Anti-Inflammatory Influences on Behavioral, But Not Cortisol, Responses During Maternal Separation

The present study examined the effect of anti-inflammatory agents on the behavioral and plasma cortisol responses of guinea pig pups during isolation in a novel environment. In Experiment 1, intracerebroventricular (ICV) administration of interleukin-10 (IL-10), across a range of doses, significantly reduced passive behaviors of isolated pups. In Experiment 2, neither ICV IL-10 nor alpha-melanocyte stimulating hormone affected the plasma cortisol response to the separation procedure. These results provide further evidence that the passive behaviors of isolated guinea pig pups are mediated by proinflammatory cytokines and indicate that cytokines do not affect behavior indirectly by means of an action on hypothalamic-pituitary-adrenal activity

Anti-Inflammatory Influences on Behavioral, But Not Cortisol, Responses During Maternal Separation

The present study examined the effect of anti-inflammatory agents on the behavioral and plasma cortisol responses of guinea pig pups during isolation in a novel environment. In Experiment 1, intracerebroventricular (ICV) administration of interleukin-10 (IL-10), across a range of doses, significantly reduced passive behaviors of isolated pups. In Experiment 2, neither ICV IL-10 nor alpha-melanocyte stimulating hormone affected the plasma cortisol response to the separation procedure. These results provide further evidence that the passive behaviors of isolated guinea pig pups are mediated by proinflammatory cytokines and indicate that cytokines do not affect behavior indirectly by means of an action on hypothalamic-pituitary-adrenal activity

Anti-Inflammatory Influences on Behavioral, But Not Cortisol, Responses During Maternal Separation

The present study examined the effect of anti-inflammatory agents on the behavioral and plasma cortisol responses of guinea pig pups during isolation in a novel environment. In Experiment 1, intracerebroventricular (ICV) administration of interleukin-10 (IL-10), across a range of doses, significantly reduced passive behaviors of isolated pups. In Experiment 2, neither ICV IL-10 nor alpha-melanocyte stimulating hormone affected the plasma cortisol response to the separation procedure. These results provide further evidence that the passive behaviors of isolated guinea pig pups are mediated by proinflammatory cytokines and indicate that cytokines do not affect behavior indirectly by means of an action on hypothalamic-pituitary-adrenal activity

The role of matrix metalloproteinases in ulcerative keratolysis associated with perioperative diclofenac use

To investigate the role of matrix metalloproteinases (MMPs) in the pathogenesis of ulcerative keratolysis associated with topical use of generic diclofenac preoperatively and postoperatively. To characterize the inflammatory response of the cornea in this case of ulcerative keratolysis. Case report with clinicopathologic correlation. Corneal culture for microbial growth. Clinical and histopathologic examinations including routine histolopathologic, immunofluorescent, and immunohistochemical studies. Microscopic examination of the corneal button disclosed fibrinous material with neutrophils and mononuclear inflammatory cells. The corneal epithelial basement membrane was irregularly thickened and patchy. Immunohistochemical staining detected weak staining of MMP-1 and a strong presence of MMP-8 in the epithelium. MMP-8 and 9 were also present in areas of leukocytic infiltration. MMP-2 appeared in a few stromal cells. Macrophages and leukocytes were the predominant infiltrating cells. A nonspecific inflammatory response occurred in this case of ulcerative keratolysis. Corneal epithelial cells are capable of secreting MMP-1 and 8 and may participate in the stromal degradation and repair process of the ulcerative keratolysis associated with topical nonsteroidol antiinflammatory use

Discovery of Novel Trace Amine-Associated Receptor 5 (TAAR5) Antagonists Using a Deep Convolutional Neural Network

Trace amine-associated receptor 5 (TAAR5) is a G protein-coupled receptor that belongs to the TAARs family (TAAR1-TAAR9). TAAR5 is expressed in the olfactory epithelium and is responsible for sensing 3-methylamine (TMA). However, recent studies showed that TAAR5 is also expressed in the limbic brain regions and is involved in the regulation of emotional behaviour and adult neurogenesis, suggesting that TAAR5 antagonism may represent a novel therapeutic strategy for anxiety and depression. We used the AtomNet® model, the first deep learning neural network for structure-based drug discovery, to identify putative TAAR5 ligands and tested them in an in vitro BRET assay. We found two mTAAR5 antagonists with low to submicromolar activity that are able to inhibit the cAMP production induced by TMA. Moreover, these two compounds also inhibited the mTAAR5 downstream signalling, such as the phosphorylation of CREB and ERK. These two hits exhibit drug-like properties and could be used to further develop more potent TAAR5 ligands with putative anxiolytic and antidepressant activity

