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PubMed1981

Immunoreactive adrenocorticotropin in the gastrointestinal tract and pancreatic islets of the rat.

Radioimmunoassayable ACTH is detectable in the gastrointestinal tract and pancreatic islets of the rat by using a midportion ACTH antiserum. Portions of the gastrointestinal tract and isolated pancreatic islets were extracted with 0.1 N HCl. Serial dilutions of the extracts resulted in inhibition curves parallel with human alpha ACTH-(1-39). The highest concentrations were in the isolated pancreatic islets (20 pg/ml protein) and the gastric antrum-pylorus (17.9 +/- 1.2 pg/ml protein). Chromatographic characterization of the stomach extract showed a main peak, with an elution constant identical to that of [125I]iodo-ACTH-(1-39) and an elution pattern identical to that of rat pituitary extracts and medium from incubated rat pituitaries.

Adrenocorticotropic Hormone↗
PubMedDate not supplied

Immediate and delayed implantation for complete restoration of the jaw following extraction of all residual teeth: a retrospective study comparing different types of serial immediate implantation.

From April 1986 to September 1993, 134 implants (97 immediate implants, 26 early secondary immediate implants, and 11 late secondary implants) were placed in 20 patients following extraction of all residual teeth. After a mean follow-up period of 27.1 months, the implants were analyzed with regard to peri-implant soft tissue parameters and bone resorption. When the examination was performed, 131 implants (97.7%) were found to be functioning successfully. The mean pocket depths varied between 1.3 and 2.6 mm, and the peri-implant bone resorption showed values between 0.5 and 1.3 mm. Fixed and removable prostheses revealed equal results with regard to the parameters examined. The results of the present study indicate that serial immediate implantation can be considered to be a very promising implant treatment modality for serial teeth extraction, if specific parameters are taken into account.

Aged↗
PubMed2006

Sterility assessment of multiple use botulinum A exotoxin vials: a prospective simulation.

BACKGROUND: A preponderance of evidence now indicates that storage of reconstituted botulinum toxin solution for longer than the 4 hours recommended by the manufacturer does not result in loss of effectiveness. OBJECTIVE: Then purpose of this study was to assess whether serial re-extraction of reconstituted botulinum toxin solution from vials undergoing prolonged storage is associated with increased risk of microbial contamination. METHODS: Consecutive 100-U vials of botulinun A exotoxin were reconstituted with 2.5 mL each of normal saline with preservative (benzyl alcohol). Within 1 week after reconstitution, each vial was used to treat 1 to 3 patients, for a total of 60 to 80 U used. Thereafter, each vial was stored in a plastic kidney basin in an unlocked, multiuse medication refrigerator. Two weeks later, a randomly assigned nurse removed the vial from the refrigerator and withdrew and discarded 0.1 mL (5 U) using the same technique used for therapeutic extractions. Iterations of this withdrawal and discarding procedure were continued until a 0.1-mL aliquot could not be withdrawn without prying off the metal cover. Once this threshold was reached, the vial was sent to the microbiology laboratory for sterility testing using a thioglycolate broth. RESULTS: In all, 127 vials were handled per protocol. On average (mean), vials underwent 4.5 access procedures, including 1.6 therapeutic extractions for a total of 76 U removed during a period of 7 weeks. Sterility analysis with thioglycolate broth indicated no evidence of contamination. LIMITATIONS: Preservative-containing saline was used for reconstitution. CONCLUSION: Routine refrigerator storage of medication vials containing reconstituted botulinum toxin does not result in microbial contamination of the contents even after serial re-extraction of solution from these vials, and after handling of such vials by multiple personnel. Storage and subsequent reuse of botulinum toxin appears safe for at least 7 weeks after reconstitution.

Botulinum Toxins, Type A↗
PubMed1989

Phylogenetic study on the immunoreactive atrial natriuretic peptide in the heart.

Using two antisera against atriopeptin III (AP III) which had different characteristics in cross-reactivities with atrial natriuretic peptide (ANP) analogs, we have measured an immunoreactive ANP (ir-ANP) in the heart extracts of several species. ir-ANP in atrial extracts showed both high and low molecular weights. Serial dilutions of atrial extracts from chickens, turtles, frogs, and fish yielded competition curves which were parallel to the standard curve of AP III with antiserum No. 4. In comparison with two serial dilution curves of atrial extracts made using two antisera, No. 4 and No. 9, we suggest that the amino acid at position 12 of ir-ANP in the atrial extracts of chicken, turtle, frog, and fish is not isoleucine but is some other amino acid.

