Metabolome and inflammasome in inflammatory bowel disease - Uncorrected Proof

Radu M. Nanau, Manuela G. Neuman — 2011-09-16

Inflammatory bowel disease (IBD) encompasses several chronic inflammatory disorders leading to the damage of the gastrointestinal tract. The 2 principal forms of these disorders are ulcerative colitis (UC) and Crohn’s disease (CD). Bacteria are involved in the etiology of IBD. Many microorganisms have been put forward as causative factors in IBD, but the primary etiologic agents are still not known. The underlying genetic, environmental, and lifestyle issues can affect the individual’s predisposition to these diseases. Immune factors identified in IBD are: dysregulation of the innate and adaptive immune system directed against luminal bacteria or their products found in the intestinal lumen and inappropriate immune responses to organisms in the intestine that normally do not elicit a response, possibly because of intrinsic alterations in mucosal barrier function. However, recent advances in basic science research revealed new insights into the role of specific immune cells and their mediators in intestinal inflammation. The inflammatory mediators known as “inflammasome” are a consequence of the metabolic products (metabolom) of cells and commensally or pathogenic bacteria. Elucidation of inflammasome and metabolom has led to the development of biomarkers specific for each disease that are involved into management strategies targeted at altering specific pathogenic mechanisms that have the potential to modify or change the natural course of these disease entities. The review discusses the potential role of biomarkers in monitoring the inflammasome and therefore the severity of intestinal damage. The microbial ecosystem in the human gut in different microhabitats and metabolic niches contribute to the bowel metabolome.In addition, this review will focus on our expanding understanding microbial factors associated with both the initiation and maintenance of IBD. New insights acquired from murine genetic models of inflammatory bowel disease will also be discussed.