You can treat the chemotherapy patient.

You can treat the chemotherapy patient. Increased numbers of cancer patients are treated with chemotherapy. Patients who receive antineoplastic agents can be at serious risk from dental infections and should be provided appropriate dental care. In many instances, indicated treatment can be accomplished by the patient's private dentist. Certain precautions, however, are necessary when treating dental patients medically compromised by chemotherapy. Dental conditions which may increase morbidity are identified and treatment recommendations are made. Guidelines for dental intervention before, during, and after chemotherapy are discussed with emphasis on the hematologic parameters necessary for safe dental care. The cyclic relationship between chemotherapy and oral complications is also reviewed.


Reimbursement for the oral health care of cancer patients. Oral conditions related to cancer therapy, which are largely preventable, continue to result in considerable morbidity, dysfunction and lost quality of life. Therefore, it is important for dentists to be knowledgeable about the problems of reimbursement for the oral health care of cancer patients and become involved in the issue of access to oral health care for cancer patients.


Birth prevalence of Down syndrome in a predominantly Latino population: a 15-year study. The birth prevalence of Down syndrome (DS), although decreasing in parts of the world, is not known in many specific subpopulations. The rate of DS for live births was determined for a 15-year period (1974-1988) at the Los Angeles County-University of Southern California Medical Center, which is a large public hospital serving a predominantly Latino population. DS was ascertained primarily by clinical monitoring of newborn infants. The overall rate of DS was 1.60/1,000 live births and had minimal fluctuation throughout the 15 years. The rate of DS for Latino births was 1.69/1,000 live births, 1.67 when adjusted for maternal age, and 1.75 when corrected for prenatal diagnosis. The high rate of DS in the Latino population was associated with advanced maternal ages and increased maternal-age specific rates for DS, especially in the age ranges of 30 to 34 and 40 to 44 years, when compared with other studies.


Modification of platelet function by isosorbide dinitrate in patients with coronary artery disease. The effect of a four weeks oral treatment with 100 mg isosorbide dinitrate (ISDN) daily on platelet function was evaluated in 40 patients (aged 40-65 years) with proven coronary artery disease. Isosorbide dinitrate decreased platelet reactivity to ADP (p less than 0.001), increased platelet sensitivity to PGI2 (p less than 0.01) while the production of TXB2 from exogenous arachidonic acid substrate and from endogenous substrate were both significantly reduced. Circulating platelet aggregates as measured by the Wu-test were markedly reduced (p less than 0.001) but there was little change in the plasma concentration of the platelet proteins beta-thromboglobulin and platelet factor 4. Overall, platelet activation correlated with smoking, hypertension and a family history of coronary artery disease. The reduced platelet activation seen during treatment with isosorbide dinitrate may contribute to the therapeutic benefit seen with this drug in patients with coronary artery disease.


Medium effects on the kinetics of human plasmin. Effects of water-miscible organic solvents added to an aqueous buffer on the activity of several serine proteinases were studied. Plasmin in particular showed a dramatic difference in activity depending on the hydrophobicity of the added organic solvent through a combination of Km and kcat effects. An inverse linear correlation between the polarity of the mixed solvent and the log (kcat/Km) of plasmin activity was observed for both H-D-Val-Leu-Lys-p-nitroanilide (S-2251) and pro-urokinase as the substrate. The activity of plasmin was less dependent on the polarity of the added solvent when other chromogenic substrates were employed that contained an arginyl residue in the P1 site.


[Down's syndrome. Medical care and rehabilitation]. The Norwegian Government has decided to reform the care of the mentally retarded by placing the responsibility for medical and social care with the local authorities. From 1 January 1991 the mentally retarded are to be integrated into the community. Persons with Down syndrome constitute 20-30% of the mentally retarded in institutions. The syndrome is the most common single cause of mental retardation. It represent both a medical challenge and a challenge in regard to home support, work, leisure activities and habilitation. Habilitation means identifying the fundamental deficiencies, incorporating this knowledge into plans for treatment and evaluating progress in social functioning and individual capacity. This presentation emphasizes clinically important information about the syndrome and medical complications both in the early stage of life, and among youths and adults in particular. Owing to difficulties in communication, there are often diagnostic problems. However, when recognized, many of the complications can be treated effectively.


Changes in lymphocyte subsets and mitogen responsiveness following open-heart surgery and possible therapeutic approaches. Septic multi-organ failure represents a common cause for operative mortality following open-heart surgery. One major reason might be the depression of cell-mediated immunity. The purpose of this prospective randomized trial was to quantify and specify the effects of open-heart surgery on cell-mediated immune mechanisms. In addition, the immunorestorative potential of a combined immunomodulatory therapy with the cyclooxygenase inhibitor indomethacin and the thymomimetic substance Thymopentin versus single drug administration of indomethacin was investigated. Twenty patients were given indomethacin for the first 5 postoperative days (group A). Another 20 patients also received Thymopentin perioperatively and on the second and fourth postoperative days (group B), while 20 patients underwent conventional therapy (group C). Cell-mediated immune response was quantified in vitro by measuring CD3+ T-lymphocytes and their subsets, CD4+ T-helper and CD8+ T-suppressor cells. Lymphocyte responsiveness to a specific (Antigen-Cocktail) and non-specific mitogen (phytohemagglutinin) provided information about the quality of cell-mediated immune response. On the first postoperative day CD3+ T-lymphocyte counts and Antigen-Cocktail-induced lymphocyte proliferation decreased significantly in all groups. The number of CD4+ T-Helper cells fell significantly only in groups A and C, while the decrease in group B was not statistically significant; the same applied to phytohemagglutinin-induced lymphocyte response. The CD4+/CD8+ ratio was significantly depressed only in group C, decreased slightly in group A and did not change as compared to baseline values in group B. All investigated parameters remained significantly depressed until the seventh postoperative day in group C.(ABSTRACT TRUNCATED AT 250 WORDS)


