Degradation of serine-containing oligopeptides by Peptostreptococcus micros ATCC 33270.

BACKGROUND/AIMS: Microorganisms of Peptostreptococcus micros are asaccharolytic, anaerobic gram-positive cocci that are frequently isolated from human oral sites such as periodontal pockets. Preliminary study showed that several amino acids, including serine, enhanced slightly the growth of P. micros. Therefore, we investigated the degradation of serine and serine-containing oligopeptides. METHODS: Metabolic end products were determined with high-performance liquid chromatography. The related enzymatic activities in cell-free extract were also assayed. RESULTS: Washed P. micros degraded serine-tripeptides (Ser-Ser-Ser), and produced formate, pyruvate, acetate, and ammonia. They also degraded serinyl-tyrosine (Ser-Tyr) to the same products. Related enzymatic activities, such as serine dehydratase, pyruvate formate-lyase, formate dehydrogenase, pyruvate oxidoreductase, phosphate acetyltransferase, and acetate kinase, were detected in the cell-free extract, indicating that the organisms produced ATP in the serine metabolism. CONCLUSION: P. micros utilized serine-containing oligopeptides as exogenous metabolic substrates rather than serine itself, and degraded Ser-Ser-Ser and Ser-Tyr to formate, pyruvate, acetate, and ammonia with ATP generation.

Perfusion imaging of meningioma by using continuous arterial spin-labeling: comparison with dynamic susceptibility-weighted contrast-enhanced MR images and histopathologic features.

PURPOSE: The goal of the present study was to determine the utility of continuous arterial spin labeling (CASL) for characterization of meningioma by MR perfusion imaging and to compare these results with those obtained from the T2 dynamic susceptibility contrast (T2DSC) method and from histopathologic examination. METHODS: Twenty-one cases of meningiomas were examined at 1.5T. CASL perfusion imaging was implemented on the basis of multisection single-shot echo-planar imaging with velocity-driven adiabatic spin-inversion preparation. T2DSC perfusion imaging was also performed by using a double-echo spoiled gradient echo sequence in a section containing the tumor. By focusing on the regions of interest, maps of % signal intensity change and cerebral blood flow (CBF) were determined from CASL and cerebral blood volume (CBV). CBF and mean transit time (MTT) were obtained from T2DSC. The microvessel area (MVA) was determined from specimens immunostained with anti-CD31 in 14 cases by measuring the total amount of staining in each histologic section. Linear regression analysis was performed for rCBF values from both perfusion methods and for % signal intensity change and MVA. RESULTS: There was a significant correlation between CBF values determined from both perfusion methods (r(2) = 0.73; P < .001); however, the slope from T2DSC to CASL was less than unity, likely because of the different vascular weighting used for each method. There was also a significant correlation between CASL-% signal intensity change and MVA determined by histopathology (r(2) = 0.91; P < .00001). Perfusion values were the greatest for angiomatous meningioma and lowest for fibrous meningioma when using either perfusion method. CONCLUSIONS: CASL and T2DSC perfusion methods are comparable in the characterization of meningiomas. Further, CASL is of use in assessing tumor microcirculation.

Blood volume of gliomas determined by double-echo dynamic perfusion-weighted MR imaging: a preliminary study.

BACKGROUND AND PURPOSE: After bolus injection, gadopentetate dimeglumine causes a T2* rate change in permeable tissue that is contaminated by the T1 shortening effect due to the leakage of contrast agent. Therefore, tumor vascularity as reported in previous single-echo perfusion-weighted MR imaging studies has been underestimated. Our aim was to quantitatively and qualitatively evaluate the degree of blood volume of glioblastoma multiformes (GBMs) underestimated by this T1 shortening effect. METHODS: We used double-echo dynamic MR imaging after a bolus injection of gadopentetate dimeglumine (double-echo perfusion-weighted MR imaging) to simultaneously determine tumor blood volume without (V(T1U)) and with (V(T1C)) T1 shortening correction. MR imaging was performed in five consecutive patients with GBMs. The ratios of V(T1U) and V(T1C) were calculated and compared by means of quantitative analysis. The degree of tumor blood volume as determined by V(T1U) and V(T1C) maps were qualitatively compared using a three-point scale. RESULTS: All GBMs showed contrast enhancement on postcontrast T1-weighted images. In all subjects, the values of V(T1U) were significantly lower than those of V(T1C) (mean +/- SD, 2.05 +/- 1.01 vs. 3.62 +/- 1.40, respectively [P <.05]), indicating that tumor blood volume obtained by double-echo perfusion-weighted MR imaging was significantly higher than that by single-echo imaging. In the qualitative analysis, tumor blood volume on the V(T1U) map was less conspicuous than that on the V(T1C) map. CONCLUSION: Careful attention should be paid to the underestimation of tumor blood volume resulting from T1 shortening effects when using single-echo perfusion-weighted MR imaging. Double-echo imaging may be more suitable for the analysis of blood volume in GBMs.

