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1.
Int J Med Sci ; 20(5): 566-571, 2023.
Article in English | MEDLINE | ID: mdl-37082737

ABSTRACT

In Japan, China, and Singapore, several studies have reported increased incidences of peripheral venous catheter-related bloodstream infection by Bacillus cereus during the summer. Therefore, we hypothesized that bed bathing with a B. cereus-contaminated "clean" towels increases B. cereus contact with the catheter and increases the odds of contaminating the peripheral parenteral nutrition (PPN). We found that 1) professionally laundered "clean" towels used in hospitals have B. cereus (3.3×104 colony forming units (CFUs) / 25cm2), 2) B. cereus is transferable onto the forearms of volunteers by wiping with the towels (n=9), and 3) B. cereus remain detectable (80∼660 CFUs /50cm2) on the forearms of volunteers even with subsequent efforts of disinfection using alcohol wipes. We further confirmed that B. cereus grow robustly (102 CFUs /mL to more than 106 CFUs /mL) within 24hours at 30°C in PPN. Altogether we find that bed bathing with a towel contaminated with B. cereus leads to spore attachments to the skin, and that B. cereus can proliferate at an accelerated rate at 30°C compared to 20°C in PPN. We therefore highly recommend ensuring the use of sterile bed bath towels prior to PPN administration with catheter in patients requiring bed bathing.


Subject(s)
Cross Infection , Sepsis , Humans , Bacillus cereus , Parenteral Nutrition Solutions , Cross Infection/epidemiology , Cross Infection/etiology , Cross Infection/prevention & control , Hospitals , Parenteral Nutrition/adverse effects , Risk Factors , Catheters
2.
Cytotechnology ; 75(1): 39-47, 2023 Feb.
Article in English | MEDLINE | ID: mdl-36713066

ABSTRACT

ß-lactoglobulin (BLG) and ε-polylysine (PL) were bound by using microbial transglutaminase. Dextran (Dex) was further conjugated to the BLG-PL conjugate by the Maillard reaction. Confirmation of conjugation was carried out by SDS-PAGE. From the results of isoelectric focusing, it was revealed that the isoelectric point of the BLG-PL conjugate was shifted to the basic side as compared with native BLG. Immunogenicity of BLG in BALB/c mice was lowered by conjugation with PL and further lowered by conjugation with Dex. By conjugation with PL and PL-Dex, antibacterial activity against Staphylococcus aureus was endowed to BLG. Because the conjugation method in this study is a safe method, it is valuable in that it can be applicable to food processing.

3.
J Water Health ; 20(7): 1150-1156, 2022 Jul.
Article in English | MEDLINE | ID: mdl-35902996

ABSTRACT

We report a patient who developed pneumonia after prolonged use of spray bottle containing green tea for hydration purposes. The cause was suspected to be a contamination of green tea because the patient's symptoms persisted and did not improve until stopping the use of the spray bottle and we also found the green tea in the spray bottle to harbor a high number of Pseudomonas aeruginosa (1.2 × 107 colony forming units (cfu)/mL). It is not uncommon to use green tea for hydration or gargling purposes in some patient care settings considering the antibacterial effects of catechins contained in green tea. Our findings suggest the importance of keeping vigilance on consuming green tea in spray bottles in hospital settings since it may readily be contaminated by pathogens such as P. aeruginosa.


Subject(s)
Pneumonia , Tea , Anti-Bacterial Agents/pharmacology , Humans , Plant Extracts , Pseudomonas aeruginosa
4.
J Water Health ; 20(1): 271-275, 2022 Jan.
Article in English | MEDLINE | ID: mdl-35100172

ABSTRACT

Bidet toilets (electric toilet seats with water spray) are increasing in popularity worldwide. However, the extent of reduction of microbial contamination of the hands with the use of bidet toilets after defecation is unclear. Microbe contamination of the hands with and without the use of bidet toilets after defecation was examined in 32 nursing students. Double gloves were worn on the dominant hand and four layers of toilet paper were used to wipe the buttocks after defecation, and microbe contamination of the second glove (outer glove) of the double gloves was examined. The volunteers were free to select the flow volume, wash time of the bidet, and the type of bidet. Without the use of a bidet toilet, the average value ± standard deviation of the number of microbes attached to the gloves was 39,499.3 ± 77,768.3 colony forming units (cfu)/glove; however, it was 4,146.9 ± 11,427.7 cfu/glove when the bidet toilet was used. The number of microbes adhering to gloves was significantly reduced when a bidet toilet was used (p < 0.00001, Wilcoxon signed-rank test).


