Urinary steroid profiling using liquid chromatography-tandem mass spectrometry for the diagnosis of canine Cushing's syndrome.

Serum cortisol measurements by chemiluminescence enzyme immunoassay (CLEIA) are widely used to diagnose hypercortisolism (HC) or Cushing's syndrome in dogs. However, they are associated with problems such as the need for multiple blood collections under stressful conditions or cross-reactivity between hormones. Therefore, a less invasive and more accurate diagnostic method is required. This study aimed to develop a urinary steroid profile analysis method using liquid chromatography-tandem mass spectrometry (LC/MS/MS) and to evaluate its clinical usefulness. Sixty-five healthy dogs and 38 dogs with suspected HC were included in the study. Using LC/MS/MS, the levels of 11 steroid hormones in the urine were determined. We established the upper limit of the reference interval for each urinary steroid-to-creatinine ratio and evaluated their diagnostic performances. The levels of the five steroid hormones were significantly higher in the 14 dogs with HC than in the 24 dogs with mimicking HC and 65 healthy dogs. The urinary corticosterone-to-creatinine ratio showed the highest diagnostic accuracy (area under the curve, 0.96). A significant correlation was seen between urinary cortisol concentrations measured by LC/MS/MS and CLEIA (rs = 0.88, P <0.001), although the CLEIA measurements were significantly higher than the LC/MS/MS measurements (P <0.001). LC/MS/MS-based urinary steroid profiles are a promising tool for diagnosing canine HC.

Spatial distribution of cingulate cells projecting to the primary, supplementary, and pre-supplementary motor areas: a retrograde multiple labeling study in the macaque monkey.

We examined the location and spatial distribution of cingulate cortical cells projecting to the forelimb areas of the primary motor cortex (MI), supplementary motor area (SMA), and pre-supplementary motor area (pre-SMA) using a multiple retrograde labeling technique in the monkeys (Macaca fuscata). The forelimb areas of the MI, SMA and pre-SMA were physiologically identified, based on the findings of intracortical microstimulation (ICMS) and single cell recording. Three different tracers, diamidino yellow (DY), fast blue (FB), and wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP), were injected into each of the three motor areas in the same monkey. Retrogradely labeled cells in the cingulate cortex were plotted with an automated plotting system. Cells projecting to the forelimb area of the MI were distributed in the two separate regions situated rostrocaudally in the dorsal and ventral banks of the cingulate sulcus, namely the rostral cingulate motor area (CMAr) and caudal cingulate motor area (CMAc). These two regions corresponded to the forelimb areas identified by the ICMS in the same animal. The distribution of projection cells to the SMA overlapped extensively with that of projection cells to the MI. Although the MI received relatively sparse inputs from the CMAr than from the CMAc, the SMA received inputs from the CMAr and its adjacent areas as much as from the CMAc. The projection cells to the pre-SMA were distributed in the anterior portion of the cingulate cortex, including the anterior part of the CMAr and in a small part of the cingulate gyrus. These findings indicate that the MI and SMA share a considerable common information from the cingulate cortex, including the CMAr and CMAc, whereas the pre-SMA receives a different set of information from the anterior part of the cingulate cortex.

Diterpenoids from the fruits of Vitex trifolia.

An abietane-type diterpene, named vitetrifolin A, and two labdane-type diterpenes, named vitetrifolins B and C, were isolated from the acetone extract of the fruits of Vitex trifolia L. (Viticis Fructus; Verbenaceae) along with three known diterpenes, rotundifuran, dihydrosolidagenone and abietatriene 3beta-ol. The structures of these compounds were elucidated on the basis of spectroscopic analysis, X-ray crystallographic analysis and chemical evidence.

[Rapid identification of Bilophila wadsworthia].

Biophila wadsworthia is a recently recognized nonspore-forming anaerobic gram-negative rod and is reported to be associated with various infections such as gangrenous perforated appendicitis, peritonitis, osteomyelitis and bacteremia. Although the isolation of B. wadsworthia seems to be facilitated by using Bacteroides bile esculin (BBE) agar, the reliable scheme of identification of this species has been published. This study was conducted to find simple, rapid, and reliable measures for identification of B. wadsworthia. A total of 32 B. wadsworthia-suspected clinical isolates as well as B. wadsworthia WAL 7959 and Desulfomonas pigra DSM 749, which is the species that shows bacteriological characteristics similar to B. wadsworthia, were used. Through the study of various biochemical and enzymatic tests and culture on selective media, it is indicated that a nonspore-forming, gram-negative anaerobic rod that is nonmotile, forms "black-eyed" colonies on BBE agar, and demonstrates strongly positive catalase test, positive acid phosphatase test, and no-growth on Bacterides medium or to be susceptible to colistin (10 micrograms disk) can be identified as B. wadsworthia.

