Dietary Supplementation With a Bacillus Superoxide Dismutase Protects Against γ-Radiation-induced Oxidative Stress and Ameliorates Dextran Sulphate Sodium-induced Ulcerative Colitis in Mice.

BACKGROUND AND AIMS: Commercial superoxide dismutase [SOD] is derived from melon extract and has a potential as a dietary supplement due to its beneficial antioxidative effects. We aimed to improve the productivity of SOD compared with plant SOD by using a generally regarded as safe [GRAS] microorganism, Bacillus amyloliquefaciens, and assess its antioxidative effect using γ-radiation- and dextransulphate sodium [DSS]-induced oxidative models in mice. METHODS: We identified the sodA gene encoding manganese-containing SODs [Mn-SOD] in B. amyloliquefaciens, constructed a Mn-SOD deficient mutant, and screened a high-SOD-producing strain. We compared the antioxidative effect of orally administered enteric-coated SOD protein partially purified from B. amyloliquefaciens with wild-type and high-SOD-producing strain spores. The effect of SOD on DSS-induced colitis was also investigated. Colonic inflammation was assessed using disease activity index, macroscopic and histological damage scores, antioxidant enzyme activities, and inflammatory cytokines. RESULTS: The SOD activity of B. amyloliquefaciens is derived from secreted Mn-SOD encoded by the sodA gene, as shown by comparing sodA knock-out mutant spores with wild-type and high-SOD-producing spores. Enteric-coated SOD of B. amyloliquefaciens appears to be effective in reducing oxidative stress in γ-radiation- and DSS-induced mouse models. Co-administration of SOD with wild-type B. amyloliquefaciens or high-SOD-producer strain spores showed a synergistic effect. SOD enzyme and B. amyloliquefaciens spores contribute to the reduction of oxidative stress and inflammatory response in DSS-induced colitis. CONCLUSIONS: Mn-SOD of B. amyloliquefaciens could be another source of SOD supplement and may be useful to prevent and treat ulcerative colitis.

Purification, Characterization, and Cloning of a Cold-Adapted Protease from Antarctic Janthinobacterium lividum.

In this study, a 107 kDa protease from psychrophilic Janthinobacterium lividum PAMC 26541 was purified by anion-exchange chromatography. The specific activity of the purified protease was 264 U/mg, and the overall yield was 12.5%. The J. lividum PAMC 25641 protease showed optimal activity at pH 7.0-7.5 and 40°C. Protease activity was inhibited by PMSF, but not by DTT. On the basis of the N-terminal sequence of the purified protease, the gene encoding the cold-adapted protease from J. lividum PAMC 25641 was cloned into the pET-28a(+) vector and heterologously expressed in Escherichia coli BL21(DE3) as an intracellular soluble protein.

Intracorneal melatonin delivery using 2-hydroxypropyl-ß-cyclodextrin ophthalmic solution for granular corneal dystrophy type 2.

Melatonin (MT), an effective antioxidant, has therapeutic implications for granular corneal dystrophy type 2 (GCD2) treatment. Eye drop formulations containing cyclodextrins (CDs) were studied with the objective of improving MT solubility, stability, and ocular absorption, while decreasing eye irritation. MT complexes with αCD, ßCD, γCD, and 2-hydroxypropyl-ßCD (HPßCD) were characterized by phase solubility studies, which demonstrated Higuchi's AL-type phase solubility profiles. The MT/HPßCD complex showed the highest MT solubility (2.75mg/mL). Ocular irritation experiments showed HPßCD inclusion alleviated irritation of the eye. After administration of MT formulations to rabbit corneas, each harvested cornea was separated into corneal epithelium, stroma, and endothelium. MT concentrations in the corneal epithelium, stroma, and endothelium for the F1-treated group were 55.5±9.24, 26.7±2.66, and 21.1±1.77µM while those for the F2-treated group were 127.2±21.01, 43.7±16.93, and 51.0±13.91µM, respectively. Stability studies for 60days showed no significant change in pH, osmolarity, and MT content. In conclusion, MT/HPßCD formulations can lower irritation, enhance MT stability, and improve therapeutic efficacy.

Supercritical fluid-mediated liposomes containing cyclosporin A for the treatment of dry eye syndrome in a rabbit model: comparative study with the conventional cyclosporin A emulsion.

BACKGROUND: The objective of this study was to compare the efficacy of cyclosporin (CsA)-encapsulated liposomes with the commercially available CsA emulsion (Restasis) for the treatment of dry eye syndrome in rabbits. METHODS: Liposomes containing CsA were prepared by the supercritical fluid (SCF) method consisted of phosphatidylcholine from soybean (SCF-S100) and egg lecithins (SCF-EPCS). An in vitro permeation study was carried out using artificial cellulose membrane in Franz diffusion cells. Dry eye syndrome was induced in male albino rabbits and further subdivided into untreated, Restasis-treated, EPCS, and S100-treated groups. Tear formation in the dry-eye-induced rabbits was evaluated using the Schirmer tear test. All formulations were also evaluated by ocular irritation tests using the Draize eye and winking methods with the determination of CsA concentration in rabbit tears. RESULTS: After the treatment, the Schirmer tear test value significantly improved in EPCS-treated (P=0.005) and S100-treated (P=0.018) groups compared to the Restasis-treated group. The AUC0₋24 h for rabbit's tear film after the administration of SCF-S100 was 32.75±9.21 µg·h/mg which was significantly higher than that of 24.59±8.69 µg·h/mg reported with Restasis. Liposomal CsA formulations used in this study showed lower irritation in rabbit eyes compared with Restasis. CONCLUSION: These results demonstrate that the novel SCF-mediated liposomal CsA promises a significant improvement in overcoming the challenges associated with the treatment of dry eyes.

