After conjugating thiol groups in the hinge region of monoclonal antibody (mAb) Fab' fragments specific for basic fibroblast growth factor (bFGF) with maleimido-horseradish peroxidase HRP) complexes synthesized by incubation of HRP with the heterobifunctional reagent N-succinimidyl-4-(maleimidomethyl)cyclohexane-1-carboxylate, we developed a fluorometric enzyme immunoassay method based on the sandwiching of the factor between anti-bFGF IgG-coated polystyrene beads and the conjugates, and also an immunohistochemical method for detection of the location of the factor. The discriminatory detection limit by the developed enzyme immunoassay (EIA) was as low as 30 pg/mL. The reproducibility of within- and between-assay series was 6.07-9.18% and 6.28-6.82%, respectively, and the recovery of exogenous bFGF from serum was approximately 98%. The curves generated by the concentrated fraction that eluted at the same position as standard bFGF by size-exclusion chromatography on a TSK 2000SW column were parallel to the curve for standard bFGF. From these results, we consider the developed EIA method to be acceptable in regard to sensitivity, precision, and specificity. Also, without the introduction of any additional signal amplification system, positive immunohistochemical reactions were successfully detected by the HRP-linked anti-bFGF mAb Fab' in fibroblastic and endothelial cells, which have already been shown to synthesize and secrete bFGF, indicating that these conjugates provide a useful means for direct immunohistochemical detection of the factor.

Antibodies, Monoclonal↗

PubMedDate not supplied

Synthesis of novel polyampholyte comb-type copolymers consisting of a poly(L-lysine) backbone and hyaluronic acid side chains for a DNA carrier.

The polyampholyte comb-type copolymers consisting of a poly(L-lysine) (PLL) main chain, a DNA binding site, and hyaluronic acid (HA) side chains, cell-specific ligands, have been prepared as the DNA carrier targeting sinusoidal endothelial cells of liver. The reducing end of HA and epsilon-amino groups of PLL were covalently coupled by reductive amination to obtain the resulting comb-type copolymers (PLL-graft-HA). The chain length of HA was controlled by the enzymatic hydrolysis of high-molecular weight HA. Since HA formed polyion complexes with PLL, the coupling reaction was carried out with high-ionic strength media to suppress polyion complex formation. The reaction proceeded in a homogeneous system, leading to a high efficiency of coupling (>70%) of HA onto the PLL backbone. By using the enzymatic hydrolysis of HA and the reductive amination reaction between HA and PLL with high-ionic strength media, it is possible to prepare the various comb-type copolymers with a defined density and a defined length of HA side chains. Furthermore, we also find that these polyampholyte comb-type copolymers vary their assembling structure in water in response to two kinds of environmental factors, i.e., ionic strength and pH. Finally, a 1H NMR study reveals that the PLL backbone efficiently interacts with DNA molecules despite the presence of HA side chains having negative charges.

Carbohydrate Sequence↗

PubMedDate not supplied

Comb-type copolymers for controlled DNA delivery.

Various comb-type copolymer containing a polycation as a main chain was design to construct delivery systems of DNAs. The comb-type copolymers having cell-specific polysaccharides were proved to be useful to deliver DNA to the target cells in vivo. Of interest, the copolymers with abundant side chains of hydrophilic polymers are capable of stabilizing DNA triplex. Further, injectable nanoparticles for controlled releases of DNAs were fabricated from the copolymer and a biodegradable polymer.

Biocompatible Materials↗

PubMedDate not supplied

Similarity of vasorelaxant effects of natriuretic peptides in isolated blood vessels of salmonids.

