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Biomedical subjects

M Ueda

Publications and source records attributed to M Ueda.

At least 235 records · Page 13Linked to original sources

Surgical treatment for carcinoma of the thoracic esophagus with major involvement in the neck or upper mediastinum.

BACKGROUND AND OBJECTIVES: In carcinoma of the thoracic esophagus, most surgeons consider that esophagectomy is contraindicated in patients with clinical evidence of major extraesophageal involvement in the lower neck or peritracheal regions. However, metastases to these regions are commonly found even in early phases of carcinoma invasion. With recent progress in preoperative assessment, operative technique and adjuvant therapy, esophagectomy could possibly benefit appropriately selected patients. METHODS: We retrospectively analyzed results in 42 patients who had major involvement in the neck or upper mediastinum and who underwent esophagectomy with systematic lymph node dissection. We operated upon patients unless lesions were assessed as definitely unresectable. Preoperatively, 32 had enlarged peritracheal nodes greater than 15 mm in diameter on computed tomography, 18 had hard unmobile tumors in the lower neck, 9 had recurrent laryngeal nerve palsy, and 10 had findings suggestive of tracheal invasion. Preoperative radiotherapy and/or chemotherapy was given to 32 low-risk patients. RESULTS: The hospital mortality rate was 4%. Bowel reconstruction was completed in all cases. No macroscopically recognizable lesion remained after operation in 35 patients. Eight patients were alive 5 years after esophagectomy, including 2 who had had tracheal invasion and 1 with recurrent nerve palsy. The cumulative 5-year survival was 38%. CONCLUSIONS: Evidence of major involvement of the neck and/or upper mediastinum does not always contraindicate resection. Aggressive esophagectomy combined with perioperative adjuvant therapy yielded acceptable palliation and occasional cure in cases with technically resectable lesions.

Antineoplastic Combined Chemotherapy Protocols↗

Brush cytology of herpes simplex virus infection in oral mucosa: use of the ThinPrep processor.

Evaluation was made of oral scraping specimen using Cytobrush from 11 patients with clinical suspicious lesions for herpes simplex virus (HSV) infection. In this study, oral brush cytology prepared using an automated smear apparatus (ThinPrep method) has been applied. The smears prepared by ThinPrep are equally well suited for screening purposes, producing highly cellular specimens with well-preserved cytoplasm and nuclei. The results obtained on the 11 patients, four cytologic specimens, showed pathognomonic viral cellular changes of HSV infection. Viral isolation studies were performed on 10 of the 11 patients and ten specimens tested positive for HSV by culture. Of the ten HSV positive isolates, six were positive by either cytology or immunocytochemical staining. Of the two smears showing HSV positive cells, both smears were also confirmed by in situ hybridization (ISH) with a biotinylated clone DNA probe. Using the ICC technique, detection of HSV antigen in oral scraping smears was of great value in the diagnosis of oral HSV infection, especially in cases of equivocal lesions. Oral cytology by means of ThinPrep method obviously deserves additional trials as an adjunct in the cytology of HSV-suspected lesions.

Cytodiagnosis↗

Inhibition of angiogenesis on glycated collagen lattices.

Advanced glycation endproduct (AGE) accumulation in extracellular matrix proteins has been demonstrated in diabetic patients with a significant correlation with the severity of diabetic complications. AGE accumulation induces matrix protein cross-link formation, resulting in an increased stiffness of matrix fibres and the reduction of the susceptibility of matrix proteins to proteolytic degradation. We examined whether glycation-induced collagen cross-linking may affect vascular endothelial cell behaviours such as invasion, proliferation and differentiation, using the in vitro angiogenesis model of capillary-like structure formation in three-dimensional matrices of collagen type I. Endothelial cells cultured on collagen gel with angiogenic factors (the combination of fibroblast growth factor-2 and vascular endothelial growth factor) invaded the underlying collagen matrix, and organized capillary-like cord structures in the gel. We found that endothelial cell invasion into glycated collagen gel was significantly attenuated without any effect on proteinase activity including cell-associated plasminogen activator and matrix metalloproteinase in the conditioned medium. In addition, subsequent capillary-like cord formation was also inhibited in glycated collagen gel. In contrast, endothelial cell proliferation was enhanced on glycated collagen gel with or without angiogenic factors compared with control collagen gel. These results suggest that the structural alterations of extracellular matrix proteins through the glycation-induced cross-link formation affect the interaction between endothelial cell and extracellular matrix, resulting in the impairment of an adequate neovascularization in diabetic patients.