Animals↗
PubMed1980

Investigation of the component of Propionibacterium acnes (Corynebacterium parvum) responsible for macrophage activation.

Mouse peritoneal macrophages, activated with Propionibacterium acnes (Corynebacterium parvum) in vivo, exhibited altered morphological characteristics, increased acid phosphatase activity, and release of [EH]thymidine-labeled DNA from tumor cells in vitro. Comparison with macrophages from mice injected with a control organism, P. jensenii, showed that the morphological changes, but not acid phosphatase, correlated with the development of tumoricidal activity. Investigation of the microbial component responsible for this activity indicated it was heat stable and refractory to extraction by methanol-chloroform, dilute acid, butanol, or a serial combination of extraction procedures. When the cells were disrupted by mechanical breakage, no activity could be found in the cell wall, soluble cytoplasm, or particulate cytoplasm. These results suggest that intact organisms are required for macrophage activation, and they may resolve conflicting reports on the nature of the immunostimulating activity of P. acnes.

Acid Phosphatase↗
PubMedDate not supplied

Oxygen transport variables in the identification and treatment of tissue hypoxia.

Critically ill patients have greater than normal oxygen demands because of enhanced energy requirements placed on them by the stress of acute illness, blood and fluid loss, surgery, wound healing, and hospitalization. Early recognition of major alterations in oxygen transport variables, oxygen delivery, oxygen consumption, and the oxygen extraction ratio, by the critical care team assists in the prevention and treatment of tissue hypoxia in seriously ill and injured patients. Supranormal levels of oxygen delivery are required to meet these increased oxygen demands, to prevent tissue hypoxia, and to correct tissue oxygen debt. The critical care team should promptly determine the patient's oxygen transport variables on admission to the intensive care unit to provide a starting point for patient resuscitation. When deviations from supranormal values of oxygen transport variables in these patients are identified, specific interventions that improve oxygen delivery to peripheral tissues should be implemented and evaluated for their effectiveness in normalizing the oxygen extraction ratio. When serial measurements of oxygen delivery, oxygen consumption, and the oxygen extraction ratio follow each therapeutic intervention that is directed at increasing oxygen delivery, the survival rate of critically ill patients is significantly improved.

Biological Transport↗
PubMed2000

Prospective comparative study between microsurgical and conventional testicular sperm extraction in non-obstructive azoospermia: follow-up by serial ultrasound examinations.

The value of testicular sperm extraction (TESE) by microdissection was evaluated according to its physiological consequences compared with open, classic surgical biopsy in the same patient. A total of 100 patients with non-obstructive azoospermia and bilateral identical testicular histology underwent bilateral diagnostic TESE via the conventional method on one side and the microsurgical method on the other side. The spermatozoa recovery rate by microdissection TESE was significantly higher than by conventional TESE (47 and 30% respectively; P < 0.05). In order to assess the safety of this new procedure, 60 patients were followed-up ultrasonographically for 1, 3 and 6 months. Acute and chronic complications were significantly lower in the microsurgical side compared with the conventional side (15 and 58.3% respectively and 3 and 30% respectively; P < 0.05). Segmental devascularization was detected in seven testes operated on conventionally, and in two testes operated on microsurgically. However, permanent devascularization could not be detected in any patient after 6 months. These findings suggest that microdissection TESE is not devoid of complications, but that it is relatively safer than the conventional technique and improves sperm yield significantly in patients with non-obstructive azoospermia.

Follow-Up Studies↗
PubMedDate not supplied

Effects of VPS extract of Coriolus versicolor on cancer of the large intestine using a serial sacrifice technique.

