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.

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.

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.

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.

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.

Cysteine proteinase gene expression in the endosperm of germinating carrot seeds.

A gene for carrot seed cysteine proteinase (CSCP; AB057371, AB057372) was cloned using PCR. The deduced amino acid sequence of CSCP had active sites for eukaryotic cysteine proteinases, putative signal sequences, and an endoplasmic reticulum targeting sequence. RNA gel blot analysis showed that CSCP transcripts appeared from dry seed, reached a maximum 3 days after imbibition (DAI), and disappeared by 5 DAI. In situ hybridization showed that CSCP transcripts accumulated in the endosperm of germinating seeds. This is the first report of the expression pattern of a cysteine proteinase gene in the endosperm of germinating dicotyledonous plants. The promoter of the CSCP gene had an endosperm motif and many other motifs also found in the promoters of endosperm-specific storage protein genes in monocotyledons. It is suggested that dicotyledons, like monocotyledons, have a temporal and spatial regulation system for endosperm-specific gene expression in germinating seeds.

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.

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.

Morphogenesis of craniopharyngeal derivatives in the neurohypophysis of Fisher 344 rats: abnormally developed epithelial tissues including parotid glands derived from the stomatodeum.

Morphogenesis of craniopharyngeal derivatives of the neurohypophysis found in 14 Fischer 344 (F344) rats was studied. The incidence of the craniopharyngeal derivatives was 0.17% in male (7 out of 4,200) and 0.16% in female (7 out of 4,450) F344 rats. Neither a sex-related difference in their incidence nor a strain-related difference in their morphological features was observed. Craniopharyngeal derivatives were composed of aberrant epithelial structures consisting of serous acinar and tubular and fusiform cell structures, and most of these derivatives were associated with Rathke's cleft cysts, which are suggestive of a congenital background. The acinar structures were positive for periodic acid-Schiff reaction and negative for Alcian blue stain. Immunohistochemically, cells forming these structures were positive for cytokeratin, and basal cells of the acinar or tubular structures and some of the fusiform cells showed positive staining for alpha-smooth muscle actin. Electron microscopically, these spindle-shaped basal cells had intracytoplasmic myofilaments with focal density in their cytoplasm, and they were regarded to be myoepithelial cells. These findings strongly indicate that the craniopharyngeal derivatives are not a neoplastic lesion but rather are a developmental aberration derived from the stomatodeum, which is known to be the origin of both nasal and oral epithelial tissues, including the parotid glands, other than Rathke's pouch.

[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.

A congenital ciliated epithelial cyst on the stomach of a B6C3F1 mouse.

Congenital anomalies of the alimentary tract are rare lesions in laboratory animals. We describe a congenital cyst attached to the greater curvature of the forestomach in a B6C3F1 mouse. The inner surface of the cyst was mostly covered with cuboidal or pseudostratified ciliated epithelium and was focally covered with the flat cuboidal epithelium. The base of the cyst appeared to be inserted between the layers of the outer longitudinal muscle layer of the forestomach, although no smooth muscle layer was evident in the free surface of the cyst wall. The cyst resembled duplication of the alimentary tract, as it was lined with ciliated epithelium and had developed at the greater curvature of the forestomach. Since the smooth muscle layer did not completely cover the whole wall and the cyst did not communicate with the gastric lumen, the cyst was not thought to be a standard duplication but rather a simple congenital cyst.

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.

Eosinophilic granulated cells comprising a tumor in a Fischer rat.

A systemic tumor developed in multiple organs, including spleen, bone marrow, lymph nodes, liver, ovaries, and thoracic and lumbar vertebrae, of a female F344Du/Crj rat. The tumor was composed of round to indented mononuclear cells containing abundant large eosinophilic granules in the cytoplasm. The peripheral blood smear revealed that the large granules in the cytoplasm of the tumor cells were stained basophilic with Giemsa, suggesting a basophil or mast cell origin. However, these granules did not show metachromasia with toluidine blue and were stained blue to dark blue with phosphotungstic acid hematoxylin. Cellular morphology and characteristics in the specific stains of tumor cells suggested the development of a tumor of globule leukocytes in a F344Du/Crj rat.

[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.

Promotive effects of the peptidyl plant growth factor, phytosulfokine-alpha, on the growth and chlorophyll content of Arabidopsis seedlings under high night-time temperature conditions.

In order to investigate the function of the peptidyl plant growth factor, phytosulfokine-alpha (PSK-alpha), in plants, we examined the effect of PSK-alpha on the growth and chlorophyll content of Arabidopsis seedlings under high night-time temperature conditions. Although exposure to high night-time temperatures markedly reduced the fresh weight and chlorophyll content of the seedlings, these parameters in the plants supplied with PSK-alpha remained at the same levels as those of non-treated controls. These effects were not apparent when [2-5]PSK, Tyr-SO3H and kinetin were similarly supplied. The results suggest that PSK-alpha not only promotes cell proliferation, but may aid plants in their tolerance of heat stress.

[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.

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.

Promotion by a Peptidyl Growth Factor, Phytosulfokine, of Chlorophyll Formation in Etiolated Cotyledon of Cucumber.

We examined the effects of a peptidyl growth factor, phytosulfokine-α (PSK-α), isolated from conditioned medium and known to induce the proliferation of single mesophyll cells of asparagus, and its derivatives on chlorophyll formation in etiolated cotyledons of cucumber after illumination. The chlorophyll content was increased by PSK-α treatment. Chlorophyll-content increases were not observed following either [2-5]PSK or Tyr-SO3H treatment. There was no difference between the PSK-α treatments and water controls on increasing rate of fresh weights of cotyledons. These results suggest that PSK-α has the effect of elevating the chlorophyll content in the cells.

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.

[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.