Joseph Goldfarb

Food Allergy Herbal Formula-2 silences peanut-induced anaphylaxis for a prolonged posttreatment period via IFN-γ-producing CD8+ T cells

BACKGROUND Food allergy is a serious and sometimes fatal condition for which there is no cure. We previously reported that Food Allergy Herbal Formula (FAHF)-2) protected peanut-allergic mice against anaphylactic reactions as long as 4 weeks posttherapy. This formula is now in clinical trials in the United States. OBJECTIVE We sought to determine whether FAHF-2-mediated protection could be extended long-term and explored the mechanisms underlying its persistent immunomodulatory effects. METHODS Peanut-allergic mice received FAHF-2 daily orally by gavage for 7 weeks, and then received 7 oral peanut challenges at intervals of 4 to 10 weeks over a period of 36 weeks. For mechanistic studies, some mice received CD4(+) or CD8(+) T-cell-depleting antibodies or IFN-gamma-neutralizing antibodies. Anaphylactic symptoms, body temperatures, and plasma histamine levels were recorded after each challenge, and peanut-specific immunoglobulin levels and cytokine profiles of splenocytes, mesenteric lymph node cells, and purified CD4(+) and CD8(+) T cells were determined. RESULTS Food Allergy Herbal Formula-2 treatment protected mice from anaphylaxis for more than 36 weeks after discontinuing treatment. Peanut-specific IgE levels were reduced as much as 50%, whereas IgG(2a) levels were increased as much as 60%, and these effects persisted over time. T(H)2 cytokine production by CD4(+) T cells from FAHF-2-treated mice was reduced as much as 75%, whereas CD8(+) T-cell IFN-gamma production was markedly increased by as much as 85% at the final challenge. Neutralization of INF-gamma and depletion of CD8(+) T cells markedly attenuated FAHF-2 efficacy. CONCLUSIONS Food Allergy Herbal Formula-2 provides long-term protection from anaphylaxis by inducing a beneficial shift in allergen-specific immune responses mediated largely by elevated CD8(+) T-cell IFN-gamma production.

Spiperone differentiates multiple 5-hydroxytryptamine responses in rat hippocampal slices in vitro.

The predominant effect of bath perfusion of 5-HT on CA1 population spikes in rat dorsal hippocampal slices in vitro is a dose dependent decrease in amplitude (Beck and Goldfarb, 1985; Rowan and Anwyl, 1985). This is often preceded by a smaller transient increase. The 5-HT induced decrease has an ECs0 of 3.2 #M and a slope index of 2.1 (Beck and Goldfarb, 1985). The effects of 5-HT agonists and antagonists, described below, suggest that the decrease in amplitude is mediated by a receptor with characteristics similar to the 5-HTaA binding site, whereas the receptor(s) mediating the increase do not correspond to any of the 5-HT~ or 5-HT 2 sites. The experimental methods were as described (Beck and Goldfarb, 1985) except that the perfusion fluid contained 24 mM NaHCO 4 and 2.5 mM CaC12. Monopolar stimulation of CA1 stratum radiatum was used. Antagonists were present in the perfusion fluid throughout the experiment, except as noted in fig. 1 The 5-HT analog 5-carboxyamidotryptamine (5-CONH2-T) has nanomolar affinity for [3H]5HT binding sites in brain membranes (Engel et al., 1983); these have been shown to be both 5-HT1A and 5-HTIB sites (Ebersole et al., in preparation). 5-CONH2-T produced only a decrease in population spike amplitude. The effect was rapidly reversible and concentration dependent (fig. 1A). The ECs0 was 20 _+ 2 nM with a slope index of 2.1 +_ 0.2 (mean _+ S.E.M. of 3 slices). 8-Hydroxy-2-(di-n-propylamine)tetralin (8OH-DPAT), a 5-HT agonist that selectively binds

Serotonin decreases population spike amplitude in hippocampal cells through a pertussis toxin substrate

Activation of the serotonin1A receptor decreases CA1 population spike amplitude and inhibits forskolin-stimulated adenylate cyclase in rat hippocampus. Pretreatment of rats with pertussis toxin blocked both responses. Because the electrophysiological and biochemical responses to serotonin were correlated after pertussis toxin treatment, we conclude that both responses are mediated by a common regulatory protein, presumably Gi.

