An MR-DTI-based fiber tracking method for the multimodal integrative study of cognitive brain functions.

We have developed a method for white matter fiber tracking inferred from magnetic resonance diffusion tensor imaging (MR-DTI) to match functional and anatomical information in the human brain. Functional information was obtained using functional magnetic resonance imaging (fMRI) and magnetoencephalography (MEG). One of the major problems in fiber tracking based on MR-DTI is the error of fiber orientation estimation at areas of fiber crossing. Here, we propose a novel fiber tracking method involving searching for similarity of direction vectors in the vicinity of fiber crossing areas. The proposed method was tested on simulation images and its utility was validated. Subsequently, the method was applied to in vivo data, and we tried to fuse the data obtained with the method with functional information obtained from a newly developed fMRI-MEG integrative method. The fMRI-MEG integrative method successfully detected dynamic cortical activities in multiple visual areas, such as V1, V2, and V5, during an apparent visual motion perception task. Our fiber tracking results demonstrate that the present method can be used to confirm connectivity among multiple cortical areas associated with higher cognitive brain functions.

[A case of female urethral diverticulum calculus associated with multiple sclerosis].

We report a case of urethral diverticulum with a calculus in a 52-year-old female. She had suffered from transverse myelopathy (flaccid paralysis) as a result of multiple sclerosis. She presented with total incontinence and urinary tract infection that did not respond to antibiotic therapy. We found a urethral diverticulum calculus by X-ray imaging and urethroscopy. Transvaginal urethral diverticulectomy with removal of stone was carried out without any complications. The removed stone was 35 x 31 x 19 mm in size and was composed of magnesium ammonium phosphate and calcium phosphate.

Gut cryptopatches: direct evidence of extrathymic anatomical sites for intestinal T lymphopoiesis.

Athymic cytokine receptor gamma chain mutant mice that lack the thymus, Peyer's patches, cryptopatches (CP), and intestinal T cells were reconstituted with wild-type bone marrow cells. Bone marrow-derived TCR(-) intraepithelial lymphocytes (IEL) first appeared within villous epithelia of small intestine overlying the regenerated CP, and these TCR(-) IEL subsequently emerged throughout the epithelia. Thereafter, TCR(+) IEL increased to a comparable number to that in athymic mice and consisted of TCRgammadelta and TCRalphabeta IEL. In gut-associated lymphoid tissues of wild-type mice, only CP harbored a large population of c-kit(high)IL-7R(+)CD44(+)Thy-1(+/-)CD4(+/-)CD25(low/-)alpha(E) beta(7)(-)Lin(-) (Lin, lineage markers) lymphocytes that included cells expressing germline but not rearranged TCRgamma and TCRbeta gene transcripts. These findings provide direct evidence that gut CP develop progenitor T cells for extrathymic IEL descendants.

Voiding dysfunction and Parkinson's disease: urodynamic abnormalities and urinary symptoms.

PURPOSE: We evaluated bladder dysfunction and Parkinson's disease in regard to disease severity and determined whether subjective patient urinary symptoms correlated with urodynamic abnormalities. MATERIALS AND METHODS: We assessed bladder dysfunction in 70 patients with Parkinson's disease and urinary symptoms using the International Prostate Symptom Score and urodynamic tests. RESULTS: Urodynamic evaluation revealed detrusor hyperreflexia in 47 patients (67%), hyporeflexia or areflexia in 11 (16%), hyperreflexia with impaired contractile function in 6 (9%), hyperreflexia with detrusor-sphincter dyssynergia in 2 (3%) and normal function in 4 (6%). The incidence of urodynamic abnormalities appeared to increase with disease severity. However, the only urodynamic parameter that correlated with disease severity was post-void residual urine volume. On the other hand, symptom index scores increased with disease severity. The irritative symptom score correlated with maximum cystometric capacity and volume at initial desire to void, whereas the obstructive symptom score correlated with post-void residual urine volume. Also, irritative and obstructive scores were good predictors of overactivity during the storage and underactivity at the voiding phases. CONCLUSIONS: Bladder function may deteriorate progressively with advancing disease. Symptom scores are fairly accurate for predicting likely urodynamic abnormalities. Our results imply that quantifying subjective urinary symptoms is useful for estimating the severity and type of bladder dysfunction.

