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Glaucoma is a heterogenous group of optic neuropathies characterized by the degeneration of optic nerve axons and the progressive loss of retinal ganglion cells (RGCs), which could ultimately lead to vision loss. Elevated intraocular pressure (IOP) is a major risk factor in the development of glaucoma, and reducing IOP remains the main therapeutic strategy. Endothelin-1 (ET-1), a potent vasoactive peptide, has been shown to produce neurodegenerative effects in animal models of glaucoma. However, the detailed mechanisms underlying ET-1-mediated neurodegeneration in glaucoma are not completely understood. In the current study, using a Seahorse Mitostress assay, we report that ET-1 treatment for 4 h and 24 h time points causes a significant decline in various parameters of mitochondrial function, including ATP production, maximal respiration, and spare respiratory capacity in cultured RGCs. This compromise in mitochondrial function could trigger activation of mitophagy as a quality control mechanism to restore RGC health. Contrary to our expectation, we observed a decrease in mitophagy following ET-1 treatment for 24 h in cultured RGCs. Using Morrison's model of ocular hypertension in rats, we investigated here, for the first time, changes in mitophagosome formation by analyzing the co-localization of LC-3B and TOM20 in RGCs. We also injected ET-1 (24 h) into transgenic GFP-LC3 mice to analyze the formation of mitophagosomes in vivo. In Morrison's model of ocular hypertension, as well as in ET-1 injected GFP-LC3 mice, we found a decrease in co-localization of LC3 and TOM20, indicating reduced mitophagy. Taken together, these results demonstrate that both ocular hypertension and ET-1 administration in rats and mice lead to reduced mitophagy, thus predisposing RGCs to neurodegeneration.

Large retrospective cohort studies have consistently shown that people who exercise regularly are at a markedly reduced risk of dementias such as Alzheimer's Disease (AD). Animal studies have also found that exercise can prevent cognitive decline, and recent studies have identified possible mechanisms. However, randomized controlled trials of exercise interventions in AD and mild cognitive impairment have not reached a consensus regarding the efficacy of this treatment, hampering clinical adoption of this technique. This review examines these randomized controlled trials to assess potential causes for the variability in the measured outcomes. We posit that great variance in the methods used in these studies may account for some of the differences seen in outcomes. We determined that aerobic exercise led to the most benefits, that many cognitive domains improve with exercise, and that aerobic exercise enhances the ability for independent living. However, cognitive improvements were more pronounced and consistent in patients with mild cognitive impairment than AD, suggesting a narrow window of opportunity for exercise intervention., Competing Interests: Declaration of Competing Interest None, (Copyright © 2024 Elsevier B.V. All rights reserved.)

Neurons in the central nervous system are among the most metabolically active cells in the body, characterized by high oxygen consumption utilizing glucose both aerobically and anaerobically. Neurons have an abundance of mitochondria which generate adequate ATP to keep up with the high metabolic demand. One consequence of the oxidative phosphorylation mechanism of ATP synthesis, is the generation of reactive oxygen species which produces cellular injury as well as damage to mitochondria. Mitochondria respond to injury by fusion which serves to ameliorate the damage through genetic complementation. Mitochondria also undergo fission to meet an increased energy demand. Loss of mitochondria is also compensated by increased biogenesis to generate new mitochondria. Damaged mitochondria are removed by mitophagy, an autophagic process, in which damaged mitochondria are surrounded by a membrane to form an autophagosome which ultimately fuses with the lysosome resulting in degradation of faulty mitochondria. Dysregulation of mitophagy has been reported in several central nervous system disorders, including, Alzheimer's disease and Parkinson's disease. Recent studies point to aberrant mitophagy in ocular neurodegenerative disorders which could be an important contributor to the disease etiology/pathology. This review article highlights some of the recent findings that point to dysregulation of mitophagy and it's underlying mechanisms in ocular neurodegenerative diseases, including, glaucoma, age-related macular degeneration and diabetic retinopathy., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Brooks, Kodati, Stankowska and Krishnamoorthy.)

The pathophysiology of asthma development is heterogenous. Although complex interactions between factors are present in any one individual, early life viral infections have been shown to play a major role through induction of epithelial damage as well as various innate and adaptive immune responses. The cytokine profile that results from epithelial damage leads to Th2 skewing of the adaptive immune response. Most asthma exacerbations in children and up to half of adults are related to viral infections that also induce epithelial damage and an acute predominately Th2 inflammatory response, leading to acute symptoms and chronic airway remodeling., Competing Interests: Disclosure M.H. Grayson is the Editor-in-Chief of the Annals of Allergy, Asthma & Immunology; has served on advisory boards for AbbVie, GSK, and Merck; has stock options in Invirsa, Inc.; serves on the Board of Directors of the American Board of Allergy and Immunology, Asthma and Allergy Foundation of America (AAFA), where he is Chair of their Medical Scientific Council; and is a member of the American Lung Association Scientific Advisory Committee. The other authors have no conflicts to declare., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Despite extensive research and a plethora of therapeutic options, hypertension continues to be a global burden. Understanding of the pathological roles of known and underexplored cellular and molecular pathways in the development and maintenance of hypertension is critical to advance the field. Immune system overactivation and inflammation in the kidneys are proposed alternative mechanisms of hypertension, and resistant hypertension. Consideration of the pathophysiology of hypertension in chronic inflammatory conditions such as autoimmune diseases, in which patients present with autoimmune-mediated kidney inflammation as well as hypertension, may reveal possible contributors and novel therapeutic targets. In this review, we 1) summarize current therapies used to control blood pressure and their known effects on inflammation; 2) provide evidence on the need to target renal inflammation, specifically, and especially when first-line and combinatory treatment efforts fail; and 3) discuss the efficacy of therapies used to treat autoimmune diseases with a hypertension/renal component. We aim to elucidate the potential of targeting renal inflammation in certain subsets of patients resistant to current therapies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Chaudhari, Pham, Brooks, Dinh, Young-Stubbs, Shimoura and Mathis.)