Systems Pharmacology of Adverse Event Mitigation by Drug Combinations

Drug combinations that mitigate adverse events were identified using the FDA Adverse Event Reporting System, and one combination, exenatide and rosiglitazone, was tested in a rodent model. Two Drugs: Better Than One? Everyone has seen the commercial, where a drug is advertised as the much-awaited treatment for a disease. At the end of the commercial, there is a long list of adverse events (or side effects) that may affect anything from your heart to your eyesight. Surprisingly, the addition of a second drug can mitigate the side effects of the first drug, such that the combination is actually safer for the patient. To search for those mitigating combinations, Zhao and colleagues devised a computational method for scanning the Food and Drug Administration’s Adverse Event Reporting System (FAERS). This database is freely available and contains millions of records of drug-induced adverse events reported by patients, doctors, hospitals, lawyers, and drug companies. Thus, it represents a potentially useful source of beneficial drug combinations. The authors focused on rosiglitazone—a drug that effectively controls blood glucose levels in diabetic patients, but has been associated with myocardial infarction (MI). By searching through FAERS data, Zhao et al. found that when rosiglitazone was prescribed in combination with exenatide—another drug for treating type 2 diabetes—there were significantly fewer reports of MI as an adverse event. A cell biological interaction network was then developed to look for mechanisms by which rosiglitazone + exenatide affected the heart. The rosiglitazone target PPARγ was plugged into this network, and plasminogen activator inhibitor-1 (PAI-1) was obtained as a potential point of convergence between the two drugs. To test this hypothesis, the authors administered drugs to a mouse model of type 2 diabetes. Mice treated with rosiglitazone alone showed a marked increase in PAI-1 levels, whereas mice treated with the drug combination had significantly decreased levels of PAI-1, similar to those found in untreated mice. FAERS is self-reported and thus may contain some inaccurate data. Nevertheless, it could be a useful tool for generating meaningful hypotheses from human data and for then testing in vivo in clinically relevant disease models, as shown by Zhao et al. Animal models can provide information about drug mechanism of action, and prospective clinical trials will confirm that such combinations can indeed be translated to people to prevent adverse events. Drugs are designed for therapy, but medication-related adverse events are common, and risk/benefit analysis is critical for determining clinical use. Rosiglitazone, an efficacious antidiabetic drug, is associated with increased myocardial infarctions (MIs), thus limiting its usage. Because diabetic patients are often prescribed multiple drugs, we searched for usage of a second drug (“drug B”) in the Food and Drug Administration’s Adverse Event Reporting System (FAERS) that could mitigate the risk of rosiglitazone (“drug A”)–associated MI. In FAERS, rosiglitazone usage is associated with increased occurrence of MI, but its combination with exenatide significantly reduces rosiglitazone-associated MI. Clinical data from the Mount Sinai Data Warehouse support the observations from FAERS. Analysis for confounding factors using logistic regression showed that they were not responsible for the observed effect. Using cell biological networks, we predicted that the mitigating effect of exenatide on rosiglitazone-associated MI could occur through clotting regulation. Data we obtained from the db/db mouse model agreed with the network prediction. To determine whether polypharmacology could generally be a basis for adverse event mitigation, we analyzed the FAERS database for other drug combinations wherein drug B reduced serious adverse events reported with drug A usage such as anaphylactic shock and suicidality. This analysis revealed 19,133 combinations that could be further studied. We conclude that this type of crowdsourced approach of using databases like FAERS can help to identify drugs that could potentially be repurposed for mitigation of serious adverse events.

Local GABAergic modulation of acetylcholine release from the cortex of freely moving rats.

Cortical perfusion with GABA agonists and antagonists modulates the spontaneous release of cortical acetylcholine and GABA in freely moving rats. Twenty‐four hours after implantation of a dialysis fibre, cerebral cortex spontaneously released acetylcholine (3.8 ± 0.2 pmol/10 min) and GABA (6.6 ± 0.4 pmol/10 min) at a stable rate. Local administration of GABA (1 or 5 mm) or the GABAA agonist muscimol (25 or 50 μm) had no effect on the spontaneous release of acetylcholine. However, bicuculline (1–25 μm), a GABAA antagonist, added to the dialysis perfusate, elicited a concentration‐dependent increase of acetylcholine release to approximately double that of control. This effect of bicuculline (25 μm) was completely prevented by coperfusion with muscimol (50 μm). Local administration of the GABAB receptor agonist baclofen (10 or 50 μm) elicited a concentration‐dependent increase in spontaneous acetylcholine release with a maximal increase of about 60%. Intracortical administration of baclofen also decreased the spontaneous release of GABA. The GABAB receptor antagonist CGP 35348 (1 mm), administered alone for 20 min through the dialysis fibre, was without effect on spontaneous acetylcholine release; however, it completely blocked both the baclofen‐induced increase in acetylcholine release and the decrease in GABA release. These results suggest that cortically released GABA exerts a tonic influence on cholinergic activity.