[Papillary adenocarcinoma of the prostate: report of 3 cases].

We report here 3 cases of papillary adenocarcinoma of the prostate. In all 3 cases, the tumors were discernible on cystourethroscopy and transurethral biopsy established the diagnosis, whereas no significant finding was found on digital rectal examination. Although androgen deprivation therapy was administered in all cases, different surgical procedures were employed according to the stage in each case. In case 1, since the papillary tumor was confined within the prostatic urethra, complete resection was accomplished by transurethral resection (TUR). In case 2, since pelvic lymph nodes metastases were found, local radiation therapy was added. In case 3, since the patient had vesical invasion of tumor total cysto-prostatectomy was performed. Papillary adenocarcinoma of the prostate originates from the prostatic duct, resulting in existence at the "central portion" of the prostate gland. Cystourethroscopy and transurethral biopsy is helpful for diagnosis of this disease, whereas rectal digital examination is useless. As a surgical procedure for the primary site, TUR may be efficient for tumors confined within the prostatic urethra, although more extensive surgery may be necessary for those with a more invasive profile.

A preliminary study on the knowledge and attitudes of physicians at two university hospitals towards the medical insurance system of Japan.

In this preliminary study, we surveyed the physicians at two academic hospitals on their knowledge of and attitudes toward the medical insurance system in Japan. Most of the physicians had not read the "Ministerial Ordinance on Insurance Medical Institutions' and Insurance Medical Doctors' Medical Treatment under Health Insurance." Of the 433 physicians who filled out the questionnaire completely, 34% had either not read or rarely read the "Medical Fee Point List." Most (89.1%) of the physicians knew that there is a stepwise reduction in the hospitalization fee as the length of a patient's hospital stay increases. However, approximately 30% did not know the stipulation of obtaining an informed consent from the patient prior to blood transfusion. As for the right of patients to see their medical care remuneration statements, which was decided by the government in 1997, 26.8% of the physicians did not know this rule. Physicians who had read the "Ministerial Ordinance on Medical Treatment," were more likely to read the "Medical Fee Point List" frequently; were more likely to know the stipulation about diminishing hospitalization fee; were more likely to know that an informed consent must be obtained prior to blood transfusion; and were more likely to know that patients had a right to see their medical care remuneration statements. The longer the clinical experience of the physician, the more likely that the physician had read the "Ministerial Ordinance on Medical Treatment" and know the other stipulations well. In these two academic hospitals, it is important to establish educational seminars for physicians on the guidelines of the medical insurance system so that physicians will become familiar with the medical insurance system quickly.

Role of gut cryptopatches in early extrathymic maturation of intestinal intraepithelial T cells.

Lympho-hemopoietic progenitors residing in murine gut cryptopatches (CP) have been shown to generate intestinal intraepithelial T cells (IEL). To investigate the role of CP in progenitor maturation, we analyzed IEL in male mice with a truncated mutation of common cytokine receptor gamma-chain (CRgamma-/Y) in which CP were undetectable. IEL-expressing TCR-gammadelta (gammadelta-IEL) were absent, and a drastically reduced number of Thy-1highCD4+ and Thy-1highCD8alphabeta+ alphabeta-IEL were present in CRgamma-/Y mice, whereas these alphabeta-IEL disappeared from athymic CRgamma-/Y littermate mice. Athymic CRgamma-/Y mice possessed a small TCR- and alphaEbeta7 integrin-negative IEL population, characterized by the disappearance of the extrathymic CD8alphaalpha+ subset, that expressed pre-Talpha, RAG-2, and TCR-Cbeta but not CD3epsilon transcripts. These TCR- IEL from athymic CRgamma-/Y mice did not undergo Dbeta-Jbeta and Vdelta-Jdelta joinings, despite normal rearrangements at the TCR-beta and -delta loci in thymocytes from euthymic CRgamma-/Y mice. In contrast, athymic severe combined immunodeficient mice in which CP developed normally possessed two major TCR-alphaEbeta7+ CD8alphaalpha+ and CD8- IEL populations that expressed pre-Talpha, RAG-2, TCR-Cbeta, and CD3epsilon transcripts. These findings underscore the role of gut CP in the early extrathymic maturation of CD8alphaalpha+ IEL, including cell-surface expression of alphaEbeta7 integrin, CD3epsilon gene transcription, and TCR gene rearrangements.