Induction of stable chimerism and transplantation tolerance to rat islet and heart allografts by ultraviolet-B modulation of bone marrow cells. Ultraviolet-B irradiation (UV-B) (700 J/m2) of BM cells prior to transplantation into lethally gamma-irradiated (1050 rads) allogeneic rats prevents the development of GVHD and results in stable chimerism. This study was developed to determine if UV-B modulation of BMT is useful for preconditioning recipients for the induction of tolerance to donor islets and heart allografts. Lethally irradiated Lewis rats that received UV-B irradiated (700 J/m2) WF BMT (10(8) BM cells) demonstrated stable chimerism without any evidence of GVHD. The stable Lewis chimeras were made diabetic with streptozotocin (STZ) at 28-35 days after BMT and subdivided into 3 experimental groups that received 1000-1200 islets from WF, Lewis, or BN (third-party), respectively. The results showed that group I diabetic Lewis chimeras accepted permanently (greater than 300 days) BM donor WF islets and became normoglycemic. When 3 of 6 Lewis chimeras transplanted with WF islets were rechallenged with WF hearts 60 days after islet grafts, they accepted both islets and cardiac allografts permanently (greater than 240 days). Similarly, the remaining 3 animals accepted Lewis cardiac allografts permanently, thus indicating tolerance to both donor and recipient alloantigens. Group II diabetic chimeras accepted permanently (greater than 300 days) recipient (Lewis) islets. In contrast, group III chimeras rejected acutely (7-8 days) third-party (BN) islets. However, when these animals that rejected BN islets and again became diabetic were retransplanted with BM donor-type (WF) islets, they became permanently normoglycemic (greater than 200 days). This finding emphasizes the specificity of the induction of tolerance in this model and the apparent lack of organ-specific sensitization. To define the underlying mechanism of tolerance, in vivo adoptive transfer of 10(8) spleen cells to naive Lewis or WF recipients, obtained from tolerant Lewis chimeras carrying donor islets and heart allografts, showed no prolongation of cardiac allografts in the unmodified syngeneic hosts, thus questioning the role of suppressor mechanisms in the tolerant rats. Furthermore, cells from the tolerant chimeras that showed no mixed lymphocyte reaction (MLR) response to Lewis or; WF alloantigens failed to suppress anti-Lewis and anti-WF MLR-response in coculture MLR. These results suggest that tolerance to donor alloantigens in the UV-B BMT model is most likely due to selective elimination of anti-BM donor helper or effector cell precursors (clonal deletion) rather than induction of suppressor cell activity. This study demonstrates that this relatively simple and effective approach to modulation of T cells in BM treatment may be potentially useful in the induction of tolerance to donor organs.


[Postoperative duplex ultrasonic surveillance of infrainguinal arterial reconstructions]. Postoperative bypass surveillance can identify graft-related stenoses in 25% of grafts during the first two years after surgery. Correction of the stenoses before they lead to graft occlusion improves the patency significantly. Clinical assessment including distal pressure measurements is of poor sensitivity. Surveillance is best performed with color duplex scanning. Ideally, grafts are examined first after one to two months, thereafter every three months and, after one year, at intervals of six months. Graft surveillance beyond two years is hardly justified, as is the case with prosthetic grafts. Symptom-producing and tight stenoses should be corrected. Percutaneous transluminal angioplasty is appropriate for short stenoses, whereas stenoses longer than 2 to 3 cm are best treated surgically.


Immune complex-mediated glomerulonephritis associated with bacterial kidney disease in the rainbow trout (Oncorhynchus mykiss). Rainbow trout (Oncorhynchus mykiss) developed a post-infectious chronic membranous glomerulonephritis 15 months after they had been experimentally infected with Renibacterium salmoninarum. Histologically, peritubular and periglomerular fibrosis, hypercellular glomeruli with occluded Bowman's space, and partial or complete adhesion to Bowman's capsule were constant features. Electron microscopy revealed thickened glomerular basement membranes with spikes accompanied by finely granular electron-dense deposits at the epithelial side and dense material in the mesangial matrix. Indirect immunofluorescence indicated linear immunoglobulin deposits along the glomerular basement membrane. The presence of R. salmoninarum was demonstrated by culture and by indirect immunofluorescence. Low serum hemagglutination-inhibiting antibody titers were demonstrated.


[Drug clearance as a decision aid for further invasive liver diagnosis--studies with hexobarbital as a model substrate]. The clearance of a drug predominantly metabolized in the liver may serve as an estimate of quantitative liver function. In 260 consecutive patients presenting with a history of liver disease and abnormal laboratory findings but without a current definite diagnosis we have measured the clearance of hexobarbital and investigated if low values in patients are able to support the decision for an invasive diagnostic procedure such as needle biopsy or laparoscopy. 250 mg of hexobarbital was given orally to the patients between 8 and 10 hrs p.m. 12 hrs later blood samples were taken. Hexobarbital was determined by gas chromatography with N-selective detection, and a single point clearance was calculated. We recommended liver biopsy or laparoscopy to all patients with a hexobarbital clearance below 2.7 ml/min/kg body weight (normal 2.66-5.34 ml/min/kg). 73 out of 260 patients showed a reduced hexobarbital clearance. In 44 patients blind liver biopsy (n = 14) or laparoscopy (n = 30) was performed, 29 patients refused an invasive diagnostic procedure. 17 out of 26 patients with the tentative diagnosis chronic hepatitis had already an incomplete or complete liver cirrhosis. In 11 out of 18 patients with the tentative diagnosis alcohol toxic liver injury we found a progressive portal fibrosis or complete liver cirrhosis. Reduced drug clearance reflecting quantitative liver function can be an indicator of advanced liver disease, thus adding substantially to the decision for further invasive diagnostic procedures.