Angiogenesis Inducing Agents↗

Mechanism of the pathogenesis of glutamate neurotoxicity in retinal ischemia.

PURPOSE: This study was carried out to examine the involvement of glutamate and nitric oxide neurotoxicity in ischemia/reperfusion-induced retinal injury in vivo. METHODS: We monitored glutamate release from in vivo cat retina during and after pressure-induced ischemia using a microdialysis technique. Morphometric studies were performed to study the effects of MK-801 (dizocilpine), L-NAME (N omega-nitro-L-arginine methyl ester), and D-NAME (N omega-nitro-D-arginine methyl ester) on the histological changes in the rat retina induced by ischemia or intravitreal injection of NMDA (N-methyl-D-aspartate; 200 nmol). RESULTS: A large release of glutamate occurred during ischemia, followed by a marked release after reperfusion. Histological changes occurred selectively in the inner part of the retina after ischemia as well as intravitreal injection of NMDA. Pretreatment with intravenous injection of MK-801 or L-NAME significantly inhibited the ischemic injury of the inner retina. Intravitreal injection of L-NAME inhibited NMDA-induced neurotoxicity in the retina. CONCLUSION: These findings indicate that nitric oxide mediates neurotoxic actions of glutamate which are responsible for ischemic injury in the retina.

Animals↗

The influence of compressive loading on growth of cartilage of the mandibular condyle in vitro.

The purpose of this study was to clarify the change in mandibular condyles under compressive loading. An organ-culture system of fetal rat mandibular condyles was used, and mechanical loading was generated by compressing the gas phase within a closed chamber. After the culture period, with compressive loading, type I collagen and fibronectin were observed in the lower half of the hypertrophic chondrocyte layer in the mandibular condyles; in contrast, without compressive loading, there was no such reaction. The size of the condyle was not increased by compressive loading. These results suggest that intermittent compressive loading could induce type I collagen and fibronectin production by chondrocytes.

Animals↗

Effect of nuclear stages during IVM on the survival of vitrified-warmed bovine oocytes.

The effect of nuclear stages during IVM on the survival of vitrified-warmed bovine oocytes was investigated. Oocytes with compact cumulus cells were cultured for 0, 6, 12 and 24 h in TCM199 supplemented with 5% fetal bovine serum (FBS) in 3% CO2 in air. The oocytes were first exposed to 20% ethylene glycol solution and were subjected to vitrification in a solution containing 40% ethylene glycol, 18% Ficoll-70 and 0.3 M sucrose. After warming in 20 degrees C water, oocytes which had been vitrified at less than 24-h of IVM were again cultured to complete the 24-h of IVM period. Oocytes were then incubated with frozen-thawed spermatozoa in Brackett and Oliphant (BO) medium containing 60 micrograms/ml heparin and 0.25% BSA for 20 h. In vitro fertilization rates of oocytes vitrified-warmed at 0, 6, 12 and 24-h IVM were 75.2, 68.0, 82.0 and 72.4%, respectively, comparable to the rates for unvitrified control oocytes (80.6%). A higher incidence of polyspermic fertilization was observed in oocytes vitrified at 24-h IVM (44.9 vs 22.6% in the control group, P < 0.05). Vitrification of oocytes at 12-h IVM seemed to be better than that of other IVM groups, since the normal fertilization rate of all treated oocytes was the highest (36.0%) among the vitrification groups. Developmental competence of the oocytes following vitrification and in vitro fertilization (12-h IVM group) was examined by cell-free culture of presumptive zygotes up to 9 d in modified synthetic oviduct fluid (mSOF) in 5% CO2, 5% O2 and 90% N2. The cleavage rate of zygotes from vitrified oocytes 48 h after insemination was 29.8%, which was lower than that of the control group (57.0%, P < 0.05). Development to blastocysts from the vitrified oocytes (4.8%) was much lower than that of the control group (27.0%, P < 0.05). These results indicate that cryopreservation of bovine oocytes by vitrification may be affected by their maturation stage in vitro, and that developmental competence to blastocysts of cleaved oocytes following vitrification may be impaired compared with unvitrified control oocytes.

