The proinflammatory cytokine human interleukin-6 (hIL-6) plays an important role in the early and late courses of inflammation, trauma, and wound healing caused by sulfur mustard (HD). Previously, we demonstrated that hIL-6 might be involved in the early event of structural changes of the signal transducer glycoprotein, which indirectly initiates the cascade of events, such as skin irritation and blister formation observed in the pathophysiology of HD injury. In this present work, we focus on the neutralization effect of IL-6 antibodies with regard to the modulation of hIL-6 secretion. Levels of secreted cytokine hIL-6 in normal human epidermal keratinocytes (NHEK) stimulated with HD (10−4M) and incubated for 24h at 37°C were determined by enzyme immunoassay, protein immunocytologic assay and reverse-transcriptase-polymerase chain reaction (RT-PCR). The ratio of HD-treated NHEK to constitutive non-stimulated NHEK controls (S/C) on the induction of hIL-6 is reported. S/C was four-fold higher than non-stimulated NHEK controls as determined by ELISA. By using a more sensitive immunocytologic assay, Luminex100™, the increment was verified. hIL-6 levels in NHEK stimulated with HD were 21±11ng/mL as measured by Luminex100™. The messenger RNA expression of the cytokine (hIL-6) gene was analyzed semiquantitatively. RT-PCR demonstrated that HD induced an increase in the transcription of hIL-6 gene. Selective immunosuppression, using IL-6 neutralizing antibodies, led to a reduction of such expression of HD-induced transcription of hIL-6 in human keratinocytes. The neutralization by pre-incubating NHEK with monoclonal anti-IL6 antibodies decreased hIL-6 secretion by 76%±1.8 (*P<0.05).

Abstract : The central nervous system (CNS) is an immuneprivileged site where the role of immune cells and mediators in brain injury caused by organophosphates (OP) is poorly understood. Many mediators have been identified in nervous system tissue. For instance, interleukin-6 (IL-6), a cytokine that acts on a wide range of tissue influencing cell growth and differentiation, is an agonist for vascular endothelial growth factor (VEGF). CNS cells producing IL-6 include astrocytes, macrophages, microglia, neurons, and brain endothelial cells. Here we describe the response of five different mediators, human interleukin-1beta (hIL-1b), hIL-6, hIL-8, tumor necrosis factor alpha (hTNF-a), and human granulocyte macrophage-colony stimulating factor (hGM-CSF) associated with human astrocytes incubated with an OP, sarin (GB). Human astrocytes (^ 106 cell density) were stimulated with a high concentration of GB (0.8 mM) for 48 hours at 37 degrees C. The expressed mediators in human astrocytes were detected by using the Luminex100 trademark protein multiplex immunoassay. Constitutive non-stimulated human astrocytes secreted hGM-CSF (0.59 0.03 ng/mL), hIL- 1b (0.33 0.05 ng/mL), hIL-6 (1.43 0.02 ng/mL) and hIL-8 (0.39 0.02 ng/mL). hTNF-a secretion was not detected or observed. GB decreased the endogenous secretion of these mediators as follows: hGM-CSF (0.51 0.02 ng/mL), hIL-1b (0.28 0.03 ng/mL), hIL-6 (1.00 0.02 ng/mL). Meanwhile, the induction chemokine hIL-8 was increased 0.42 0.03 ng/mL by GB as measured by Luminex100 trademark. Up and down-regulation of these mediators in human astrocytes promises a nonintrusive mechanism for assessment of the role of individual mediators in brain cell development, function and response to insult, such as that caused by the OP. Many neurodegenerative disorders are associated with inflammatory processes in the central nervous system (CNS).

