Methemoglobin (Met-Hb) is very unstable and, during the storage of blood samples, Met-Fib can be both formed and reduced. To prevent changes in Met-Hb concentrations in blood samples during storage, we have devised two methods, one in which samples are stored at -30°C with a cryoprotectant and another in which samples are stored at -80°C or -196°C without any further additions. Both methods work well for samples of human blood stored for less than 30 days and prepared in vitro. In the present study, we examined the possibility of using these two methods to store samples for longer (e.g. up to 180 days) in the case of untreated, heated, or nitrite-treated human blood samples, as well as blood samples obtained from nitrite-treated and untreated rats. There was a marked increase in Met-HI, concentrations as a result of autoxidation in samples of untreated or heated human blood stored without a cryoprotectant at -80°C over a period of 180 days; however, Met-Hb levels were relatively stable in the same sorts of samples at -196°C over the same period. For all three types of human blood sample stored with a cryoprotectant, Met-Hb levels were relatively stable at -30°C, but even more so at -80°C and -196°C, for 180 days. When blood samples from nitritetreated or untreated rats were stored without a cryoprotectant, marked autoxidation occurred at -80°C and at -196°C, although the degree of autoxidation was less at the lower temperature. When blood samples from rats were stored with a cryoprotectant, Met-Fib levels were relatively stable at -30°C, and even more so at -80°C and -196°C, for 90 days regardless of the initial concentration of Met-Fib in the sample. From these results, it appears that, for Met-Hb determination, storage of human blood samples with a cryoprotectant at temperatures of -30°C or lower and storage without any further additions at -196°C are both suitable. However, frozen storage of blood samples from rats requires the addition of a cryoprotectant. Storage of blood samples from rats without a cryoprotectant at -80°C and -196°C resulted in Met-Fib formation by autoxidation.

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The sensorimotor area, supplementary motor area, and cingulate cortex of the frontal medial wall are important regions for the generation and control of movement. Traditionally, investigations into the control of movement were performed using neuroimaging and electroencephalography recordings with subdural electrodes. Recent advances in electroencephalograph analysis with dipole-tracing analysis incorporating a realistic three-layer head model (scalp-skull-brain head model) provide a non-invasive method for the detection of dipoles in the millisecond range. These advances allow investigations into the underlying processing of cognitive function and movement execution. In the present study, we constructed a scalp-skull-brain head model from Montreal Neurological Institute standard brain images and detected dipole localizations in the millisecond range from the grand-averaged negative slope to motor potentials during a simple pinching movement. We used dipole analysis of the grand average for all subjects, incorporating the Montreal Neurological Institute standard brain images, and show the involvement of general locations related to the task. For left hand pinching, the electroencephalograph revealed the order as Bereitschafts potential, negative slope, motor potential. The dipoles were converged in the same order in the supplementary motor area proper, cingulate motor area, and in the sensorimotor area, area for hand motor area. A similar pattern was observed for right hand pinching movement. We show that the dipole converges in the contralateral side of the hippocampus and amygdala before the onset of the Bereitschafts potential. Since the hippocampus and amygdala are associated with the limbic cycles, our findings suggest that both the hippocampus and amygdala are required for the function of the cortico-basal ganglionic thalamocortical loop. Together, this study suggests that strong emotional factors must exist before intentional movement, and before activation of the higher cortical centers.

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To evaluate the image quality of the urinary tract using oral administration of a combination of both water and a negative gastrointestinal contrast agent, Bothdel. The study was divided into two parts: the phantom study and the healthy volunteers study. The imaging sequences used were the rapid acquisition with relaxation enhancement (RARE) and the half-Fourier acquisition single-shot turbo spin-echo (HASTE) techniques. Magnetic resonance Urography (MRU) was performed in 13 healthy volunteers using the RARE imaging sequences. After collection of the pre-contrast images, post-contrast MRU was obtained 30 min and 60 min after the oral administration of the contrast agent. Image assessment was based on the contrast effect, the image effect, and the opacification score. Statistical analysis was performed using the Wilcoxon signed-rank test. The signal intensity using 100%-12.5% Bothdel was no different to the background noise using the RARE sequence. At concentrations below 12.5%, there were statistically significant improvements in the contrast, the image effect, and the opacification score between pre-and post-contrast images. No significant difference was observed between the 30 min and 60 min post-contrast images. In contrast, the effect and pre-contrast images were graded as poor in seven cases, whereas no post-contrast images were graded as poor. There was a particularly significant improvement in the opacification score with the distal ureter being the most difficult segment to opacify. Oral administration of both water and Bothdel effectively removed the bowel signal and improved visualization of the urinary tract.

