Nontoxic crystal protein from Bacillus thuringiensis demonstrates a remarkable structural similarity to beta-pore-forming toxins.
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Bacillus thuringiensis↗
Biomedical subjects
Kazuhiko Higuchi
Publications and source records attributed to Kazuhiko Higuchi.
Total anomalous pulmonary venous return in an adult.
A 47-year-old man with total anomalous pulmonary venous return met all the conditions necessary for long survival. In particular, his pulmonary vascular resistance was almost normal because of right ventricular outflow tract obstruction. Surgical treatment was performed with a good result.
Humans↗
Efficient solubilization, activation, and purification of recombinant Cry45Aa of Bacillus thuringiensis expressed as inclusion bodies in Escherichia coli.
A cytotoxic protein Cry45Aa of Bacillus thuringiensis expressed as inclusion bodies in Escherichia coli was solubilized in 10 mM HCl. Protein concentration of saturated solution of the recombinant Cry45Aa in 10 mM HCl was about 25 times higher than that in the buffer of previous method (in 50 mM sodium carbonate buffer, pH 10.5, containing 1 mM EDTA, and 10 mM dithiothreitol). The Cry45Aa solubilized in the acidic solution was activated by pepsin as an alternative to proteinase K in the previous method. Cytotoxic activity against CACO-2 cells of the pepsin-treated Cry45Aa was almost identical to the proteinase K-treated protein. The pepsin-treated Cry45Aa was purified by cation-exchange chromatography. The concentration of the purified protein was 539 microg/ml, which was 27-fold higher than that of the activated Cry45Aa by the previously method. The cytotoxic activity of the purified protein was stable in broad pH region (pH 2.0-11.0) for 3 days, and 97% cytotoxic activity remained after incubation at 30 degrees C for 360 min.
Bacillus thuringiensis Toxins↗
Use of an aortic connector system for vein grafting: intermediate outcomes.
OBJECTIVE: The aortic connector system was used to minimize cerebrovascular complications when performing the proximal anastomosis of vein grafts during coronary artery bypass grafting (CABG). The goal of this study was to investigate the intermediate outcomes of patients undergoing CABG with the aortic connector system. METHODS: The aortic connector was used on nine patients undergoing CABG between November 2002 and July 2003. Intermediate outcomes of the patients were examined, and the results of coronary angiography, which were performed before patient discharge and at least 6 months after discharge, were evaluated. RESULTS: There were no operative deaths or cerebrovascular accidents. One patient died 9 months after discharge, one patient had angina, and the remaining seven patients were asymptomatic. When evaluating the results of angiography performed before patient discharge, two of the 21 distal vein graft anastomoses were occluded (patency rate, 90.5%), but there was no stenosis or occlusion at the proximal anastomoses sites that were performed using the aortic connector. When evaluating the results of the second angiography performed after patient discharge, four of the eight proximal anastomoses were patent, one was completely occluded, two had 90% stenosis and one had 75% stenosis. Further, four of the 18 distal anastomoses were occluded (patency rate, 77.8%). There was no significant difference in graft flow or device size when comparing patients with patent vein grafts and those with stenotic or occluded vein grafts. CONCLUSION: Intermediate outcomes of vein grafting using the aortic connector were suboptimal. Long-term outcome data are forthcoming.
Aged↗
Characterization of surfactant-induced skin damage through barrier recovery induced by pseudoacylceramides.
BACKGROUND AND OBJECTIVE: To determine how much barrier-reinforcing effects of ceramides contribute to prevent the surfactant-induced cutaneous deterioration. METHODS: We compared the effects of topical application of two types of pseudoceramides on cutaneous deterioration induced by sodium dodecyl sulfate (SDS) treatment for 10 days in association with alterations of barrier function. RESULTS: Daily application of pseudoacylceramide immediately after each SDS treatment significantly prevented the marked elevation of transepidermal water loss, which was accompanied by a marked abrogation of the increased expression in intercellular adhesion molecule 1 by epidermal cells as well as by suppressed epidermal hyperplasia. In contrast, topical application of a nonacylated pseudoceramide with poor barrier-reinforcing potential showed less preventive effects on cutaneous deterioration. CONCLUSION: These results strongly suggest that perturbation of the skin barrier is a causative factor in surfactant-induced cutaneous changes and that reinforcing the barrier function by ceramide application is effective in preventing the surfactant-induced skin deterioration.