Studies were conducted to examine the effect of two vesicant chemical warfare agents (VCWA), one of them an arsenical, on cytokine gene expression in normal human epidermal keratinocyte (NHEK) cells. We tested 2,2′-dichlorethylsulfide (sulfur mustard, military designation HD) and 2,chlorovinyldichloroarsine (Lewisite, military designation L), which have significant differences in their chemical, physical, and toxicological properties. Human tumor necrosis factor-alpha (hTNF-α) cytokine was detected by using the enzyme-linked immunosorbent assay, a protein multiplex immunoassay, Luminex 100 ™ , and reverse transcription-polymerase chain reaction (RT-PCR). The messenger RNA expression of hTNF-α was determined to provide a semi-quantitative analysis. HD-stimulated NHEK induced secretion of hTNF-α in a dose-dependent manner. Dose response effect of Lewisite decreased hTNF-α levels. Time-response data indicated that the maximum response for HD occurred at 24 h with an associated cytotoxic concentration of 10 –4 mol/L. NHEK cells stimulated with 10 –4 mol/L HD for 24 h at 37°C increased detectable levels of hTNF-α from 5 to 28 ng/ml at an index of cell viability between 85 to 93% as detected by Luminex 100 ™ . Our results indicated that the increased levels of hTNF-α by HD are dependent on the primary cultures, cell densities, and chemical properties of the stimulation. Lewisite under the same conditions as HD caused a reduction of hTNF-α from control levels of 1.5 ng/ml to 0.3 ng/ml after stimulation (10 –4 mol/L), with an index of cell viability of ∽34%. We analyzed the transcriptional of hTNF-α gene and found that HD (10 –6 to 10 –4 mol/L) activates hTNF-α gene in cultured NHEK and that L at 10 –6 to 10 –4 mol/L markedly reduces hTNF-α gene. We conclude that the pro-inflammatory mediator, hTNF-α, could be a potential biomarker for differentiating between exposure of HD or L.

The authors applied in vitro models of controlled damage to human epidermal keratinocytes (HEKs), human skin fibroblasts (HSFs), and human breast skin tissue (HBST) to examine the mechanism responsible for sulfur mustard (HD)-induced interleukin-6 (IL-6) alterations. Treatment with 100 microM HD for 24 hours resulted in a significant increased amount of IL-6 being secreted by HEKs (HD-exposed to control ratio [E/C] = 4.15 +/- 0.07) and by HSFs (E/C = 7.66 +/- 0.04). Furthermore, the HD-induced secretion of IL-6 in HEKs was neutralized with monoclonal human IL-6 antibodies. The secretion of IL-6 in HBST supernatant exposed to HD produced conflicting results. Although an increase of IL-6 was observed in control superfusion media from HBST, IL-6 levels were observed to decrease as the concentration of HD increased. Time course of IL-6 mRNA levels were performed using a competitive polymerase chain reaction (PCR) and human IL-6 mRNA assay detection kit in control and HD (100 microM)-treated HEKs cells. IL-6 mRNA transcripts in HD-exposed HEKs were first observed within 2 hours, dropped at 5 to 6 hours, and increased by approximately 2.2-fold and 8.5-fold at 24 to 48 hours after HD exposure, respectively, as detected by the Xplore mRNA Quantification System. Surface-enhanced laser desorption ionization (SELDI) mass spectrometry was also applied to study the secretion pattern of IL-6 on lysate preparations of HBST. A peak in the area of 23,194 to 23,226 Da was detected using antibody coupled to the chip. This peak was assigned to correspond to the mass of the IL-6 glycoprotein. Recombinant human IL-6 (rhIL-6) exposed to HD lacked the second disulfide bridge and was partially unfolded, as determined by nuclear magnetic resonance-nuclear Overhauser enhancement and exchange spectroscopy (NMR-NOESY). The disappearance of the resonance peak at 3.54 ppm and the appearance of a new chemical shift at 1.85 ppm suggested that a change in structure had occurred in the presence of HD. From the data, the possibility cannot be excluded that IL-6 might be involved in the early event of structural changes of the signal transducer glycoprotein that indirectly initiates the cascade of events such as skin irritation and blister formation observed in the pathophysiology of HD injury.