Natriuretic peptides (NPs) have been implicated in cardiovascular regulation in rainbow trout (Oncorhyncus mykiss), and it has been observed that the vasorelaxant activity of distinct trout and human NPs is similar in isolated trout arteries. This study characterizes the response of a variety of vessels from rainbow trout and other salmonids to different NPs. The effects of heterologous (rat atrial and human atrial) and homologous (rainbow trout atrial and rainbow trout ventricular) NPs were examined in precontracted efferent branchial arteries from rainbow trout (O. mykiss, Kamloops strain), lake whitefish (Coregonus clupeaformis), and in rainbow trout celiacomesenteric arteries and anterior cardinal veins. The response to mammalian NPs was also examined in efferent branchial arteries from the steelhead (O. mykiss, Skamania strain), coho salmon (Oncorhyncus kisutch), brook trout (Salvelinus fontinalis), and brown trout (Salmo trutta). In general, there were relatively few differences that were species, peptide, or vessel specific. There was no difference in the sensitivity (concentration producing a half-maximal response EC(50)) or efficacy (percent relaxation, i.e., E(max)) of trout or whitefish efferent branchial arteries to any NP, except human NP, which was significantly less effective (greater EC(50) and lower E(max)) in whitefish arteries. There were no differences in E(max) of mammalian NPs in efferent branchial arteries from any species, and only coho and brook trout had significantly different EC(50)'s (coho, 1.0+/-0.2 nM; brook trout, 4. 2+/-0.6 nM; and other species, from 1.9 to 3.5 nM). Rainbow and coho anterior cardinal veins were less sensitive than arteries to mammalian NPs (EC(50)'s; 8.8+/-2.0, 2.0+/-0.1 vs. 3.0+/-0.9, 1.0+/-0. 2, respectively), whereas brown trout veins were more sensitive (1. 0+/-0.2, 3.5+/-1.3, respectively). Sodium nitroprusside (SNP), which activates soluble guanylate cyclase, was vasodilatory, albeit significantly less potent than all NPs, in efferent branchial arteries of all species. SNP was significantly more potent in trout than whitefish efferent branchial arteries, whereas it was equally efficacious in these vessels. These results demonstrate that multiple vessels from various salmonids are similarly responsive to the vasorelaxant effects of a variety of NPs and that the salmonid NP receptor has relatively little ability to discriminate between homologous and heterologous peptides. We conclude that the vascular NP receptor complex is highly conserved among salmonids. Further, salmonids utilize cyclic guanosine monophosphate (cGMP) elevations for reductions of vascular tonus by both particulate and soluble guanylate cyclase pathways.

Animals↗

PubMedDate not supplied

Effects of loxoprofen sodium, a newly synthesized non-steroidal anti-inflammatory drug, and indomethacin on gastric mucosal haemodynamics in the human.

Non-steroidal anti-inflammatory drugs (NSAID) are, and have been, frequently used for alleviation of pain in patients; however, they are known to cause gastric mucosal injury in experimental animals and in humans. A decrease in the gastric mucosal blood flow also plays an important role in the aetiology of acute gastric mucosal injury, as we previously reported. This study investigated the effect of a newly synthesized NSAID, loxoprofen sodium (sodium 2[p-2 oxocyclopentylmethyl) phenyl]propionate dihydrate, on gastric mucosal haemodynamics using a reflectance spectrophotometry system. Both single and cross-over methods were used in five volunteer subjects. Loxoprofen sodium 60 mg (one tablet) or indomethacin 25 mg (one tablet), was diluted in 10 mL water at 25 degrees C and sprayed on the gastric mucosa via a polyethylene tube inserted into the biopsy channel of an endoscope. After drug administration, reflectance spectra were taken every 5 min for 30 min. The indices of mucosal haemoglobin content (IHb) and oxygen saturation of haemoglobin (ISO2) were determined by the method previously reported by the authors. Indomethacin administration produced a significant decrease in both IHb and ISO2 values, indication ischaemia. Loxoprofen sodium, however, showed no significant differences in either of the parameters. Haemorrhagic erosions were evident after indomethacin administration, but none were found after loxoprofen sodium administration. The conclusion reached on the basis of this evidence is that one-time topical application of loxoprofen sodium is safer than indomethacin.

Administration, Oral↗

PubMedDate not supplied

A phase II study of 72-hour continuous infusion consisting of cisplatin and 5-fluorouracil for treatment of non-small cell lung cancer.