VPS, a hot water extract of the Coriolus versicolor mushroom, was given at a 2% dose level in the diet of female Swiss Webster CFW outbred mice in a serial sacrifice experiment. The mice were also administered either 1,2-dimethylhydrazine dihydrochloride (1,2-DMH) as ten weekly subcutaneous (s.c) injections of 20 microg/g body weight or physiological saline (PS) as ten weekly (s.c) injections of 0.01 ml/g body weight. The animals were sacrificed at 26 weeks or 35 weeks after the first injection of 1,2-DMH or PS. The number of mice with large intestinal tumors and the total number of these tumors were: Group I (1,2-DMH), 29 and 438; Group 2 (VPS + 1,2-DMH), 29 and 344; Group 3 (VPS + PS), 0 and 0; and Group 4 (PS), I and 1, in the mice sacrificed at 26 weeks. The corresponding tumor incidences in mice sacrificed at 35 weeks were: Group 1 (1,2-DMH), 30 and 323; Group 2 (VPS + 1,2-DMH), 29 and 521; Group 3 (VPS + PS), 1 and 2; and Group 4 (PS), 0 and 0. Histopathologically, the tumors were diagnosed as polypoid adenomas and adenocarcinomas of the cecum, colon and rectum. Contrary to expectations, the VPS treatment enhanced the development of large intestinal tumors induced by 1,2-DMH in animals sacrificed at 35 weeks after the first injection of the carcinogen.

1,2-Dimethylhydrazine↗
PubMed2000

Quantitative determination of a chemically modified hammerhead ribozyme in blood plasma using 96-well solid-phase extraction coupled with high-performance liquid chromatography or capillary gel electrophoresis.

Versatile bioanalytical assays to detect chemically stabilized hammerhead ribozyme and putative ribozyme metabolites from plasma are described. The extraction protocols presented are based on serial solid-phase extractions performed on a 96-well plate format and are compatible with either IEX-HPLC or CGE back-end analysis. A validation of both assays confirmed that both the HPLC and the CGE methods possess the required linearity, accuracy, and precision to accurately measure concentrations of hammerhead ribozyme extracted from plasma. These methods should be of general use to detect and quantitate ribozymes from other biological fluids such as serum and urine.

Animals↗
PubMed1995

Poly(vinyl chloride) formulations: acute toxicity to cultured human cell lines.

Two quantitative cytotoxicity assay methods (cytoplasmic retention of carboxyfluorescein and mitochondrial cleavage of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT)) have been used to evaluate the response of two cultured human cell lines; HepG2 (hepatoma) and W138va13 (transformed lung fibroblasts) to extracts of a range of poly(vinyl chloride) (PVC) formulations. Two plasticizers; di(2-ethylhexyl)phthalate (DEHP) and di-isooctyl phthalate and a range of tin and non-tin stabilizers were incorporated in the study. Only those formulations containing both a plasticizer and a tin-based stabilizer produced extracts which were toxic. Extracts of those formulations which contained both plasticizer and dibutyl tin dimaleate stabilizer were toxic to both cell lines in both assay methods. Extracts of a formulation containing plasticizer and a dioctyl tin mercaptide were toxic to both cell lines in the carboxyfluorescein assay but were only toxic to the WI38va13 cells in the MTT assay. The WI38va13 cells were generally more sensitive to the extracts than the HepG2 cells. When serial dilutions of the extracts were evaluated, the carboxyfluorescein assay proved to be the more sensitive of the two. The acute toxicity of extracts of these PVC formulations cannot be directly attributed to the plasticizers or to the tin stabilizers. It is likely that a synergistic mechanism, such as plasticizer facilitated extraction of the tin stabilizer, exists.

Carcinoma, Hepatocellular↗
PubMed1986

Development of cholecystokinin radioimmunoassay using synthetic CCK-10 as immunogen.

Using an antiserum generated against synthetic CCK-10, we have developed a radioimmunoassay specific for the carboxyl-terminus of cholecystokinin (CCK). Three rabbits were immunized with synthetic sulfated carboxy-terminal CCK decapeptide (CCK-10) conjugated to keyhole limpet hemocyanin. Using 125I-CCK-39 prepared by the Iodogen method as a tracer, we found that all immunized rabbits produced antibodies against the conjugate. Antiserum R016 had the highest titer (1:225,000 after four immunizations) and was studied most extensively. R016 recognizes all molecular forms of CCK, including unsulfated and oxidized forms, but has negligible cross-reactivity with gastrin and other peptides. Using CCK-8 as a standard, the assay has a minimum detection limit of 0.5 pM and an ED50 of 11.5 pM. Serial dilutions of water/acid extracts of canine intestine were parallel to serial dilutions of sulfated CCK-8, CCK-33 and CCK-39. The assay was used to measure CCK concentrations in canine plasma after C18 Sep-Pak extraction; the concentration of immunoreactive CCK increased from a basal value of 7.8 +/- 1.0 to 9.5 +/- 1.2 and 11.1 +/- 1.2 pM 30 and 60 min postprandially (P less than 0.05 by paired analysis). This sensitive and uniquely specific CCK radioimmunoassay should be useful in characterizing several aspects of CCK physiology and the method for generating CCK antisera should be of value to other investigators.