Postoperative analgesia for major abdominal surgery with continuous thoracic epidural infusion of bupivacaine with sufentanil, versus bupivacaine with morphine. A randomized double blind study. Forty-six patients undergoing major abdominal surgery were given postoperative epidural analgesia for four days with bupivacaine-sufentanil or bupivacaine-morphine. Both groups received a bolus of 8 ml bupivacaine 0.5% followed after 30 minutes by an infusion of 20 ml/h bupivacaine 0.1%. The sufentanil group (group A: 21 patients) received a loading dose of 50 micrograms sufentanil and a continuous infusion of 5 micrograms/h sufentanil. The morphine group (group B: 25 patients) received no loading dose of morphine but only a continuous infusion of 0.5 mg/h morphine. Both regimens provided excellent analgesia, but the frequency of respiratory depression was much greater in group A (33% versus 4% in group B). This respiratory depression occurred in the first hours postoperatively, when the patients were still in the post-anesthesia care unit. There was also a high incidence of hypotension after the loading dose of bupivacaine 0.5%. Although we noticed a large incidence of pruritus, no patient needed naloxone reversal. In view of these side effects we recommend a lower loading dose of both bupivacaine and sufentanil.


Interactions between epidurally and intrathecally administered sufentanil and bupivacaine in hydroxypropyl-beta-cyclodextrin in the rat. The interaction between inactive doses of the opioid sufentanil and the local anesthetic bupivacaine after complexation in various concentrations of hydroxypropyl-beta-cyclodextrin (HP-B-CD) were studied after epidural and intrathecal administration in the rat. Whereas 0.125 microgram sufentanil and 80 micrograms bupivacaine produced only limited effects, a combination of the two compounds resulted in a profound surgical analgesia in all rats tested after epidural administration. Also intrathecally, a clear potentiation was present. The complexation of the same doses of sufentanil plus bupivacaine in HP-B-CD increased the duration of analgesia. Epidurally, a maximal potentiation of analgesia was present at 0.125 microgram sufentanil plus 80 micrograms bupivacaine in 20% HP-B-CD. Intrathecally, maximal potentiation of the analgesic activity was present starting from a complexation of both drugs in 10% hydroxypropyl-beta-cyclodextrin. Higher concentrations of hydroxypropyl-beta-cyclodextrin did not significantly further increase the duration of analgesia. These results indicate that for both routes of spinal administration, the complexation of sufentanil plus bupivacaine in HP-B-CD can produce a longer analgesia than either plain sufentanil, sufentanil combined with bupivacaine or sufentanil alone in HP-B-CD. The complexation of sufentanil plus bupivacaine in HP-B-CD did not diminish the increased safety observed to occur after complexation of sufentanil in HP-B-CD and after combination of sufentanil plus bupivacaine.


Surgical treatment of acute type V acromioclavicular injuries. A prospective study. In a prospective study, 28 consecutive patients with an acute Type V acromioclavicular sprain were treated with a coraco-clavicular repair using a double velour Dacron graft. All patients were reviewed after a mean follow-up period of 5.1 years (range: 1 to 9 years). At follow-up, 20 patients (71.4%) showed good or excellent results, according to the Imatani evaluation system, and 8 patients (28.6%) demonstrated a fair or poor result according to the same system. Loss of reduction was encountered in 11 shoulders (40%), despite an initial anatomical reduction. No correlation was seen between the overall scores at follow-up and the degree of residual dislocation, between the overall scores and the presence of coraco-clavicular calcifications or ossifications, between the overall scores and the development of posttraumatic arthritic changes, or between the overall scores and the presence of osteolysis of the distal clavicle.


Sleep apnea syndrome in children--secondary to adenotonsillar hypertrophy? In this study we investigated the relationship between the size of adenoids and/or tonsils and obstructive sleep apnea syndrome (OSAS) in 19 children aged 3 to 7 years. After clinical examination by an ENT specialist, electrocardiogram (ECG), vectorcardiogram (VCG) and echocardiogram (EC) were performed while the patients were awake, and static charge sensitive bed (SCSB) method recording (noninvasive monitoring of ballistocardiogram, respiration and body movements) while asleep. After adenoidectomy and/or tonsillectomy, the volume of the tonsils and the adenoids was determined. RVH was found in 4 out of 19 children (21%). RVH findings correlated highly with the number of apneas but, surprisingly, not with measured adenotonsillar size in those children. However, at 6 months of follow-up, the VCG and EC changes had returned to normal. These results suggest that RVH is more common in children suffering from upper airway obstruction than previously believed, and that factors other than adenotonsillar size also have an influence on upper airway obstruction.


Receptor binding properties of the new and specific thromboxane receptor antagonist Bay U 3405. Human platelet membranes were used to characterize the receptor binding properties of the specific thromboxane receptor antagonist 3H-SQ 29548 and the displacement of 3H-SQ 29548 from its binding site by the new thromboxane receptor antagonist Bay u 3405. The specific binding of 3H-SQ 29548 was saturable with an association rate constant of 1 x 10(-11) mol-1 min-1 and a dissociation rate constant of 0.032 min-1. Nonspecific binding of 3H-SQ 29548 was below 10%. When Scatchard plot analysis was performed on equilibrium saturation binding the kD was 69 nmol/l and the Bmax was calculated as 3.9 pmol/mg membrane protein. 3H-SQ 29548 was dose dependently displaced from its binding site by addition of increasing concentrations of Bay u 3405 yielding an IC50 value of 68 +/- 12 nmol/l, being not significantly different from the IC50 of nonlabelled SQ 29548 (38 +/- 13 nmol/l). The results show that Bay u 3405 is a potent and specific thromboxane receptor antagonist, displacing 3H-SQ 29548 from its binding site on human platelet membranes with IC50 values being not significantly different. These receptor binding properties and the long biological half life reported in vivo make Bay u 3405 a promissing compound for the treatment of human cardiovascular diseases.


pH-dependent binding of the TXA2/PGH2-receptor of human platelet membranes to various ligands. [3H]-BAY U 3405 was used to characterize the pH-dependency of the binding of various ligands to the TXA2/PGH2-receptor of human platelet membranes. Maximum binding of [3H]-BAY U 3405 is achieved at pH 5.8. In inhibition studies the ligands Daltroban, CTA2, and U 46619 also show a higher affinity at pH 5.8 compared to pH 7.4. In contrast, the ligands I-PTA-OH and GR 32191 have a higher affinity at pH 7.4. No difference is seen with SQ 29548. The ligands I-PTA-OH, GR 32191, and SQ 29548 have a second protonable group in common, which is thought to be the reason for the different pH-dependent binding.