[Collecting duct carcinoma (Bellini duct carcinoma) of the kidney with tumor extension into the inferior vena cava].

We report a case of collecting duct carcinoma (Bellini duct carcinoma) of the left kidney accompanied with a tumor thrombus in the inferior vena cava and the lymph node metastasis. A 69-year-old male presented with gross hematuria and left flank dullness. Computed tomography revealed an isodensity tumor in the left kidney with tumor extension into the inferior vena cava and the regional lymph node swelling. The T1-weighted magnetic resonance image displayed a slightly heterogeneous low-intensity-mass. Renal angiography revealed a hypervascular tumor. We performed left radical nephrectomy with tumor thrombectomy and regional lymphadenectomy. Histopathological examination revealed a collecting duct carcinoma (pT3bN1M0V2a). Seven months after surgery, multiple metastates in bone and liver developed. Then we performed systemic chemotherapy consisting of methotrexate and cisplatin. However, the patient died from the carcinoma 10 months postoperatively.

Lifestyles and sleep disorders among the Japanese adult population.

To clarify the effects of daily stress, habitual exercise, drinking and smoking on the prevalence of sleep disorders, we selected 4000 residents (> or =20 years) in Japan by stratified random sampling and carried out structured interviews (response rate 75.8%). Frequencies of sleep disorders (difficulty initiating sleep: DIS; difficulty maintaining sleep: DMS; early morning awakening and hypnotic medication use) were treated as dependent variables. Significant effects of stress were prevalent in all sleep disorders. Habitual exercise had significant negative association with DIS and DMS. Drinking and smoking did not affect sleep disorders.

Laparoscopic treatment for common bile duct stones by transcystic papilla balloon dilatation technique.

The laparoscopic transcystic common bile duct (CBD) approach is becoming increasingly more refined as an ideal technique to deal with gallbladder stones (GBS) and common bile duct stones (CBDS) during a single operation. Our method, transcystic CBD exploration and papilla balloon dilatation (PBD), is an easier, safer, and less invasive technique than the transcystic approaches that have previously been reported. With our method, a sheath is introduced through the cystic duct into the CBD in order to allow catheter exchange, and the CBDS is flushed out through the papilla into the duodenum after PBD. We applied our new technique, without complication, to a patient with GBS and CBDS. Our technique is one of the safest, easiest, and least invasive methods for the treatment of patients with GBS and CBDS.

New gut associated lymphoid tissue "cryptopatches" breed murine intestinal intraepithelial T cell precursors.