Vascular permeability: quantitative measurement with double-echo dynamic MR imaging--theory and clinical application.

Double-echo dynamic magnetic resonance (MR) imaging was used to evaluate both vascularity and permeability of tissues simultaneously. Vascularity was evaluated on the basis of the T2*-shortening effect due to the intravascular fraction of the contrast agent and permeability on the basis of the T1-shortening effect due to the extravascular fraction. Meningioma was characterized on the basis of higher vascularity and neurinoma on the basis of higher permeability. The proposed method enables better tissue characterization.

Coregistration of FDG PET and MRI of the head and neck using normal distribution of FDG.

UNLABELLED: For better localization of head and neck structures by PET with 2-(18)F-2-deoxy-D-glucose (FDG), direct incorporation of anatomical information from MRI by the coregistration of FDG PET and MRI without external markers is proposed. METHODS: Seventeen patients with neoplasms and 16 normal subjects who had both FDG PET and MRI were studied. First, the three-dimensional normal distribution of FDG was evaluated, and then the structures of the head and neck regions with normal distribution patterns of FDG were used as internal markers for the coregistration of PET and MRI. The effectiveness of the coregistration was evaluated using focal neoplasms that were identified by both PET and MRI as fiducial internal markers. RESULTS: The normal structures selected as internal landmarks for coregistration were the tonsils, salivary glands, mucosal layers of the oral cavity and pharynx, spinal cord, inferior portion of the frontal lobe, cerebellum and nasal turbinates. These structures were more easily observed in sagittal or coronal sections than in transaxial sections. All primary neoplasms were delineated by PET, whereas 4 were missed by MRI. Thirteen primary tumors and 7 cervical lymph node metastases coregistered well, with a center-of-mass distance of <2 mm, whereas 10 lymph node metastases were slightly misregistered, with a center-of-mass distance of 7.8+/-6.5 mm (mean+/-s.d.), probably due to differences in neck positions. CONCLUSION: Normal distribution of FDG uptake in the head and neck regions delineated by multidirectional sections is important for effective coregistration of FDG PET with MRI.

Visualization of the cervical spinal cord with FDG and high-resolution PET.

PURPOSE: Our aim was to evaluate the visibility of the cervical spinal cord with [18F]2-fluoro-2-deoxyglucose (FDG) and a high-resolution PET scanner and to quantify the glucose utilization by the cervical cord. METHOD: Twenty-one normal subjects and three cervical myelopathy patients were studied. The visibility of the cervical spinal cord in sagittal and coronal sections was evaluated. The metabolic rate of glucose (MRGlu) and standardized uptake value (SUV) of FDG in the cord were calculated. RESULTS: The entire cervical spinal cord was clearly visualized in 57% of the subjects: the upper cord in 81%, the middle cord in 73%, and the lower cord in 57%. The MRGlu of the normal cord was 1.93 +/- 0.37 mg/100 g/min. SUV was constant across all the vertebral levels and negatively correlated with subject age. In the myelopathy patients, the SUV of the entire cervical cord was lower than in the age-matched normal subjects. CONCLUSION: These preliminary results indicate that the cervical spinal cord can be visualized as a normal structure in routine head and neck PET imaging and that FDG-PET may provide quantitative information about spinal cord disorders.

Non-invasive estimation of the net influx constant using the standardized uptake value for quantification of FDG uptake of tumours.

To reduce the variability of the standardized uptake value (SUV) which is widely used to evaluate 2-[18F]fluoro-2-deoxy-d-glucose (FDG) uptake by neoplasms, net influx constant (Ki) was derived from SUV. The relationship Ki=SUV.kp.V0, where kp is the plasma clearance rate and V0 is the initial distribution volume of FDG, was utilized. A total of 71 plasma input functions were measured up to 60 min after intravenous injection of FDG in 55 patients and were analysed to obtain kp and V0. SUV and V0 were calculated based on either body weight or body surface area. To validate the Ki estimation, another group of eight patients with squamous cell carcinoma of the head and neck was included. Parametric images of the net influx constant were obtained by Patlak graphical analysis of dynamic positron emission tomography (PET) data and measured plasma input function. V0 based on body weight was 0.1627+/-0.0329 (ml/g) and showed a weak negative correlation with body weight (y=0.23356-0.00138x, r=0.591). V0 based on body surface area was 5540+/-871 (ml/m2) and had no significant correlation with body weight. kp at 50 min post injection was 0. 03272+/-0.00243 (1/min), and had no correlation with the plasma glucose concentration. A highly significant positive correlation was noted between true Ki and estimated Ki based on both body weight (y=0.0033+1.0371x, r2=0.897), and body surface area (y=0.0033+1. 0351x, r2=0.926). Ki, a better indicator of FDG uptake by tumour than SUV, is derivable non-invasively. Quantification of FDG uptake by Ki will aid standardization of diagnostic criteria of FDG PET oncology.

