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1.
Clin Transplant ; 35(8): e14385, 2021 08.
Article in English | MEDLINE | ID: mdl-34132442

ABSTRACT

INTRODUCTION: Kidney transplant (KT) directors are general surgeons or urologists. All KT centers must meet established performance standards. However, it has not been established if general surgery and urology led programs have disparate outcomes. METHODS: Transplant outcomes and donor-recipient characteristics by director training were investigated. Organ Procurement and Transplantation Network (OPTN) directory, program websites were analyzed for surgical director demographics. Scientific Registry of Transplant Recipients (SRTR) 1-year kidney survival and deceased donor (DD) wait-time rankings were evaluated. A retrospective analysis of 142 157 KT recipients from 2010 to 2019 was performed using the United Network for Organ Sharing (UNOS) database. RESULTS: One hunderd and seventy three (90.6%) KT programs were led by general surgeons. There were no significant differences in gender, ethnicity, region, credentials, or fellowship completion. Recipients undergoing KT with urology led programs were older (P = .002) and had longer wait-times (P < .001). These centers used higher KDPI (.47 vs. .45, P < .001) and higher HLA mismatch (3.92 vs. 3.89, P = .02) kidneys. Urology led centers utilized living donors less frequently (32.1% vs. 35.8%, P < .001) and had longer CIT (15.44 vs. 12.21, P < .001). Both had similar SRTR ranking of 1-year survival and DD wait-time. CONCLUSION: Most directors were general surgeon. Patient outcomes did not differ by transplant director training. Urologists represent a viable option for KT leadership and recruitment should be encouraged.


Subject(s)
Kidney Transplantation , Surgeons , Humans , Living Donors , Retrospective Studies , Urologists
2.
Clin Radiol ; 76(7): 550.e9-550.e17, 2021 07.
Article in English | MEDLINE | ID: mdl-33691950

ABSTRACT

AIM: To clarify the utility of contrast-enhanced ultrasonography (CEUS) for interim evaluation of response to chemotherapy in lymphoma treatment. MATERIALS AND METHODS: CEUS was performed both before (day 0) and after the treatment (7 and/or 14 days), and a time-intensity curve was obtained. The patients were divided into two groups (complete remission [CR] group and non-CR group) according to the results of conventional response evaluation, and peak enhancement (PE), time to peak enhancement, perfusion index (PI), the total area under the curve during wash-in (AUC-in), and the total AUC were compared between the groups. RESULTS: Among 27 patients with various types of lymphoma, the median change ratio of PE and PI at day 7 evaluation were significantly different between the CR group and the non-CR group (0.81 versus 1.39, p=0.017 for PE and 0.92 versus 2.09, p=0.010 for PI). The change ratio of PE < 1.09 (specificity: 86%; sensitivity, 88%) and PI < 1.65 (specificity: 86%; sensitivity: 94%) distinguished CR from non-CR. Patients who achieved a PE change ratio <1.09 or a PI change ratio <1.65 had significantly better estimated progression-free survival (p<0.001). CONCLUSION: The present study demonstrated that changes in tumour perfusion parameters evaluated with CEUS at 1 week after the treatment initiation were significantly different between lymphoma patients in CR group and non-CR group. Alterations in perfusion parameters evaluated via CEUS could impact the prognosis of lymphoma patients.


Subject(s)
Induction Chemotherapy , Lymphoma/diagnostic imaging , Lymphoma/drug therapy , Neovascularization, Pathologic/diagnostic imaging , Ultrasonography/methods , Aged , Contrast Media , Female , Fluorocarbons , Humans , Image Interpretation, Computer-Assisted , Male , Pilot Projects , Prognosis , Prospective Studies
3.
Transpl Immunol ; 76: 101774, 2023 02.
Article in English | MEDLINE | ID: mdl-36528248