Email represents a useful organizational tool that can facilitate rapid and flexible communication between organizations, managers, and employees regardless of their physical location (e.g., office, home, on vacation). However, despite the potential benefits of email, its usage is a double-edged sword that also has the potential to negatively affect its users. To advance knowledge and inform both researchers and practitioners of such negative outcomes, we integrate the job demands-resources model with spillover theory to investigate email as a potential job demand and explore how it may relate to employees' job tension and work-family conflict. Using an interval-contingent experience sampling methodology with respondents from two separate organizations ( n = 134) providing 704 observations across 6 days of surveys, we hypothesize that, as a job demand, email can have negative consequences on the job that can spill over into the home. Furthermore, we also examine an individual trait (i.e., trait self-regulation) as a potential boundary condition that moderates the extent to which experienced tension from email demands spills over into home life. Finally, theoretical and practical implications are also discussed., (© The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2021.)

Background: Lung cancer is the number one cancer killer worldwide. A major drawback in the lung cancer treatment field is the lack of realistic mouse models that replicate the complexity of human malignancy and immune contexture within the tumor microenvironment. Such models are urgently needed. Mutations of the tumor protein p53 are among the most common alterations in human lung cancers., Methods: Previously, we developed a line of lung cancer mouse model where mutant human TP53-273H is expressed in a lung specific manner in FVB/N background. To investigate whether the human TP53 mutant has a similar oncogenic potential when it is expressed in another strain of mouse, we crossed the FVB/N-SPC-TP53-273H mice to A/J strain and created A/J-SPC-TP53-273H transgenic mice. We then compared lung tumor formation between A/J-SPC-TP53-273H and FVB/N-SPC-TP53-273H., Results: We found the TP53-273H mutant gene has a similar oncogenic potential in lung tumor formation in both mice strains, although A/J strain mice have been found to be a highly susceptible strain in terms of carcinogen-induced lung cancer. Both transgenic lines survived more than 18 months and developed age related lung adenocarcinomas. With micro CT imaging, we found the FVB-SPC-TP53-273H mice survived more than 8 weeks after initial detection of lung cancer, providing a sufficient window for evaluating new anti-cancer agents., Conclusions: Oncogenic potential of the most common genetic mutation, TP53-273H, in human lung cancer is unique when it is expressed in different strains of mice. Our mouse models are useful tools for testing novel immune checkpoint inhibitors or other therapeutic strategies in the treatment of lung cancer.

Preoperative radiation is associated with an increased risk of wound complications. However, the influences of radiation on musculofascial wound healing remains unclear. The purpose of the study was to investigate the short-term effects of preoperative local radiation on the musculofascial healing of laparotomy incisions in a rat model. Eighteen Fischer 344 rats received radiation doses of 0, 10, or 20 Gy to the abdominal wall and underwent laparotomy 4 weeks later. Two weeks after laparotomy, samples of irradiated muscle were harvested for mechanical tests, histological (Hematoxylin & Eosin, and Masson's Trichrome) and immunohistochemical analyses using KI67, CD31, TGF-β, and MYOD1 antibodies. The elastic modulus (EM), maximum strain (MS), and ultimate tensile strength (UTS) in the 20-Gy group were significantly weaker than those in the 0-Gy group. The EM and UTS in the 20-Gy group were significantly lower than those in the 10-Gy group. The UTS and MS in the 10-Gy group were significantly lower than those in the 0-Gy group. The mean number of inflammatory cells per mm 2 in the 20-Gy group was significantly larger than those in the 10- and 0-Gy groups. The mean numbers of CD31-, KI67-, and MYOD1-positive cells, the optical density of TGF-β, and the microvessel density in the 20-Gy group were significantly smaller than those in the 10- and 0-Gy groups. These results indicated that radiation delays musculofascial healing and decreases mechanical strength of the laparotomy incision by creating a chronic inflammatory environment, inhibiting cell proliferation, angiogenesis, granulation maturation, collagen deposition, and muscular regeneration in a dose-dependent manner. The impaired biomechanical, histological and molecular properties may be associated with the higher risk of wound complications in patients who undergo radiotherapy prior to laparotomy.

