Computerized training of cryosurgery - a system approach.

The objective of the current study is to provide the foundation for a computerized training platform for cryosurgery. Consistent with clinical practice, the training process targets the correlation of the frozen region contour with the target region shape, using medical imaging and accepted criteria for clinical success. The current study focuses on system design considerations, including a bioheat transfer model, simulation techniques, optimal cryoprobe layout strategy, and a simulation core framework. Two fundamentally different approaches were considered for the development of a cryosurgery simulator, based on a finite-elements (FE) commercial code (ANSYS) and a proprietary finite-difference (FD) code. Results of this study demonstrate that the FE simulator is superior in terms of geometric modeling, while the FD simulator is superior in terms of runtime. Benchmarking results further indicate that the FD simulator is superior in terms of usage of memory resources, pre-processing, parallel processing, and post-processing. It is envisioned that future integration of a human-interface module and clinical data into the proposed computer framework will make computerized training of cryosurgery a practical reality.

Clinical effects of combined Lactobacillus paracasei and kestose on canine atopic dermatitis.

Probiotics and prebiotics are viable bacteria with beneficial effects on the host and components that selectively act on the beneficial commensal bacteria, respectively. The combined use of probiotics and prebiotics is termed synbiotics. Probiotic intake improves dysbiosis in the intestinal microbiota and can positively affect canine atopic dermatitis (CAD). However, clinical studies on improvements in CAD using synbiotics remain limited. In this study, 15 dogs with CAD who received prednisolone, a synthetic glucocorticoid (GC) used in the treatment of CAD, for more than 90 days were continuously treated with Lactobacillus paracasei M-1 from fermented food as a probiotic, and trisaccharide kestose as a prebiotic, for 90 days to determine their synbiotic effects on CAD. The CAD symptoms were evaluated using the canine atopic dermatitis lesion index (CADLI) and pruritus visual analog scores (PVAS) at 30, 60 and 90 days after synbiotic administration. The total prednisolone use for 90 days pre- and post-administration was also evaluated. Synbiotic administration significantly reduced the CADLI (pre: median, 28.0 [22.0-32.0]; 30 days: median, 20.0 [20.0-28.0]; 60 days: median, 20.0 [10.0-21.0]; 90 days: median, 12.0 [10.0-19.0]) and PVAS (pre: median, 6.0 [5.0-7.0]; 30 days: median, 3.0 [3.0-3.5]; 60 days: median, 3.0 [3.0-3.5]; 90 days: median, 2.0 [2.0-3.5]) scores, and reduced the total prednisone use over 90 days (pre: 112.0 [25-450] mg; post: 80.0 [18.-300.0] mg; p⟨0.001) in the 15 dogs. Thus, the synbiotic activity of L. paracasei M-1 and trisaccharide kestose can improve CAD.

Preservation of ovarian function by ovarian shielding when undergoing total body irradiation for hematopoietic stem cell transplantation: a report of two successful cases.

The purpose of this study was to evaluate the possibility of preserving ovarian function by ovarian shielding to reduce the irradiation dose in total body irradiation (TBI). The subjects in the study were females aged less than 40 years, who were undergoing allogeneic hematopoietic stem cell transplantation using a TBI-based regimen and who desired to have children after transplantation. For ovarian shielding, abdominal computed tomography (CT) and skin marking were performed in both the supine and prone positions, prior to the TBI. A pair of columnar blocks was placed just above the patient's body. Thus far three patients have been treated. The serum estradiol level decreased to an undetectable level (<8.5 pg/ml) after transplantation and the follicle-stimulating hormone (FSH) level increased above 90 mIU/ml in all patients and they became amenorrheic. However, regular menstruation recovered in patients no. 1 and 2 about 800 and 370 days after transplantation, respectively, with a decrease in the serum FSH level. Menstruation did not recover in patient no. 3, and serum estradiol was transiently detected above 20 pg/ml. The preservation of ovarian function was made possible by ovarian shielding. However, a longer follow-up is needed to know if normal pregnancy and delivery can occur.

Evolutionary aspects of the S-related genes of the Brassica self-incompatibility system: synonymous and nonsynonymous base substitutions.

