Response of normal human keratinocytes to sulfur mustard: cytokine release.

Cytokines play a major role in both acute and chronic inflammatory processes, including those produced by sulfur mustard (2,2'-dichlorodiethyl sulfide, HD). This study describes responses of normal human epidermal keratinocytes (NHEK) to HD, defined by interleukin-1beta (IL-1beta), IL-6, IL-8 and tumor necrosis factor alpha (TNF-alpha) release. Commercially available enzyme-linked immunosorbent assay (ELISA) kits were used to measure the cytokine release in NHEK during exposure to 100 and 300 microM of HD. Exposure to 100 microM HD increased the release of cytokines. The amounts of IL-8 and TNF-alpha present in cell suspensions increased up to 59-fold and 4-fold, respectively, above control levels when NHEK were exposed to 300 microM HD. Exposure of NHEK to 300 microM HD had a highly variable effect on the release of IL-1beta, where sometimes the secretion of IL-1beta increased above baseline level and at other times it decreased in cell suspensions. Supernatants were collected from cell culture flasks 24 h after exposure of 100 and 300 microM HD and significantly increased levels of IL-6 were observed. Interleukin-6 was released in a concentration-dependent manner, 3.6-fold up to 8.4-fold, respectively, in supernatant. These pro-inflammatory mediators IL-1beta, IL-8, TNF-alpha and IL-6 may play an important role in HD injury. The present findings suggest that the cytokine changes detected could be used as potential biomarkers of cutaneous vesicant injury.

Reactivity of chloroethyl sulfides in the presence of a chlorinated prophylactic: a kinetic study by EPR/spin trapping and NMR techniques.

This study reports the kinetic reaction of a chlorinated glycoluril, 1,3,4,6-tetrachloro-7, 8-diphenyl-2,5-diimino glycoluril, also known as S-330, with butyl 2-chloroethyl sulfide (half-sulfur mustard, H-MG) and bis-(2-chloroethyl) sulfide (sulfur mustard, HD) using electron paramagnetic resonance (EPR)/spin trapping and nuclear magnetic resonance (NMR) techniques. Both H-MG and HD are highly reactive in water and are capable of alkylating a variety of critical target molecules. It is well known that compounds containing reactive chlorine are useful neutralizers of HD and other vesicating agents. Organic compounds containing a chloroamide group are generally preferred. Currently, the reactive mechanism of this chlorinated glycoluril with these chloroethyl sulfides has not been documented. The kinetic experiments were performed by adding the monofunctional sulfur mustard (H-MG) directly to the spin trap agent alpha-phenyl-N-tert-butylnitrone (PBN, pH 7.1). The intensity of the EPR spectra obtained from the resulting spin adduct (hyperfine coupling constants aN = 1.45 mT and abetaH = 0.225 mT) was sensitive to the rate at which the spin adduct was formed. Different concentrations of the chloroamide were added to the reaction mixtures of PBN and H-MG. The EPR spectra of separate identical reaction mixtures were recorded with the spectrometer set for kinetic experiments. The rate constant determined by EPR was 1.78 +/- 0.14 x 10(7) M(-1)s(-1). It was found that S-330 reacts 55 times faster than PBN. The results obtained for S-330 by EPR indicate that S-330 is an efficient scavenger of H-MG. Furthermore, a 13C-NMR chemical shift of 0.903 +/- 0.002 ppm was observed for the Cl-N-C-N-Cl carbon in S-330 after exposure to HD (1 mM). In addition, the decay of 13C-NMR resonance at 91.7 ppm chemical shift was observed in the presence of HD. The 13C-NMR data showed that the formation of the ethylene sulfonium ion usually found in the case of HD was not observed in the presence of S-330.

Response of normal human keratinocytes to sulfur mustard (HD): cytokine release using a non-enzymatic detachment procedure.