ABSTRACT

BACKGROUND: Belatacept is employed alongside calcineurin inhibitor (CNI) therapy to prevent graft rejection in kidney transplant patients who are Epstein-Barr virus (EBV) seropositive. Preliminary data suggested that rates of post-transplant lymphoproliferative disorder (PTLD) were higher in individuals treated with belatacept compared to CNI therapy alone. METHODS: The records of 354 adults who underwent kidney only transplantation from January 2015 through September 2021 at one medical center were evaluated. Patients underwent treatment with either low-doses of mycophenolate, tacrolimus and sirolimus (B0, n = 235) or low-doses of mycophenolate, tacrolimus and belatacept (B1, n = 119). All recipients underwent induction with antithymocyte globulin and a rapid glucocorticosteroid taper. Relevant donor and recipient information were analyzed and endpoints of PTLD were assessed. RESULTS: There were no cases of PTLD in either cohort within the study period. Recipients in the belatacept cohort experienced lower estimated glomerular filtration rates at 12 months (B0: 67.48 vs. B1: 59.10, p = 0.0014). Graft failure at 12 (B0: 1.28% vs. B1: 0.84%, p = 1.0) and 24 months (B0:2.55% vs. B1: 0.84%, p = 0.431) were similar. There was no difference in rejection rates at 12 (B0: 1.27% vs. B1: 2.52%, p = 0.408) or 24 months (B0: 2.12% vs. B1: 2.52%, p = 1.000). Both groups had similar rates of malignancy, mortality and CMV/BK viremia. CONCLUSION: Non-belatacept (MMF, tacrolimus and sirolimus) and belatacept-based (MMF, tacrolimus and belatacept) regimens do not appear to pose any increased risk of early onset PTLD. Both cohorts benefited from low rates of rejection, malignancy, mortality and graft failure. Recipients will continue to be monitored as PTLD can manifest as a long-term complication.


Subject(s)
Epstein-Barr Virus Infections , Kidney Transplantation , Lymphoproliferative Disorders , Neoplasms , Adult , Humans , Tacrolimus/therapeutic use , Kidney Transplantation/adverse effects , Epstein-Barr Virus Infections/epidemiology , Epstein-Barr Virus Infections/drug therapy , Herpesvirus 4, Human , Immunosuppressive Agents/therapeutic use , Abatacept/therapeutic use , Calcineurin Inhibitors/therapeutic use , Sirolimus/therapeutic use , Lymphoproliferative Disorders/etiology , Lymphoproliferative Disorders/chemically induced , Neoplasms/drug therapy , Graft Rejection/prevention & control , Graft Survival
4.
Int Angiol ; 28(3): 232-7, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19506543

ABSTRACT

AIM: A newly-designed bifurcated graft with the distal end larger than the conventional type has been developed. The purpose of this study was to evaluate the early results of graft replacement using this new graft, and to compare whether the new graft is more advantageous than the conventional graft in terms of peripheral blood flow and arterial stiffness. METHODS: Records of 36 patients who underwent bifurcated graft replacement for infrarenal abdominal aortic aneurysm (AAA), from May 2003 to September 2006 were reviewed after excluding peripheral arterial disease (ABI > 0.9). Subjects were divided into two groups: group C (N.=20), with implantation of the conventional type and group N (N.=16), with implantation of the new type. We investigated changes in brachial-ankle pulse wave velocity (baPWV) and ankle-brachial pressure index (ABI), measurements being performed preoperatively and postoperatively. RESULTS: baPWV in the postoperative group as a whole was significantly higher than in the preoperative group (P<0.05), while ABI in the postoperative group was lower than in the preoperative group (P<0.05). In group C, baPWV increased (P<0.05) and ABI decreased (P<0.05) after bifurcated graft replacement, whereas in group N, there were no significant differences in changes of baPWV and ABI. CONCLUSIONS: This study shows that the new graft reduces the development of arterial stiffness postoperatively compared with the conventional type. These results may predict the new type graft decrease in the risk of morbidity and mortality caused by atherosclerotic disease.


Subject(s)
Ankle Brachial Index , Aortic Aneurysm, Abdominal/surgery , Blood Pressure , Blood Vessel Prosthesis Implantation/instrumentation , Blood Vessel Prosthesis , Aged , Aged, 80 and over , Aortic Aneurysm, Abdominal/physiopathology , Blood Flow Velocity , Blood Vessel Prosthesis Implantation/adverse effects , Chi-Square Distribution , Elasticity , Female , Humans , Japan , Male , Middle Aged , Plethysmography , Prosthesis Design , Regional Blood Flow , Time Factors , Treatment Outcome
5.
Med J Malaysia ; 63 Suppl A: 42, 2008 Jul.
Article in English | MEDLINE | ID: mdl-19024975

ABSTRACT

The cultivated epithelial transplantation is a new surgical modality for treating a variety of severe ocular surface disorders. This type of tissue-engineered epithelial sheet provides a rapid epithelial coverage on the corneal surface that reduces inflammation and postoperative complications. Although cultivated corneal epithelial transplantation is an effective surgical strategy, autologous transplantation is limited to unilateral cases. Autologous cultivated oral mucosal epithelial transplantation (COMET) enables surgeons to reconstruct the ocular surface using autologous, non-ocular surface cells, and has opened a new pathway for treating severe, bilateral ocular surface disorders.