Subject(s)
Bathroom Equipment , Defecation , Humans , Toilet Facilities , Water
5.
Plant Biotechnol (Tokyo) ; 38(3): 379-386, 2021 Sep 25.
Article in English | MEDLINE | ID: mdl-34782826

ABSTRACT

Lignocellulosic materials are potential renewable sources of fermentable sugars for bioethanol production. In this study, we used the CcAbf62A gene encoding CcAbf62A, a putative extracellular α-L-arabinofuranosidase, cloned from the mycotrophic basidiomycete Coprinopsis cinerea. CcAbf62A acts on arabinoxylan, the major hemicellulose of grasses, releasing arabinose. CcAbf62A was introduced into rice with the aim of enhancing delignification efficiency and the availability of lignocellulosic materials without reducing lignin content. Among the 32 lines of regenerated transgenic rice, 13 exhibited markedly disrupted elongation growth and excessive tillering (dwarf), seven showed delayed elongation growth (retarded-growth), and 12 showed phenotypes similar to those of control plants (normal). Additionally, the dwarf lines showed reduced acclimation. RT-PCR analysis revealed that dwarf lines had higher levels of CcAbf62A expression than retarded-growth and normal lines. Although the lignin content of transgenic rice plants expressing CcAbf62A did not differ significantly from that of control rice plants, dwarf lines were characterized by delayed deposition of lignin in the culms compared with the controls. The reduced acclimation ability of dwarf lines is believed to be associated with increased water loss and reduced water conductivity concomitant with delayed lignin deposition. Contrary to expectations, the alkaline delignification rates of dwarf and retarded-growth Abf lines were slightly lower than those of control rice plants. Our findings indicate that CcAbf62A reduces ferulate-lignin cross-links by detaching arabinose side chains from arabinoxylan and increases the relative abundance of alkaline-resistant benzyl ether cross-links. CcAbf62A is anticipated to provide new approaches for breeding plants containing altered lignocellulosic materials or lodging-resistant crops.

6.
Plant Cell Rep ; 31(9): 1573-80, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22547095

ABSTRACT

UNLABELLED: Secondary xylem is composed of daughter cells produced by the vascular cambium in the stem. Cell proliferation of the secondary xylem is the result of long-range cell division in the vascular cambium. Most xylem cells have a thickened secondary cell wall, representing a large amount of biomass storage. Therefore, regulation of cell division in the vascular cambium and differentiation into secondary xylem is important for biomass production. Cell division is regulated by cell cycle regulators. In this study, we confirm that cell cycle regulators influence cell division in the vascular cambium in tobacco. We produced transgenic tobacco that expresses Arabidopsis thaliana cyclin D2;1 (AtcycD2;1) and AtE2Fa-DPa under the control of the CaMV35S promoter. Each gene is a positive regulator of the cell cycle, and is known to influence the transition from G1 phase to S phase. AtcycD2;1-overexpressing tobacco had more secondary xylem cells when compared with control plants. In order to evaluate cell division activity in the vascular cambium, we prepared a Populus trichocarpa cycB1;1 (PtcycB1;1) promoter containing a destruction box motif for ubiquitination and a ß-glucuronidase-encoding gene (PtcycB1;1pro:GUS). In transgenic tobacco containing PtcycB1;1pro:GUS, GUS staining was specifically observed in meristem tissues, such as the root apical meristem and vascular cambium. In addition, mitosis-monitoring plants containing AtcycD2;1 had stronger GUS staining in the cambium when compared with control plants. Our results indicated that overexpression of AtcycD enhances cell division in the vascular cambium and increases secondary xylem differentiation in tobacco. KEY MESSAGE: We succeeded in inducing cell proliferation of cambium and enlargement of secondary xylem region by AtcycD overexpression. We also evaluated mitotic activity in cambium using cyclin-GUS fusion protein from poplar.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Cyclins/metabolism , Nicotiana/cytology , Nicotiana/genetics , Xylem/cytology , Arabidopsis Proteins/genetics , Cambium/cytology , Cambium/metabolism , Cell Proliferation , Cyclins/genetics , E2F Transcription Factors/genetics , E2F Transcription Factors/metabolism , G1 Phase , Gene Expression Regulation, Plant , Glucuronidase/metabolism , Mitosis , Plant Stems/cytology , Plant Stems/metabolism , Plant Vascular Bundle/cytology , Plant Vascular Bundle/metabolism , Plants, Genetically Modified , Reverse Transcriptase Polymerase Chain Reaction , S Phase , Staining and Labeling , Transfection , Xylem/metabolism
7.
Plant Cell Environ ; 35(11): 2031-44, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22574770