Concomitant aspirin treatment abolishes the antiatherosclerotic effects of the calcium channel blocker isradipine mediated by PGI2. 108 male rabbits, aged 6 months, with experimental hypercholesterolemia and experimental abdominal aortic lesioning received different regimen of antiatherosclerotic treatment; 36 of them were treated with isradipine, a dihydropyridine calcium antagonist (0.3 mg/kg/daily), 36 with isradipine in combination with aspirin whereas 36 animals received placebo. The entry of 125I-radiolabelled LDL into the aorta was demonstrated to be significantly diminished in isradipine-treated rabbits as well as positive Sudan-III-staining and aortic cholesterol content were in comparison to placebo. This benefit was almost completely abolished by concomitant aspirin-treatment. The notable increase in vascular prostacyclin (PGI2) is supposed to mediate the strong antiatherosclerotic effect of isradipine resulting in an inhibition of LDL-entry and vascular cholesterol accumulation. Aspirin almost totally blocked the raise in PGI2-synthesis by the inhibition of cyclooxygenase detected in isradipine-treated animals. It can be concluded, that aspirin-treatment may minimize the antiatherosclerotic actions of calcium antagonists which are mediated by the PG-system.


Myocyte cellular hypertrophy and hyperplasia contribute to ventricular wall remodeling in anemia-induced cardiac hypertrophy in rats. To determine the effects of chronic anemia on the functional and structural characteristics of the heart, 1-month-old male rats were fed a diet deficient in iron and copper, which led to a hemoglobin concentration of 4.63 g/dl, for 8 weeks. At sacrifice, under fentanyl citrate and droperidol anesthesia, systolic, diastolic, and mean arterial blood pressures were decreased, whereas differential pressure was increased. Left ventricular systolic pressure and the ventricular rate of pressure rise (mmHg/s) were reduced by 9% and 14%, respectively. Moreover, developed peak systolic ventricular pressure and maximal dP/dt diminished 14% and 12%. After perfusion fixation of the coronary vasculature and the myocardium, at a left ventricular intracavitary pressure equal to the in vivo measured end diastolic pressure, a 10% thickening of the left ventricular wall was measured in association with a 13% increase in the equatorial cavitary diameter and a 44% augmentation in ventricular mass. The 52% hypertrophy of the right ventricle was characterized by an 11% thicker wall and a 37% larger ventricular area. The 33% expansion in the aggregate myocyte volume of the left ventricle was found to be due to a 14% myocyte cellular hypertrophy and a 17% myocyte cellular hyperplasia. These cellular parameters were calculated from the estimation of the number of myocyte nuclei per unit volume of myocardium in situ and the evaluation of the distribution of nuclei per cell in enzymatically dissociated myocytes. Myocyte cellular hyperplasia provoked a 9% increase in the absolute number of cells across the left ventricular wall. In contrast, myocyte cellular hypertrophy (42%) was responsible for the increase in myocyte volume of the right ventricle. The proliferative response of left ventricular myocytes was not capable of restoring diastolic cell stress, which was enhanced by the changes in ventricular anatomy with anemia. In conclusion, chronic anemia induced an unbalanced load on the left ventricle, which evoked a hyperplastic reaction of preexisting myocytes, in an attempt to normalize diastolic wall and myocyte stress.


Endotoxin-induced cytokine gene expression in vivo. IV. Expression of interleukin-1 alpha/beta and interleukin-1 receptor antagonist mRNA during endotoxemia and during endotoxin-initiated local acute inflammation. After the intravenous (IV) injection of endotoxin, (lipopolysaccharide [LPS]), in the rat, interleukin-1 alpha/beta (IL-1 alpha/beta) mRNA expression peaks at 1 hour in whole organ RNA preparations of the lung, liver, spleen, and bowel. Interleukin-1 receptor antagonist (IL-1ra) mRNA peaks at 2 to 4 hours, consistent with the hypothesis that IL-1ra acts as an endogenous negative feedback mechanism to downregulate the proinflammatory effects of IL-1. After the intratracheal (IT) injection of LPS, however, IL-1 and IL-1ra mRNA levels in whole lung peak at 6 hours, concurrent with the maximum influx of neutrophils (PMNs) into the bronchoalveolar space. To address the cellular source of IL-1 and IL-1ra mRNA in the lung during acute pneumonitis, mRNA levels were studied in bronchoalveolar lavage (BAL) macrophages incubated with LPS in vitro for 6 hours as compared with BAL cells (95% PMNs) obtained 6 hours after IT injection of LPS. A much greater expression of IL-1 and IL-1ra mRNA was observed in PMN-rich BAL cells obtained after IT injection of LPS, suggesting that PMNs contribute substantially to IL-1 and IL-1ra mRNA expression. Fractionation of alveolar macrophage-enriched and PMN-enriched subpopulations from the BAL cells obtained at 6 hours after IT injection of LPS confirmed that neutrophils are a source of IL-1 and IL-1ra mRNA. The difference in the kinetics of IL-1 and IL-1ra mRNA expression in whole lung RNA preparations after IV and IT injections of LPS is due to the contribution of PMNs that appear in the lung in large numbers after IT injection. Finally, human peripheral blood PMNs were found to express IL-1ra mRNA and protein after in vitro incubation with LPS. PMNs may contribute to the up- and downregulation of their own accumulation by expressing both IL-1 and IL-1ra.


Comparison of the effects of loratadine and astemizole in the treatment of children with seasonal allergic rhinoconjunctivitis. The efficacy and safety of a once-daily dose of loratadine 10 mg (5 mg in patients whose body weight was less than or equal to 30 kg) and astemizole 2 mg/10 kg body weight o.d. were compared in a 14-day third-party blind study. Forty-one children (30 boys and 11 girls, aged 6-14 years) with seasonal allergic rhinoconjunctivitis entered the study. The pollen count was monitored throughout the study. A significant improvement (p less than 0.01) in allergy symptoms was observed from the third day for both drugs; there was no significant difference between drugs, although loratadine led to a greater reduction in symptoms. In the astemizole group, the apparently rapid onset of drug action might by explained by reduced pollen exposure. Therapeutic response was excellent/good in 83.3% and 58.8% of the loratadine and astemizole groups, respectively. The results reported by the patients/parents were similar. Nine of the children on astemizole and four children on loratadine complained of side effects; three patients in the astemizole group were withdrawn from treatment because of adverse effects. No abnormal changes in lab values were observed in either group.