Effects of topical nonsteroidal antiinflammatory drugs on the expression of matrix metalloproteinases in the cornea

To assess the effects of nonsteroidal antiinflammatory drug (NSAID) eyedrops on the expression of matrix metalloproteinases in corneal tissue. Ocular Microbiology and Immunology Laboratory, Refractive Surgery Research Laboratory, The Wilmer Eye Institute, Johns Hopkins University, Baltimore, Maryland, USA. Seventy rats were divided into 2 groups: intact and debrided epithelium. Uniform central corneal epithelial defects were created in the right eye of the debrided corneal group. Each group was divided into 4 subgroups, each receiving 1 of the following eyedrops or artificial tears: The 3 NSAIDs were diclofenac sodium 0.1% (Falcon® or Voltaren®) and preservative-free ketorolac 0.5% (Acular® PF). The artificial tears were carboxymethylcellulose sodium 0.5% (Refresh Plus® PF). The eyedrops were administered 4 times a day for 1 week. The rats were killed on days 2 and 7. The corneas were excised and processed for immunohistochemical staining, Western blot assay, and zymography studies to determine the localization of the production of the following matrix metalloproteinases (MMPs): MMP-1, MMP-2, MMP-8, and MMP-9. Matrix metalloproteinase-1, MMP-8, and MMP-2 were detected in rat corneas at 48 hours in the debrided and intact epithelium groups treated with NSAID eyedrops. The MMP-1 and MMP-8 expression levels were higher in intact corneas in the diclofenac sodium groups than in the ketorolac and artificial tears groups. The expression was localized mostly in the epithelial cells and occasionally in keratocytes. This study provides preliminary evidence that topical application of some NSAIDs can induce the early expression of MMP-1, MMP-2, and MMP-8 in the cornea, suggesting that MMPs play a role in the corneal cytotoxicity of certain NSAIDs

Predicting Productive Binding Modes for Substrates and Carbocation Intermediates in Terpene SynthasesBornyl Diphosphate Synthase As a Representative Case

Terpene synthases comprise a family of enzymes that convert acyclic oligo-isoprenyl diphosphates to terpene natural products with complex, polycyclic carbon backbones via the generation and protection of carbocation intermediates. To accommodate this chemistry, terpene synthase active sites generally are lined with alkyl and aromatic, i.e., nonpolar, side chains. Predicting the correct, mechanistically relevant binding modes for entire terpene synthase reaction pathways remains an unsolved challenge. Here, we describe a method for identifying such modes: <i>TerDockin</i>, a series of protocols to predict the orientation of carbon skeletons of substrates and derived carbocations relative to the bound diphosphate group in terpene synthase active sites. Using this recipe for bornyl diphosphate synthase, we have predicted binding modes that are consistent with all current experimental observations, including the results of isotope labeling experiments and known stereoselectivity. In addition, the predicted binding modes recapitulate key findings of a seminal study involving more computationally demanding QM/MM molecular dynamics methods on part of this pathway. This work illustrates the value of the <i>TerDockin</i> approach as a starting point for more involved calculations and sets the stage for the rational engineering of this family of enzymes

Multicenter, Randomized, Double-Blind, Placebo-Controlled Study on the Effect of Oral Tolvaptan on Left Ventricular Dilation and Function in Patients With Heart Failure and Systolic Dysfunction