Amino Acid Sequence↗
PubMedDate not supplied

Identification of immunoreactive somatostatin in the rat harderian gland: regulation of its content by growth hormone, beta-adrenergic agonists and calcium channel blockers.

Immunoreactive somatostatin (IRS) was identified in the male rat Harderian gland (HG) by radioimmunoassay. Tissue was extracted and a displacement curve performed; there were no significant differences between values obtained with serial dilutions of extracted tissue and those from purified somatostatin standard used in the radioimmunoassay. Basal values of HG-IRS were found to be in the nanomolar range (10.8 +/- 3.5 ng IRS/mg protein). Hypophysectomy did not change the HG-IRS but, in vivo growth hormone (GH) treatment led to a dramatic increase (6-7-fold) in the levels of IRS in the HG. Isoproterenol, a beta-adrenergic agonist, when administered in vivo significantly decreased the HG-IRS content. The effect of two different calcium channel blockers on the isoproterenol-induced decrease of HG-IRS was studied; no changes were observed with nifedipine but verapamil, injected one hour after isoproterenol administration, prevented the drop in HG-IRS levels. These data demonstrate the existence of IRS in a new location, the rat Harderian gland, and support a classical endocrine regulation for its tissue concentration.

Animals↗
PubMed1981

Effect of ether, ethanol and aqueous extracts of ginseng on cardiovascular function in dogs.

Ether, ethanol and aqueous extracts of ginseng were serially prepared from Korean ginseng plants. Each extract in the dose of 40 mg/kg was administered intravenously to ten dogs under light halothane anesthesia while 11 cardiovascular variables were compared during the ensuing two hours. The variable included cardiac output, stroke volume, heart rate, mean arterial pressure, pulse pressure, central venous pressure, total peripheral resistance, pH, PaCO2, PaO2 and base deficit. Following the administration of the ether extract (40 mg/kg) the heart rate and the central venous pressure decreased significantly. The administration of ethanol extract (40 mg/kg) caused a significant decrease in the heart rate and the mean arterial pressure. After the administration of the aqueous extract (40 mg/kg) the cardiac output, stroke volume and central venous pressure were significantly decreased, while the total peripheral resistance was significantly increased.

Animals↗
PubMed1980

Location of Streptococcus mutans in the dentinal tubules of open infected root canals.

Seventy-six teeth from open canals were extracted to prepare serial longitudinal sections. The sections were made from apical portions of the teeth and stained. Ninety similar teeth were extracted to prepare dentinal splinters with files from the enlarged infected canal. The splinters were spread on a selected medium to grow S. mutans. S. mutans was detected in 48.7% of the 76 teeth examined. The distance of invasion of S. mutans in the dentinal tubules revealed by immunofluorescence averaged 509 micrometer from the canal wall and reached 1150 micrometer, depending on the serogroups of S. mutans. Unidentified germs in the sections which were demonstrated by Gram's stain invaded further than S. mutans. The frequency of appearance of the serogroups of S. mutans was 32.6% (d), 27.9% (c), 24.4% (a), and 15.5% (b).

Adolescent↗
PubMed1997

Glutaraldehyde-containing dentin bonding agents are mutagens in mammalian cells in vitro.

The mutagenic potential of glutaraldehyde-containing dentin bonding agents was shown in previous studies using a bacterial gene mutation assay, the Ames test. However, current strategies of genotoxicity testing and regulatory requirements for the biological evaluation of medical devices recommend a battery of tests that indicate induced mutations in prokaryotic and eukaryotic cells. Accordingly, the mutagenicity of three glutaraldehyde-containing bonding agents (Syntac adhesive, Prisma Universal Bond 3 adhesive, and Gluma 3) was investigated using a quantitative mammalian cell gene mutation assay (V79/HPRT test) in the present investigation. The materials were extracted in dimethyl sulfoxide (0.1 g/2 mL) for 24 h and original extracts were then serially diluted in cell culture medium before exposure to V79 cells. Cytotoxic and mutagenic effects were observed with identical concentrations of extracts of the different test materials. There was a moderate decrease of the number of surviving cells immediately after the end of exposure. Mutagenicity at the hprt locus in V79 cells was found with all materials tested, and the increases in the absolute numbers of mutants were dose dependent. The mutant frequencies were about 15- (Syntac adhesive and Gluma 3) to 20-fold (Prisma UB3 adhesive) higher than solvent control values. Since other substances than glutaraldehyde may be responsible for the mutagenic effects in mammalian cells in this study, work is currently in progress to identify the individual mutagenic compounds of dentin adhesives and related composite materials.