Cytokines play a major role in both acute and chronic inflammatory processes, including those produced by sulfur mustard (HD). This study describes responses of normal human epidermal keratinocyte (NHEK) cells to 2,2'-dichlorodiethyl sulfide, sulfur mustard (HD), defined by interleukin-1 beta (IL-1beta), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF-alpha) release. A new method for detaching cell to cell adhesion between keratinocytes has been applied. This method permits the characterization of endogenous fluid from cellular content that could be applied for the development of therapeutic intervention. NHEK (typical average cell density 4.4 x 10(6) cells/mL) were exposed to HD (100 and 300 microM) in keratinocyte growth medium (KGM) for 24 h at 37 C in humidified air. Commercially available enzyme-linked immunosorbent assay (ELISA) kits were used to measure the cytokine release in NHEK during exposure to 100 and 300 microM of HD. Exposure to 100 microM HD increased release of cytokines. IL-1beta (exposed: 1.41 x 10(-5) pg/ cell+/-1.60 x 10(-6) pg/cell: control 7.10 x 10(-6) pg/ cell+/-1.20 x 10(-6) pg/cell), TNF-alpha (exposed: 1.06 x 10(5) pg/cell+/-7.3 x 10(-7)pg/cell; control: 4.04 x 10(-6)+/-2.80 x 10(-7) pg/cell) and IL-8 (exposed: 3.71 x 10(-5) pg/ cell+/- 3.26 x 10(-6) pg/cell; control: 2.99 x 10(-6) pg/cell+/-8.80 x 10(-7) pg/cell) were significantly enhanced when NHEK cells were detached from culture flasks by non-enzymatic procedures. Cell suspensions of NHEK released low amounts of IL-6 when exposed to 100 microM for 24 h (exposed: 1.47 x 10(-6)+/-1.60 x 10(-7) pg/cell; control: 1.28 x 10(-6)+/-8.40 x 10(-8) pg/cell). However, cell suspensions of NHEK increased levels of IL-6 after exposure to 300 microM HD (4.67 x 10(-5) pg/cell+/-3.90 x 10(-6) pg/cell; control: 3.99 x 10(-6) pg/cell+/-5.50 x 10(-7) pg/cell). The amount of IL-8 and TNF-alpha present in cell suspensions increased up to 59-fold and fourfold, respectively, above control levels when NHEK cells were exposed to 300 microM HD. Exposure of NHEK to 300 microM HD had a highly variable effect on the release of IL-1beta, where sometimes the secretion of IL-1beta increased above baseline level and other times decreased in cell suspensions. Supernatants were collected from cell culture flasks 24 h after exposure of 100 and 300 microM and significantly increased levels of IL-6 were observed. IL-6 was released in a concentration-dependent manner, 3.6-fold up to 8.4-fold, respectively, in supernatant. These pro-inflammatory mediators IL-1beta, IL-8, TNF-alpha and IL-6 may play an important role in HD injury. The present findings suggest that cytokine changes detected could be used as potential biomarkers of cutaneous vesicant injury.

[Extramedullary tumor formation by acute myeloid leukoses in children (author's transl)]. / Extramedulläre Tumorbildung bei akuten myeloischen Leukosen im Kindesalter.

In rare cases, leukoses can produce tumors before general symptoms and changes in the blood picture appear. Two boys were reported in whom tumors of the eye socket were removed, the tumors were histologically classified as myelosarcoma. During the course of several months, the clinical picture of an acute leukemia developed. The difficulties involved in the exact histologic recognition of myelosarcoma, its localization, the differential diagnosis of tumors of the eye socket in children as well as the therapy and prognosis for children with myelosarcoma were discussed.

[Unusual generalized granulomatosis. Two case reports (author's transl)]. / Uber zwei ungewöhnliche Granulomatosen

Report about two cases of unusual generalized granulomatosis in males, with multiple lymphomas and involvement of internal organs and brain, leading soon to death. The diagnosis was malignant histiocytosis and disseminated eosinophilic granulomatosis. The malignant histiocytosis is differentiated from malignant fibroxantoma (malignant histiocytoma). The unusual cerebral features in the second case - diagnosed as disseminated eosinophilic granulomatosis - seem to show some relations to histiocytosis X. The problems in diagnosis of generalized granulomatosis are pointed out.

[Midline malignant reticulosis (so-called lethal midline granuloma) (author's transl)]. / Granuloma gangraenescens als maligne Retikulose

Case report about a 30-year old man with midline malignant reticulosis. Diagnosis was not successful by means of biopsy but given by microscopic examination of autopsy material afterwards. So-called lethal midline granuloma is a clinical term. After excluding specific infections, carcinomas or other rare diseases, it is caused by midline malignant reticulosis or by Wegener's granulomatosis. Frequently diagnosis is made by the pathologist at autopsy.