The novel inhibitor 7-N-phenylcarbamoylamino-4-chloro-3-propyloxyisocoumarin (ICD 1578) was tested for its ability to antagonize the zinc metalloprotease activity of botulinum toxin B (BoNT/B). The efficacy of this compound was tested in a cell-free system using a 50-mer synaptobrevin peptide as substrate. The peptide, designated as [Pya88] S 39-88, had a fluorescent amino acid analog, L-pyrenylalanine (Pya), substituted for the normal Phe88 of synaptobrevin-2. Cleavage by BoNT light chain yielded fragments of 38 and 11 amino acids, respectively. The smaller fragment, containing the Pya fluorophore, was readily separated and quantified by fluorescence spectroscopy at 377 nm. In the presence of 7-200 microM ICD 1578, cleavage of [Pya88] S 39-88 was progressively reduced (IC50 = 27.6 microM), and 100 microM ICD 1578 produced >95% inhibition. For comparison, captopril, a well-known zinc metalloprotease inhibitor, generated less than 10% inhibition at a concentration of 5 mM. ICD 1578 is the most potent antagonist of BoNT/B light chain thus far described.

Due to a clerical error, Fabrice Cornille was omitted as an author on the above publication. The correct list of authors is provided above.

The efficacies of atropine (ATR)/ 2-PAM and ATRl HI-6 in treating male rhesus monkeys injected with the neurotoxic organophosphonate (OP) agents GA, GB, GD, GF, or VX were compared. Experiments were conducted using no more than 8 monkeys per OP and treatment regimen. Doses were selected using a modified up-and-down experimental design, challenging one monkey per day per OP and treatment. Results were used to approximate the median lethal dose (MLD) for groups of treated monkeys or monkeys given only a vehicle following challenge with an OP. Mortality and signs of intoxication with each treatment were statistically compared. Doses of 2-PAM (25.7 mgl kg) or HI-6 (50 mgl kg) and atropine (0.4 mg free base! kg) were given in a single intramuscular (IM) injection 1 min following challenge with an OP. Strong and well-defined relationships between agent dose and 10-h lethality were observed in untreated animals. The 10-h IM OP MLDs for untreated monkeys were estimated to be 80, 43, 8.0, 22, and 7.4 μg/kg for GA, GB, GD, GF, and VX, respectively. No statistical differences w ere found between AT Rl 2-PAM and AT Rl HI-6 treatment efficacies in preventing lethality for any of the OPs. Both oxime combinations appear to provide protection against a 2 × 10-h MLD of GA, GF, or VX; only

: Studies on the characterization of wastewaters from Army manufacture of trinitrotoluene (TNT), glyceryl trinitrate (trinitroglycerine, TNG) and nitrocellulose (NC) are described. The studies were undertaken to provide information to be used in design of toxicological experiments for the purpose of assigning water quality guidelines. The complexity of the chemical characterization became evident by the fact that in TNT wastewaters alone (toward which most of the effort was directed), more than 40 organic compounds were detected by GLC and TLC in addition to inorganic materials and organic aggregates not resolved by TLC. Extraction methods were developed for separation of the organic materials contained in NG wastewaters. Results of GC and TLC showed a complex mixture of mononitrate, dinitrate and trinitrate esters of glycerol and a number of unidentified compounds, two containing the carbonyl function. Products were a function of time from sampling, of pH, and of amount of exposure to light. Lyophilization of samples containing high concentrations of salts appears to be feasible without risk of explosion. Polarized light and electron microscopy of nitrocellulose wastewaters, showed fibers from a few to several hundred nm in length and of a varying degree of nitration, together with fine particulate material, brown amorphous slime, and a crystalline precipitate. Suspended particles could be coagulated by either dialysis or addition of ionic solutes. Further characterization of nitrocellulose wastes were deemphasized after preliminary toxicological results indicated low toxicity.

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTThe synthesis of oximes. IIBrennie E. Hackley Jr. and Francis A. DaniherCite this: J. Org. Chem. 1967, 32, 8, 2624–2625Publication Date (Print):August 1, 1967Publication History Published online1 May 2002Published inissue 1 August 1967https://pubs.acs.org/doi/10.1021/jo01283a060https://doi.org/10.1021/jo01283a060research-articleACS PublicationsRequest reuse permissionsArticle Views226Altmetric-Citations9LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InRedditEmail Other access optionsGet e-Alertsclose Get e-Alerts