Increasing the activity of affinity-purified DNA-binding proteins by adding high concentrations of nonspecific proteins. A large decrease in the activity of two sequence-specific DNA-binding proteins implicated in transcription control was seen when these were affinity purified and assayed under standard conditions in electrophoretic mobility shift assays. Increasing the concentration of bovine serum albumin in the reaction mixtures from 0.1 to 5 mg/ml stimulated the DNA-binding activity of these affinity-purified proteins, human CREB (cyclic AMP response element binding protein) and MDBP (methylated DNA-binding protein), approximately 5-to more than 20-fold. In the case of affinity-purified MDBP, adding back the affinity flow-through fraction to the assay mixture gave similar extents of stimulation at much lower final protein concentrations. The specific DNA-binding activity of the affinity-purified CREB, but not that of MDBP, was also increased by adding a nonionic detergent to the binding reaction buffer although not as much. The large increase in the amount of MDBP.DNA complex seen upon supplementation of the affinity-purified MDBP with the affinity flow-through fraction or 5 mg/ml of BSA was shown to be due to stimulation, by nonspecific proteins, of specific complex formation and not to prevention of activity losses by adsorption or denaturation during the assay.


Effects of hypocarbia on the pharmacodynamics of sufentanil in humans. Descriptors of power and frequency derived from power spectral analysis of the electroencephalogram (EEG) were used to determine the effects of low-dose sufentanil (0.1 micrograms/kg) on brain activity. The effects of hypocarbia alone and of hypocarbia with sufentanil in patients receiving a N2/O2 (70%:30%) anesthetic were also studied. Hypocarbia alone caused changes in most EEG descriptors from both the anterior (F3-C3) and posterior (P3-O1) EEG montages. All EEG descriptors in both hypocarbic and normocarbic patients significantly changed when sufentanil was administered, reflecting a shift of power into the lower frequency ranges. When the anterior EEG montages from the two groups that received sufentanil were compared, the delta power band, spectral edge 50 (median power frequency), and the relative power in the delta power band divided by the alpha plus beta power bands [D/(A + B)] in the hypocarbic group exhibited a significantly greater shift of power into the lower frequency range. It is concluded that (a) power spectral analysis is a sensitive measure of the effects of hypocarbia and small doses of sufentanil on the brain; (b) the power spectral analysis descriptors--delta power band, spectral edge 50, and [D/(A + B)]--are statistically the most sensitive to EEG changes induced by sufentanil; and (c) hypocarbia intensifies patient EEG response to sufentanil, as judged by changes in EEG descriptors.


Ionic modulation of the effects of heparin on plasminogen activation by tissue plasminogen activator: the effects of ionic strength, divalent cations, and chloride. Ionic strength, divalent cations, and Cl- modulate the ability of the glycosaminoglycan heparin to stimulate the activation of human plasminogen (Pg) by tissue-type Pg activator. Kinetic analysis of Pg activation indicates that heparin is inhibitory, stimulatory, or nonstimulatory as a function of ionic strength. While increasing ionic strength inhibits Pg activation in the absence of heparin, in it presence an activation phase followed by an inhibitory phase is observed. Divalent cations, inhibitors of activation in the absence of heparin, increase the rate of activation in its presence. Kinetic analysis demonstrates that divalent cations augment the heparin stimulatory effect a maximum of 60-fold due to increases in kcat without changes in Km of the reaction. This effect is heparin-specific, since activation is not affected by Ca2+ in the presence of heparan sulfate or de-N-sulfated heparin. Also, Cl- inhibits Pg activation in the presence of heparin by acting as a competitive inhibitor (Kic of 100 mM). Furthermore, inhibition by Cl- reduces the overall magnitude of heparin stimulation of Pg activation. These results suggest that physiologic ions in combination with heparin may be significant effectors of Pg activation in the vascular microenvironment.


Coupled expression of Ca2+ transport ATPase and a dihydrofolate reductase selectable marker in a mammalian cell system. Stable expression of a full-length cDNA encoding chicken fast muscle Ca2+ transport ATPase was obtained in a Chinese hamster lung cell line (DC-3F), using a dual-promoter expression vector (pH beta FCaA3) in which the ATPase was cloned downstream of a human beta-actin gene promoter, and a mutant dihydrofolate reductase cDNA (A3/DHFR) was cloned downstream of an SV40 promoter-enhancer. Owing to its essentially normal catalytic activity and modest (20-fold) resistance to the antifolate methotrexate (MTX), the A3/DHFR mutant enzyme served as an efficient dominant selection marker in transfected cell populations challenged with MTX and, within a broad range of drug concentrations, allowed subsequent amplification and overexpression of vector sequences. In stable transfectants, the expressed ATPase was targeted to intracellular membranes, and the microsomal fractions from those cells exhibited high rates of Ca2+ transport. In comparative experiments using transient expression in COS1 cells, the level of ATPase per transfected cell was greater, but less than 5% of the transfected population exhibited ATPase expression. Furthermore, as opposed to the stable lines, the transiently expressing cells could not be propagated. Overall, the yield of ATPase was 12-16 and 4-6 micrograms per milligram of microsomal protein in the stable and the transient expression systems, respectively. The advantages of the stably transfected cell lines therefore lie in the homogeneity of ATPase expression and its distribution in cells and microsomes, in the large yield of microsomes obtained by continuous cell propagation, and in the reproducible functional characteristics of the microsomes. Moreover, the microsomes derived from stably transfected cell lines provide a convenient system for studies of Ca2+ transport and ATPase partial reaction, eliminating the need to conduct repetitive transient transfections to obtain sufficient amounts of enzyme for functional studies.