In the Brassicaceae, self-vs. nonself-recognition in self-incompatibility is controlled by sporophytic S-alleles. Haplotypes specifying both SRK (S-receptor kinase) and SLG (S-locus glycoprotein) are considered to play an important role in the recognition reactions. We compared the nucleotide sequences of SRK9(Bc) and SRK6(Bo). The number of nonsynonymous substitutions per site (Pn) was lower, constrained, in the kinase than the receptor domain, while the numbers of synonymous substitutions (Ps) in the two domains were largely comparable. Pairwise values for Ps and Pn were calculated among 17 operational taxonomic units, including eight SLGs, the receptor domains of two SRKs, four SRAs (S-related A) and three SRBs (S-related B), which have high homologies with each other. The values of Ps and Pn of SLG were mostly comparable to those of the receptor domain of SRK. Dendrograms constructed on the basis of Pn and Ps indicated that SRA differentiated first, followed by SRB. The differentiation of SLG alleles is one of prerequisite factors for the establishment of self-incompatibility, and the allelic differentiation has occurred more than tens of million years ago.

7-Hydroperoxycholesterol as a marker of oxidative stress in rat kidney induced by paraquat.

The in vivo paraquat-induced oxidative stress in rat tissue was studied by analyzing cholesterol-derived hydroperoxide as an index of lipid peroxidation. Paraquat (10 mg/kg) was administered i.p. to rats. Rats were sacrificed and lung, liver, and kidney were collected 2, 24 h, and 5 d after paraquat injection. Lipids were extracted and analyzed by HPLC with post-column chemiluminescence. We found that two cholesterol-derived hydroperoxides, 7alpha-hydroperoxycholest-5-en-3beta-ol (7alpha-OOH) and 7beta-hydroperoxycholest-5-en-3beta-ol (7beta-OOH) were present in lungs of control animals (0.06 and 0.06 nmol/g, respectively), in livers (6.5 and 15.8 nmol/g, respectively) and in kidneys (3.7 and 8.9 nmol/g, respectively). In liver paraquat increased lipid peroxidation approximately by 60% over the levels of control animals only at 2 h after paraquat treatment. In kidney, augmented lipid peroxidation, 7alpha-OOH and 7beta-OOH (by 70% and 147%, respectively) above levels was found at 2 h after paraquat treatment. Interestingly, these increase remained in kidney of rats 5 d after a single dose of paraquat. In contrast, cholesterol-derived hydroperoxides were not affected in lung of paraquat dosed rats. This is the first report on 7alpha-OOH and 7beta-OOH accumulations in rat liver and kidney, and it seems to reflect greater oxidative stress in the pathology of kidney of rats treated with acute paraquat at low dose.

The SLGs corresponding to the same S24-haplotype are perfectly conserved in three different self-incompatible Brassica campestris L.

We have identified three strains of the same S24-haplotype in self-incompatible B. campestris L. Two of them, S-12j and 1-1j, have been derived from Japanese populations and one, 27-1t, from Turkish one. The cDNA clones of SLG24 (S24-locus glycoprotein), which linked to the S-locus, were isolated from each strain, and sequenced. Each clone isolated from S-12j, 1-1j and 27-1t, was designated as SLG24 (S-12j), SLG24 (1-1j) and SLG24 (27-1t), respectively. Their nucleotide sequences were completely identical in coding region, 5' non-coding region, and 3' non-coding region, though the position of the polyadenylation site was slightly different among the cDNA clones. This result suggests that the origin of S24-haplotype in these three strains might be common, and that the nucleotide sequences of SLG24 of the same S24-haplotype are completely conserved among different populations. The high conservation of the SLG24 nucleotide sequences is probably essential for the recognition of self or non-self of self-incompatibility in this S24-haplotype.

Comparative studies of MRI and operative findings in rotator cuff tear.