Using a perfusion decellularization protocol, we developed a decellularized skin/adipose tissue flap (DSAF) comprising extracellular matrix (ECM) and intact vasculature. Our DSAF had a dominant vascular pedicle, microcirculatory vascularity, and a sensory nerve network and retained three-dimensional (3D) nanofibrous structures well. DSAF, which was composed of collagen and laminin with well-preserved growth factors (e.g., vascular endothelial growth factor, basic fibroblast growth factor), was successfully repopulated with human adipose-derived stem cells (hASCs) and human umbilical vein endothelial cells (HUVECs), which integrated with DSAF and formed 3D aggregates and vessel-like structures in vitro. We used microsurgery techniques to re-anastomose the recellularized DSAF into nude rats. In vivo, the engineered flap construct underwent neovascularization and constructive remodeling, which was characterized by the predominant infiltration of M2 macrophages and significant adipose tissue formation at 3months postoperatively. Our results indicate that DSAF co-cultured with hASCs and HUVECs is a promising platform for vascularized soft tissue flap engineering. This platform is not limited by the flap size, as the entire construct can be immediately perfused by the recellularized vascular network following simple re-integration into the host using conventional microsurgical techniques., Statement of Significance: Significant soft tissue loss resulting from traumatic injury or tumor resection often requires surgical reconstruction using autologous soft tissue flaps. However, the limited availability of qualitative autologous flaps as well as the donor site morbidity significantly limits this approach. Engineered soft tissue flap grafts may offer a clinically relevant alternative to the autologous flap tissue. In this study, we engineered vascularized soft tissue free flap by using skin/adipose flap extracellular matrix scaffold (DSAF) in combination with multiple types of human cells. Following vascular reanastomosis in the recipient site, the engineered products successful regenerated large-scale fat tissue in vivo. This approach may provide a translatable platform for composite soft tissue free flap engineering for microsurgical reconstruction., (Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Insufficient neovascularization is associated with high levels of resorption and necrosis in autologous and engineered fat grafts. We tested the hypothesis that incorporating angiogenic growth factor into a scaffold-stem cell construct and implanting this construct around a vascular pedicle improves neovascularization and adipogenesis for engineering soft tissue flaps. Poly(lactic-co-glycolic-acid/polyethylene glycol (PLGA/PEG) microspheres containing vascular endothelial growth factor (VEGF) were impregnated into collagen-chitosan scaffolds seeded with human adipose-derived stem cells (hASCs). This setup was analyzed in vitro and then implanted into isolated chambers around a discrete vascular pedicle in nude rats. Engineered tissue samples within the chambers were harvested and analyzed for differences in vascularization and adipose tissue growth. In vitro testing showed that the collagen-chitosan scaffold provided a supportive environment for hASC integration and proliferation. PLGA/PEG microspheres with slow-release VEGF had no negative effect on cell survival in collagen-chitosan scaffolds. In vivo, the system resulted in a statistically significant increase in neovascularization that in turn led to a significant increase in adipose tissue persistence after 8 weeks versus control constructs. These data indicate that our model-hASCs integrated with a collagen-chitosan scaffold incorporated with VEGF-containing PLGA/PEG microspheres supported by a predominant vascular vessel inside a chamber-provides a promising, clinically translatable platform for engineering vascularized soft tissue flap. The engineered adipose tissue with a vascular pedicle could conceivably be transferred as a vascularized soft tissue pedicle flap or free flap to a recipient site for the repair of soft-tissue defects., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

ACHN-490 is an aminoglycoside with activity against multidrug-resistant pathogens, including those resistant to currently used aminoglycosides. Two randomized, double-blind, placebo-controlled clinical studies investigated the pharmacokinetics (PK), safety, and tolerability of ACHN-490 injection in healthy subjects. Study 1 used a parallel-group design with escalating single (SD) and multiple doses (MD). Study 2 explored a longer duration of the highest dose tolerated in the first study. Subjects were randomly assigned to receive either ACHN-490 injection or a placebo administered by a 10-min intravenous infusion. Study 1 enrolled 39 subjects (30 active and 9 placebo) and consisted of a single dose of 1 mg/kg body weight followed by ascending SD and MD cohorts of 4, 7, 11, and 15 mg/kg for 10, 10, 5, and 3 days, respectively. Study 2 enrolled 8 subjects (6 active and 2 placebo) who received 15 mg/kg for 5 days. Safety was assessed from adverse event (AE) reporting, standard clinical laboratory procedures, and testing for renal, cochlear, and vestibular function. ACHN-490 exhibited linear and dose-proportional PK, with agreement between the studies for PK parameters assessed. The 15-mg/kg dose did not accumulate with repeated dosing over 5 days. Mean steady-state (±standard deviation) area under the concentration-time curve from 0 to 24 h (AUC(0-24)), maximum concentration of drug in serum (C(max)), half-life (t(1/2)), clearance, and volume of distribution at steady state (V(ss)) for the 15-mg/kg, day 5 dose were 239 ± 45 h·mg/liter, 113 ± 17 mg/liter, 3 ± 0.3 h, 1.1 ± 0.1 ml/min/kg, and 0.24 ± 0.04 liters/kg, respectively. AEs were mild to moderate and rapidly resolved. No evidence of nephrotoxicity or ototoxicity was observed.

A Q-switched microchip laser generating 1064 nm wavelength, subnanosecond pulses at a 13.4 kHz repetition rate was used to seed a dual-stage amplifier featuring a 40 microm core Yb-doped photonic-crystal fiber (PCF) as the power amplifier. From this source, we obtained diffraction-limited (M2 = 1.05), approximately 450 ps pulses of energy > 0.7 mJ, peak power in excess of 1.5 MW, and an average power of approximately 9.5 W. By further amplifying the PCF output in a multimode 140 microm core Yb-doped fiber, we generated a peak power in excess of 4.5 MW, the highest obtained in a fiber source to our knowledge.