Numerous mouse intraepithelial T cells (IEL) bearing either TCR-alphabeta or TCR-gammadelta have been shown to develop somewhere in the intestinal mucosa without passing through the thymus. However, just where these T cells develop has been much less clear and has remained an open question to date. In an effort to investigate this issue, we carried out immunohistochemical study on the murine gastrointestinal tract and identified numerous tiny lymphoid tissues (approximately 1,650 tissues/intestine) in the cryptal region of the small and large intestinal mucosa except for the stomach in which clusters of c-kit+ IL-7R+ Thy1+ lympho-hemopoietic progenitors accumulated (cryptopatches). The cryptopatch cells isolated from the small intestine, which were c-kit positive (c-kit+) but lineage marker negative (Lin-), gave rise to TCR-alphabeta and TCR-gammadelta IELs following in vivo transfer or tissue engraftment into 2 Gy-irradiated severe combined immunodeficient mice. In contrast, cells isolated from Peyer's patches and mesenteric lymph nodes, which belong in the same intestinal immune compartment but lack c-kit+Lin- cells, failed to do so. These results in conjunction with the findings of electron microscopic analysis provide direct evidence of a local intestinal T cell precursor that develops in the cryptopatches.

Expression of CD44 in rat liver allografts during rejection.

As CD44 is believed to be a homing receptor involved in lymphoid trafficking and inflammatory responses, it is expected to be closely linked to transplant rejection. In this study, the expression of CD44 during liver transplant rejection was compared with the expression of lymphocyte-function associated antigen-1 (LFA-1) and intercellular adhesion molecule-1 (ICAM-1), which play an essential role in cell interactions and the initiation of immune responses. Male Brown Norway (BN) and Lewis (LEW) rats were used as donors and recipients, respectively. Orthotopic liver transplantation (OLTX) was done using the cuff technique of Kamada and Calne. Animals were killed on days 3, 5, and 7 after OLTX, and a piece of tissue from each of the liver grafts was obtained. Immunohistochemical staining was used to investigate the expression of CD44, ICAM-1, and LFA-1. CD44 was strongly expressed in portal areas of the rejected liver, and LFA-1 and ICAM-1 were expressed mainly on sinusoids and hepatocytes. These findings indicate that CD44 is closely involved in lymphocyte infiltration, which is dominant in portal areas, and that lymphocyte infiltration during the rejection process may involve a homing mechanism.

Fluorescence lifetime imaging microscopy (flimscopy). Methodology development and application to studies of endosome fusion in single cells.

A new method of fluorescence microscopy for cell imaging has been developed that takes advantage of the spatial variations of fluorescence lifetimes in single cells as a source of image contrast, and thus it is named "fluorescence lifetime imaging microscopy (flimscopy)". Since time-resolved fluorescence measurements are sensitive to molecular dynamics and interactions, flimscopy allows the molecular information to be visualized in single cells. In flimscopy measurements, several (nanosecond) time-resolved fluorescence images of a sample are obtained at various delay times after pulsed laser excitation of the microscope's entire field of view. Lifetimes are calculated pixel-by-pixel from these time-resolved images, and the spatial variations of the lifetimes are then displayed in a pseudocolor format (flimscopy image). The total data acquisition time needed to obtain a flimscopy image with the diffraction-limited spatial resolution (approximately 250 nm) is decreased to just approximately 30 s for approximately 300 fluorescent molecules/micron2. This was achieved by developing a high-frequency (400 kHz) nanosecond-gating (9 ns full width at half height)-signal accumulation system. This technique allows the extent of resonance energy transfer to be visualized in single living cells, and is free from the errors due to variations in path length, light scattering, and the number of fluorophores that necessitate complex corrections in steady-state microfluorometry and fluorescence ratio imaging microscopy. Flimscopy was applied here to observe the extent of fusion of individual endosomes in single cells. Results revealed the occurrence of extensive fusion between primary endocytic vesicles and/or sorting endosomes, thereby raising the possibility that the biogenesis of sorting endosomes involves multiple fusions of primary endocytic vesicles.

Preparation and characterization of oligonucleotides containing S-[2-(N7-guanyl)ethyl]glutathione.