A prospective study was performed to determine the accuracy of magnetic resonance imaging (MRI) compared with operative findings in the evaluation of patients associated with rotator cuff tears. Fifty-four of 60 shoulders (58 patients) examined by MRI were confirmed as full-thickness tears and 6 as partial-thickness tears at the time of surgery. The oblique coronal, oblique sagittal, and axial planes of T2-weighted images with the 0.5 tesla MRI system were obtained preoperatively and compared with operative findings. MRI correctly identified 46 of 54 full-thickness rotator cuff tears and 5 of 6 partial-thickness tears. A comparison of MRI and operative findings in full-thickness cuff tears showed a sensitivity of 85%, a specificity of 83%, and a positive prospective value (PPV) of 99%. A comparison of partial-thickness tears showed a sensitivity of 83%, a specificity of 85%, and PPV of 39%. Linear regression analysis showed an excellent correlation between the MRI assessment and measurement at the time of surgery (r = 0.90, P < 0.01). MRI was useful in evaluating large and medium-sized rotator cuff tears, but less useful in distinguishing small full-thickness tears from partial-thickness tears.

[Antigenicity studies of prulifloxacin (NM441)].

Antigenicity of prulifloxacin, a new antibacterial agent, and that of its active metabolite (NM394) were investigated using guinea pigs and mice in this study. In the study with guinea pigs, the animals were immunized with prulifloxacin by oral administration, the predetermined clinical route, and with prulifloxacin alone or emulsified with Freund's complete adjuvant (FCA) by s.c. When active systemic anaphylaxis (ASA) test and passive cutaneous anaphylaxis (PCA) test of these animals were conducted by eliciting with NM394 alone or protein conjugate (NM394-GPSA) of NM394 and guinea pig serum albumin (GPSA), no ASA and PCA reactions were observed. When guinea pigs were immunized subcutaneously with protein conjugate (NM394-BSA) of NM394 and bovine serum albumin (BSA) together with FCA, ASA and PCA reactions were positive by eliciting with NM394-GPSA or NM394-BSA, but were negative by eliciting with NM394 alone. In the study with mice, the animals were immunized orally with prulifloxacin and intraperitoneally with prulifloxacin alone or suspended with aluminum hydroxide gel (alum). When rat PCA test of sera from these mice was conducted by eliciting with NM394 or NM394-GPSA, no positive PCA reaction was observed. When mice were immunized intraperiotneally with NM394-BSA together with alum, PCA reactions were positive by eliciting with NM394-GPSA or NM394-BSA, but were negative by eliciting with NM394 alone. These results show that prulifloxacin has no immunogenicity to guinea pigs and mice, and NM394 as its active form has no eliciting potential to anti-NM394-BSA antibody.

Nine cases of granular cell tumors in B6C3F1 mice.

The histological characteristics of 9 cases of granular cell tumors (GCTs) observed in B6C3F1 mice were examined to determine their cellular origin. Seven of the 9 cases were found in the uterus and other 2 cases were in the subcutaneous tissue. Tumor cells had abundant granules in the cytoplasm which were stained with PAS and were resistant to diastase treatment. Ultrastructurally, the granules were identified as lysosomes. The cell surface had cytoplasmic processus showing interdigitation with adjacent cells. A character feature of the tumor cells was the presence of a desmosome-like structure on their cell surface but no basal lamina was demonstrated. Although GCTs have been considered to be derived from Schwann cells on the basis of their ultrastructural features and S-100 protein-immunopositive findings, the absence of basal lamina in the present cases may raise a controversy as to their origin.

A case of multiple osteochondroma in the rat.

The occurrence of spontaneous osteochondroma in rats is extremely rare and only a few cases have been reported. Spontaneous multiple osteochondroma was found in a male SD strain rat, at the age of 58 weeks from the control group in a toxicity study. Histological findings of both a walnut-sized mass involving the humerus and scapula of right forelimb and a soybean-sized mass adjacent to the costochondrial junction of the right posterior rib were similar in appearance. The outer layer of the tumors consisted of hyaline cartilage and the core of the tumors consisted of trabecular bone with abundant fatty bone marrow. The periosteum of the surface was continuous with that of the parent bone, and cortical bone and the medullary cavity of the parent bone communicated with those of the tumors. Because of showing progressive enlargement, multiple osteochondromas in the rat were considered to be neoplastic in nature.