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Non-functioning pituitary adenomas (NFPA) can be classified as either null cell adenomas or gonadotropin adenomas. NFPAs show few clinical signs and symptoms reflecting hormone production and do not manifest clinically as easily recognizable pituitary hypersecretory syndromes. MRI scans are the best way to detect NFPAs, but unfortunately most of the patients who undergo MRI generally have neuroophthalmological or neurological symptoms. It is therefore necessary to confirm the diagnosis of NFPAs using other methods such as gene expression profiling. However, the expression profiles of NFPA transcripts have been unclear to date. To overcome these diagnostic obstacles and to identify NFPA molecular markers, we used a combined approach involving microdissection, immunostaining and cDNA microarray analysis. Samples were collected from patients clinically diagnosed with NFPA by transsphenoidal resection. The control pituitary gland used in this study was freshly dissected from a patient with a brain tumor. The amount of RNA extracted did not correlate with the immunostaining results, but microarray analysis revealed overexpression of candidate molecules in NFPAs. In particular, SLITRK4 was found to have a specific expression pattern in sub-classified NFPA, and was overexpressed specifically in null cell adenomas. SLITRK4 expression was inversely correlated with the MIB-1 index. Our results suggested that SLITRK4 was expressed while the cells of the pituitary gland were undergoing tumorigenic transformation. Expression of SLITRK4 aggravated the development of NFPA, and facilitated tumor generation.

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Complications such as malnutrition and diabetes often result from dysfunctional pancreatic endocrine and exocrine systems in pancreatitis. These systems also differ functionally during healing following pancreatitis. This study examined pancreatic regeneration in both patients with autoimmune pancreatitis (AIP) during treatment with prednisolone, and in two different rat models of acute pancreatitis. Human pancreatic tissue from AIP patients was immunostained to identify pancreatic cells in the phase of regeneration by using an anti-Ki-67 (a proliferation marker) antibody. One rat model used caerulein (CAE) to generate acute edematous pancreatitis (AEP), and the second involved administration of DL-ethionine to generate acute necrotizing pancreatitis (ANP) . A labeling index (LI) was established as the ratio of proliferating cell nuclear antigen (PCNA) -labeled acinar cells to total acinar cells, while the ratio of acinar cells showing mitosis to the total number of acinar cells was calculated as the mitotic index (MI) . Pancreatic progenitor cells were identified using an anti-pancreatic/duodenal homeobox-1 (PDX-1; an transcription activator) antibody. Additionally, anti-hairy/enhancer-of-split homologue-1 (HES-1) antibody was used to detect immature pancreatic exocrine cells. Human pancreatic tissue with AIP was immunostained. Ki-67-positive cells coincided with acinar cells and ductal cells, and PDX-1-positive cells coincided with pancreatic ductal-type cells. Some α-amylase-positive cells and most insulin- and glucagon-positive cells were PDX-1-positive. The severity of AEP was evident 4-12h after initial injection of CAE in the first rat model. The PCNA LI gradually increased to a maximum at 120h, while the MI peaked at 96 h. In rats with ANP, necrosis and inflammation were prominent but diminished during healing, while the PCNA-LI increased to a peak at 12 days after initial injection of ET, and the MI maximum occurred at 10 days. PDX-1 was strongly expressed only in islet cells after AEP, while expression following ANP was intense in islet cells, several ductal-type cells, and a few acinar cells. Few HES-1-positive cells were observed among acinar or ductal cells. This study clarified the temporal nature of pancreatic regeneration in two rat models of acute pancreatitis. The results indicated that regeneration after AEP in rats might not involve pancreatic progenitor cells. However, regeneration after ANP in rats and AIP in humans involves pancreatic progenitor cells that probably derive from the dedifferentiation of acinar and ductal cells affected by inflammation.

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Two cases of putative benign transitional epithelial (urothelial) cyst of the ovary are reported. A 45-year-old woman (case 1) presented with torsion of an ovarian cyst and a 70-year-old woman (case 2) presented with a large multi-locular ovarian cyst. The inner surface of the thickened cystic wall in both cases was covered by a multilayered transitional epithelium without atypia, and in each case the epithelium had a smooth luminal border. Immunohistochemically, the epithelial component of the resected cystic lesions was positive for uroplakin III in case 1 and negative in case 2. Both cases were positive for cytokeratin (CK) 7, and negative for CK 20. In these cases, the epithelial components resembled the urothelium of the bladder and the epithelial element of Brenner tumors, both histologically and immunohistologically. In conclusion, we propose that there are two sub-classifications of benign transitional cell tumors, a common (mainly solid) Brenner tumor and a transitional epithelial (urothelial) cyst.

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