ABSTRACT

Rice internodes are vital for supporting high-yield panicles, which are controlled by various factors such as cell division, cell elongation and cell wall biosynthesis. Therefore, formation and regulation of the internode cell-producing intercalary meristem (IM) are important for determining the shape of internodes. To understand the regulation of internode development, we analysed a rice dwarf mutant, dwarf 50 (d50). Previously, we reported that parenchyma cells in the elongated internodes of d50 ectopically deposit cell wall phenolics. In this study, we revealed that D50 encodes putative inositol polyphosphate 5-phosphatase (5PTase), which may be involved in phosphoinositide signalling required for many essential cellular functions, such as cytoskeleton organization, endocytosis and vesicular trafficking in eukaryotes. Analysis of the rice genome revealed 20 putative 5PTases including D50. The d50 mutation induced abnormally oriented cell division, irregular deposition of cell wall pectins and thick actin bundles in the parenchyma cells of the IM, resulting in abnormally organized cell files of the internode parenchyma and dwarf phenotype. Our results suggest that the putative 5PTase, encoded by D50, is essential for IM formation, including the direction of cell division, deposition of cell wall pectins and control of actin organization.


Subject(s)
Meristem/genetics , Oryza/enzymology , Phosphoric Monoester Hydrolases/physiology , Plant Proteins/physiology , Actins/metabolism , Actins/ultrastructure , Amino Acid Sequence , Cell Division/genetics , Cell Shape , Cell Wall/metabolism , Chromosome Mapping , Cloning, Molecular , Inositol Polyphosphate 5-Phosphatases , Microtubules/metabolism , Microtubules/ultrastructure , Molecular Sequence Data , Mutation , Oryza/genetics , Oryza/growth & development , Pectins/metabolism , Phosphoric Monoester Hydrolases/genetics , Phosphoric Monoester Hydrolases/metabolism , Phylogeny , Plant Proteins/genetics , Plant Proteins/metabolism , Sequence Alignment
8.
Plant Cell Rep ; 30(12): 2195-205, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21800099

ABSTRACT

Rice internodes must have the proper shape to support high-yielding panicles. The shape of internodes is controlled by various factors involved in their formation, such as developmental patterns, cell division, cell elongation, and cell wall biosynthesis. To understand the regulation of internode development, we screened dwarf mutants to identify those with a phenotype of ectopic deposits of phenolic components in parenchyma cell walls of internodes. We named these mutants ectopic deposition of phenolic components1 (edp1). Two alleles were identified, edp1-1 and edp1-2. Furthermore, these mutants showed disordered cell files in internode parenchyma. These abnormal phenotypes were very similar to that of a previously reported dwarf50 (d50) mutant. Genetic analyses of edp1 mutants revealed that the edp1 loci are distinct from d50. Our results indicate that analyses of edp1 mutants as well as the d50 mutant will be useful for understanding the molecular mechanisms behind ectopic deposition of cell wall phenolic components in internode parenchyma cells and the regulation of internode development.