Bacterial flora detected in the uterine endometrial cavity of normal puerperae on the puerperal first day and on the puerperal fifth day after incidental use of cefpodoxime proxetil.

In Japan, oral antimicrobial agents are prophylactically used with oxytocics after normal delivery to prevent puerperal infections. The present study was designed to investigate bacterial floras in the endometrial cavity immediately after normal delivery and the effect of prophylactic use of anti-microbial agents on those floras. Sixty-six puerperae who underwent uneventful courses of pregnancy and delivery were subjected for this study. Intrauterine contents were collected on the first day and the fifth day of the puerperium and submitted to microbiological examinations. Cefpodoxime proxetil (CPDX-PR) was orally given to the puerperae for prophylaxis for 5 days after the initial sampling. On the puerperal first day, a total of 98 strains (71 strains of aerobic bacteria, 27 strains of anaerobic bacteria) was detected in the uteri of the 66 subjects. The incidences of aerobic Gram-positive cocci, aerobic Gram-negative bacilli and anaerobic bacteria were 59.2%, 12.2%, 27.6% of the 98 strains, respectively. On the puerperal fifth day, a total of 82 strains (51 strains of aerobic bacteria and 31 strains of anaerobic bacteria) were detected in the uteri of the 66 subjects. The incidences of aerobic Gram-positive cocci, aerobic Gram-negative bacilli and anaerobic bacteria were 52.5%, 8.6% and 37.7% of 82 strains, respectively.

[The frequency of Streptococcus pneumoniae strains and sensitivity surveillance for several antibiotics in Gifu Prefecture].

The frequency and the antibacterial sensitivity of Streptococcus pneumoniae strains isolated from 6 key hospitals (in 5 areas) and 1 otorhinolaryngology clinic in Gifu Prefecture from February to March, 1999, were investigated with several antibiotics. A total of 128 strains of Streptococcus pneumoniae were isolated throughout the study: 47 strains (36.7%) of penicillin-susceptible S. pneumoniae (PSSP), 51 strains (39.8%) of penicillin-intermediate S. pneumoniae (PISP), and 30 strains (23.4%) of penicillin-resistant S. pneumoniae (PRSP); the resistant bacteria being relatively prominent. In these hospitals, PSSP was isolated by 38.8% in all the key hospitals and by 30% in the otolaryngology clinic with almost no discernible difference. PISP was isolated by 63.3%, higher in the otolaryngology clinic and PRSP by 28.6%, higher in the key hospitals conversely. The MIC90s in PISP and PRSP were determined with the antibiotics. In result, only cefditoren (CDTR) showed favorable antibacterial activities with the MIC90 of 0.78 microgram/ml among penicillins or oral cephems. The MIC90s of carbapenems such as imipenem (IPM), meropenem (MEPM), and panipenem (PAPM) were less than 0.39 microgram/ml; particularly, PAPM showed the highest antibacterial activities. Among new quinolones such as tosufloxacin (TFLX), levofloxacin (LVFX), sparfloxacin (SPFX), and ciprofloxacin (CPFX), TFLX showed the highest antibacterial activities with the MIC90 of 0.39 microgram/ml. Other agents showed very low antibacterial activities as the MIC90s were 25 micrograms/ml in minocycline (MINO) and more than 100 micrograms/ml in clarithromycin (CAM) and clindamycin (CLDM).

[Mineralization patterns in the dentine of the floor of the pulp chamber of porcine molars].

