Biodegradable citrate-based polyesters with S-nitrosothiol functional groups for nitric oxide release.

Nitric oxide (NO) is a biologically-active free radical involved in numerous physiological processes such as regulation of vasodilation, promotion of cell proliferation and angiogenesis, and modulation of the inflammatory and immune responses. Furthermore, NO has demonstrated the ability to mitigate the foreign body response that often results in the failure of implanted biomedical devices. Although NO has promising therapeutic value, the short physiological half-life of exogenous NO complicates its effective delivery. For this reason, the development of NO-releasing materials that permit the localized delivery of NO is an advantageous method of utilizing this molecule for biomedical applications. Herein, we report the synthesis and characterization of biodegradable NO-releasing polyesters prepared from citric acid, maleic acid, and 1,8-octanediol. NO release was achieved by incorporation of S-nitrosothiol donor groups through conjugation of cysteamine and ethyl cysteinate to the polyesters, followed by S-nitrosation with tert-butyl nitrite. The extent of NO loading and the release properties under physiological conditions (pH 7.4 PBS, 37 °C) were determined by chemiluminesence-based NO detection. The average total NO content of poly(citric-co-maleic acid-co-1,8-octanediol)-cysteamine was determined to be 0.45 ± 0.07 mol NO g-1 polymer, while the NO content for poly(citric-co-maleic acid-co-1,8-octanediol)-ethyl cysteinate was 0.16 ± 0.04 mol NO g-1 polymer. Continuous NO release under physiological conditions was observed for at least 6 days for the cysteamine analog and 4 days for the ethyl cysteinate analog. Cell viability assays and morphological studies with human dermal fibroblasts indicated an absence of toxic leachates at a cytotoxic level, and suggested that these citrate-based polyesters may be suitable for future biomedical applications.

Different mitogen-activated protein kinase signaling pathways cooperate to regulate tumor necrosis factor alpha gene expression in T lymphocytes.

Tumor necrosis factor a (TNF-alpha) is a potent proinflammatory cytokine and plays a crucial role in early events of inflammation. TNF-alpha is primarily produced by monocytes and T lymphocytes. In particular, T-cell-derived TNF-alpha plays a critical role in autoimmune inflammation and superantigen-induced septic shock. However, little is known about the intracellular signaling pathways that regulate TNF expression in T cells. Here we show that extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38-mitogen-activated protein kinase (MAPK) pathways control the transcription and synthesis of TNF-alpha in A3.01 T cells that produce the cytokine upon T cell activation by costimulation with 12-O-tetradecanoylphorbol-13-acetate (TPA) and ionomycin. Selective activation of each of the distinct MAPK pathways by expression of constitutively active kinases is sufficient for TNF-alpha promoter induction. Furthermore, blockage of all three pathways almost abolishes TPA/ionomycin-induced transcriptional activation of the TNF-alpha promoter. Selective inhibition of one or more MAPK pathways impairs TNF-alpha induction by TPA/ionomycin, indicating a cooperation between these signal transduction pathways. Our approach revealed that the MAPK kinase 6 (MKK6)/p38 pathway is involved in both transcriptional and posttranscriptional regulation of TNF expression. Moreover, analysis of the progressive 5' deletion mutants of the TNF-alpha promoter indicates that distinct promoter regions are targeted by either ERK-, JNK-, or p38-activating pathways. Thus, unlike what has been reported for other TNF-alpha-producing cells, all three MAPK pathways are critical and cooperate to regulate transcription of the TNF-alpha gene in T lymphocytes, suggesting a T-cell-specific regulation of the cytokine.

The stress inducer arsenite activates mitogen-activated protein kinases extracellular signal-regulated kinases 1 and 2 via a MAPK kinase 6/p38-dependent pathway.

Cell response to a wide variety of extracellular signals is mediated by either mitogenic activation of the Raf/MEK/ERK kinase cascade or stress-induced activation of the mitogen-activated protein kinase (MAPK) family members c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) or p38. We have examined communications between these stress- and mitogen-induced signaling pathways. We show here that the stress cascade activator arsenite activates extracellular signal-regulated kinase (ERK) in addition to p38 albeit with different kinetics. Whereas p38 is an early response kinase, ERK activation occurs with delayed time kinetics at 2-4 h. We observed activation of ERK upon arsenite treatment in many different cell lines. ERK activation is strongly enhanced by overexpression of p38 and mitogen-activated protein kinase kinase 6 (MKK6) but is blocked by dominant negative kinase versions of p38 and MKK6 or the specific p38 inhibitor SB203580. Arsenite-induced ERK activation is mediated by Ras, Raf, and MEK but appears to be independent of de novo protein synthesis. These data provide the first evidence for a p38 dependent activation of the mitogenic kinase cascade in stress-stimulated cells.

Sclerosing lipogranuloma of the scalp. A report of two cases.

The authors report two cases of sclerosing lipogranuloma of the scalp diagnosed postmortem. The first was suspected to have acquired the lipid material from a grease gun injury to the scalp, while the second had received injections of an unidentified lipid for treatment of baldness. In each, the resultant sclerosing lipogranuloma extended laterally beneath the scalp from the vertex of the skull. The histologic reaction was round to oval cystic spaces within a dense collagenous stroma typical of the "swiss cheese" pattern of sclerosing lipogranuloma. Punctate, ringlike, or globoid densities were seen on roentgenograms of a 5-mm-thick transmural section of scalp in the first case.

Myofibrillar protease activity in muscle tissue from patients in catabolic conditions.

Myofibrillar alkaline protease activity was shown to be present in human skeletal muscle. Endogenous myofibrillar proteins and 14C-labelled, exogenous haemoglobin were both active as substrates, and the enzymic activity appeared to be similar to the myofibrillar protease previously described in rodent muscles. The activity of this enzyme was determined in patients undergoing surgery for a variety of diseases. Significant elevations in proteolytic activity were found in the abdominal wall muscle of patients in wasting conditions as compared with non-catabolic diseases. In cachectic patients on total parenteral nutrition the protease activity was similar to reference values. The results imply that increased activity of the myofibrillar alkaline protease plays a role in the development of cachexia in human wasting diseases by prompting degradation of muscle proteins.