Subject(s)
Cell Wall/metabolism , Lignin/metabolism , Mesophyll Cells/metabolism , Oryza/growth & development , Phenols/metabolism , Alleles , Crosses, Genetic , Gene Expression Regulation, Plant , Mutation , Oryza/genetics , Plant Stems/anatomy & histology
9.
Tree Physiol ; 28(1): 21-8, 2008 Jan.
Article in English | MEDLINE | ID: mdl-17938110

ABSTRACT

We report the isolation and characterization of CjNdly, a homolog in Japanese cedar (Cryptomeria japonica D. Don) of the FLORICAULA/LEAFY (FLO/LFY) genes. We determined the entire nucleotide sequence of CjNdly, including short 5'- and 3'-untranslated regions. The deduced amino acid sequence was similar to those of the products of the FLO/LFY genes from other species. The nucleotide sequence showed the closest homology to that of the NEEDLY gene in Pinus radiata D. Don. Although no proline-rich region has been reported previously in homologous gene products from gymnosperms, we found such a region at the amino-terminal end of the deduced amino acid sequence encoded by CjNdly. We detected the expression of CjNdly in both reproductive and vegetative tissues and organs of C. japonica. Heterologous expression of CjNdly in transgenic tobacco plants induced precocious flowering of regenerating shoots on agar-solidified medium and flowers with an abnormal phenotype, namely, petal-like stamens. Our findings suggest that the CjNdly gene may have important roles in flower development in Japanese cedar, resembling those of its angiosperm homologs.


Subject(s)
Cryptomeria/genetics , Genes, Plant , Plant Leaves/genetics , 3' Untranslated Regions/genetics , 5' Untranslated Regions/genetics , Amino Acid Sequence , Conserved Sequence , Cryptomeria/classification , DNA Primers , DNA, Plant/genetics , Molecular Sequence Data , Phylogeny , Plant Proteins/chemistry , Plant Proteins/genetics , RNA, Plant/genetics
10.
Eur Spine J ; 16(11): 1951-7, 2007 Nov.
Article in English | MEDLINE | ID: mdl-17549525

ABSTRACT

No clinical diagnostic support tool can help identify patients with LSS. Simple diagnostic tool may improve the accuracy of the diagnosis of LSS. The aim of this study was to develop a simple clinical diagnostic tool that may help physicians to diagnose LSS in patients with lower leg symptoms. Patients with pain or numbness of the lower legs were prospectively enrolled. The diagnosis of LSS by experienced orthopedic specialists was the outcome measure. Multivariable logistic regression analysis identified factors that predicted LSS; a simple clinical prediction rule was developed by assigning a risk score to each item based on the estimated beta-coefficients. From December 2002 to December 2004, 104 orthopedic physicians from 22 clinics and 50 hospitals evaluated 468 patients. Two items of physical examination, three items of patients' symptom, and five items of physical examination were included in the final scoring system as a result of multiple logistic regression analysis. The sum of the risk scores for each patient ranged from -2 to 16. The Hosmer-Lemeshow statistic was 11.30 (P = 0.1851); the area under the ROC curve was 0.918. The clinical diagnostic support tool had a sensitivity of 92.8% and a specificity of 72.0%. The prevalence of LSS was 6.3% in the bottom quartile of the risk score (-2 to 5) and 99.0% in the top quartile (12 to 16). We developed a simple clinical diagnostic support tool to identify patients with LSS. Further studies are needed to validate this tool in primary care settings.


Subject(s)
Diagnostic Techniques and Procedures , Lumbar Vertebrae/pathology , Spinal Stenosis/diagnosis , Adult , Aged , Aged, 80 and over , Demography , Female , Humans , Incidence , Japan/epidemiology , Magnetic Resonance Imaging , Male , Middle Aged , Risk Factors , Software Design , Spinal Stenosis/epidemiology
12.
Clin Orthop Relat Res ; (436): 208-15, 2005 Jul.
Article in English | MEDLINE | ID: mdl-15995443