We report a large-core, Yb-doped photonic crystal fiber amplifier generating diffraction-limited (M2 = 1.04), approximately 0.45 ns, approximately 8 GHz linewidth pulses with energies of 540 microJ, peak power in excess of 1.1 MW, and average power of approximately 7.2 W at a repetition rate of 13.4 kHz. The pulse peak spectral brightness exceeds 10 kW/(sr Hz cm2) and is the highest generated by a fiber source, to our knowledge.

Objective: Sulfasalazine (SSZ) has regulatory approval for treatment of inflammatory bowel disease in children and adults, and for use as a slow acting agent in adult rheumatoid arthritis (RA). This report surveys the literature for experience with SSZ in juvenile RA., Methods: Medline, Excerpta Medica, and Derwent were searched under the terms juvenile, rheumatoid, arthritis, and sulfasalazine., Results: The search found reports of experience in 550 patients, of whom about half had pauciarticular and nearly one-third polyarticular disease. The studies generally reported at least some drug associated benefit in all subtypes. Some identified late onset pauciarticular disease as most responsive, but others reported poly- and pauciarticular response rates about the same. Systemic onset disease responded poorly and showed a substantial incidence of intolerance in the form of serum sickness. Most studies showed useful disease control in spondylitis. Overall, the patterns of toxicity and intolerance were close to those seen in adult SSZ recipients, with the possible exception of the serum sickness-like response., Conclusion: SSZ has demonstrated useful antirheumatic activity and can contribute to the care of selected patients.

Symmetrical bis(quinolylmethoxyphenyl)alkylcarboxylic acids were investigated as inhibitors of leukotriene biosynthesis and 4, 4-bis(4-(2-quinolylmethoxy)phenyl)pentanoic acid sodium salt (47.Na) met our design parameters for a drug candidate (ABT-080). This compound was readily synthesized in three steps from commercially available diphenolic acid. Against intact human neutrophils, 47.Na inhibited ionophore-stimulated LTB(4) formation with an IC(50) = 20 nM. In zymosan-stimulated mouse peritoneal macrophages producing both LTC(4) and PGE(2), 47.Na showed 9000-fold selectivity for inhibition of LTC(4) (IC(50) = 0.16 nM) over PGE(2) (IC(50) = 1500 nM). Preliminary pharmacokinetic evaluation in rat and cynomolgus monkey demonstrated good oral bioavailability and elimination half-lives of 9 and 5 h, respectively. Pharmacological evaluation of leukotriene inhibition with oral dosing was demonstrated in a rat pleural inflammation model (ED(50) = 3 mg/kg) and a rat peritoneal passive anaphylaxis model (LTB(4), ED(50) = 2.5 mg/kg; LTE(4), ED(50) = 1.0 mg/kg). In a model of airway constriction induced by antigen challenge in actively sensitized guinea pigs, 47.Na dosed orally blocked bronchoconstriction with an ED(50) = 0.4 mg/kg, the most potent activity we have observed for any leukotriene inhibitor in this model. The mode of inhibitory action of 47.Na occurs at the stage of 5-lipoxygenase biosynthesis as it blocks both leukotriene pathways leading to LTB(4) and LTC(4) but not PGH(2) biosynthesis. However, 47.Na does not inhibit 5-lipoxygenase catalysis in a broken cell enzyme assay; therefore it is likely that 47.Na acts as a FLAP inhibitor.

A novel series of heteroarylmethoxyphenylalkoxyiminoalkylcarboxylic acids was studied as leukotriene biosynthesis inhibitors. A hypothesis of structure-activity optimization by insertion of an oxime moiety was investigated using REV-5901 as a starting point. A systematic structure-activity optimization showed that the spatial arrangement and stereochemistry of the oxime insertion unit proved to be important for inhibitory activity. The promising lead, S-(E)-11, inhibited LTB(4) biosynthesis in the intact human neutrophil with IC(50) of 8 nM and had superior oral activity in vivo, in a rat pleurisy model (ED(50) = 0.14 mg/kg) and rat anaphylaxis model (ED(50) = 0.13 mg/kg). In a model of lung inflammation, S-(E)-11 blocked LTE(4) biosynthesis (ED(50) of 0.1 mg/kg) and eosinophil influx (ED(50) of 0.2 mg/kg). S-(E)-11 (A-93178) was selected for further preclinical evaluation.

An in vitro glucuronidation assay was used to optimize a series of N-hydroxyurea-containing 5-lipoxygenase inhibitors for metabolic stability. The glucuronidation of these compounds in cynomolgus monkey microsomes followed Michaelis-Menten kinetics allowing calculation of V(max) and K(M). The V(max) values ranged from 0.02 to 7.9 nmol/min/mg microsomal protein, a 400-fold difference, whereas K(M) ranged from 204 to 2500 microM, only a 12-fold difference. In vitro intrinsic clearance values (CL(int) were calculated for 18 compounds tested in the kinetic assay and compared with the in vivo plasma clearance (CL(p)) calculated from intravenous studies done in cynomolgus monkeys. These initial results suggested a relationship between the in vitro CL(int) and in vivo duration as defined by CL(p). A more rapid in vitro assay was developed in a 96-well format using a single concentration of substrate (100 microM) from which a glucuronidation rate was calculated. The results from this assay for 40 compounds correlated with in vivo plasma clearance (r = 0.57). This more efficient assay was used to test more than 100 compounds and develop structure-metabolism relationships based on metabolic stability and improved duration. The culmination of this effort contributed to the discovery of ABT-761, a 5-lipoxygenase inhibitor with in vivo duration in monkey improved 40-fold over thefirst generation inhibitor. Further studies performed in human liver microsomes demonstrated a similar trend that was corroborated by the 8-fold increase in duration after oral dosing in humans observed with ABT-761.