Assessment of hepatic portal perfusion using T2 measurements of Gd-DTPA.

Recently, perfusion imaging has been of increasing interest in MRI. We applied this method for semiquantitative evaluation of hepatic parenchymal portal blood flow in patients with diffuse liver damage. Twenty patients with diffuse hepatic damage were divided according to the Child's Classification and studied. Gadolinium-diethylenetriamine pentaacetic acid (Gd-DTPA) was administered into the superior mesenteric artery (SMA), and a dynamic series of T2*-weighted fast low angle shot (FLASH) images was obtained. We evaluated relative regional portal blood volume (rrPBV), mean transit time (MTT), and relative regional portal blood flow (rrPBF). The relationship between the rrPBV, rrPBF, and plasma indocyanine green retention rate test at 15 minutes (ICGR15 was also evaluated in 12 patients. Both rrPBF and rrPBV are significantly decreased in Child B & C patients compared with Child A patients. On the other hand, the MTT is significantly prolonged in Child B & C patients compared with Child A patients. Significant correlations were also noted between rrPBV and ICGR15 and between rrPBF and ICGR15. By means of selective catheterization into the SMA, we were able to estimate rrPBV, rrPBF, and MTT. This method may play a clinical role for assessment of regional portal perfusion in various diseases with diffuse liver damage.

Multiple single sections Turbo FLASH MR arterial portography in the detection of hepatic neoplasms.

OBJECTIVE: To determine the sensitivity of multiple single sections Turbo FLASH MR arterial portography (MRAP) in the detection of hepatic neoplasms. METHODS AND PATIENTS: Twelve patients with hepatic mass underwent MRAP prior to hepatic resection. Findings of MRAP were compared with surgical specimen and intra-operative ultrasonography (US). RESULTS: A total of 19 separate malignant neoplastic nodules were identified in the resected specimens or intra-operative ultrasonography. The sensitivity was 89.5% (17/19) for MRAP. MRAP depicted all neoplasms more than 1.0 cm in diameter. Two lesions not depicted on MRAP had a diameter of 5 and 9 mm, respectively. One lesion identified by MRAP was confirmed to false positive lesion by intra-operative US. CONCLUSION: Multiple single sections Turbo FLASH MRAP may be a valuable adjunct for pre-operative detection of malignant hepatic neoplasms.

Fluorine-18-fluorodeoxyglucose PET versus thallium-201 scintigraphy evaluation of thyroid tumors.

UNLABELLED: To determine whether PET could help differentiate malignant from benign thyroid tumors, 18F-fluorodeoxyglucose (FDG) accumulation and 201Tl scintigraphy were examined relative to histological diagnosis. METHODS: Nodular thyroid lesions in 11 patients were evaluated before surgical resection. Static PET scanning with 370 MBq FDG was done for 20 min (from 40 to 60 min postinjection) in all patients, and standardized uptake values (SUVs) in these lesions were calculated. In addition, eight patients were evaluated with dynamic PET scan up to 60 min postinjection, and the lesions were further evaluated using graphical analysis. Thallium-201 delayed images were visually evaluated in 10 patients. RESULTS: Four of 11 nodules were well-differentiated papillary carcinoma, another five were benign follicular adenomas, one was a multinodular goiter and another a case of chronic thyroiditis that was proved not to contain a nodule. Time-activity curves of FDG uptake showed different patterns in malignant and benign tumors. In the malignant tumors, FDG uptake increased with time after the tracer injection. By contrast, FDG uptake in benign tumors gradually decreased. With use of a cutoff value of 5.0 mg/ml for SUV and 10 microl x min(-1) x ml(-1) for Kc (K complex value determined using the linear fitting of the time-activity curve of FDG accumulation), all of the four malignant nodules and the six benign nodules were separated correctly. Chronic thyroiditis had high SUV in the malignant range. Of the four patients with thyroid carcinoma, the delayed 201Tl images revealed a slightly higher or equal uptake to background activity. In a patient with chronic thyroiditis, the delayed 201Tl images revealed diffuse accumulation higher than background activity. CONCLUSION: FDG-PET is superior to 201Tl in differentiating malignant from benign tumors. Both SUVs and Kc values were useful indexes for this discrimination. Although careful evaluation is needed for chronic inflammatory lesions, this technique appears to be useful in evaluating thyroid nodules.