ABSTRACT

UNLABELLED: This study was done to assess the long-term functional outcome of very elderly patients with hip fractures, to determine whether bone mineral density and prevalent vertebral fractures could affect mortality and ambulatory status, and to examine which patient characteristics reported in the literature are predictive of patient mortality and ambulatory status. Seventy-four patients 90 years and older with hip fractures were analyzed and followed up for at least 4 years or until death. The mean age of the patients was 92.8 years and all were treated surgically. Walking ability before injury was better than at discharge; walking ability decreased during the first year after discharge, but thereafter reached a plateau. The predictors of survival were the preoperative American Society of Anesthesiologists score, walking ability, fracture type, type of surgery, and the number of prevalent vertebral fractures on admission. Dementia and the number of prevalent vertebral fractures were predictors of the recovery of walking ability. Type of surgery and fracture type are collinear variables, and because it is difficult to separate the effects of one versus the other, additional well-designed, randomized studies on the effect of the type of surgery and fracture type on outcome are needed. LEVEL OF EVIDENCE: Prognostic study, Level II-1 (retrospective study). See the Guidelines for Authors for a complete description of levels of evidence.


Subject(s)
Activities of Daily Living , Frail Elderly , Health Status Indicators , Hip Fractures/mortality , Walking , Absorptiometry, Photon , Aged , Aged, 80 and over , Bone Density , Bone and Bones/diagnostic imaging , Bone and Bones/metabolism , Comorbidity , Female , Hip Fractures/metabolism , Hip Fractures/physiopathology , Humans , Japan/epidemiology , Male , Osteoporosis/mortality , Retrospective Studies , Risk Factors , Spinal Fractures/metabolism , Spinal Fractures/mortality , Survival Rate
13.
Br J Plast Surg ; 58(4): 533-40, 2005 Jun.
Article in English | MEDLINE | ID: mdl-15897039

ABSTRACT

We studied the effects of peripheral nerve adhesion in a rabbit sciatic nerve model. After nerve exposure, its adventitial layer was sutured with 8-0 nylon to the nerve bed, which had been cauterised to promote adhesion. Nerve kinematics, electrophysiology, blood flow and histology were assessed. Rabbits in which Fontana's bands were visible as normal through the epineurium, classified as a nonadhesion group (group I), lacked intraneural fibrosis. In this group, nerve conduction and nerve blood flow were well maintained. Rabbits in which Fontana's bands could not be seen were classified as the adhesion group (group II). This group was classified into two levels pathologically; thickening of epineurium and perineurium was observed but no endoneurial fibrosis (group IIa), and endoneurial fibrosis (Wallerian degeneration, myelin sheath thinning and fibrosis between nerve fibers) was noted (group IIb). Compound muscle action potentials (CMAP) were reduced in amplitude and blood flow was significantly decreased at adhesion sites in group IIb. In conclusion, adhesion of peripheral nerve to surrounding tissues results in fibrosis in the nerve that contributes to peripheral nerve dysfunction.


Subject(s)
Nerve Compression Syndromes/surgery , Peripheral Nerves/pathology , Peripheral Nervous System Diseases/surgery , Action Potentials , Adhesiveness , Animals , Cicatrix/etiology , Cicatrix/pathology , Disease Models, Animal , Fibrosis , Male , Movement , Neural Conduction , Peripheral Nerves/physiopathology , Postoperative Complications , Rabbits , Regional Blood Flow , Sciatic Nerve/blood supply , Sciatic Nerve/pathology , Sciatic Nerve/physiopathology , Sciatic Nerve/surgery , Severity of Illness Index , Tissue Adhesions/etiology , Tissue Adhesions/pathology , Tissue Adhesions/physiopathology
14.
J Surg Res ; 124(1): 38-44, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15734477