Subject(s)
Cornea/pathology , Corneal Diseases/surgery , Epithelial Cells/cytology , Epithelium/transplantation , Eye Diseases/surgery , Eye/pathology , Mouth Mucosa/transplantation , Ophthalmologic Surgical Procedures/methods , Tissue Engineering/methods , Cells, Cultured , Cornea/cytology , Corneal Diseases/pathology , Epithelial Cells/transplantation , Eye Diseases/therapy , Humans , Inflammation , Pilot Projects , Plastic Surgery Procedures , Transplantation, Autologous
7.
J Med Microbiol ; 67(9): 1249-1252, 2018 Sep.
Article in English | MEDLINE | ID: mdl-30024367

ABSTRACT

Leptospirosis is an endemic infectious disease causing considerable morbidity and mortality in Sri Lanka; however, reports on the isolation of Leptospira from infected patients in Sri Lanka have been largely unavailable since the 1970s. Two isolates were obtained and characterized from 100 blood cultures from leptospirosis-suspected patients. Phylogenic analysis of partial flaB gene sequences identified the isolates as Leptospira interrogans. The patient serum samples from which Leptospira was isolated reacted with the Leptospira serogroups Sejroe and Canicola at a titre of 1 : 200. Exposure to domestic sewage and gutters filled with muddy water was suspected to be the source of infection in these two culture-positive patients. This study reports the successful isolation of pathogenic Leptospira from two patients in Western Province, Sri Lanka.


Subject(s)
Leptospirosis/microbiology , Adult , Aged, 80 and over , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , DNA, Bacterial/genetics , Female , Humans , Leptospira interrogans/classification , Leptospira interrogans/genetics , Leptospira interrogans/isolation & purification , Male , Phylogeny , Polymerase Chain Reaction , Prospective Studies , Sri Lanka
8.
Transplant Proc ; 48(7): 2392-2395, 2016 Sep.
Article in English | MEDLINE | ID: mdl-27742306

ABSTRACT

OBJECTIVE: The purpose was to review the increase of minority organ donation. METHODS: The methodology was based on the efforts of the DC Organ Donor Program and the Dow Take Initiative Program that focused on increasing donors among African Americans (AAs). From 1982 to 1988, AA donor card signings increased from 20/month to 750/month, and Black donations doubled. A review of the data, including face-to-face grassroots presentations combined with national media, was conducted. Gallup polls in 1985 and 1990 indicated a tripling of black awareness of transplantation and the number of blacks signing donor cards. Based on the applied successful methodologies, in 1991, the National Minority Organ Tissues Transplant Education Program was established targeting AA, Hispanic, Asian, and other ethnic groups. A review of the United Network for Organ Sharing (UNOS) database from 1990 to 2010 was accomplished. RESULTS: Nationally, ethnic minority organ donors per million (ODM) increased from 8-10 ODM (1982) to 35 ODM (AA and Latino/Hispanics) in 2002. In 1995, ODMs were white 34.2, black 33.1, Hispanic 31.5, and Asian 17.9. In 2010, Black organ donors per million totaled 35.36 versus white 27.07, Hispanic 25.59, and Asian 14.70. CONCLUSIONS: Based on the data retrieved from UNOS in 2010, blacks were ranked above whites and other ethnic minority populations as the number one ethnic group of organ donors per million in the US.


Subject(s)
Black or African American/statistics & numerical data , Health Education/methods , Minority Groups/statistics & numerical data , Tissue Donors/statistics & numerical data , Tissue and Organ Procurement/trends , Black or African American/education , Asian/education , Asian/statistics & numerical data , Ethnicity/education , Ethnicity/statistics & numerical data , Health Promotion , Hispanic or Latino/education , Hispanic or Latino/statistics & numerical data , Humans , Mass Media , Minority Groups/education , Power, Psychological , Tissue Donors/education , United States , White People
9.
Eur J Surg Oncol ; 42(8): 1236-46, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27055944

ABSTRACT

BACKGROUND: The establishment of a precise and rapid method to detect metastatic lymph nodes (LNs) is essential to perform less invasive surgery with reduced gastrectomy along with reduced lymph node dissection. We herein describe a novel imaging strategy to detect 5-aminolevulinic acid (5-ALA)-induced protoporphyrin IX (PpIX) fluorescence in excised LNs specifically with reduced effects of tissue autofluorescence based on photo-oxidation of PpIX. We applied the method in a clinical setting, and evaluated its feasibility. METHODS: To reduce the unfavorable effect of autofluorescence, we focused on photo-oxidation of PpIX: Following light irradiation, PpIX changes into another substance, photo-protoporphyrin, via an oxidative process, which has a different spectral peak, at 675 nm, whereas PpIX has its spectral peak at 635 nm. Based on the unique spectral alteration, fluorescence spectral imaging before and after light irradiation and subsequent originally-developed image processing was performed. Following in vitro study, we applied this method to a total of 662 excised LNs obtained from 30 gastric cancer patients administered 5-ALA preoperatively. RESULTS: Specific visualization of PpIX was achieved in in vitro study. The method allowed highly sensitive detection of metastatic LNs, with sensitivity of 91.9% and specificity of 90.8% in the in vivo clinical trial. Receiver operating characteristic analysis indicated high diagnostic accuracy, with the area under the curve of 0.926. CONCLUSIONS: We established a highly sensitive and specific 5-ALA-induced fluorescence imaging method applicable in clinical settings. The novel method has a potential to become a useful tool for intraoperative rapid diagnosis of LN metastasis.