Animals↗
PubMed1993

Functional mapping of flow and back-diffusion rate of N-isopropyl-p-iodoamphetamine in human brain.

Iodine-123-labeled N-isopropyl-p-iodoamphetamine (IMP) has been reported to be an excellent tracer for mapping cerebral blood flow with single-photon emission computed tomography (SPECT). Clinical interpretation of these SPECT images, however, requires further understanding of the kinetics of IMP in the human brain. In order to evaluate the kinetic behavior of IMP in normal and diseased areas, we measured flow and back-diffusion rates with serial dynamic SPECT scans following an intravenous bolus injection of IMP using a multi-detector SPECT scanner. Arterial input function was determined by octanol extracted radioactivity of serial arterial blood samples. Average values for influx rate (K1) and back-diffusion rate (k2) were 0.43 ml/g/min and 0.014 min-1 in the normal cerebral cortex, 0.43 and 0.013 in the basal ganglia, 0.28 and 0.012 in the white matter and 0.48 and 0.016 in the cerebellar hemisphere. The partition coefficient (K1/k2 ratio) was 32.4 ml/g in the cerebral cortex, 35.3 in the basal ganglia, 24.7 in the white matter and 30.4 in the cerebellum. The K1-to-k2 ratio in the infarcted and ischemic regions as well as in the tumor was smaller than that of the normal cortex. Accurate measurement of local cerebral blood flow (LCBF) based on the microsphere model was possible only on the early SPECT images, but a relative pattern of LCBF can be assessed with SPECT images obtained within 1 hr after injection except for tumors.

Adolescent↗
PubMed1979

Widespread distribution of a chorionic gonadotropin-like substance in normal human tissues.

Recent studies have demonstrated that the normal human testes, colon, and liver contain a substance that resembles hCG. To extend these findings, we examined aqueous extracts of a variety of normal human tissues for the presence of this material. The beta-hCG RIA, rat Leydig cell radioreceptor assay, and a newly developed, highly specific hCG RIA were used to measure hCG activity in a serial dilutions of the extracts. Detectable concentrations of the hCG-like material were found in 146 of the 149 individual tissue samples studied. Parallelism was noted between the hCG standard and serial dilutions of extracts of testis, ovary, pituitary, lung, liver, kidney, spleen, stomach, placenta, and some small intestinal tissue samples in the beta-hCG RIA, radioreceptor assay, and the highly specific hCG RIA. An absence of parallelism was found between extracts of nonpituitary tissues and LH in the beta-LH RIA. Pancreatic extracts altered the [125I]hCG used as the labeled ligand in these assays, which led to spurious results. Chromatography of the extracts on Concanavalin A-Sepharose columns revealed that the hCG-like materials from different tissues varied widely in their adsorbtion to Concanavalin A, possibly reflecting differences in their carbohydrate contents. These results indicate that an hCG-like substance is widely distributed throughout normal human tissues and further supports the concept that the fetal genome responsible for hCG production is not completely suppressed in adult tissues.

Aging↗
PubMed1995

Expression in Escherichia coli and affinity purification of a CKS-troponin I fusion protein.

The human cardiac troponin I gene was subcloned and expressed at high levels in Escherichia coli as a fusion protein to CMP-KDO synthetase (CKS). Expression levels of the CKS-troponin I fusion were 8% of total cellular protein 4 h after induction with IPTG. The fusion was expressed primarily as an insoluble protein as shown by SDS-PAGE analysis. Expressed CKS-troponin I fusion from a crude lysate was antigenic against anti-CKS and anti-troponin I monoclonal antibodies in Western blots. The fusion was affinity-purified over a TnC affinity column using a urea-solubilized extract of a crude cell lysate. Serial dilutions of crude soluble extracts of the troponin I fusion were assayed in several microparticle enzyme immunoassays and found to exhibit similar immunogenic responses relative to cardiac troponin I isolated from human heart tissue.

Biomarkers↗