Administration, Topical↗
Crystallization of parasporin-2, a Bacillus thuringiensis crystal protein with selective cytocidal activity against human cells.
Bacillus thuringiensis is a valuable source of protein toxins that are specifically effective against certain insects and worms but harmless to mammals. In contrast, a protein toxin obtained from B. thuringiensis strain A1547, designated parasporin-2, is not insecticidal but has a strong cytocidal activity against human cells with markedly divergent target specificity. The 37 kDa inactive protein is proteolytically activated to a 30 kDa active form. The active form of the recombinant protein toxin was crystallized in the presence of ethylene glycol and polyethylene glycol 8000 at neutral pH. The crystals belong to the hexagonal space group P6(1) or P6(5), with unit-cell parameters a = b = 134.37, c = 121.24 A. Diffraction data from a native crystal were collected to 2.75 A resolution using a synchrotron-radiation source.
Bacillus thuringiensis↗
Urgent off-pump coronary artery bypass grafting in a patient with HIV.
We report a case of successful urgent off-pump coronary artery bypass grafting (CABG) for unstable angina pectoris in a 54-year-old human with human immunodeficiency virus (HIV) infection. We studied the changes in CD4+ cells and HIV-RNA copy during the perioperative period. The results showed that off-pump CABG did not reduce the CD4+ cell count and did not affect the condition of HIV infection in this patient.
Angina, Unstable↗
Left-sided cervical aortic arch aneurysm: case report.
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Adolescent↗
Sphingosylphosphorylcholine is a Melanogenic Stimulator for Human Melanocytes.
As lysosphingolipids have multiple bio-modulator functions in various types of cells, we measured the biological effects of sphingosylphosphorylcholine (SPC) on cultured human melanocytes to determine whether these lysosphingolipids have the potential to activate these cells. The addition of SPC to cultured human melanocytes significantly stimulated DNA synthesis assessed by [3H]thymidine and melanogenesis assessed by the release of [3H]H2O (tyrosinase activity), the incorporation of [14C]thiouracil (melanin synthesis) and dopa-oxidase activity. Reverse transcription-polymerase chain reaction of RNA isolated from human melanocytes exposed to SPC revealed an upregulation of mRNA transcripts for tyrosinase, microphthalmia-associated transcription factor-M, endothelin B receptor and the stem cell factor receptor, c-kit. An increase in expression of tyrosinase and c-kit proteins was also demonstrated by Western blot analysis. This stimulation of melanogenesis by SPC was associated with a marked increase in the phosphorylation of extracellular signal-regulated kinase 1/2. These results suggest that SPC may be a melanogenic stimulator of human melanocytes inducing the coordinated upregulated expression of various melanogenic molecules, including c-kit.
Cell Division↗
Surgical repair of a distal arch aneurysm with a stent-graft.
This study evaluates the effectiveness and potential complications of stent-grafting for the treatment of distal arch aneurysms using profound hypothermia and circulatory arrest with retrograde cerebral perfusion. Between December 1998 and December 2001, 9 consecutive patients with a distal arch aneurysm (6 men and 3 women, mean age 71 years) underwent surgical repair using a stent-graft. Profound hypothermic circulatory arrest and retrograde cerebral perfusion were performed in all patients. Endovascular leakage was screened postoperatively using three-dimensional computerized tomography. The mean follow-up period was 27.4 months. Thirty day mortality was 0%. One patient died 3 months after stent-grafting due to proximal leakage into her aneurysm. The mean postoperative extubation period was 2.1 days. No patients suffered cerebral infarction or paraplegia. Although preliminary outcomes using this technique were good, endovascular leakage is a concern. We suggest that, if major proximal leakage is recognized postoperatively, re-intervention should be performed as soon as possible. Endovascular stent-grafting appears to be a good alternative treatment for distal arch aneurysms, although longer follow-up is necessary to more comprehensively evaluate this procedure.
Aged↗