Subject(s)
Adenocarcinoma/pathology , Aminolevulinic Acid , Light , Lymph Nodes/pathology , Photosensitizing Agents , Protoporphyrins , Spectrometry, Fluorescence/methods , Stomach Neoplasms/pathology , Adenocarcinoma/surgery , Adult , Aged , Aged, 80 and over , Cell Line, Tumor , Female , Humans , Image Processing, Computer-Assisted , In Vitro Techniques , Lymph Node Excision , Lymph Nodes/diagnostic imaging , Lymphatic Metastasis , Male , Middle Aged , Neoplasm Staging , Optical Imaging , Oxidation-Reduction , Stomach Neoplasms/surgery
10.
Biochim Biophys Acta ; 1568(1): 13-20, 2001 Nov 07.
Article in English | MEDLINE | ID: mdl-11731080

ABSTRACT

One of the hurdles to adenovirus (Ad)-mediated gene transfer is that Ad vectors mediate inefficient gene transfer into cells lacking in the primary receptors, Coxsackievirus and adenovirus receptor (CAR). We previously developed a fiber-mutant Ad vector containing the Arg-Gly-Asp (RGD)-containing peptide motif on the HI loop of the fiber knob, and showed that the mutant vector had enhanced gene transfer activity to human glioma cells, which showed little CAR expression, compared to the vector containing wild type fiber. In this study, the feasibility of the Ad vector containing RGD peptide on the fiber knob was examined in a wide variety of cell types: CAR-positive or -negative human tumor cells, mouse cells, and leukemia cells. The mutant vector infected the cells, which lacked CAR expression but showed alpha(v) integrin expression, about 10-1000 times more efficiently than the vector containing wild type fiber via an RGD-integrin (alpha(v)beta3 and alpha(v)beta5)-dependent, CAR-independent cell entry pathway. The results of this study indicate that Ad vector containing RGD peptide on the fiber knob could be of great utility for gene therapy and gene transfer experiments.


Subject(s)
Adenoviridae/genetics , Gene Transfer Techniques , Oligopeptides/genetics , Animals , Cell Line , Genetic Therapy , Genetic Vectors , Humans , Integrins/analysis , Mice , Receptors, Virus/analysis , Receptors, Vitronectin/analysis , Tumor Cells, Cultured
11.
Biochim Biophys Acta ; 1444(1): 131-7, 1999 Jan 18.
Article in English | MEDLINE | ID: mdl-9931470

ABSTRACT

Bacillus thuringiensis Cry1Aa toxin binds to a 120 kDa putative receptor protein in the Bombyx mori midgut. Recently, this protein was purified and identified as glycosyl-phosphatidylinositol (GPI) anchored aminopeptidase N (APN). In this study, a full-length cDNA thought to encode this 120 kDa APN was isolated and sequenced. It has a 2958 bp ORF encoding 986 amino acids. In the deduced amino acid sequence, we identified GPI-anchor and zinc-metallopeptidase signals, which are the same as those of APNs of other insects that are reported to be putative Cry1 toxin receptors. The B. mori APN amino acid sequence also has a high similarity with those of the other APNs. Subsequently, the recombinant APN was expressed by Escherichia coli and its Cry1Aa toxin binding ability was analyzed. Ligand blotting showed that Cry1Aa toxin bound to the recombinant APN.


