Vascular density of superficial esophageal squamous cell carcinoma determined by direct observation of resected specimen using narrow band imaging with magnifying endoscopy.

Observation of the microvasculature using narrow band imaging (NBI) with magnifying endoscopy is useful for diagnosing superficial squamous cell carcinoma. Increased vascular density is indicative of cancer, but not many studies have reported differences between cancerous and noncancerous areas based on an objective comparison. We observed specimens of endoscopic submucosal dissection (ESD) using NBI magnification, and determined the vascular density of cancerous and noncancerous areas. A total of 25 lesions of esophageal squamous cell carcinoma that were dissected en bloc by ESD between July 2013 and December 2013 were subjected to NBI magnification. We constructed a device that holds an endoscope and precisely controls the movement along the vertical axis in order to observe submerged specimens by NBI magnification. NBI image files of both cancerous (pathologically determined invasion depth, m1/2) and surrounding noncancerous areas were created and subjected to vascular density assessment by two endoscopists who were blinded to clinical information. The invasion depth was m1/2 in 20, m3/sm1 in four and sm2 in one esophageal cancer lesion. Mean vascular density was significantly increased in cancerous areas (37.6 ± 16.3 vessels/mm2) compared with noncancerous areas (17.6 ± 10.0 vessels/mm2) (P < 0.05). The correlation coefficients between vascular density determined by two endoscopists were 0.86 and 0.81 in cancerous and noncancerous areas, respectively. Receiver operating curve (ROC) analysis revealed that the area under the curve (AUC) of vascular density was 0.895 (95% CI, 0.804-0.986). For this ROC curve, sensitivity was 78.3% and specificity was 87.0% when the cutoff value of vascular density was 26 vessels/mm2. NBI magnification confirmed significant increases in vascular density in cancerous areas compared with noncancerous areas in esophageal squamous cell carcinoma. The rates of agreement between vascular density values determined by two independent operators were high.

Absence of IgD-CD27(+) memory B cell population in X-linked hyper-IgM syndrome.

The present study analyzed peripheral blood B cell populations separated by IgD and CD27 expression in six males with X-linked hyper-IgM syndrome (XHIM). Costimulation of mononuclear cells from most of the patients induced no to low levels of class switching from IgM to IgG and IgA with Staphylococcus aureus Cowan strain (SAC) plus IL-2 or anti-CD40 mAb (anti-CD40) plus IL-10. Measurable levels of IgE were secreted in some of the patients after stimulation with anti-CD40 plus IL-4. Costimulation with SAC plus IL-2 plus anti-CD40 plus IL-10 yielded secretion of significant levels of IgG in addition to IgM, but not IgA. The most striking finding was that peripheral blood B cells from all of the six patients were composed of only IgD+ CD27(-) and IgD+ CD27(+) B cells; IgD- CD27(+) memory B cells were greatly decreased. IgD+ CD27(+) B cells from an XHIM patient produced IgM predominantly. Our data indicate that the low response of IgG production in XHIM patients is due to reduced numbers of IgD- CD27(+) memory B cells. However, the IgG production can be induced by stimulation of immunoglobulin receptors and CD40 in cooperation with such cytokines as IL-2 and IL-10 in vitro.

Imbalance among T-cell subsets in patients with coronary arterial aneurysms in Kawasaki disease.

The populations of T cells were studied in 46 patients with Kawasaki disease, separated into 2 groups: group I--11 patients with coronary aneurysms; and group II--35 patients with normal coronary arteries. Patients from both groups with early acute illness, before day 5, had a significant reduction in the population of OKT3+ (p less than 0.001), OKT4+ (p less than 0.02) and OKT8+ cells (p less than 0.002), but normal OKT4/OKT8 ratios compared with age-matched control subjects. These abnormal values quickly returned to normal levels during week 2 in patients with normal coronary arteries. In contrast, patients in whom coronary aneurysms developed within 3 weeks of the onset had an imbalance between OKT4 and OKT8 during week 2, characterized by a decrease in the number of OKT8+ cells and an increase in the number of OKT4+ cells, resulting in a high OKT4/OKT8 ratio (p less than 0.01). Three patients in whom large coronary aneurysms developed had ratios higher than 4.50. Follow-up analysis of T-cell subsets from individual patients with coronary aneurysms showed that the OKT4/OKT8 ratio during the acute stage was reduced during the convalescent stage (p less than 0.005). In contrast, the ratio in patients with normal coronary arteries was normal during the course of the illness. These observations suggest that an immune regulatory process operating in coronary aneurysm formation is present.