The relationship between the arrangement of the matrix fibre bundles and the shape of the mineralization front (mineralization pattern) were observed in the dentine of the subpulpal wall of porcine molars. The observations were made using mainly light microscopy and scanning electron microscopy on the primary dentine excluding the secondary dentine. The results obtained shows that the dentine of the subpulpal wall consists of two parts, a surface layer and the circumpulpal dentine. The surface layer can be divided into two sublayers according to the arrangement of it's matrix fibres. The mineralization pattern was also observed to have another two sublayers which did not correspond with the two fibrous subdivisions. Adjacent to the dentine-cement junction, there is a layer, measuring about 3 microns in thickness, in which the matrix fibres run parallel to the junction. The second layer is characterized by two types of fibre bundles which run perpendicularly to the dentine-cement junction. These outer radially oriented fibres are about 1-2 microns in diameter, and the inner radially oriented fibre bundles are coarse, 10 microns in diameter. In the outermost 10 microns, mineralization begins as an aggregation of mineral granules, and forms a hypermineralized layer which overlays the junctional region between the fibres which are oriented parallel and perpendicular to the dentine-cement junction. Except for the most-superficial layer, mineralizing cones surround the radial fibre bundles at the mineralizing front. The circumpulpal dentine, which forms the floor of the pulp chamber, can also be divided into two layers by the differences in their fibrous matrix. The fibres of both of these layers lay in the plane of the predentine surface. However, in the outer layer the fibres of the matrix are randomly arranged around and between the dentinal tubules, whereas in the inner layer the fibres have a preferred orientation extending radially from a central region between the roots. In the outer layer, calcospherites, which are the units of the mineral aggregation, appear at the mineralizing front. In the inner layer, at the mineralizing front, ridges, measuring approximately 10-15 microns in width, 10 microns in height and 100 microns in length, also have a preferred orientation, 90 degrees to the fibre direction. Microradiographs show that a higher mineralized "lamellae-like' region exists in the center of each ridge-crest. X-ray diffraction indicates two preferred crystallite orientation, one is parallel to the matrix fibres and the other perpendicular to the predentine surface. The latter seems to be situated in the highly mineralized lamellae. In the circumpulpal dentine, therefore the mineralizing units have a spheritic shape in the outer layer, and a columner shape in the inner layer. As far as the author is aware the columnar mineralization unit in mammalian dentine has not been described before.

Differential effects of prostaglandin E1 and prostaglandin E2 on growth and differentiation of murine myeloid leukemic cell line, M1.

Effects of prostaglandins (PGs) of the E series on growth and differentiation of murine myeloid leukemic cell line M1 were studied. PGE1, but not PGE2, inhibited the growth of M1 cells. PGE2 neither inhibited nor augmented the antiproliferative effect of PGE1. PGE1 augmented the differentiation of M1 cells into macrophage-like cells induced by interleukin 6. PGE2, however, did not exhibit any effect on the differentiation. PGE1 caused a marked increased in intracellular cAMP level in M1 cells, whereas PGE2 had no effect. These results indicate that M1 cells are able to respond only to PGE1. Radiolabeled PGE1 binding experiments, however, revealed that there was no specific binding in M1 cells, suggesting that the cells express low numbers of receptors or very low affinity receptors specific for PGE1. Stable agonists of PGI2, iloprost, cicaprost or carbacyclin, also potently inhibited the growth of M1 cells. These findings suggest that PGE1 as well as PGI2 may play a role in the differentiation of monocyte-macrophage lineage cells.

Entrapment neuropathy due to bilateral epitrochleoanconeus muscles: a case report.

A case of bilateral entrapment of the ulnar nerves due to accessory epitrochleoanconeus muscles was observed in a young athlete. Hypertrophy of this muscle may be one of the causes of cubital tunnel syndrome.

Studies on 67Ga uptake by mouse granuloma tissues.

In connection with the uptake of 67Ga into the inflammatory tissues, such as granuloma tissues produced with turpentine oil, the influence of Fe3+ on the uptake of 67Ga into mouse granuloma and normal tissues and on the uptake of 125I-labeled transferrin and 59Fe were investigated. Fe3+ decreased the uptake of 67Ga into the liver and spleen, but had no influence on the uptake of 67Ga into the granuloma tissues. The uptake patterns of 125I-labeled transferrin and 59Fe in the granuloma tissues were not consistent with that of 67Ga at all. These results show that the uptake of 67Ga into the granuloma tissues occurs in a free, transferrin-unbound form, but into the liver and spleen in a transferrin-bound form.

Effect of human leukocyte interferon on the metastatic lung tumor of osteosarcoma: case reports.

Three patients with pulmonary metastases of osteosarcoma underwent treatment with intravenous or intramuscular administration of human leukocyte interferon. In 2 cases, the size of the metastasized tumor mass diminished temporarily six or eight months after interferon treatment, and serum alkaline phosphatase levels decreased to normal. In 1 case, the tumor cell from the pleural exudate 1 could be isolated and cultivated in vitro. When 2,000 IU of interferon was added to the tumor cell culture, marked inhibition of tumor cell growth resulted. In the other case, interferon had no effect on the metastasized lung tumor.