Subject(s)
Bacillus thuringiensis/genetics , Bacterial Proteins/genetics , Bacterial Toxins , Bombyx/genetics , CD13 Antigens/genetics , DNA, Complementary/isolation & purification , Endotoxins/genetics , Amino Acid Sequence , Animals , Bacillus thuringiensis/metabolism , Bacillus thuringiensis Toxins , Bacterial Proteins/chemistry , Bacterial Proteins/metabolism , Base Sequence , Bombyx/metabolism , CD13 Antigens/biosynthesis , CD13 Antigens/metabolism , Cloning, Molecular , DNA, Complementary/metabolism , Endotoxins/chemistry , Endotoxins/metabolism , Escherichia coli/metabolism , Glycosylphosphatidylinositols/metabolism , Hemolysin Proteins , Molecular Sequence Data
12.
Plant Physiol ; 121(3): 813-820, 1999 Nov.
Article in English | MEDLINE | ID: mdl-10557229

ABSTRACT

WPK4, a gene encoding a putative protein kinase, was initially identified in wheat (Triticum aestivum) and shown to be up-regulated by light, nutrient deprivation, and cytokinins. To confirm that WPK4 has protein kinase activity, the protein was produced in Escherichia coli as a fusion protein with glutathione S-transferase. The purified protein exhibited autophosphorylation activity and phosphorylated both myelin basic protein and a peptide fragment of rice 3-hydroxy-3-methylglutaryl-coenzyme A reductase. Levels of WPK4 transcripts in wheat seedlings were increased and decreased by the removal and addition of sucrose (Suc), respectively, to the culture medium. The introduction of the N-terminal kinase region of WPK4 into the yeast snf1 mutant cells, which cannot utilize Suc as a carbon source, rescued growth in Suc-containing medium. Cytokinins up-regulated the accumulation of WPK4 transcripts, but their effects were cancelled by the addition of Suc. Our results suggest that Suc negatively regulates the signaling pathway in which transcriptional activation of WPK4 is mediated by cytokinins.

13.
Placenta ; 26(10): 729-34, 2005 Nov.
Article in English | MEDLINE | ID: mdl-16226122

ABSTRACT

The transfer of genes of interest is a useful method for studying placental biology. Recombinant adenovirus (Ad) vector is an efficient vector for transgene expression. An interaction between the fiber of Ad and the coxsackievirus and adenovirus receptor on the cell membrane is the first step in infection. We previously developed fiber-modified Ad vectors and showed that they improved transgene activity in several cell lines when compared to wild-type vector. In the present study, we examined the ability of three fiber-modified Ad vectors to transduce human choriocarcinoma cell lines (JEG-3, JAR and BeWo) and rat trophoblast cell lines (Rcho-1, TR-TBT 18d-1 and TR-TBT 18d-2). We compared the transgene efficacy of wild-type Ad-L2 vector, Ad-RGD(HI)-L2 vector containing an Arg-Gly-Asp motif, Ad-K7(C)-L2 vector containing a 7-tandem lysine motif, and Ad-RGD(HI)K7(C)-L2 vector containing both motifs in the fiber. We used the luciferase gene as a reporter gene. In the human and rodent trophoblast cell lines, Ad-RGD(HI)-L2 had the greatest infectious potential, followed by Ad-RGD(HI)K7(C)-L2, Ad-K7(C)-L2 and Ad-L2. Compared to the amount of luciferase produced by wild-type vector, Ad-RGD(HI)-L2 mediated 8.1-fold the amount of luciferase in JEG-3 cells, 13.5-fold in JAR cells, 76.8-fold in BeWo cells, 5.0-fold in Rcho-1, 19.4-fold in TR-TBT 18d-1 and 15.0-fold in TR-TBT 18d-2. These results indicate that Ad-RGD(HI) is a potential recombinant Ad vector for transgene expression in some trophoblast cell lines.


Subject(s)
Adenoviridae/genetics , Genetic Vectors/genetics , Transduction, Genetic/methods , Transgenes/genetics , Trophoblasts/virology , Animals , Cell Line, Tumor , Choriocarcinoma/genetics , Choriocarcinoma/metabolism , Choriocarcinoma/virology , Coxsackie and Adenovirus Receptor-Like Membrane Protein , Female , Humans , Integrins/biosynthesis , Integrins/genetics , Integrins/metabolism , Pregnancy , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Rats , Receptors, Virus/biosynthesis , Receptors, Virus/genetics , Receptors, Virus/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Trophoblasts/metabolism , Trophoblasts/physiology
14.
J Neuropathol Exp Neurol ; 36(6): 964-76, 1977 Nov.
Article in English | MEDLINE | ID: mdl-925721

ABSTRACT

A 68-year-old man, after having been diagnosed as having hepatic disease at about the age of 41 years, had been hospitalized frequently until his death. Blood sugar, iron, and copper had not increased during his illness. Although the diagnosis of liver cirrhosis had been made and he had been receiving therapy, various neurologic symptoms without disturbances of consciousness appeared six months before his death. Autopsy revealed hemochromatosis, liver cirrhosis, and pancreatic fibrosis. A large amount of iron had accumulated in the liver, the pancreas, and the thyroid gland, while considerable numbers of ceroid and lipofuscin pigment granules had accumulated diffusely in the brain. Abnormal astrocytes of the Alzheimer II type were diffusely distributed in the brain and contained no intranuclear glycogen which stained positive with the carmine stain. No spongy changes were seen in the deeper layers of the cerebral cortex. Chemical analyses for trace metals in the brain, liver, and kidneys revealed a large amount of iron and increased copper in the liver, and considerable quantities of copper, manganese, calcium, and mercury in the brain. Because of changes in the erythrocyte sedimentation rate and marked thymol turbidity seen before and after the occurrence of the neurologic symptoms, this man was suspected of having disorders of the trace-metal binding proteins and/or of their polymers.