Tracheal allotransplantation in beagle dogs without immunosuppressants.

BACKGROUND: The antigenicity of tracheal grafts is still unclear. We investigated the possibility of performing tracheal allotransplantation without immunosuppressants. METHODS: Intrathoracic five-ring tracheal replacements were performed in beagle dogs without immunosuppressants (n = 18). The dogs were divided into 9 pairs, and grafts were exchanged within the pairs. In group 1 (n = 6), the paired dogs were blood relatives, whereas in group 2 (n = 12), the paired dogs were not related. Full-thickness skin transplantation was also performed in both groups. RESULTS: In group 1, 5 animals survived uneventfully for more than 3 months. No stenosis was observed in any of the dogs. In group 2, the grafts were incorporated by the host trachea in 2 dogs. Four animals died of airway obstruction within 3 months. Moderate or slight airway stenosis was observed in 6 dogs. Rejection was confirmed by histologic examination. In both groups, all of the skin allografts were destroyed within 2 weeks. CONCLUSIONS: After tracheal allotransplantation, long-term survival was achieved, especially in recipient dogs that were blood relatives of donors. We conclude that it is possible to perform tracheal allotransplantation using histocompatible matched grafts without immunosuppressants.

Growth suppression and induction of heat-shock protein-70 by 9-cis beta-carotene in cervical dysplasia-derived cells.

The aim of the present study was to determine to what extent 9-cis beta-carotene, one of the most abundant naturally-occurring cis-isomers of beta-carotene, can inhibit the growth of cervical dysplasia-derived cells in comparison with all-trans beta-carotene. We found that 9-cis beta-carotene was dose-dependently more effective than all-trans beta-carotene. Both carotenes induced the intracellular accumulation of heat-shock protein-70 (HSP70), and the treated cells showed morphological changes indicative of apoptosis. The results of the present study strongly suggest that the induction of HSP70 by beta-carotene might be involved in beta-carotene-mediated suppression of the cell growth through apoptosis.

Immunofluorescence microscopic studies on distribution of Lactobacillus kefiranofaciens and Lactobacillus kefir in kefir grains.

Immunofluorescence staining was applied to observe distribution of two Lactobacillus species in kefir grains with cauliflower floret forms. Kefiran-producing, encapsulated Lactobacillus kefiranofaciens was located all over the grain and increased toward the center, while Lactobacillus kefir populated only a small region at the surface layers.

Distribution of microorganisms with particular reference to encapsulated bacteria in kefir grains.

Propagable and non-propagable kefir grains in a form resembling cauliflower florets were observed by scanning electron microscopy (SEM) and light microscopy. In propagable grains short and long rod-shaped bacteria and yeasts formed separate colonies on the outside surface and inside. Internally, filaments which derived from capsules around the cells extended radially from a population of long rod-shaped bacteria. In non-propagable grains long rod-shaped bacteria with filamentous appendages were not observed, but only short rod-shaped bacteria and yeasts. Indian ink preparations showed presence of encapsulated bacteria in propagable grains and absence of these in non-propagable ones. The above results suggest that encapsulated bacteria are responsible for propagation of kefir grains.

Capsular polysaccharide of a slime-forming Lactococcus lactis ssp. cremoris LAPT 3001 isolated from Swedish fermented milk 'långfil'.