S-[2-(N7-Guanyl)ethyl]glutathione is the major adduct derived from modification of DNA with 1,2-dibromoethane in biological systems and is postulated to be a mutagenic lesion [Humphreys, W. G., Kim, D.-H., Cmarik, J. L., Shimada, T., & Guengerich, F. P. (1990) Biochemistry 29, 10342-10350]. Oligonucleotides containing this modified base were prepared by treatment of oligonucleotides with S-(2-chloroethyl)glutathione and purified by chromatography. The self-complementary oligonucleotide d(ATGCAT), when thus modified at the single guanine, appeared to associate with itself as judged by UV measurements, but CD and NMR measurements indicated a lack of hybridization, with a decrease in the melting temperature of greater than 10 degrees C. The same lack of self-association was noted when d(ATGCAT) was modified to contain an N-acetyl-S-[2-(N7-guanyl)ethyl]cysteine methyl ester moiety. The oligomer d-(C1A2T3G4C5C6T7) was modified to contain a single S-[2-(N7-guanyl)ethyl]glutathione moiety at the central position, and UV, CD, and 1H NMR studies indicated that this oligomer hybridized to its normal complement d(A8G9G10C11A12T13G14), although the binding was considerably weakened by adduction (imino proton NMR spectroscopy in the presence of H2O indicated that the hydrogen bond signals seen in the oligomer were all broadened upon modification). All proton resonances were identified using two-dimensional 1H NMR spectroscopy. Adduct formation affected the chemical shifts of the base and 1', 2', and 2" protons of T3 and C5, the 2" proton of C6, and the 8 and 1' protons of C11, while little effect was observed on other protons. No cross-peaks were detected between the glutathione and oligomer moieties in two-dimensional nuclear Overhauser enhanced NMR studies. These results suggest that a rather local structural perturbation occurs in the DNA oligomer upon modification and that the glutathione moiety appears to be relatively unperturbed by its placement in the duplex. When the cytosine in the normal d(AGGCATG) complement to d-(CATGCCT) was changed to each of the other three potential bases at the central position, no hybridization with the oligomer d(CATGCCT) containing S-[2-(N7-guanyl)ethyl]glutathione was detected. We conclude that these N7-guanyl derivatives destabilize hybridization and that bases other than cytosine do not appear to show preferential thermodynamic bonding to these adducts, at least in the sequences examined to date.(ABSTRACT TRUNCATED AT 400 WORDS)

Effects of adduct formation derived from ethylene dibromide on DNA conformation.

Spectral properties of DNA oligomers containing the single modified guanine, S-[2-(N7-guanyl)ethyl]-glutathione, the major adduct derived from 1,2-dibromoethane, were investigated using UV, CD, and NMR. Two palindromic hexamers, d(ATGCAT) and d(ATCGAT), did not form a duplex with guanine bases modified. When the non self-complementary heptamer, d(CATGCCT), was modified at the single guanine, it formed a duplex with its normal complement d(AGGCATG), although the melting temperature was lowered. However, no duplex formation was observed when a non complementary base other than cytosine was placed in d(AGGXATG), suggesting that non Watson-Crick type base pairs are not stabilized by formation of this adduct.

Direct radioimmunoassay for haloperidol in human serum.

A direct radioimmunoassay for the accurate determination of haloperidol in human serum has been developed. Based on recent information about the metabolism of haloperidol, a new haloperidol hapten, in which a (3-carboxypropionyl)methylamino group was attached as a bridge in the place of fluorine atom, was synthesized and coupled to bovine serum albumin through the bridge to provide a new immunogen. Guinea pigs were used for the immunization. Since the antisera obtained by the new immunogen still cross reacted greater than 10% with reduced haloperidol, the immunological tolerance to reduced haloperidol was induced by administration of a copolymer of D-glutamic acid and D-lysine linked with reduced haloperidol. This gave an antiserum in guinea pigs which was highly specific for unchanged haloperidol with negligible cross reactivity (less than or equal to 1.0%) to any haloperidol metabolites including the newly found ones. With the newly developed antiserum and [3H]haloperidol, serum haloperidol levels can be determined over the concentration range from 0.3 to 20 ng/mL, using 0.1 mL of human serum, without an extraction procedure.