Animals↗

Cryopreservation of in vitro-derived bovine blastocysts microinjected with foreign DNA at the pronuclear stage.

Days 6 and 7 bovine blastocysts derived from in vitro-fertilized and DNA-injected zygotes (day of IVF = Day 0) were cryopreserved either by conventional two-step freezing or by vitrification. Foreign DNA used for microinjection was the green fluorescent protein gene under the control of the immediate early promoter of human cytomegalovirus. All blastocysts were produced by an in vitro system and were harvested on Days 6 and 7. The proportion of DNA-injected zygotes developing into blastocysts on Days 6 and 7 (total 8%) was lower than that of nontreated zygotes (total 19%; P < 0.01). After cryopreservation in 1.5 M ethylene glycol, the survival rates of DNA-injected blastocysts assessed by re-expansion at 24 h of culture (Day 6: 59%, Day 7: 71%) were comparable with those of nontreated blastocysts (Day 6: 76%, Day 7: 71%). The post-thaw hatching rate within 72 h of culture of DNA-injected Day 7 blastocysts (38%) was not different from that of nontreated Day 7 blastocysts (40%), but the hatching rate of DNA-injected Day 6 blastocysts (23%) was lower than that of nontreated Day 6 blastocysts (47%; P < 0.05). After vitrification in 7.2 M ethylene glycol, 0.0026 M Ficoll-70 and 0.3 M sucrose, the survival and hatching rates of DNA-injected Day 7 blastocysts (61 and 28%, respectively) were similar to those of nontreated Day 6 (71 and 33%, respectively) and Day 7 (75 and 36%, respectively) blastocysts. However, the post-warming survival rate of DNA-injected Day 6 blastocysts was only 30%, and none of the blastocysts hatched (P < 0.01). The mean cell number of DNA-injected Day 6 blastocysts (100.3 +/- 36.4 cells) was lower than that of nontreated Day 6 blastocysts (130.5 +/- 37.1 cells; P < 0.01), while those of DNA-injected and nontreated Day 7 blastocysts were not different (111.2 +/- 42.8 and 119.6 +/- 31.4 cells, respectively). These results indicate that Day 7 IVMFC bovine blastocysts derived from DNA-injected zygotes can be successfully cryopreserved by conventional two-step freezing or vitrification.

Animals↗

Natural hepatocyte growth factor (HGF) from human serum and a bound form of recombinant HGF with heparan sulfate are indistinguishable in their physicochemical properties.

Natural hepatocyte growth factor (nHGF) purified from human serum showed a difference in molecular mass (Mr) between SDS PAGE (76-90 kDa) and gel filtration chromatography on a Sephadex G-200 column (> 200 kDa), whereas nHGF or recombinant HGF (rHGF) from cell culture medium did almost the same Mr (74-100 kDa). A bound form of rHGF with heparan sulfate (or heparin), and an aggregate form of rHGF itself showed a homogeneous band with a Mr of 76-90 kDa on SDS PAGE, but showed a Mr larger than 200 kDa on a Sephadex G-200 column. Both nHGFs, rHGF and the bound form were basic, but the aggregate form was acidic in ionic nature. No significant difference was found in affinity for heparin among these HGF preparations. The bound form treated by the procedures for purification of nHGF from human serum still showed a larger molecular form. The bound form mimicked physicochemical properties of nHGF purified from human serum. These results suggest that a possible form of nHGF in human serum may be a bound form with heparin-like molecules such as heparan sulfate, which are found in the circulation and on cell surface, and purified as the bound form.

Animals↗

Isolation of nucleation-competent centrosomes from Dictyostelium discoideum.