A combination chemotherapy consisting of a 72-hour continuous infusion of cisplatin (CDDP; 100 mg/m2/72 h) and 5-fluorouracil (5-FU; 3 g/m2/72 h) was conducted for inoperable non-small cell lung cancer (NSCLC). Sixty patients were accepted for this study between June 1988 and December 1990. Forty-seven patients were male (median age 68). Thirty-four patients had stage III and 26 had stage IV disease. The response rate was 25.0% (95% confidence interval, CI, 14.0-36.0%), median survival was 15.7 months. In squamous cell carcinoma, the response rate was 35.5% (95% CI, 18.7-52.3%) and median survival was 15.1 months. In non-squamous cell carcinoma, the response rate was 13.8% (95% CI, 1.2-26.4%) and median survival was 17.7 months. There was a significant difference in response rate (p < 0.01), but no significant difference in survival (p = 0.36). Grade 3 leukopenia was 11.7%, grade 3 and 4 thrombocytopenia was 13.3%. Grade 3 nausea and vomiting were 8.3%. One patient had grade 4 renal toxicity. However, there was no treatment-related death. This regimen was well tolerated. In multivariate analysis, the significant parameters were CEA, performance status and response. In conclusion, 72-hour continuous infusion of CDDP and 5-FU for treatment of NSCLC provides similar response and toxicity as previously reported regimens using CDDP.

Adult↗

PubMedDate not supplied

CYFRA 21-1: an indicator of survival and therapeutic effect in lung cancer.

CYFRA 21-1 is a new tumor marker using two different monoclonal antibodies which recognize the divergent epitope on the N- or C-terminal region of domain 2 of cytokeratin 19 fragment, respectively. In this study, we investigated the relationship between levels of CYFRA 21-1 and survival duration, as well as the efficacy of chemotherapy associated with changes in CYFRA 21-1. Serum samples were obtained from 87 patients with nonoperable lung cancer (35 cases with squamous-cell carcinoma, 33 with adenocarcinoma, 3 with large-cell carcinoma, and 16 with small-cell carcinoma). The cutoff point was set at 3.5 ng/ml. In a CYFRA 21-1 assay, significantly more patients with squamous-cell carcinoma and adenocarcinoma were positive compared to patients with small-cell and large-cell carcinomas (p = 0.0017). Following chemotherapy, blood levels of CYFRA 21-1 decreased significantly in responders versus nonresponders (p = 0.0246). A significant correlation was noted between survival periods and pretreatment levels of CYFRA 21-1 (p = 0.0036). The present study suggests that CYFRA 21-1 might be useful as a possible indicator of survival and therapeutic effect for lung cancer.

Aged↗

PubMedDate not supplied

Roles of head, gaze, and spatial orientation in the production of oscillopsia.

To clarify the factors causing oscillopsia, we investigated head movement, gaze stability, and perception under various situations. High-frequency head movements, whether they were horizontal rotations or passively induced vertical oscillations, produced blurred vision and gaze fluctuations in patients with labyrinthine loss. However, this sensation differed from the oscillopsia perceived during walking, as it did not involve a sensation of oscillation of the surrounding space or a loss of body balance. Although patients with labyrinthine loss showed large irregular head perturbations during stepping, the resultant retinal velocity slips seemed too small to explain oscillopsia. Walking while wearing horizontal reversing prisms produced loss of spatial orientation, dysequilibrium, and instability of vision in normal subjects, which resembled the symptoms found in patients with oscillopsia. The present study suggests that oscillopsia represents a perceptual inability to detect spatial orientation during head or body movements rather than a mere blurring of vision caused by deficient compensation.

Adult↗

PubMedDate not supplied

Intracranial ganglioglioma: MR, CT, and clinical findings in 18 patients.