The discovery of second generation N-hydroxyurea 5-lipoxygenase inhibitors was accomplished through the development of a broad structure-activity relationship (SAR) study. This study identified requirements for improving potency and also extending duration by limiting metabolism. Potency could be maintained by the incorporation of heterocyclic templates substituted with selected lipophilic substituents. Duration of inhibition after oral administration was optimized by identification of structural features in the proximity of the N-hydroxyurea which correlated to low in vitro glucuronidation rates. Furthermore, the rate of in vitro glucuronidation was shown to be stereoselective for certain analogs. (R)-N-[3-[5-(4-Fluorophenoxy)-2-furyl]-1-methyl-2-propynyl]-N-hydroxyure a (17c) was identified and selected for clinical development.

Aims: The aim of this study was to establish whether pharmacokinetic differences between two pro-drugs of methylprednisolone (MP) are likely to be of clinical significance., Methods: This study was a single-blind, randomized, crossover design comparing the bioequivalence of MP released from the pro-drugs Promedrol (MP suleptanate) and Solu-Medrol (MP succinate) after a single 250 mg (MP equivalent) intramuscular injection to 20 healthy male volunteers. Bioequivalence was assessed by conventional pharmacokinetic analysis, by measuring pharmacodynamic responses plus a novel approach using pharmacokinetic/pharmacodynamic modeling. The main measure of pharmacodynamic response was whole blood histamine (WBH), a measure of basophil numbers., Results: The MP Cmax was less for MP suleptanate due to a longer absorption halflife of the prodrug from the intramuscular injection site. The bioavailability of MP was equivalent when based on AUC with a MP suleptanate median 108% of the MP succinate value (90% CI: 102-114%). For Cmax the MP suleptanate median was 81% of the MP succinate value (90% CI: 75-88%). The tmax for MP from MP suleptanate was delayed relative to MP succinate. The median difference was 200% (90% non-parametric CI: 141-283%). The area under the WBH effect-time curve (AUEC) and the maximum response (Emax) were found to be equivalent (90% CI: 98-113% and 93-109% respectively). The maximum changes in other white blood cell counts, blood glucose concentration and the parameters of the pharmacodynamic sigmoid Emax model (EC50, Emax and gamma) were also not significantly different between prodrugs., Conclusions: MP suleptanate is an acceptable pharmaceutical alternative to MP succinate. The use of both pharmacokinetic and pharmacodynamic response data together gives greater confidence in the conclusions compared with those based only on conventional pharmacokinetic bioequivalence analysis.

Our primary goal has been to discover leukotriene biosynthesis inhibitors with characteristics that are appropriate for use as clinical agents. The success of the use of zileuton in the treatment of asthma led us to explore further the use of the N-hydroxyurea class of 5-lipoxygenase inhibitors as longer-acting compounds with good lung penetration. A variety of in vitro and in vivo methods were used to evaluate a large number of compounds, from which ABT-761 [(R)-N-(3-(5-(4-fluorophenylmethyl)thien-2-yl)-1-methyl-2-pr opynyl)-N-hydroxyurea] was selected for study. ABT-761 exhibited potent and selective inhibition of leukotriene formation both in vitro and in vivo. More importantly, the compound potently inhibited antigen-induced bronchospasm in guinea pigs when given either prophylactically or therapeutically. In addition, ABT-761 was a potent inhibitor of eosinophil influx into the lungs of Brown Norway rats. These data provide added support for the role of leukotrienes in both bronchospasm and eosinophilic inflammation and characterize ABT-761 as a particularly potent inhibitor of leukotrienes formed in pulmonary tissues. These data combined with the excellent pharmacokinetic characteristics of the compound indicate its potential use in the treatment of leukotriene-dependent human disease.

A series of substituted indolylalkoxyiminoalkylcarboxylates were found to be potent leukotriene biosynthesis inhibitors. The structure-activity relationships were investigated. Representative potent inhibitors identified were the quinolyl 3a (A-86885) and pyridyl 3b (A-86886) congeners with in vitro IC50s of 21 and 9 nM and in vivo leukotriene inhibition in the rat with oral ED50s of 0.9 and 1.7 mg/kg, respectively.

This study examined the effects of low dose systemic corticoid (methylprednisolone, MP), standard dose antihistamine (terfenadine, TF) or the combination on response to out-of-season acute allergen challenge. We feel that a single dose challenge delivered to the nose may represent real disease imperfectly and in this study used two doses given 1 hour apart, hoping to approximate better the circumstances of natural allergen stimulation. The study used clinical endpoints only: measured nasal airway resistance (NAR), sneeze count, and weight of blown nasal secretions. Subjects showed similar NAR, sneezing, and secretion response to both challenges. With placebo treatment, NAR rose after the first allergen provocation and returned to baseline about 30 minutes later. Antihistamine pretreatment appeared to delay but did not prevent this rise; low dose corticoid partially inhibited it, and the combination totally ablated the response. All active treatments suppressed sneezing and secretion better than placebo. Combination corticoid/antihistamine treatment showed no greater effect on sneeze/ secretion than did antihistamine alone; this differs from our findings in separate studies comparing analogous drug combinations in naturally-acquired ragweed hayfever.