A case of acute pericarditis with hemophagocytic syndrome, cytomegalovirus infection and systemic lupus erythematosus.

Hemophagocytic syndrome, cytomegalovirus infection and systemic lupus erythematosus (SLE) would each be critical diseases separately. Viral infections, autoimmune diseases or malignancies can complicate the Hemophagocytic syndrome. Cytomegalovirus infection is known to be prevalent in immune compromised hosts, and can exacerbate the symptoms of systemic lupus erythematosus. A 25-year-old man presented with fever and acute pericarditis with the hemophagocytic syndrome, cytomegalovirus infection and systemic lupus erythematosus; all developed concurrently at the onset of illness. With treatment, using ganciclovir and glucocorticoid medication the patient improved. Here we report this rare case and review the medical literature.

Therapeutic experience of Bing-Neel Syndrome associated with Waldenstrom's macroglobulinemia.

Waldenstrom's macroglobulinemia is an uncommon low-grade B-cell lymphoproliferative disorder in which monoclonal immunoglobulin M is produced. Neurological symptoms due to hyperviscosity are frequent manifestations of Waldenstrom's macroglobulinemia. However, central nervous system infiltration by plasmacytoid lymphocytes (Bing-Neel syndrome) has only rarely been reported. We report a case of a 51-yr-old woman suffering from Waldenstrom's macroglobulinemia who complained of persistant headache. Brain magnetic resonance imaging revealed an extra-axial soft tissue mass along the left cavernous sinus, left tentorium, right tentorium, and falx cerebri. A stereotactic biopsy of dural tissue from the falx was performed and showed plasmacytoid lymphocyte infiltration. The patient became symptom- free with irradiation of the whole brain followed by chemotherapy with fludarabine.

A case of Behçet's disease with pericarditis, thrombotic thrombocytopenic purpura, deep vein thrombosis and coronary artery pseudo aneurysm.

Behçet's disease with concomitant thrombotic thrombocytopenic purpura (TTP), coronary artery stenosis and coronary artery pseudo aneurysm is rare. Here we report a case of Behçet's disease with several cardiovascular complications, namely: pericarditis, deep vein thrombosis (DVT), TTP, coronary artery stenosis, and a coronary artery pseudo aneurysm. A 37-year-old female presented with sudden dyspnea and syncope at our emergency room and underwent pericardiectomy and pericardial window formation for the diagnosis of cardiac tamponade with acute hemorrhagic pericarditis. Thereafter, TTP and DVT complicated her illness. After confirmation of Behçet's disease on the basis of a history of recurrent oral and genital ulcers and erythema nodosum, remission was achieved after treatment with methylprednisolone pulse therapy, colchicine, catheter directed thrombolysis and thrombectomy. However, whilst maintaining anticoagulation therapy, a newly developed pericardial aneurysmal dilatation was noted on follow-up radiologic evaluation. Further evaluation revealed right coronary artery stenosis and a left coronary artery pseudo aneurysm; these additional problems were treated with the nonsurgical insertion of an endovascular graft stent. At the time of writing three months later after stent insertion, the aneurysm has continued to regress and no additional complications have intervened with combined immunosuppressive therapy.

Effect of PEG-PLLA diblock copolymer on macroporous PLLA scaffolds by thermally induced phase separation.

A regular and highly interconnected macroporous poly(L-lactic acid) (PLLA) scaffold was fabricated from a PLLA-dioxane-water ternary system with added polyethylene glycol (PEG)-PLLA diblock using thermally induced phase separation (TIPS). The morphology of the scaffold was investigated in detail by controlling the following TIPS parameters: quenching temperature, aging time, polymer concentration, molecular structure, and diblock concentration. The phase diagram was assessed visually on the basis of the turbidity. The cloud-point curve shifted to higher temperatures with increasing PEG content in the additives (PEG-PLLA diblocks), due to a stronger interaction between PEG and water in solution. The addition of diblock series (0.5 wt% in solution) stabilized interconnections of pores at a later stage without segregation or sedimentation. The pore size of the scaffold could be easily controlled in the range 50-300 microm. A macroporous PLLA scaffold was used to study an MC3T3-E1 cell (an osteoblast-like cell) culture. The cells successfully proliferated in the PLLA scaffold in the presence of added PEG-PLLA diblock for 4 weeks.