Glycyrrhiza uralensis flavonoids present in anti-asthma formula, ASHMI™, inhibit memory Th2 responses in vitro and in vivo

Allergic asthma is associated with Th2‐mediated inflammation. Several flavonoids were isolated from Glycyrrhiza uralensis, one of the herbs in the anti‐asthma herbal medicine intervention. The aim of this investigation was to determine whether Glycyrrhiza uralensis flavonoids have inhibitory effects on memory Th2 responses in vitro and antigen‐induced Th2 inflammation in vivo. The effects of three Glycyrrhiza uralensis flavonoids on effector memory Th2 cells, D10.G4.1 (D10 cells), were determined by measuring Th2 cytokine production. Isoliquiritigenin, 7, 4′‐dihydroxyflavone (7, 4′‐DHF) and liquiritigenin significantly suppressed IL‐4 and IL‐5 production in a dose‐dependent manner, 7, 4′‐DHF being most potent. It was also evaluated for effects on D10 cell proliferation, GATA‐3 expression and IL‐4 mRNA expression, which were suppressed, with no loss of cell viability. Chronic treatment with 7, 4′‐DHF in a murine model of allergic asthma not only significantly reduced eosinophilic pulmonary inflammation, serum IgE levels, IL‐4 and IL‐13 levels, but also increased IFN‐γ production in lung cell cultures in response to antigen stimulation. Copyright

Pharmacology and immunological actions of a herbal medicine ASHMITM on allergic asthma

Allergic asthma is a chronic and progressive inflammatory disease for which there is no satisfactory treatment. Studies reported tolerability and efficacy of an anti‐asthma herbal medicine intervention (ASHMI) for asthma patients, developed from traditional Chinese medicine. To investigate the pharmacological actions of ASHMI on early‐ and late‐phase airway responses (EAR and LAR), Ovalbumin (OVA)‐sensitized mice received 6 weeks of ASHMI treatment beginning 24 h following the first intratracheal OVA challenge. EAR were determined 30 min following the fourth challenge and LAR 48 h following the last challenge. ASHMI effects on cytokine secretion, murine tracheal ring contraction and human bronchial smooth muscle cell prostaglandin (PG) production were also determined.

The Sophora flavescens flavonoid compound trifolirhizin inhibits acetylcholine induced airway smooth muscle contraction.

Asthma is a serious health problem worldwide, particularly in industrialized countries. Despite a better understanding of the pathophysiology of asthma, there are still considerable gaps in knowledge as well as a need for classes of drugs. ASHMI™ (Anti-asthma Herbal Medicine Intervention) is an aqueous extract of Ganoderma lucidum (Fr.) P. Karst (Ling Zhi), Sophora flavescens Aiton (Ku Shen) and Glycyrrhiza uralensis Fisch. ex DC (Gan Cao). It prevents allergic asthma airway hyper-reactivity in mice and inhibits acetylcholine (ACh) induced airway smooth muscle (ASM) contraction in tracheal rings from allergic asthmatic mice. The purpose of this research was to identify individual herb(s) and their active compound(s) that inhibit ASM contraction. It was found that S. flavescens, but not G. lucidum or G. uralensis aqueous extracts, inhibited ASM contraction in tracheal rings from asthmatic mice. Bioassay-guided isolation and identification of flavonoid fractions/compound(s) via methylene chloride extraction, preparative HPLC fractionation, and LC-MS and NMR spectroscopic analyses showed that trifolirhizin is an active constituent that inhibits acetylcholine mediated ASM contraction or directly relaxes pre-contracted ASM independent of β2-adrenoceptors.

Chronic estrogen effects on 5-hydroxytryptamine-mediated responses in hippocampal pyramidal cells of female rats

Intracellular recording techniques were used to assess the effect of chronic estrogen treatment of ovariectomized (OVX) rats on CA1 pyramidal cell properties and serotonin (5-HT)-mediated responses in the dorsal hippocampus. The magnitude of the 5-HT1A-mediated hyperpolarization and concomitant change in membrane resistance elicited by 15 microM 5-HT was greater in pyramidal cells from OVX rats treated with estrogen (OVX + ES) than in pyramidal cells from OVX rats. Estrogen treatment did not alter the cellular membrane properties or the reduction in AHP amplitude elicited by 15 microM 5-HT. The modulation of 5-HT neurotransmission by estrogen may contribute to variations in mood which are associated with the menstrual cycle.

De Minimis Risk: A Proposal for a New Category of Research Risk

De Minimis Risk: A Proposal for a New Category of Research Risk Rosamond Rhodes a , Jody Azzouni b , Stefan Bernard Baumrin c , Keith Benkov a , Martin J. Blaser d , Barbara Brenner a , Joseph W. Dauben c , William J. Earle c , Lily Frank c , Nada Gligorov a , Joseph Goldfarb a , Kurt Hirschhorn a , Rochelle Hirschhorn d , Ian Holzman a , Debbie Indyk a , Ethylin Wang Jabs a , Douglas P. Lackey c , Daniel A. Moros a , Sean Philpott e , Matthew E. Rhodes f , Lynne D. Richardson a , Henry S. Sacks a , Abraham Schwab g , Rhoda Sperling a , Brett Trusko a & Arnulf Zweig h a Mount Sinai School of Medicine b Tufts University c The Graduate Center, CUNY d New York University Medical School, CUNY e Union Graduate College f Pennsylvania State University g Indiana University, Purdue h University of Oregon (Emeritus)