Glutamine prevents DMBA-induced squamous cell cancer.

The etiology of oral squamous cell carcinoma has been linked to environmental carcinogens, such as activated aromatic heterocyclic radicals and epoxides. Our previous work on implantable and 7,12-dimethylbenz[a]anthracene (DMBA)-induced breast cancer showed that oral glutamine (GLN) inhibited tumor growth possibly through stimulation of host - and selective inhibition of tumor glutathione (GSH). This finding was associated with up-regulation of NK cell activity, decreased IGF-1 and TGF-beta in the circulation and downregulation of PI-3K/Akt antiapoptotic signaling in tumors. The present study was designed to investigate the effect of topically applied GLN on DMBA-induced hamster buccal pouch squamous cell carcinoma. Histopathological alterations in buccal pouches were studied by light microscopy. GLN and GSH levels in blood and buccal mucosa were determined using specific enzyme assays. The protein expression of bax, bcl-2 and PARP was determined by western blotting. H-ras and p53 genes were examined for presence of mutations using direct DNA sequencing. Fourteen weeks after DMBA application none of the GLN-supplemented animals developed tumors, while all of the control animals had well developed squamous cell carcinomas. The inhibition of DMBA-carcinogenesis by GLN application was associated with increased arterial GLN and GSH, elevated buccal mucosa GSH as well as induction of bax and PARP, and inhibition of bcl-2. H-ras and p53 were wild type. The results from this study in combination with our previous data suggest that the chemopreventive effects of GLN are exerted by enhancing the antioxidant status of the body and activation of apoptosis.

Partial characterization of serine proteinases secreted by adult Trichinella spiralis.

Serine proteinases secreted by adult Trichinella spiralis were isolated from excretory/secretory products (ES) of in-vitro-cultured parasites by affinity chromatography with p-benzamidine-celite. The purified enzymes had molecular weights of approximately 18, 40, and 50 kDa and displayed enzyme activity against a range of low-molecular-weight substrates, gelatin, and azocasein. The antigenicity of these parasite proteinases was demonstrated by the inhibition of enzymatic activity with IgG purified from infected hosts. The inactivation of major secreted proteinases of adult T. spiralis by immune antibody could presumably contribute to impairment of the survival of the parasite in sensitized hosts.

Proteolytic enzymes secreted by larval stage of the parasitic nematode Trichinella spiralis.

Excretory/secretory products (ES), collected from in vitro cultures of muscle larvae (L1) of Trichinella spiralis (Owen, 1835) were examined for the presence of proteolytic enzymes. Several discrete proteinases in the size range of 25-55 kDa were identified by substrate gel electrophoresis and were characterised according to pH optima, substrate specificity and inhibitor sensitivity using azocasein assay. Serine, cysteine and metalloproteinases active at pH 5-7 were identified. The serine proteinases were found to predominate and some of them were found to be specific for the larval stage of the parasite. The results from the substrate analysis indicated the presence of collagenolytic and elastolytic activities. The proteinase activity was inhibited by IgG isolated from T. spiralis-infected mice, an observation of relevance to understanding host/parasite interactions and, ultimately, the development of anti-Trichinella vaccine.

Proteinases in the excretory/secretory products (ES) of adult Trichinella spiralis.

Adult Trichinella spiralis were maintained in vitro using defined media and the material excreted/secreted (ES) during this time examined for proteolytic enzyme (proteinase) activity using an azocasein assay and gelatin-substrate gels. Several discrete proteinases in the size range 14-100 kDa were observed with optimal activity at pH 7.5. The use of a class-differentiating panel of proteinase inhibitors indicated that serine proteinases were predominant although some inhibition was evident in the presence of cysteine and metalloproteinase inhibitors. Of a panel of potential natural protein substrates tested, ES proteinases only degraded fibrinogen and plasminogen and degradation was, in part, susceptible to the action of serine, cysteine and aspartyl proteinase inhibitors. In addition, antibody harvested from immune but not normal mice inhibited ES proteinase activity, an observation of relevance to the immunobiology of Trichinosis.