Subject(s)
Blood Proteins/metabolism , Brain Diseases/pathology , Hemochromatosis/pathology , Liver Cirrhosis, Alcoholic/pathology , Metals/metabolism , Trace Elements/metabolism , Aged , Astrocytes/pathology , Brain Chemistry , Calcium/analysis , Copper/analysis , Humans , Iron/analysis , Kidney/analysis , Liver/analysis , Liver/pathology , Male , Manganese/analysis , Mercury/analysis , Polymers/metabolism , Protein Binding , Syndrome
15.
FEBS Lett ; 443(2): 139-43, 1999 Jan 25.
Article in English | MEDLINE | ID: mdl-9989592

ABSTRACT

We recently isolated and characterized the lipopolysaccharide (LPS)-binding protein, BmLBP, from the larval hemolymph of the silkworm Bombyx mori. BmLBP is a pattern recognition molecule that recognizes the lipid A portion of LPS and participates in a cellular defense reaction. This paper describes the cDNA cloning of BmLBP. The deduced amino acid sequence of BmLBP revealed that BmLBP is a novel member of the C-type lectin superfamily with a unique structural feature that consists of two different carbohydrate-recognition domains in tandem, a short and a long form.


Subject(s)
Acute-Phase Proteins , Carbohydrate Metabolism , Carrier Proteins/metabolism , Hemocytes/metabolism , Lectins/metabolism , Membrane Glycoproteins , Amino Acid Sequence , Animals , Base Sequence , Blotting, Southern , Bombyx , Carrier Proteins/chemistry , Cloning, Molecular , DNA, Complementary , Gene Expression Regulation , Lectins/chemistry , Molecular Sequence Data , Sequence Homology, Amino Acid
16.
FEBS Lett ; 463(3): 221-4, 1999 Dec 17.
Article in English | MEDLINE | ID: mdl-10606725

ABSTRACT

The Bacillus thuringiensis Cry1Aa toxin-binding region of Bombyx mori aminopeptidase N (APN) was analyzed, to better understand the molecular mechanism of susceptibility to the toxin and the development of resistance in insects. APN was digested with lysylendopeptidase and the ability of the resulting fragments to bind to Cry1Aa and 1Ac toxins was examined. The binding abilities of the two toxins to these fragments were different. The Cry1Aa toxin bound to the fragment containing 40-Asp to 313-Lys, suggesting that the Cry1Aa toxin-binding site is located in the region between 40-Asp and 313-Lys, while Cry1Ac toxin bound exclusively to mature APN. Next, recombinant APN of various lengths was expressed in Escherichia coli cells and its ability to bind to Cry1Aa toxin was examined. The results localized the Cry1Aa toxin binding to the region between 135-Ile and 198-Pro.


Subject(s)
Bacterial Proteins/chemistry , Bacterial Toxins/chemistry , Bombyx/enzymology , CD13 Antigens/chemistry , Endotoxins/chemistry , Insect Proteins , Amino Acid Sequence , Animals , Bacillus thuringiensis , Bacillus thuringiensis Toxins , Binding Sites , Endopeptidases , Hemolysin Proteins , Insecticide Resistance , Receptors, Cell Surface/chemistry , Recombinant Proteins/chemistry
17.
Invest Ophthalmol Vis Sci ; 41(9): 2506-13, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10937561

ABSTRACT

PURPOSE: Surgery to reconstruct the ocular surface is greatly facilitated by the use of amniotic membrane, either as a biologic drape or, more recently, as a substrate for the transplantation of cultivated corneal epithelial cells. This study was designed to compare the usefulness of intact and denuded human amniotic membranes as a substrate for corneal epithelial cell culture. METHODS: Small (3-mm-diameter) biopsy specimens of superficial cornea including epithelium were excised from the central and limbal regions in rabbits. They were cultured on human amniotic membrane with or without amniotic epithelial cells and examined by light, scanning electron, and transmission electron microscopy. RESULTS: Cellular outgrowth from the central explants (n = 10) after 14 days in culture measured 1.82 +/- 2.62 mm2 on intact amniotic membrane and 131.83 +/- 28.31 mm2 on denuded amniotic membrane. In contrast, outgrowths from the limbal explants (n = 10) at the same time measured 4.58 +/- 4.56 and 505.39 +/- 134.20 mm2 on intact and denuded amniotic membranes, respectively. The leading edges of the outgrowths on intact amniotic membrane were much less uniform than those on denuded amniotic membrane, and, in the former, corneal epithelial cells appeared to migrate over the top of amniotic epithelial cells. Limbal cells cultivated on denuded amniotic membrane formed a nicely stratified layer that adhered well to the underlying amniotic membrane. CONCLUSIONS: Denuded amniotic membrane appears to be an excellent substrate for the cultivation of corneal epithelial cells, with a view to transplantation.