Representative nonsteroidal anti-inflammatory drug (NSAID) cyclooxygenase inhibitors such as ibuprofen, naproxen, and indomethacin were used as orally bioavailable scaffolds to design selective 5-lipoxygenase (5-LO) inhibitors. Replacement of the NSAID carboxylic acid group with a N-hydroxyurea group provided congeners with selective 5-LO inhibitory activity.

Representative examples of NSAID cyclooxygenase inhibitors such as naproxen, ibufenac, ibuprofen, and butibufen have been transformed into 5-lipoxygenase inhibitors by replacement of the carboxylic acid moiety with a 4-hydroxythiazole group.

Synthetic routes were developed to access a variety of novel 1-aryl-2H,4H-tetrahydro-1,2,4-triazin-3-one analogs which were evaluated as 5-lipoxygenase (5-LO) inhibitors. The parent structure, 1-phenylperhydro-1,2,4-triazin-3-one (4), was found to be a selective inhibitor of 5-LO in broken cell, intact cell, and human blood assays with IC50 values of 5-21 microM. In a rat anaphylaxis model, 4 blocked leukotriene formation with an ED50 = 7 mg/kg when administered orally. Compound 4 exhibited selectivity for inhibition of 5-LO with little activity against related enzymes: 12-LO from human platelets, 15-LO from soybean, and cyclooxygenase (COX) from sheep seminal vesicle. In pilot subacute toxicity testing, 4 did not produce methemoglobinemia in rats (400 mg/kg po daily for 9 days) or in dogs (200 mg/kg po daily for 28 days). These results indicated that the triazinone structure provided a 5-LO inhibitor template devoid of the toxicity problems observed in the related phenidone (1) and pyridazinone (3) classes of 5-LO inhibitors. The parent compound 4 is a selective, orally bioavailable 5-LO inhibitor which can serve as a useful reference standard for in vivo pharmacological studies involving leukotriene-mediated phenonmena.

RAW 264.7 macrophages respond to lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma) by producing large amounts of nitric oxide (NO) and prostaglandin E2 (PGE2), with maximal production 18-24 h after treatment. Following stimulation with the calcium inophore A23187, cultures of RAW cells also produce modest amounts of leukotrienes. However, the capacity of these cells to produce leukotrienes is transient, beginning 2 h after vehicle or LPS/IFN-gamma treatment, peaking by 4-6 h and absent by 8 h. A-79175, (R(+) N-[3-[5-(4-Fluorophenoxy)-2-furanyl]-1-methyl-2-propynyl]-N-hydroxyurea) a specific inhibitor of 5-lipoxygenase (5-LO), abolished leukotriene production by RAW cells in a dose-dependent, non-cytotoxic fashion while having no effect on PGE2 or NO production. By contrast, nordihydroguaiaretic acid (NDGA) inhibited production of leukotrienes, PGE2 and NO only at doses that were cytotoxic to the RAW cells. Exogenous leukotriene B4 (LTB4) had no effect on either NO or PGE2 production. An inhibitor of NO production, L-N-5-(1-iminoethyl) ornithine HCl (NIO) also did not affect leukotriene or PGE2 production, while dexamethasone blocked PGE2 and NO production, but did not affect leukotriene production in these cells. Taken collectively, these results indicate that there is no interaction between the pathways for leukotriene and nitric oxide production in RAW 264.7 macrophages.

Structure-activity optimization of inhibitory potency and duration of action of N-hydroxyurea containing 5-lipoxygenase inhibitors was conducted. The lipophilic heteroaryl template and the link group connecting the template to the N-hydroxyurea pharmacophore were modified. Inhibition of 5-lipoxygenase was evaluated in vitro in a human whole blood assay. An in vitro assay using liver microsomes from monkey was used to evaluate congeners for comparative rates of glucuronidation. (3-Heteroaryl-1-methyl-2-propynyl)-N-hydroxyureas were found to be more resistant to in vitro glucuronidation. The promising inhibitor N-[3-[5-(4-fluorophenoxy)-2-furyl]-1-methyl-2-propynyl]-N- hydroxyurea (6) was found to have stereoselective glucuronidation in monkey and man. The R enantiomer 7 provided longer duration of inhibition as evaluated by an ex vivo whole blood assay. Further optimization of the lipophilic template led to the discovery of (R)-(+)-N-[3-[5-[(4-fluorophenyl)methyl]-2- thienyl]-1-methyl-2-propynyl]-N-hydroxyurea (11) with more effective and prolonged inhibition of leukotriene biosynthesis than zileuton (1) and 7 in monkey and man. The optimized 5-lipoxygenase inhibitor 11 was selected for development as an investigational drug for leukotriene-mediated disorders.

Acute nasal allergen challenge produces airway obstruction which varies in amount and timing with the allergen dose delivered. To see whether different mechanisms might contribute variably to mucosal swelling with different amounts of allergen, we challenged sensitive volunteers with threshold and 10-times threshold allergen doses, with and without topical vasoconstrictor pre-treatment. The vasoconstrictor effectively eliminated obstruction at both allergen dose levels, suggesting that acute vascular changes were responsible for all the measurable obstruction seen with acute allergen provocation. Alpha-adrenergic vasoconstrictor pre-treatment was associated with increased weight of secretion and numbers of sneezes.