The alteration of nasal resistance before and after local exposure to heated aerosol in perennial allergic rhinitis.

To determine the patho-physiological effects of heated vapour to the normal or allergic nasal mucosa, we measured the nasal resistance before and after a 10 min. exposure of hyperthermal (43.0 degrees C) aerosol to the nasal mucosa in normal subjects and perennial allergic rhinitis patients. In the allergic patients the mean nasal resistances after hyperthermal stimulation were significantly higher than those resistances without stimulation, both in expiration or inspiration. No significant differences of nasal resistances in normal individuals during the whole schedule with and without heated aerosol stimulation were found on expiration or inspiration. The local heated aerosol exposure increases the nasal resistance in nasal allergic patients while in normal subjects no changes were found, and the reaction may have arisen from a non-specific hypersensitivity of the susceptible allergic nasal mucosa.

[A case of acute pancreatitis that occurred after an operation of the lumbar spine].

A 56-year-old female, who had been suffering from heart failure and diabetes mellitus, underwent posterior instrumentation in the prone position and anterior interbody fusion in the right lateral decubitus position for pyogenic spondylitis between the fourth and fifth lumbar spine under general and epidural anesthesia. We induced hypotensive anesthesia by using continuous infusion of dopamine, prostaglandin E1 and nitroglycerin in order to prevent heart failure and reduce the blood loss. After the operation, the patient complained of upper abdominal pain, nausea and vomiting. We found high levels of serum amylase and other pancreatic enzymes. The massive gas of small intestine was pooled in abdominal X-P, and the pancreatic head was slightly swollen in abdominal CT and US. Therefore we came to the diagnosis of postoperative acute pancreatitis. We administered a single bolus intravenous infusion of ulinastatine and continuous venous infusion of gabexate mesilate. As the serum amylase level gradually decreased, the patient improved. We suspect that postoperative pancreatitis was due to invasive anesthetic and surgical stress on the patient who had had pancreatitis in the preoperative period.

[Superior vena cava reconstruction combined with resection of mediastinal-pulmonary malignant tumors].

From 1988 to 1997, we experienced 5 cases of the superior vena cava (SVC) replacement with expanded polytetrafluoroethylene (ePTFE) grafts combined with resection of mediastinal or pulmonary malignant tumors. Two patients had lung cancer and three had invasive thymoma. Resection and reconstruction of the superior vena cava (SVC) were performed by application of a bypass graft between the innominate vein and the right atrium in two cases and a temporary bypass using a heparin-coated tube in three cases. Except in one patient who died of acute respiratory failure, no complication or occlusive symptom were observed postoperatively. Two patients remain healthy for 5 years 4 months and 2 years 7 months after operation. Three died 9 years, 5 months, and 110 days after operation respectively. In conclusion, ePTFE graft replacement or patch angioplasty of the SVC should be part of planning and execution of radical excision with curative intent of mediastinal or pulmonary malignant tumors.

[Probability of leaving cancer tissue unresected by limited operation in patients with peripheral, stage I non-small cell lung cancer].

To study the probability of leaving metastatic lymph nodes or intralobar metastasis unresected by limited surgery, we reviewed histopathologically 189 patients who had major pulmonary resection and complete lymph adenectomy for peripheral, p-T 1 or T 2 non-small cell lung cancer from 1975 to 1997 at our hospital. Lymph node involvements and or intralobar metastasis were found in 25 (26.0%) of 96 cases with tumor smaller than 3.0 cm and 44 (47.3%) of 93 cases with tumor larger than 3.0 cm. There was no difference between histological types of lung cancer. From this result, it was suggested that the risk of leaving cancer by limited operation would not be low. In conclusion, we believe that limited operation is not an alternative to the standard resection in patients with peripheral, stage I non-small cell lung cancer.

Recovery of normal photosynthesis and respiration in common wheat with Agropyron cytoplasms by telocentric Agropyron chromosomes.