Subject(s)
Amnion , Cell Culture Techniques/methods , Epithelium, Corneal/cytology , 3T3 Cells/cytology , Amnion/cytology , Amnion/ultrastructure , Animals , Coculture Techniques/methods , Epithelium, Corneal/ultrastructure , Humans , Mice , Microscopy, Electron, Scanning , Rabbits
18.
Biochem Pharmacol ; 58(2): 335-41, 1999 Jul 15.
Article in English | MEDLINE | ID: mdl-10423176

ABSTRACT

Osaterone acetate (17alpha-acetoxy-6-chloro-2-oxa-4,6-pregnadiene-3,20-dione, OA) is a new steroidal antiandrogen. There is a marked species difference in the metabolism of OA in that 11beta-hydroxylated metabolites are found in the plasma, feces, and urine of mice after oral administration of OA, but there is very little metabolism in rats and humans. OA reduces the adrenal gland weight in mice, but not in rats, and this effect in mice might be explained by the species difference in 11beta-hydroxylation activity. The objectives of this study were to elucidate the enzyme(s) involved in this particular oxidation and to explain the species difference observed. Mouse hepatic microsomes oxidize OA to 11beta-OH OA, and this oxidation requires NADPH as a cofactor. The use of various competitive and allosteric inhibitors of cytochrome P450 and flavin-containing monooxygenase (i.e. CO, N-octylamine, and methimazole) showed that the oxidation of OA was catalyzed by cytochrome P450. In microsomes from mice pretreated with phenobarbital (a CYP2B-selective inducer), 3-methylcholanthrene (a CYP1A-selective inducer), pregnenolone-16alpha-carbonitrile (a CYP3A-selective inducer), and EtOH (a CYP2E-selective inducer), an increase in the rates of oxidation was seen only in microsomes from EtOH-treated animals. However, metyrapone, a selective inhibitor for enzymes of the cytochrome P45011B and P4502B family, inhibited mouse hepatic microsomal 11beta-hydroxylation by < 30%. The results obtained showed that the production of 11beta-OH OA may be catalyzed by a novel cytochrome P450 in mouse liver.


Subject(s)
Androgen Antagonists/metabolism , Chlormadinone Acetate/analogs & derivatives , Microsomes, Liver/enzymology , Steroid 11-beta-Hydroxylase/metabolism , Animals , Catalysis , Chlormadinone Acetate/metabolism , Enzyme Induction , Humans , Hydroxylation , In Vitro Techniques , Kinetics , Liver/enzymology , Male , Mice , Microsomes, Liver/metabolism , Rats , Steroid 11-beta-Hydroxylase/antagonists & inhibitors , Steroid 11-beta-Hydroxylase/biosynthesis , Tissue Distribution
19.
Environ Health Perspect ; 28: 223-43, 1979 Feb.
Article in English | MEDLINE | ID: mdl-114386