The authors compared the effect of several doses an oral corticosteroid on symptom profile and severity in ragweed hay fever. Thirty-one patients were randomized to receive 0, 6, 12, or 24 mg methylprednisolone (Medrol Tablets [MP], Upjohn, Kalamazoo, MI). A baseline week in which no treatment was given preceded the treatment comparison. At the end of this week, symptom diaries showed that most of the subjects were experiencing moderate or severe symptoms. The corticoid produced dose-related reduction in all symptoms. The difference between placebo and 24 mg MP was significant for all the symptoms monitored, except itching, which benefited marginally. With 6 mg MP, congestion, drainage, and eye symptoms showed significant drug-placebo differences but itching, running/blowing, and sneezing did not. Not all rhinitis symptoms responded equally to corticoid treatment. Those that responded least could reflect histamine effect, which was not effectively suppressed by low-dose, short-term corticoid treatment.

We examined unilaterality of obstruction after acute bilateral nasal allergen provocation in two groups of pollen-sensitive volunteers studied out of season. One group was challenged on one occasion with a threshold allergen dose and on another with placebo. We measured nasal airway resistance (NAR) unilaterally for 3.5 hr before the challenges and for 40 min after. Most subjects' noses had marked asymmetry of response. Over half showed marked obstruction on one side and none at all on the other side. The side which showed higher resistance and greater lability before challenge was typically more obstructed after. In a second group we compared responses to threshold and x 10 threshold doses. Threshold challenge produced results similar to those seen with the first group. After the higher allergen dose, there was some obstruction in the less responsive side and the rate of rise was much slower. Obstructive response after acute threshold allergen challenge is typically one-sided. This pattern may be related to the stage of the nasal cycle in which the challenge was delivered. Higher allergen doses produce more obstruction in the less responsive side but the response is still asymmetrical.

Data from 58 premarketing studies of the nonsteroidal antiinflammatory drug flurbiprofen were pooled for analyses of adverse drug reactions (ADRs). These studies included 5602 patients treated with flurbiprofen (N = 4123), aspirin (N = 1033), or placebo (N = 446) for varying durations. Diagnoses included rheumatoid arthritis, osteoarthritis, and other painful musculoskeletal conditions. In these studies serious upper gastrointestinal ADRs occurred in flurbiprofen-treated patients at less than one half the rate seen in aspirin-treated patients. The incidence of serious urinary tract ADRs was lower with flurbiprofen than with aspirin. The flurbiprofen group had no serious clinical ADRs related to the hemic/lymphatic system. The most common laboratory abnormality was a decrease in hematocrit, which occurred less often than in the aspirin group. We also evaluated serious flurbiprofen-related ADRs in 4370 patients in a variety of other studies and reviewed published reports of flurbiprofen clinical trials and case reports. These reviews showed no additional, unanticipated patterns of intolerance. These clinical safety data indicate that in the doses studied, flurbiprofen is a well tolerated agent for patients requiring nonsteroidal antiinflammatory drug therapy.

Forty-two ragweed hay fever patients participated in a study which examined the profile of symptom relief provided by terfenadine, and the relative adequacy of symptom control with the drug given from the beginning of the season compared with treatment started after symptoms were well established. Compared with placebo, terfenadine effectively relieved sneeze, itch and eye symptoms. It had no effect on running, blowing and drainage. Subjectively perceived congestion benefited marginally. When the drug was begun after symptoms were well established, sneezing responded quickly and maximally. Eye discomfort lessened but not to the level experienced by those dosed from the beginning of the season. The pattern in other symptom categories was less clear. Overall, terfenadine improved all rhinitis symptoms except those related to hypersecretion. Some symptoms appeared to respond better when drug dosing commenced at the beginning of the season.

Allergic rhinitis patients were challenged with intranasal allergen aerosols after pretreatment with hydroxyzine and phenylpropanolamine, singly and in combination. Hydroxyzine protected against itching, sneezing and hypersecretion but aggravated obstruction. Phenylpropanolamine had a subtle but measurable anti-congestive effect. This model quantitates and confirms the complementary effects of combined antihistamine-decongestant therapy.

This study evaluated the relative contributions of vascular engorgement and less reversible components of inflammation (edema, cellular infiltration) to production of allergic nasal congestion. We established the minimum nasal airway resistance (NAR) inducible by oxymetazoline spray in 29 asymptomatic patients. During ragweed season, we restudied them with the same decongestant regimen before and after 1 week of treatment with oral methylprednisolone (Medrol tablets, Upjohn) or a matched placebo. It was not possible, in the conditions of this study, to decongest symptomatic patients to baseline using only the topical vasoconstrictor. This could be accomplished in those receiving methylprednisolone. Corticoid therapy relieved a non-vascular component of congestion or rendered vasoconstrictor therapy more effective.

Though cromolyn sodium exhibits tachyphylaxis in laboratory models of allergy, this pattern has not been identified in clinical use or in human asthma provocation models. We looked for it using a treadmill exercise challenge scheme that had previously confirmed cromolyn's protective effect. Twelve exercise-sensitive asthmatic volunteers received in random order, 12 20-mg cromolyn inhalations, 12 placebo inhalations, and 11 placebo inhalations with a final 20-mg dose of cromolyn. Each regimen was given over a three-day period, with the final dose administered 30 minutes before exercise. Subjects' respiratory functions were measured before and repeatedly after exercise with body plethysmography and peak flow rates. Beginning six minutes after the end of exercise, parameters measured after single-dose cromolyn and placebo administration differed in all endpoints at most of the measurement intervals. Results with 12-dose cromolyn pretreatment were significantly different from those seen with placebo only at 40 minutes after exercise, whereas single- and multiple-dose cromolyn results differed at 20 and 30 minutes. Cromolyn accelerated recovery from exercise-induced falls in lung function; this effect was attenuated with multiple dosing. These results are consistent with development of tachyphylaxis.