Effect of education, occupation and some lifestyle factors on common rheumatic complaints in a Swedish group aged 50-70 years. The relation between common rheumatic diseases such as osteoarthrosis, arthralgia without definite signs of osteoarthrosis, subacromial shoulder pain, different forms of tendinitis, low back pain and neck pain, and the level of formal education, occupational workload and some lifestyle factors were examined in 502 of 900 randomly selected subjects aged 50-70 years. The group with rheumatic complaints had a higher proportion of subjects with a lower level of formal education (less than or equal to eight years) by bivariate analysis. In multivariate analysis, the major risk factors were: a self rated heavy workload (odds ratio (OR) 6.4), sleep disturbance (OR 3.6), and advanced age (OR 2.0 per five year increase) for osteoarthrosis; a self rated heavy workload for subacromial shoulder pain (OR 5.4) and low back pain (OR 4.8); and a self rated heavy workload (OR 8.0) and female sex (OR 4.8) for neck pain. A self rated heavy workload was strongly correlated with a low level of formal education. A heavy workload (i.e. previous or present principal occupation) could only be confirmed in the groups with neck pain and low back pain on the basis of available occupational classification data. Neck pain was thus associated with occupations entailing repetitive tasks and awkward posture with respect to the neck, shoulders, and back. Low back pain was associated with occupations entailing awkward posture with respect to the neck, shoulders, and back, and occupations entailing exposure to vibration and heavy manual work. It is concluded that, in a cross sectional sample of an elderly population, a low level of formal education and self rated heavy physical work are associated with the occurrence of adult rheumatic complaints, though the self rated heavy workload could only be verified in the groups with neck pain and low back pain. There correlations between heavy work and low back pain, and especially neck pain, suggest that successful prevention would mean a substantial economic gain to the community. Whether the level of education is a marker of risk factors other than a heavy occupational workload needs further evaluation.


Pityriasis rosea and discoid eczema: dose related reactions to treatment with gold. Sixteen cases of either a pityriasiform or discoid eczematous rash occurring in patients with rheumatoid arthritis receiving treatment with gold (sodium aurothiomalate and auranofin) were studied. The results suggest that this is a dose related, not allergic, reaction to gold. The development of this rash is not an absolute indication to stop treatment with gold. Control can often be effected with potent topical steroids or a reduction in the dose or frequency of treatment with gold.


Laparoscopic appendectomy. Initial experience in a teaching program. From February 1990 to December 1991, 16 laparoscopic procedures were performed for right lower quadrant pain. There were nine men and seven women, aged 16 to 47 years (mean, 27.2 years). All procedures were performed by surgical chief residents with prior experience in laparoscopic cholecystectomy, first-assisted by an attending surgeon. The appendix was visualized and a definitive diagnosis was made in all patients. One patient with acute salpingitis underwent diagnostic laparoscopy only; two patients underwent laparotomy (perforated appendicitis, perforated diverticulitis). A fourth patient had an acute torsion of an ovarian cyst managed laparoscopically. Laparoscopic appendectomy was successfully performed in 12 patients (acute appendicitis, 9; fibrosis or chronic inflammation, 2; normal appendix, 1). Mean operative time for laparoscopic appendectomy was 95.7 minutes, and mean postoperative stay was 2.5 days. The authors conclude that operative time, diagnostic accuracy, and complication rates for laparoscopic appendectomy are acceptable. Within the context of a training program, laparoscopic appendectomy provides an opportunity for surgical residents to expand laparoscopic skills.


Selective cholangiography. Current role in laparoscopic cholecystectomy. The initial 22-month experience with laparoscopic cholecystectomy in 400 patients employing an algorithm of selective cholangiographic evaluation is reported. Preoperative or postoperative endoscopic retrograde cholangiography was performed whenever stones were suspected clinically. Preoperative endoscopic retrograde cholangiography was performed in 44 patients (11%), in whom 14 (3.5%) had an endoscopic sphincterotomy with extraction of common bile duct stones. Intraoperative cholangiography was performed in only eight patients (2%) almost exclusively to acquire experience with the technique, and all cholangiograms were normal. Laparoscopic cholecystectomy was successfully completed in 96% of the patients. There were no deaths in this series, and major complications occurred in only 5% of patients. Two patients (0.5%) had a significant common bile duct injury that was recognized and successfully repaired at the initial operation. No late common bile duct strictures have been recognized. Six patients (1.5%) underwent postoperative endoscopic retrograde cholangiography for suspected common bile duct stones, with three patients requiring endoscopic sphincterotomy and stone extraction. This experience suggests that the use of preoperative and postoperative endoscopic retrograde cholangiography can be based on clinical presentation and laboratory evaluation and does not need to be performed routinely. Routine intraoperative cholangiography is not necessary in most patients undergoing laparoscopic cholecystectomy. The authors conclude that laparoscopic cholecystectomy can be performed safely with the selective use of cholangiography.


Endovascular surgery for peripheral arterial occlusive disease. A critical review. Endovascular surgery is a new multidisciplinary field that applies the recently innovated techniques of angioscopy, intraluminal ultrasound, balloon angioplasty, laser, mechanical atherectomy, and stents. This field can be defined as a diagnostic and therapeutic discipline that uses catheter-based systems to treat vascular disease. As such, it integrates the subspecialties of vascular surgery, interventional radiology, interventional cardiology, and biomedical engineering for the common purpose of improving arterial hemodynamics. Endovascular surgery offers many potential benefits: long incisions are replaced with a puncture wound, the need for postoperative intensive care is significantly reduced, major cardiac and pulmonary complications from general anesthesia are side stepped, and the dollar savings could be dramatic as the need for intensive care unit and in-hospital stay diminishes. Despite these technological advancements, endovascular surgery is still in its infancy and currently has limited applications. This review provides an updated summary of endovascular surgery today and addresses some of the obstacles still preventing its widespread use.