Multicenter, Randomized, Double-Blind, Placebo-Controlled Study on the Effect of Oral Tolvaptan on Left Ventricular Dilation and Function in Patients With Heart Failure and Systolic Dysfunction James E. Udelson, Frank A. McGrew, Enrique Flores, Hassan Ibrahim, Stewart Katz, Gregory Koshkarian, Terrence O’Brien, Marvin W. Kronenberg, Christopher Zimmer, Cesare Orlandi, Marvin A. Konstam In a multicenter, randomized, double-blind, placebo-controlled trial in patients with heart failure and reduced left ventricular systolic function, 1 year of administration of tolvaptan 30 mg/day was studied to evaluate the effect on reducing left ventricular (LV) end-diastolic volume compared with placebo by using quantitative radionuclide ventriculography. A total of 240 patients were randomized. In a well-treated population of stable HF patients, LV volumes were stable during 1 year of follow-up. There was no significant effect of tolvaptan therapy on LV volumes observed during 1 year of therapy. Nonprespecified natural history data favored therapy with tolvaptan, with a reduction in the combined end point of mortality and heart failure hospitalization observed over the course of the trial. This study sought to examine the effects of vasopressin V2receptor antagonism with tolvaptan on the changes in left ventricular (LV) volumes over time. Vasopressin levels may be increased in patients with heart failure (HF) and may be a factor driving the progression of HF. This was a multicenter, randomized, double-blind, placebo-controlled trial conducted to evaluate the effect of long-term administration of the vasopressin V2-receptor antagonist tolvaptan (30 mg/day) on reducing left ventricular end-diastolic volume (LVEDV) compared with placebo in patients with HF and reduced systolic function, using quantitative radionuclide ventriculography at baseline, repeated after 1 year of therapy, and repeated again approximately 1 week after withdrawal of study drug. A total of 120 patients were randomized to tolvaptan and 120 were randomized to placebo. In the placebo group, there was no change in LVEDV over the course of follow-up (change of 0.0 ± 10.0 ml/m2). After 1 year of tolvaptan, there was a small reduction in LV volume (decrease of 1.8 ± 10.7 ml/m2); the between-group difference was not significant (p = 0.21). During the course of the trial, there were 6 deaths (5%) and 21 HF hospitalizations (18%) in the tolvaptan group, compared with 11 deaths (9%) and 34 HF hospitalizations (28%) in the placebo group. In a time-to-event analysis, there was a significant favorable effect of tolvaptan on the composite of mortality or heart failure hospitalization (p < 0.03 by log-rank test). In a well-treated population of stable HF patients, there was no significant effect of tolvaptan therapy on LV volumes observed during 1 year of therapy. Nonprespecified natural history data favored therapy with tolvaptan, with a reduction in the combined end point of mortality and heart failure hospitalization observed. (Multicenter, Randomized, Double-Blind, Placebo Controlled, Efficacy Study on the Effects of Tolvaptan on Left Ventricular Dilatation in Congestive Heart Failure Patients; http://clinicaltrials.gov/ct/show/NCT00043758?order=1; NCT00043758

A Multicenter, Randomized, Double-blind, Placebo-controlled Study of Tolvaptan Monotherapy Compared to Furosemide and the Combination of Tolvaptan and Furosemide in Patients With Heart Failure and Systolic Dysfunction

Increased vasopressin levels may be present in patient with chronic heart failure (HF) and contribute to pathophysiology through effects on the vasopressin V2 receptor. The presence of background diuretic therapy may confound evaluations of vasopressin receptor antagonists (VRA). Eligible patients had HF (New York Heart Association Class II-III), systolic dysfunction (left ventricular ejection fraction ≤0.40) and signs of congestion (eg, edema, rales). At screening, patients were removed from baseline diuretic therapy and placed on a low-sodium diet (2 g/day). After a 2-day run-in period, 83 patients were randomized to placebo (n = 21), monotherapy with the vasopressin V2 receptor antagonist tolvaptan (TLV) 30 mg (n = 20), monotherapy with furosemide 80 mg (FURO, n = 22) or both TLV 30 mg and FURO 80 mg (n = 20) once daily for 7 days. Patients were on standard background therapy and not fluid-restricted throughout the study. A decrease in body weight of −1.37 ± 1.61, −0.54 ± 1.59, and −1.13 ± 1.49 kg was observed versus baseline for TLV, FURO, and TLV+FURO, respectively, at day 8. At the same point, the placebo group showed a body weight increase of +0.72 ± 2.42 kg versus baseline ( P = .0006 for TLV versus placebo). Increases in urine volume from baseline were greater with TLV alone (2646 ± 1503 mL/24 hours) than with FURO (894 ± 853 mL/24 hours, P < .001), or PLC (423 ± 786 mL/24 hours, P < .001), and similar to TLV+FURO (2585 ± 2119 mL/24 hours). An increase in serum sodium within the normal range was also observed in TLV-treated patients ( P < .02 versus placebo; P < .01 versus FURO). No changes in serum potassium, other laboratory values, or blood pressure were observed. TLV therapy was well tolerated. In patients with HF and signs of volume overload, TLV monotherapy without concomitant loop diuretic therapy reduced body weight when compared to placebo without adverse changes in serum electrolytes, during a sodium restricted diet while on background medications including angiotensin-converting enzyme inhibitors and β-blockers