Alloplasmic common wheat (Triticum aestivum L. cultivais 'Penjamo 62' and 'Siete Cerros 66') with cytoplasms of wheatgrass (Agropyron trichophorum and Ag. glaucum) showed two aberrant phenotypes, i.e., gross reduction in plant vigor and male sterility. Plant vigor and male fertility were restored by cytoplasm-specific telocentric chromosomes (telosomes). Studies on carbon assimilation and consumption and on oxygen evolution and uptake showed that maximum rates of apparent photosynthesis were significantly lower in the alloplasmic lines than in their corresponding euplasmic lines and that the telosomes restored a normal level of photosynthesis. The decreased apparent photosynthetic rates in the alloplasmic lines were shown to be not due to decreased rates of true photosynthesis but to increased rates of dark respiration in the green leaves. In contrast, dark respiration in the roots was significantly low in the alloplasmic lines. The alloplasmic lines also showed decreased rates of respiratory consumption of new photosynthates. These results suggest that growth depression and male sterility in the alloplasmic lines are related to aberrant mitochondrial function, which is compensated for by cytoplasm-specific telosomes.

Development of a Simple Cell Invasion Assay System.

Cell invasion assay is important for studying various biological events, such as inflammation, cancer metastasis, and angiogenesis. In this study, we developed a simple method for the quantification of cell invasion by using a culture insert with fluorescence blocking micropore membrane (FBM). Fluorescence labeled cells were simply added to a culture insert with a 8 micrompore FBM precoated with Matrigel and incubated for an appropriate duration. Then, the FBM was examined under a fluorescence microscope to count the invaded cell number. By this method, accurate invasion assay is easily performed without the steps of fixation and staining of cells and removal of cells which do not invade.

Root-specific expression of genes for novel glycine-rich proteins cloned by use of an antiserum against xylem sap proteins of cucumber.

Candidate cDNAs for xylem sap proteins (XSPs) were isolated by screening of a cucumber cDNA library prepared from poly(A)+ RNA of cucumber roots with an antiserum raised against the XSPs of cucumber. The patterns of expression of corresponding transcripts and the sequences of the cDNAs were examined. Two cDNAs with a glycine-rich domain in each deduced amino acid sequence and whose transcripts were expressed predominantly in roots were selected for further analysis. Both cDNAs encoded proteins with a putative signal sequence, with similar non-glycine-rich domains at the amino terminal of the encoded polypeptides. The corresponding mRNAs accumulated concurrently with the formation of adventitious roots from cuttings of cucumber hypocotyls and the development of vascular tissues, and strong expression of the mRNAs was detected in the root-hair zone of tap roots. These data suggest that the production of the two novel glycine-rich proteins, which are putative XSPs, might be associated with the functions of the vascular tissues in roots.

A high degree of homology exists between the protein encoded by SLG and the S receptor domain encoded by SRK in self-incompatible Brassica campestris L.

The sporophytic self-incompatibility system in Brassica campestris is controlled by multiple alleles of a single locus, S. We isolated and characterized cDNA clones that correspond to genes for S-glycoprotein (SLG) and S-receptor kinase (SRK) from an S9-homozygous strain. An SRK9 fragment was amplified by the polymerase chain reaction (PCR) by use of primers that corresponded to regions of the gene for the extracellular receptor domain (S-domain) and the kinase domain. The fragment was used as a probe to isolate an SRK9 cDNA clone from the cDNA library of stigmas of an S9-homozygote. An SLG9 cDNA clone was also isolated from the library by use of SLG8 cDNA as a probe. Restriction fragment length polymorphism (RFLP) linkage analysis of the F2 plants from an S8S9 hybrid demonstrated that the SRK and SLG genes were tightly linked to one another and were also linked to S-genotypes, as determined by pollination tests. The transcripts of SRK9 and SLG9 were detected in stigmas, but not in anthers or leaves. The cytoplasmic kinase domain encoded by SRK9 contained amino acid sequences that are conserved in serine/threonine kinases. The nucleotide sequence encoding the S-domain of SRK9 was 98.4% homologous to that of SLG9 at the nucleotide level. This observation suggests the existence of a mechanism for maintaining a high degree of similarity between SLG and the region that encodes the S-domain of SRK.