The centrosome of Dictyostelium discoideum is a box-shaped, layered core structure surrounded by a corona which is made up of dense nodules embedded in amorphous material. It is also known as nucleus-associated body. Because of its tight association with the nucleus the centrosome has resisted so far all attempts for isolation in sufficient purity and quantity for biochemical analysis. Here we report on the large-scale isolation of D. discoideum centrosomes after treatment of nucleus-centrosome complexes with a buffer containing sodium pyrophosphate. Following heparin treatment and a filtration step, centrosomes were further purified by density gradient centrifugation. Immunofluorescence analysis of the isolated centrosomes revealed the presence of the D. discoideum 350-kDa antigen, a centrosomal marker protein, gamma-tubulin, and the D. discoideum homologues of pericentrin, Spc110p, and Cdc31p. The structural integrity of the isolated centrosomes was demonstrated by confocal laser microscopy and electron microscopy. Microtubule nucleation assays with purified pig brain tubulin showed that the isolation procedure did not only preserve the structure but also the functionality of the isolated centrosomes. D. discoideum centrosomes should now become an attractive new model system in addition to, and for comparison with, centriolar centrosomes and yeast spindle pole bodies.

Animals↗

Immunization with the "immunogenic peptide" of TSH receptor induces oligoclonal antibodies with various biological activities.

Balb/c mice were immunized with a synthetic peptide (P354-14) corresponding to "the immunogenic peptide" of human thyrotropin receptor (hTSH-R). Through screening for binding to the peptide, we obtained several monoclonal antibodies with various biological activities: thyroid stimulation (SAb), inhibition of TSH stimulation (BAb) and no significant effect on cAMP production. One of the stimulatory clones was further studied. This clone enhanced cAMP production in Cos-7 cells transformed with the truncated TSH-R cDNA deleting the immunogenic peptide. These results indicated that the immunogenic peptide of the TSH-R induces oligoclonal anti-TSH-R antibodies, although the region is not essential for the functional epitope.

Animals↗

Segmental mandibular reconstruction by distraction osteogenesis under skin flaps.

In five adult dogs, molars were extracted and skin flaps from the neck prepared for delayed transplantation. Two weeks later, a 25-mm segment of the mandible was excised with surrounding periosteum and gingiva. The mandible was stabilized with a reconstruction plate and the intraoral defect repaired with a pedicled skin flap. A proximal transport segment was created and an external distraction device was applied. After one week, distraction of the transport segment was begun at a rate of 1 mm/day. After distraction was completed, the lengthening appliance was left in place for 12 weeks until the dogs were killed. Radiologic and histologic examination revealed new bone at the distraction site. The intraoral skin flap was pushed out of the defect as distraction progressed. Bony union of the transport segment to the distal stump was not achieved due to intervening soft tissue. These results suggest that it is feasible to bridge a mandibular defect that is covered with a skin flap, with distraction osteogenesis.

Animals↗

Influence of food consistency and dental extractions on the rat mandibular condyle: a morphological, histological and immunohistochemical study.

The purpose of this study was to investigate the relationship between food consistency and the growth of the mandibular condyle in rats. Secondly, the effect of dental extractions on cartilage of the mandibular condyle was examined in young adult rats fed foods of varying consistency. Thirty-six male Wistar rats were divided into four groups: (A) Solid diet--non-Extraction (non-Ext.) group; (B) Solid diet--Extraction (Ext.) group; (C) Powder diet--non-Ext. group; and (D) Powder diet--Ext. group. Extractions were performed at 12 weeks of age. The mandibular condyles were removed at 1, 4, and 8 weeks after the extractions. The shape of the mandibular condyles in the powder diet groups (C and D) was significantly narrower. In the Ext. groups (B and D), the thickness of the hypertrophic zone was reduced one week after the extractions. In the powder diet groups (C and D), the intensity of the staining of fibronectin decreased in the proliferative zone regardless of the extractions. In group B, a decreased intensity of this reaction was observed one week after the extractions. From these results, it appeared that food consistency and/or dental extractions affected the morphology of the mandibular condyle and the histological characteristics of the mandibular condylar cartilage.

Age Factors↗