Contribution of baroreceptor reflexes to blood pressure and sympathetic responses to cholecystokinin and vasoactive intestinal peptide in anesthetized dogs.

The effects of synthetic vasoactive intestinal peptide (VIP) and cholecystokinin (CCK) on systemic blood pressure and renal nerve activity were studied before and after cervical vagotomy, and sino-aortic denervation with vagotomy in anesthetized dogs. Intravenous injection of VIP (5 micrograms/kg) in animals with an intact neuraxis produced a significant decrease in systemic blood pressure and a significant increase in renal nerve activity. These responses to VIP did not change after vagotomy and after complete denervation, VIP did not cause any change in renal nerve activity, even during hypotension. The level of hypotension after complete denervation was greater than that under other conditions. These results indicate that the cardiovascular effects of VIP are reduced by activation of the systemic baroreceptors. Intravenous injection of CCK (10 micrograms/kg) in animals with an intact neuraxis produced significant decreases in blood pressure and renal nerve activity. These responses to CCK were abolished in animals with cervical vagotomy only. However, following complete denervation of the carotid sinus and total section of the vagal nerves, CCK caused a significant increase in blood pressure and renal sympathetic nerve activity. These results indicate that the sympathetic depressor effect of CCK may be mediated by activation of the vagal afferents, and that the sympathetic pressor effect may be due to a direct action of CCK on the central nervous system. Thus, each gastrointestinal peptide may regulate the cardiovascular system through a different mechanism.

[A case report of reflex sympathetic dystrophy treated with nifedipine].

Reflex sympathetic dystrophy (RSD) refers to a symptom complex observed after nerve injury and consists primarily of severe burning pain associated with sensory, vasomotor and trophic phenomena. A 54-year-old male had undergone nephrectomy. At surgery left XIth intercostal nerve had been injured by cautery. After a few weeks following surgery, the patient developed progressive deep burning pain, stabbing sensation and dysesthesia in the left abdominal region. Analgesics and narcotics were ineffective. We diagnosed his case as RSD. He received nifedipine 10mg sublingually. Pain relief was obtained within 10min and lasted for 6hs. Consequently, nifedipine therapy was started at a daily dose of 30 to 60mg. His symptoms were markedly improved within 4 weeks. After 3 months his pain resolved. At the present, some pain often returns, but nifedipine is effective. Nifedipine may be useful as a drug for the management of reflex sympathetic dystrophy.

Electron microscopic visualization of cathepsin D using mercury-labeled pepstatin as an enzyme inhibitor.

Pepstatin, a specific inhibitor of pepsin, cathepsin D (E), renin, etc., was used in a new method to demonstrate the sites of enzymes. Pepstatin (Pst) was covalently attached to glutathione (GSH), using a dicyclohexylcarbodiimide, and CH3Hg+ was then put in the residue of SH for electron microscopic observation. Pst-GS-HgCH3 thus obtained was nonisotopical, had a very low molecular weight (about 1,200 daltons), and still almost completely retained its inhibitory activity. Sections of rat liver (less than 1 mm3 thick), prefixed by glutaraldehyde, were incubated in the prepared reaction medium. Cathepsin D, located in the lysosomes, was demonstrated by this compound, but when the sections were preincubated with pepstatin, this was not the case. These results demonstrate that mercury-labeled pepstatin is a useful reagent for the histochemical staining of acid proteases for electron microscope.

The role of macrophages in demyelination in experimental allergic neuritis.

The role of macrophages and serum factors in demyelination in experimental allergic neuritis (EAN) was examined by a simple in vitro method. Cultivated rabbit peritoneal macrophages, preincubated with serum obtained from rabbit EAN produced by sensitization with bovine spinal nerve roots, could agglutinate and phagocytize purified bovine or rabbit peripheral nerve myelin. Sera from normal animals or from controls given adjuvant alone could not. Adhesion and phagocytosis were inhibited if EAN sera were absorbed with peripheral nerve myelin. Rabbit red blood cells were not phagocytized by macrophages exposed to EAN serum. Concomitant to these observations, three lysosomal acid hydrolases: acid proteinase, acid phosphatase and beta-glucuronidase, were assayed with respect to their topographical and chronological distribution. In the group examined at clinical onset, increases in the specific activities were 1.5-3.0-fold in the spinal roots and 1.0-1.5-fold in the sciatic nerves compared with control. The degree of increase in total activities per whole root or sciatic nerve was much higher for specific activities. The topographical distribution of the increase closely corresponded to the histological distribution of EAN lesions. These observations suggested that the increased lysosomal activity originated from lysosomal-rich infiltrating cells. These observations strongly indicated the significant role of macrophages activated by EAN serum in the demyelination of EAN.