The sarcoplasmic reticulum Ca(2+)-ATPase, SERCA1a, contains endoplasmic reticulum targeting information. The fast-twitch skeletal muscle Ca(2+)-ATPase isoenzyme, SERCA1a, is localized in chick skeletal myotubes to both the sarcoplasmic reticulum (SR) and to the nuclear envelope, an extension of the endoplasmic reticulum (ER). The ER labeling remained after cycloheximide treatment, indicating that it did not represent newly synthesized SERCA1a in transit to the SR. Expression of the cDNA encoding SERCA1a in cultured non-muscle cells led to the localization of the enzyme in the ER, as indicated by organelle morphology and the co-localization of SERCA1a with the endogenous ER luminal protein, BiP. Immunopurification analysis showed that SERCA1a was not bound to BiP, nor was any degradation apparent. Thus, the SR Ca(2+)-ATPase appears to contain ER targeting information.


The human complement C4B/steroid 21-hydroxylase (CYP21) and complement C4A/21-hydroxylase pseudogene (CYP21P) intergenic sequences: comparison and identification of possible regulatory elements. We determined the 1.8 kb intergenic sequences between the human complement C4B gene and the active steroid 21-hydroxylase gene in two subjects, and between the C4A gene and the steroid 21-hydroxylase pseudogene in one subject. Comparison of these sequences with each other and with published homologues revealed no differences which were unique to either intergenic region. Sequence analysis revealed two copies of an AGGTCA motif in all sequences. This motif is common to steroidogenic enzyme gene promoters and to the response elements for nuclear hormone receptors. Similarities with human enhancers were also found.


Morphological and biochemical evidence for partial nuclear localization of annexin 1 in endothelial cells. Using immunofluorescence, an affinity-purified anti-annexin-1 polyclonal antibody showed both cytoplasmic and nuclear staining, whereas antibodies against annexins 2, 5 and 6 labelled almost exclusively the cytoplasm of cultured endothelial cells. This was further confirmed by immunogold labelling and electron microscopy using a monoclonal antibody, annexin 1 being detected close to the plasma membrane, in the cytoplasm, as well as inside the nucleus. Finally, using immunoblotting, purified nuclei were shown to contain annexin 1, which was not removed by EDTA treatment. These data open some new perspectives in the understanding of annexin function, including possible involvement in nucleoskeleton dynamics and regulation of proliferation through cell signalling.


Effects of ethanol and its fluorinated analogues on the calcium ATPase of sarcoplasmic reticulum. The effects of ethanol, monofluoro- and trifluoroethanol on the hydrolysis of ATP and coupled Ca2+ translocation by sarcoplasmic reticulum Ca(2+)-ATPase (EC 3.6.1.38) were measured. All three alcohols had parallel effects on the enzyme activities at concentrations up to 200 mM, suggesting a similar mechanism of action. 19F nuclear magnetic resonance spectroscopy was used to investigate their site of action in the purified enzyme.


Interconversion of tetrahydrofolate cofactors to dihydrofolate induced by trimetrexate after suppression of thymidylate synthase by fluorodeoxyuridine in L1210 leukemia cells. Previous studies from this laboratory demonstrated that marked suppression of thymidylate synthase activity is required to slow the rate of interconversion of tetrahydrofolate cofactors to dihydrofolate when dihydrofolate reductase is blocked by an antifolate. This finding is due to the high catalytic activity of thymidylate synthase within cells in comparison to the tetrahydrofolate cofactor pool size. In the present study, we assessed the rate of resumption of thymidylate synthase catalytic activity in terms of [3H]deoxyuridine incorporation into DNA and dihydrofolate generation from tetrahydrofolate cofactors following exposure of cells to fluorodeoxyuridine. Log phase L1210 leukemia cells, incubated with fluorodeoxyuridine to abolish thymidylate synthase catalytic activity, were suspended into drug-free medium. Resumption of [3H]deoxyuridine incorporation into DNA was negligible; by 4 hr enzyme activity was still inhibited by approximately 98%. However, this was sufficient to interconvert all available tetrahydrofolate cofactors to dihydrofolate (T1/2 approximately 2 hr) when dihydrofolate reductase was inhibited by the lipophilic antifolate trimetrexate. Interconversion of tetrahydrofolate cofactors to dihydrofolate correlated with a decline, then cessation, of purine synthesis as measured by the incorporation of [14C]formate into purine bases. These data suggest that an earlier than previously expected depletion of tetrahydrofolate cofactors with consequent inhibition of purine and other folate-dependent synthetic processes is likely to occur when antifolates are administered after a fluoropyrimidine.


The effect of glycerol and 4-nitroquinoline 1-oxide on active oxygen formation in sub-cellular fractions of lung tissue. Glycerol enhances pulmonary tumorigenesis in mice treated with 4-nitroquinoline 1-oxide (4NQO). The present study shows that active oxygen formation by glycerol may be responsible for this enhanced tumorigenesis. Male ddY mice were treated with 4NQO and given a 5% glycerol solution instead of drinking water for up to 4 weeks after 4NQO injection. There was no difference in NADH-dependent active oxygen formation of the mitochondria between the 4NQO- and 4NQO-plus-glycerol-treated groups but the ratios of NADH- and NADPH-dependent active oxygen formation of the microsomes and nuclei of the 4NQO-plus-glycerol-treated group to the 4NQO-treated group increased with increasing time after 4NQO injection. Significant differences in the maximum NADH- and NADPH-dependent active oxygen formation were observed 2 or 4 weeks after 4NQO injection.