Slime-forming Lactococcus lactis ssp. cremoris strain LAPT 3001 isolated from Swedish ropy sour milk 'långfil' was investigated for the chemical nature of its capsule. The capsular material purified by gel filtration chromatography and ion-exchange chromatography consisted of rhamnose, glucose, galactose, glycerol and phosphorus. It is most likely a deacylated lipoteichoic acid.

Enhancement of antigen-specific antibody production by extracellular slime products from slime-forming Lactococcus lactis subspecies cremoris SBT 0495 in mice.

The effect of extracellular slime products (ESP) produced by Lactococcus lactis subspecies cremoris SBT 0495 on antigen specific antibody production was studied in mice. ESP contained 48.5% protein, 15.4% neutral sugar, and 1.1% of phosphorus. The optimum dose of ESP was between 100 to 500 micrograms per mouse. ESP administered intraperitoneally (200 micrograms per mouse) enhanced the production of specific antibody in mice. These results indicate that ESP may act as an adjuvant.

Scanning electron microscopic and texture studies on characteristic consistency of Nordic ropy sour milk.

The characteristic consistency of Nordic ropy sour milk was studied. Skim milk, reconstituted from non-fat dry milk, was fermented at 20 degrees C for 24 h by addition of 5% (v/v) inoculum of slime-producing (ropy) strain of Lactococcus lactis ssp. cremoris SBT 0495, isolated from Finnish ropy sour milk 'viili' starter culture, and its non-ropy variant SBT 1275. Measurements of texture showed that milk gel prepared by the ropy strain exhibited remarkably increased adhesiveness as compared to that by the non-ropy variant. Milk gel prepared by the ropy strain also exhibited decreased syneresis (wheying-off) as compared to that by the non-ropy variant. Scanning electron micrographs of milk gel prepared by the ropy strain showed that slime was in the form of a network attaching the bacterial cells to the protein matrix. A thick network of slime attached the casein micelle clusters to each other to make casein conglomerates, which is likely to result in the characteristic consistency of 'viili'.

Some taxonomical characteristics of encapsulated Lactobacillus sp. KPB-167B isolated from kefir grains and characterization of its extracellular polysaccharide.

A capsular polysaccharide-producing strain KPB-167B isolated from kefir grains was identified as a homofermentative Lactobacillus. The carbohydrate fermentation pattern and DNA base composition of the strain were different from those of other capsular Lactobacillus species previously isolated from kefir grains. The polysaccharide produced by Lactobacillus sp. KPB-167B was found similar to kefiran by 13C-NMR and methylation analysis. Lactobacillus sp. KPB-167B could grow and produce capsular polysaccharide in MRSL medium with better yield than L. kefiranofaciens, which suggested that it is suitable for kefiran production.

Structure of the extracellular polysaccharide from slime-forming Lactococcus lactis subsp. cremoris SBT 0495.

The extracellular polysaccharide obtained from slime-forming Lactococcus lactis subsp. cremoris SBT 0495 is composed of D-glucose, D-galactose, L-rhamnose, and phosphate. Methylation analysis of the native and dephosphorylated polysaccharides provided information on the linkage of the sugar residues and the location of the phosphate group. N.m.r. spectroscopy confirmed the structure of the polysaccharide, which is assigned the following repeating-unit: [formula: see text]

Effect of hyaluronic acid on the excursion resistance of tendon grafts. A biomechanical study in a canine model in vitro.

The excursion resistance between the tendon and pulley is an important factor contributing to the limitation of function after surgery to the hand. The administration of hyaluronic acid (HA) in the early rehabilitation after tendon grafting may help to prevent adhesions. We evaluated changes in the excursion resistance between potential sources of flexor tendon grafts and the annular pulley in a canine model after administration of HA. The intrasynovial and extrasynovial tendons were soaked in 10 mg/ml of HA for five minutes. The excursion resistance between these tendons and the annular pulley of an intact proximal phalanx and that of the same tendons of the opposite foot without administration of HA were evaluated. The tendon of flexor digitorum profundus of the second toe without administration of HA was used as a control. The gliding resistance of canine tendons was significantly decreased after the administration of HA especially in the extrasynovial tendons. Our findings suggest that the administration of HA may improve the gliding function of a flexor tendon graft.