ABSTRACT

BACKGROUND: The movement of cells from a transplanted tissue into the host organs, the so-called systemic chimerism, is a phenomenon known to occur and be associated with the development of immunologic tolerance in allotransplantation cases. The purpose of this study was to identify donor cell engraftment in recipient lymphoid tissues after performing rat hind limb allograft. MATERIALS AND METHODS: Fifty-five whole-limb allotransplantations were performed in sex-mismatched pairs of rats. Syngeneic male Lewis and allogeneic Dark Agouti donors were transplanted to female Lewis recipients. FK506 was used for immunosuppression. Donor male cells could be identified in the recipient female tissues by semiquantitative polymerase chain reaction analysis for a Y chromosome-specific DNA sequence. Chimerism was assessed at 1, 24, and 48 weeks after transplantation. RESULTS: There was no rejection episode in any of the limb grafts. Although levels of chimerism were highly variable in each lymphoid tissue, a gradual increase was noticed in all during the course of time. At 1 week after the transplant period, only intrasplenic chimerism was at high level (1%) in three groups. At 48 weeks after the transplant, all recipients with allografts showed very high level (10%) of chimerism in the bone marrow. Two, two, and two of six recipients showed very high levels in the spleen, lymph node, and liver, respectively, at 48 weeks. Intrathymic chimerism was higher at 24 weeks after transplant rather than at 48 weeks. CONCLUSION: We demonstrated donor cell engraftment into recipient lymphoid tissues after successful whole limb transplantation. We conclude that limb allograft can work as a vascularized carrier for the bone marrow transplantation, provide a continuous source of donor cells and contribute to chimerism in the recipient.


Subject(s)
Cell Movement/immunology , Extremities/transplantation , Lymphoid Tissue/immunology , Tissue Transplantation , Transplantation Tolerance/immunology , Animals , Chimerism , Female , Immunosuppressive Agents/immunology , Male , Models, Animal , Rats , Rats, Inbred Strains , Tacrolimus/immunology , Time Factors , Transplantation, Homologous/immunology
15.
Am J Med ; 117(8): 549-55, 2004 10 15.
Article in English | MEDLINE | ID: mdl-15465502

ABSTRACT

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editor in Chief because of the stated concerns listed below. This article was accepted for publication by a previous editor and editorial board, at a time when submissions and documentation were in paper form, prior to the transition of The American Journal of Medicine to a digital submission and review process. These records are no longer extant and consequently we are unable to review the comments of the reviewers and editors involved at that time. We have attempted to contact the authors regarding these concerns and received no response. We are therefore retracting this article since the evidence presented below strongly argues for scientific misconduct. The integrity of these publications is severely compromised by wide-ranging and serious concerns about governance, ethics, authorship, implausible study conduct, implausible workload, discrepant participant numbers and treatment groups, impossible data, implausible data, implausible outcome data, and discrepant methodology.


Subject(s)
Adjuvants, Immunologic/therapeutic use , Bone Density/drug effects , Calcitonin/therapeutic use , Estrogen Replacement Therapy , Etidronic Acid/therapeutic use , Hydroxycholecalciferols/therapeutic use , Osteoporosis, Postmenopausal/drug therapy , Spinal Fractures/prevention & control , Vitamin K/therapeutic use , Aged , Female , Humans , Japan , Middle Aged , Osteoporosis, Postmenopausal/complications , Spinal Fractures/etiology
16.
Clin Rheumatol ; 23(5): 467-9, 2004 Oct.
Article in English | MEDLINE | ID: mdl-15459820

ABSTRACT

Subcutaneous rheumatoid nodules occur commonly in advanced cases of rheumatoid arthritis and are the most common extra-articular lesion of this disease. We present a case of a very unusual giant rheumatoid nodule that developed on the lateral side of a knee. The case was devoid of systemic symptoms of arthritis and the lesion was limited to a rheumatoid nodule. The nodule was successfully treated by surgical excision. However, other new nodules developed in her hand. Her clinical course has not been satisfactory.


Subject(s)
Rheumatoid Nodule/pathology , Aged , Aged, 80 and over , Female , Granuloma Annulare/pathology , Granuloma Annulare/surgery , Humans , Knee/pathology , Magnetic Resonance Imaging , Recurrence , Rheumatoid Nodule/surgery , Treatment Outcome
17.
Biochem Biophys Res Commun ; 322(3): 918-22, 2004 Sep 24.
Article in English | MEDLINE | ID: mdl-15336551

ABSTRACT

Bone marrow stromal cells (MSCs) can be expanded rapidly in vitro and have the potential to be differentiated into neuronal, glial and endodermal cell types. However, induction for differentiation does not always have stable result. We present a new method for efficient induction and acquisition of neural progenitors, neuronal- and glial-like cells from MSCs. We demonstrate that rat MSCs can be induced to neurospheres and most cells are positive for nestin, which is an early marker of neuronal progenitors. In addition, we had success in proliferation of these neurospheres with undifferentiated characteristics and finally we could obtain large numbers of neuronal and glial phenotypes. Many of the cells expressed beta-tubulin III when they were cultivated with our method. MSCs can become a valuable cell source as an autograft for clinical application involving regeneration of the central nervous system.