We dosed eight normal volunteers with single doses of probenecid alone and with aspirin or ibuprofen. Urinary urate clearance was increased by probenecid. This increase was partially inhibited by aspirin but not by ibuprofen.

To find whether patients with chronic rhinitis might be congested because of hyporesponsiveness to adrenergic vasoconstrictive influences, we measured nasal airway resistance (NAR) in normals and allergic and non-allergic rhinitics during intravenous infusion of graded doses of phenylephrine. All responded with decreases in NAR and first evidences of NAR fall appeared no later in those with rhinitis than in normals. Nasal congestion and response were asymmetrical; absolute NAR in the low resistance side was similar in all groups and there was little response to phenylephrine. In the high resistance side, NAR reached its minimum by the time the total infused dose was 1400 mcg, indicating maximum response to drug was achieved within the dose range studied. Minimum NAR achieved on the high resistance side was higher in rhinitics suggesting residual vascular engorgement resistant to phenylephrine or non-vascular mucosal swelling. Resistance to adrenergic vasoconstriction does not appear to be the primary contributor to mucosal swelling in chronic rhinitis.

To test whether prostaglandins contribute to the nasal allergic reaction, we subjected allergic rhinitis patients to nasal allergen challenge after treatment with flurbiprofen, a cyclooxygenase inhibitor. We challenged 18 patients after two pretreatment doses of flurbiprofen, 75 mg, chlorpheniramine, 6 mg, both drugs combined, and placebo. All patients received all treatments. None of the treatments affected measured nasal airway resistance. Nasal secretion weight, number of sneezes, and overall subjective severity scores for all active treatments differed significantly from placebo treatment. Flurbiprofen proved nearly as effective as chlorpheniramine in reducing severity of induced rhinitis. Some mechanism that can be influenced by flurbiprofen (possibly prostaglandin synthesis) contributes to the acute, induced allergic reaction.

Twenty-eight ragweed-allergic patients with hay fever participated in a clinical trial that examined the pattern of symptom relief resulting from addition of a cyclooxygenase-inhibiting drug to standard antihistamine therapy. In the first week antihistamine use by the subjects was standardized, and subjects were oriented to use of the hay fever symptom diary. They were then allocated, according to symptom severity, to two treatment groups. One group received 120 mg of terfenadine and 300 mg of flurbiprofen (a cyclooxygenase-inhibiting drug) per day. Members of the other group received terfenadine and a placebo that looked like flurbiprofen. This treatment lasted 1 week. Subjects recorded severity of congestion, drainage, running, nose blowing, itching, and sneezing four times each day. The flurbiprofen-treated patients experienced less severe symptoms, demonstrating maximum benefit 3 to 5 days into the drug trial with some loss of effect thereafter. Secretion-related symptoms appeared to benefit most. Combined blockade of histamine and cyclooxygenase products appears to offer improved symptom control in ragweed hay fever.

Sixty-two volunteer hay fever patients participated in a 2-week trial which examined the protective effect of the cyclooxygenase-inhibiting drug flurbiprofen. The drug suppressed symptom severity significantly, though it was not as protective as the antihistamine also employed in the trial. Flurbiprofen's effects suggests that prostaglandin release may contribute to the symptoms of allergic rhinitis.

Eye evaluations before and after 24 weeks of treatment with either ibuprofen (Motrin) or aspirin did not show eye toxicity with either drug in a double-blind study of 78 patients with osteoarthritis. The possibility of eye complications resulting from ibuprofen therapy suggested in two earlier reports has not been confirmed in clinical usage in other countries or in clinical trials in the United States.

Ibuprofen is a nonsteroidal drug with analgesic, antipyretic, and anti-inflammatory properties that was recently introduced for use in antiarthritis therapy in the United States. In a year-long double-blind multiclinic trial in 885 patients with rheumatoid arthritis, ibuprofen was at least as satisfactory as aspirin, considering both efficacy and tolerance. In the majority of patients, daily doses ranged from 800 to 1,600 mg of ibuprofen and 3 to 6 gm of aspirin. The drugs did not differ greatly in providing relief from arthritis symptoms, but ibuprofen was definitely better tolerated, especially in regard to gastrointestinal complaints. Seven percent of the ibuprofen group dropped out of the study because of adverse reactions, as compared with 16% of the aspirin group; 17% of the ibuprofen group and 31% of the aspirin group had gastrointestinal symptoms.

Both radiolabeled and nonlabeled drug have been used to study the pharmacokinetics of flurbiprofen (Ansaid, Upjohn). Drug absorption is rapid, drug disappearance half-life is independent of oral dose, and the area under the plasma drug concentration versus time curve increases with increasing oral dose. Elimination of intact drug from the peripheral circulation is biphasic and rapid. Following a single oral dose of 100 mg of flurbiprofen, drug bioavailability is equivalent using regimens of four 25-mg tablets, two 50-mg tablets, or one 100-mg tablet once daily. Long-term administration of flurbiprofen appears neither to inhibit nor induce the drug's metabolism.