Regeneration of the junctional epithelium and connective tissue after transplantation of detergent-processed allo-teeth.

The authors have developed a new artificial dental implant and evaluated it in a dog model in terms of its potential to produce: I) regeneration of junctional epithelium; II) regeneration and attachment of connective tissue. The implants were constructed from allo-teeth. We removed the cell components from the periodontal ligaments of these teeth with a detergent (1% TritonX-100); the remaining acellular periodontal ligament acted as an extracellular matrix upon which regeneration and attachment could proceed. We placed 10 of these implants in the just-extracted sites of three beagle dogs. We observed regeneration of both junctional epithelium and connective tissue at all implant sites after 3 months. The connective tissue was attached in all cases. Use of the acellular periodontal ligament as an extracellular matrix may facilitate regeneration of host periodontal ligament tissue, thus contributing to recovery of host immunological defense and long-term oral function.

Evaluation of peripheral nerve regeneration across an 80-mm gap using a polyglycolic acid (PGA)--collagen nerve conduit filled with laminin-soaked collagen sponge in dogs.

We evaluated peripheral nerve regeneration using a novel artificial nerve conduit. The conduit was made of a polyglycolic acid(PGA) - collagen tube filled with laminin- soaked collagen sponge. We implanted this nerve conduit across an 80mm gap in the peroneal nerve of dogs. Histological observation 12 months after implantation showed numerous unmyelinated and myelinated nerve fibershad regenerated beyond the gap. Neurofilaments were widely observed immunohistochemically in the regenerated nerve segments. These findings indicated that newly regenerated axons had extended across the gap and connected into the distal nerve segments. Compound muscle action potentials(CMAPs) and somatosensory evoked potentials (SEPs) were recorded in all dogs. At 12 months, the CMAPs indicated complete recovery, while the SEPs showed incomplete but substantial recovery. Walking patterns had returned to near-normal 12 months after implantation. Use of this nerve conduit can lead to peripheral nerve elongation and favorable functional recovery across a wider nerve gap.

Regenerative repair of the mandible using a collagen sponge containing TGF-beta1.

INTRODUCTION: Alveolar bone resorption and atrophy of the mandible are a major challenge for regeneration medicine. In the present investigation, a collagen sponge that contained TGF-beta1 was placed at a mandibular defect and the osteogenic effects of collagen-TGF-beta1, complex were evaluated. MATERIAL AND METHODS: The Pm2, Pm3, and Pm4 teeth on both sides of the mandibles of 12 adult beagle dogs (9.0-12.0 kg) were extracted. After the extraction-site wounds healed, a bone defect (10.0 x 15.0 mm-wide, 10.0 mm-deep or 10.0 x 10.0 mm-wide, 10.0 mm-deep) was created on the mandible. A collagen sponge (10.0 x 10.0 x 10.0 mm) that contained TGF-beta1 (1.0 microg, 5.0 microg, or 10.0 microg, in physiological saline) was placed at the bottom of the defect and the overlying mucous membrane was sutured with 4-0 prolene. As a control, a collagen sponge that contained physiological saline only was placed in a defect on the opposite side. Two weeks after the surgery the wounds above the bone defects on both the control and TGF-beta1-treated sides had healed completely. RESULTS: At four, six, or eight weeks post-operatively animals were killed. Soft X-ray and bone-salt measurement analyses confirmed clearly that there was greater calcified bone formation in the defects into which TGF-beta1 had been incorporated than with the control defects. The implanted collagen sponges were fully resorbed and the bone tissue had regenerated from the bottom of the defects on the TGF-beta1, side by four weeks. On the control side, no such regeneration was observed. CONCLUSIONS: These results indicate that TGF-beta1, released slowly from a collagen sponge was effective in promoting bone remodeling when applied to mandibular defects in adult dogs.