ABSTRACT

Ten male rhesus monkeys, each weighing 3.5 kg, were divided into four groups of 3, 3, 2, and 2, and were fed daily with 100 g pelleted food containing 300, 30, 3, and 0 ppm cadmium, respectively. Urine samples were collected every 2 weeks and blood samples every 4 weeks. One monkey each of the 300 and 30 ppm groups was autopsied for pathological examination and tissue cadmium determination at the week 24 of the experiment; the remaining 8 animals were killed after 55 weeks. The lowest exposed group (3 ppm) did not show any specific biological response to cadmium over a period of 55 weeks. In the 30 ppm group, no significant changes were observed for up to 24 weeks, although cadmium concentration in the renal cortex and urine at 24 weeks were 300 mug/g wet weight and 18 mug/l., respectively. Plasma urea nitrogen and urine protein (quantitative determination) increased after 30 and 36 weeks. At 55 weeks of the experiment, qualitative tests were negative for low molecular weight proteinuria and glycosuria, and the results remained normal for renal and liver function tests and blood analysis, although cadmium concentrations in the renal cortex of two monkeys were 460 and 730 mug/g wet weight and those in the liver were 110 and 160 mug/g wet weight, respectively. In the highest exposure group (300 ppm), urine cadmium increased to 250 mug/l. by 11 weeks, and urine retinol-binding protein, plasma GOT, GPT, and LDH increased after 12 weeks. Proteinuria (quantitative determination), glycosuria, aminoaciduria (panaminoaciduria), and erythrocytopenia were observed after 16 weeks, when urine cadmium was 500-900 mug/l. Hypohemoglobinopathy and proteinuria (qualitative determination) were observed after 20 and 24 weeks, while cadmium concentrations in the renal cortex and the liver were 760 and 430 mug/g wet weight at 24 weeks, respectively. Slightly depressed tubular reabsorption of phosphate, increased urine beta(2)-microglobulin, increased plasma urea nitrogen, and increased plasma alpha(2)-globulin fraction (electrophoresis) were observed between 28 and 30 weeks of the experiment. Creatinine clearance and plasma cholinesterase decreased after 47 and 54 weeks, respectively. Cadmium concentrations in the renal cortex and the liver of two monkeys at 55 weeks were 350 and 580 mug/g wet weight and 410 and 630 mug/g wet weight, respectively. Pathological examinations revealed denaturation, destruction, and regeneration of the epithelial cells in renal proximal tubules, but no pathological changes in osseous tissues. Critical cadmium concentration in the renal cortex was estimated to be 380 mug/g wet weight for low molecular weight proteinuria and 470 mug/g wet weight for proteinuria, glycosuria, and aminoaciduria. Critical concentration in the liver was also estimated to be 210 mug/g wet weight. The apparent biological half-time of cadmium in monkeys at autopsied stage was calculated to be 0.66, 6.4, 5.2, and 22.4 years for the 300, 30, 3, and 0 ppm groups, respectively.


Subject(s)
Cadmium/toxicity , Kidney/drug effects , Animals , Blood Pressure , Body Weight , Cadmium/metabolism , Diet , Dose-Response Relationship, Drug , Electrocardiography , Haplorhini , Kidney/pathology , Kidney Tubules/metabolism , Kidney Tubules/pathology , Macaca , Male , Phosphates/metabolism , Tissue Distribution
20.
J Thorac Cardiovasc Surg ; 115(4): 811-8, 1998 Apr.
Article in English | MEDLINE | ID: mdl-9576215

ABSTRACT

OBJECTIVE: To predict spinal cord ischemia after endovascular stent graft repair of descending thoracic aortic aneurysms, temporary interruption of the intercostal arteries (including the aneurysm) was performed by placement of a novel retrievable stent graft (Retriever) in the aorta under evoked spinal cord potential monitoring. METHODS: From February 1995 to October 1997, endovascular stent graft repair of descending thoracic aortic aneurysms was performed in 49 patients after informed consent was obtained. In 16 patients with aneurysms located in the middle and distal segment of the descending aorta, the Retriever was placed temporarily before stent graft deployment. The Retriever consisted of two units of self-expanding zigzag stents connected in tandem with stainless steel struts. Each strut was collected in a bundle fixed to a pushing rod, and the stent framework was lined with an expanded polytetrafluoroethylene sheet. The Retriever was delivered beyond the aneurysm through a sheath and was retracted into the sheath 20 minutes later. A stent graft for permanent use was deployed in patients whose predeployment test results with the Retriever were favorable. Evoked spinal cord potential was monitored throughout placement of the Retriever and stent grafting until the next day. RESULTS: The Retriever was placed in 17 aneurysms in 16 patients. There were no changes in amplitude or latency of evoked spinal cord potential records obtained before or during Retriever placement. After withdrawal of the Retriever, all aneurysms were excluded from circulation immediately after permanent stent grafting. There were no changes in evoked spinal cord potential, nor were neurologic deficits seen after stent graft deployment in any patient. CONCLUSIONS: These results suggest that predeployment testing with the Retriever under evoked spinal cord potential monitoring is promising as a predictor of spinal cord ischemia in candidates for stent graft repair of thoracic aortic aneurysms.


Subject(s)
Aortic Aneurysm, Thoracic/surgery , Blood Vessel Prosthesis Implantation , Ischemia/epidemiology , Monitoring, Intraoperative/instrumentation , Spinal Cord/blood supply , Stents , Aged , Equipment Design , Evoked Potentials , Female , Humans , Ischemia/diagnosis , Ischemia/prevention & control , Male , Monitoring, Intraoperative/methods , Postoperative Complications/epidemiology , Postoperative Complications/prevention & control , Postoperative Period , Predictive Value of Tests , Spinal Cord/physiology
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