Subject(s)
Astrocytes/cytology , Bone Marrow Cells/cytology , Cell Differentiation/physiology , Neurons/cytology , Oligodendroglia/cytology , Stromal Cells/cytology , Animals , Animals, Genetically Modified , Gene Expression Regulation/genetics , Gene Expression Regulation/physiology , Immunohistochemistry , Rats , Reverse Transcriptase Polymerase Chain Reaction
18.
J Cell Biochem ; 93(3): 454-62, 2004 Oct 15.
Article in English | MEDLINE | ID: mdl-15372628

ABSTRACT

Pluripotent embryonic stem (ES) cells have the capability to differentiate to various cell types and may represent an alternative cell source for the treatment of cartilage defects. Here, we show that differentiation of ES cells toward the chondrogenic lineage can be enhanced by altering the culture conditions. Chondrogenesis was observed in intact embryoid body (EB) cultures, as detected by an increase in mRNA levels for aggrecan and Sox9 genes. Collagen IIB mRNA, the mature chondrocyte-specific splice variant, was absent at day 5, but appeared at later time points. Dexamethasone treatment of alginate-encapsulated EB cultures did not have a strong chondrogenic effect. Nor was chondrogenesis enhanced by alginate encapsulation compared to simple plating of EBs. However, disruption of day 5 EBs and culture as a micromass or pelleted mass, significantly enhanced the expression of the cartilage marker gene collagen type II and the transcription factor Sox9 compared to all other treatments. Histological and immunohistochemical analysis of pellet cultures revealed cartilage-like tissue characterized by metachromatically stained extracellular matrix and type II collagen immunoreactivity, indicative of chondrogenesis. These findings have potentially important implications for cartilage tissue engineering, since they may enable the increase in differentiated cell numbers needed for the in vitro development of functional cartilaginous tissue suitable for implantation.


Subject(s)
Cell Differentiation/physiology , Chondrogenesis/physiology , Dexamethasone/pharmacology , Stem Cells/cytology , Aggrecans , Alginates/metabolism , Animals , Cell Culture Techniques , Cell Differentiation/drug effects , Cells, Cultured , Chondrocytes/cytology , Chondrocytes/drug effects , Chondrocytes/metabolism , Chondrogenesis/drug effects , Collagen Type II/metabolism , Extracellular Matrix Proteins/metabolism , Glucuronic Acid/metabolism , Hexuronic Acids/metabolism , High Mobility Group Proteins/metabolism , Lectins, C-Type , Mice , Proteoglycans/metabolism , SOX9 Transcription Factor , Stem Cells/drug effects , Stem Cells/metabolism , Transcription Factors/metabolism
20.
Article in English | MEDLINE | ID: mdl-15259672

ABSTRACT

We reviewed 12 patients who had had vascularised tissue transfer after oncological resection in the upper extremity. All patients had immediate reconstruction, and one had a double tissue transfer. Tissues used were skin flaps, free muscles, and vascularised fibulas, for resurfacing the wound, motor recovery, and reconstruction of large bony defects, respectively. All the patients returned early to their daily activities. Although local recurrences were encountered in two patients, they were again rendered disease-free by salvage operations. Two patients had secondary reconstructions using pedicle latissimus dorsi flaps for late problems, including one of the two patients who developed a local recurrence. The mean functional score using the system devised by the Musculoskeletal Tumor Society was 84%. All the upper extremities were successfully rescued with satisfactory function. Vascularised tissue transfer was invaluable for achieving both curative resection of the tumour and a useful upper extremity.


Subject(s)
Arm/surgery , Neoplasms/surgery , Surgical Flaps/blood supply , Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Plastic Surgery Procedures/adverse effects , Plastic Surgery Procedures/methods
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