Regeneration of hypogastric nerve using a polyglycolic acid (PGA)-collagen nerve conduit filled with collagen sponge proved electrophysiologically in a canine model.

The hypogastric nerve (HGN) is a sympathetic nerve in the peritoneal cavity and controls urinary and seminal functions. In this study, the regeneration of HGN was determined by using a new type of an artificial nerve conduit, polyglycolic acid (PGA)-collagen nerve conduit filled with collagen sponge in two dogs. A PGA-collagen nerve conduit (diameter=2 mm) was interposed in a 10 mm gap of the right HGN. The regeneration of the HGN was evaluated electrophysiologically 8 months after the operation. The intraluminal pressure of spermatic duct and the bladder neck were elevated 80 mmHg and 25 mmHg respectively by the stimulation across the regenerated HGN. The prostate contraction was also elicited. The responses diminished after the excision of the regenerated portion of HGN. These results proved the regeneration of HGN and this nerve conduit will be great help for patients who suffer from urinary and seminal disturbances.

Artificial trachea and long term follow-up in carinal reconstruction in dogs.

We have already reported successful carinal reconstruction of the trachea with an observation period of 1 - 2 years. In this study, we evaluate the long-term safety and efficacy of the reconstruction after 5-years of follow-up. The Y-shaped Marlex mesh tube was reinforced with a polypropylene spiral and coated with atelocollagen made from porcine skin. The prosthesis was 60 mm long with an outer diameter of 18 mm. Replacement of the tracheobronchial bifurcation was preformed through a right thoracotomy in a beagle dog. Bronchoscopical examination and sampling of the tracheal epithelium was performed periodically to check the function of cilia. The implanted prothesis was promptly infiltrated by the surrounding connective tissue and completely incorporated by the host trachea and bronchus. Bronchoscopically, sufficient epithelization was confirmed from the upper to the lower site of anastomosis. After 5 years neither stenosis nor dehiscence was observed. In spite of there being mesh-exposure at the luminal surface, the dog had no clinical symptoms until sacrifice for pathological examination. The bent frequency of the cilia was maintained within the normal range, indicating functional recovery of the regenerating airway. Our tracheal prosthesis is promising for clinical repair of the tracheobronchial bifurcation.

Subfascial temporalis dissection preserving the facial nerve in pterional craniotomy--technical note.

The subfascial temporalis dissection and reconstruction of the temporalis muscle for pterional craniotomy are described. These procedures preserve the frontotemporal branch of the facial nerve and increase exposure along the sphenoid ridge. A good cosmetic appearance and good temporalis muscle function are achieved postoperatively.

Postoperative outcome of lumbar spinal canal stenosis after fenestration: correlation with changes in intradural and extradural tube on magnetic resonance imaging.

PURPOSE: To evaluate the serial changes in clinical results and the intradural and extradural spaces on magnetic resonance imaging (MRI) after bilateral fenestration in 48 patients with lumbar spinal canal stenosis (LSCS). METHODS: A prospective interventional study was performed to study the clinical results, magnetic resonance imaging scans among patients who were followed up for more than 3 years. RESULTS: All patients showed improvement in clinical symptoms after operation, but clinical results deteriorated in 9 (19%) patients. Postoperative MRI scans showed that poor dural tube expansion, grouping of the cauda equina, and decrease in the cross-sectional area of the dural tube were factors associated with poor outcomes. The cross-sectional area of the dural tube and images of the cauda equina observed by MRI, before and after fenestration and during follow-up, reflected changes in clinical symptoms involving decompressed segments. CONCLUSION: Serial changes in the cross-sectional area of the dural tube and images of the cauda equina observed preoperatively, postoperatively, and on follow-up by MRI may be useful when evaluating patients' condition before and after operation. It is also useful for predicting outcomes.