Eighteen cases of pathologically proved intracranial gangliogliomas were reviewed to determine their MR, CT, and clinical characteristics. Seventeen patients were evaluated with contrast-enhanced CT and 14 were studied by MR imaging. Eight tumors were predominantly cystic; half of these demonstrated some contrast enhancement, and five contained calcifications. These cystic gangliogliomas were located, in order of decreasing frequency, in the cerebellum, temporal, frontal, and parietal lobes. Ten tumors were solid; eight of these showed contrast enhancement, and only one contained calcifications. Small cysts were present in one solid mass. Solid gangliogliomas occurred preferentially in the temporal lobes. On MR, the findings were nonspecific and reflected the CT findings. In one patient who received gadolinium-DTPA the lesion did not enhance. Clinically, all patients presented with nonfocal long-standing symptoms and all but three were alive an average of 18 months after the initial diagnosis. Pathologists are recognizing ganglioglioma with increasing frequency, and although its radiographic characteristics vary, it should be included in the differential diagnosis when the above-described findings are encountered.

Adolescent↗

PubMedDate not supplied

Primary cerebral neuroblastoma: CT and MR findings in 12 cases.

A retrospective CT, MR, and clinical study was performed in 12 patients, five children and seven adults, with histologically proved primary CNS neuroblastoma. The CT and MR appearances of these neoplasia were more variable than generally recognized. Although seven tumors were predominantly intraparenchymal masses with calcification and cyst formation, five were intra- or juxtaventricular. CT was preferable to noncontrast MR both at initial diagnosis and follow-up for identification of calcification, recurrent tumor at surgical sites, and leptomeningeal disease. Noncontrast MR was useful primarily for localization of peri- and intraventricular lesions. We conclude that primary CNS neuroblastoma has a more variable radiographic appearance than is generally recognized, and that an intra- or periventricular epicenter is common.

Adult↗

PubMedDate not supplied

Gadolinium-DTPA and MR imaging of pituitary adenoma: a preliminary report.

Gadolinium-DTPA MR imaging (Gd-MR), unenhanced MR imaging, and contrast-enhanced CT studies were compared prospectively in six patients with surgically confirmed pituitary adenomas and three patients without sellar pathology to determine the utility of Gd-MR in the diagnosis of pituitary adenoma. In normal patients, the pituitary gland, cavernous sinus, and infundibulum enhanced by T1 shortening after gadolinium. In adenoma patients, two of four focal lesions identified with contrast-enhanced CT were identified with Gd-MR, and one was identified with unenhanced MR. The earliest short repetition-time sequence performed after gadolinium injection was best for focal lesion detection. Normal cavernous sinus enhancement by gadolinium made identification of cavernous sinus extension of adenoma difficult. Infundibulum displacement was better seen with contrast-enhanced CT (two vs one); however, unenhanced and Gd-MR were better than contrast-enhanced CT for demonstrating chiasmal compression (four vs three). Contrast-enhanced CT, Gd-MR, and plain MR were equally able to identify gland enlargement, sellar floor erosion, and abnormalities of the diaphragma sellae. In this preliminary series, we found Gd-MR to be promising for imaging adenomas; however, modifications in Gd-MR technique including thinner slices and immediate scanning after gadolinium injection are necessary for the best detection of focal lesions.

Adenoma↗

PubMedDate not supplied

Astrocytic tumorette: microscopic to minute foci of glioma unexpectedly found in autopsy or surgical specimens.

Four examples of astrocytic tumorettes (microscopic to minute foci of glioma) are described herein. They include one malignant astrocytoma and three low grade astrocytomas. The first patient, who died of heart failure, was found incidentally to havour a small malignant astrocytoma at the time of autopsy. The other three patients with astrocytomas of low grade in malignancy, ranged from 18 to 25 years in age, and presented with intractable seizures. Electroencephalography defined a temporal lobe focus in all three patients. Subsequently, all three underwent a unilateral temporal lobectomy with resection of the epileptic focus. Careful histological examinations on the removed tissues from each patient revealed that each of them had a minute astrocytoma. The histogenesis of benign and malignant astrocytomas and the importance of surgical exploration in the management of the patients with intractable seizure disorders are discussed.

Adolescent↗