Fatty acid conjugates of 2'-deoxy-5-fluorouridine as prodrugs for the selective delivery of 5-fluorouracil to tumor cells. We have prepared a novel class of prodrugs by coupling 2'-deoxy-5-fluorouridine (5dFU) to oleic (18:1) and docosahexaenoic (22:6) acids, respectively. The cytotoxic activity of the drug and its conjugates (5dFU-18:1 and 5dFU-22:6) has been assayed in vitro upon HT-29, a colon carcinoma cell line of human origin. After short term (2-hr) treatments with the drugs, both fatty acid conjugates of 5dFU showed cytotoxic activity in a dose-dependent way, while 5dFU alone was devoid of toxic effects within the whole range of concentrations (10-200 microM) tested. Following long term (24- or 48-hr) incubations only a fraction of the HT-29 cell population was sensitive to 5dFU, the rest of the population being resistant even at the highest concentration tested (200 microM). In contrast, 5dFU-oleic acid and, particularly, 5dFU-docosahexaenoic acids appeared toxic for the whole population of HT-29 cells under the same experimental conditions. The considerable gain in cell toxicity and, to a lesser extent, in selectivity resulted from the conjugation since the toxic effect of the drug alone was not modified when equimolar mixtures of 5dFU and fatty acids were assayed. These results confirm a previous study on the cytotoxicity of fatty acid derivatives of chlorambucil toward malignant lymphoblastoid cells and reinforce the potential use of fatty acid conjugates as efficient anti-tumor prodrugs.


Identification of novel reduced pyridinium derivatives as synthetic co-factors for the enzyme DT diaphorase (NAD(P)H dehydrogenase (quinone), EC 1.6.99.2). The enzyme DT diaphorase (NAD(P)H dehydrogenase (quinone), EC 1.6.99.2) is unusual in that it can utilize either NADH or NADPH as a co-factor for the reduction of its substrates. We have shown that the intact NAD(P)H molecule is not required and that other reduced pyridinium compounds can also act as co-factors for DT diaphorase. The entire adenine dinucleotide portion of NAD(P)H can be dispensed with entirely and the simplest quaternary (and therefore reducible) derivative of nicotinamide, 1-methylnicotinamide, was as effective as NAD(P)H as a co-factor for the reduction of the quinone, menadione. Nicotinamide 5'-O-benzoyl riboside was also as effective a co-factor as NAD(P)H, whilst nicotinamide ribotide and riboside have a higher Km, and decreased the kcat of DT diaphorase. Nicotinic acid derivatives had little activity. Kinetic analysis indicated that both nicotinamide ribotide and riboside may be interacting with the menadione binding site rather than the NAD(P)H site. Irrespective of the differences between the various reduced pyridinium derivatives in their ability to act as co-factors for the reduction of menadione by DT diaphorase, all the compounds that showed activity in this assay were equally effective co-factors for the reduction of the nitrobenzamide, CB 1954 (5-(aziridin-1-yl)-2,4-dinitrobenzamide). The apparent Km of DT diaphorase for all these co-factors approached zero. It was concluded that co-factor binding is not a rate-limiting step in the nitroreductase activity of DT diaphorase.


Lipoprotein(a) at birth, in blacks and whites. It has been shown that blacks have considerably higher concentrations than whites of lipoprotein(a) (Lp(a)), which has been identified as an independent risk factor for coronary heart disease, vein graft restenosis, and cerebrovascular disease. Smaller differences in Lp(a) concentrations have been noted between males and females. To examine whether gender and race differences are already detectable at birth, we examined Lp(a) concentrations in cord blood samples of 109 black (49 male and 60 female) and 123 white (67 male and 56 female) newborns. Maternal age, gestational age, fetal maturity indicators, weight, height and head circumference were analyzed as covariates. For race and sex combined, the mean Lp(a) concentration was 4.0 mg/dl (S.D. of 3.94 mg/dl), approximately 5-fold lower than that observed in adults. No statistically significant differences were found between race or gender groups. The cross-sectional examination of serum Lp(a) concentrations of 221 infants, children and adults showed a gradual increase in Lp(a) concentrations from birth to adult values by the second year of life. We conclude that the system responsible for the production and control of Lp(a) concentration is not yet mature--or has not yet been challenged--at birth.


Lipoprotein(a) in subjects with familial defective apolipoprotein B100. The plasma lipoprotein(a) (Lp(a)) concentration and apolipoprotein(a) (apo(a)) phenotype were determined in the members of two families affected with familial defective apo B100 (FDB), resulting from the Arg3500----Gln mutation in apo B that disrupts binding to LDL receptors. Eleven different phenotypic species of apo A were identified, five of which were present in both families. Although there was a general increase in Lp(a) concentration as the size of the predominant apo(a) component decreased, there was considerable variability and in three clear instances the concentration of an inherited phenotypic species was atypically low. In five cases where a direct comparison could be made, the plasma Lp(a) concentration was significantly higher in heterozygous FDB subjects than in their non-FDB siblings or close relatives with the same phenotype. However, in vitro competition studies using purified Lp(a) that had been reduced with dithiothreitol to remove the apo(a) component, indicated that the Lp(a) from FDB heterozygotes contained a smaller proportion of defective particles than their LDL. Lp(a) particles containing normal and binding-defective apo B were present at approximately the same concentration, suggesting that the increase in Lp(a) concentration observed in FDB subjects could not be explained by the inability of the particles containing the defective apo B100 to be cleared through LDL-receptor mediated processes.


Multivariate genetic analysis of high density lipoprotein particles. The aim of this study was to investigate the effect of genetic factors on three components of plasma high density lipoproteins, HDL-cholesterol (HDL-C), apolipoprotein A-I (apo A-I) and lipoprotein particle Lp A-I (Lp A-I), which contains apo A-I but not apo A-II. These analyses were carried out on 106 nuclear families with one or more children (407 subjects) who volunteered for health screening at the Center for Preventive Medicine, Vandoeuvre, France. After adjustment by stepwise multiple linear regression analysis for age, gender, weight, height, ponderosity, alcohol consumption, smoking habits, and hormonal treatment in females, a multifactorial model (considering the effect of polygenes, individual, specific, environmental and common household factors) was fitted to each variable separately. The hypothesis of no common household effects was accepted for each of the traits. The contribution of genetic factors to inter-individual variance was larger than the contribution of environmental factors for apo A-I (h2 = 0.81) and Lp A-I (h2 = 0.63) but not for HDL-C (h2 = 0.44). Bivariate analyses were carried out by parameterizing covariance components between traits. The genetic correlations were always significantly different from zero. They were estimated to be 0.73 between HDL-C and apo A-I, 0.40 between HDL-C and Lp A-I, 0.51 between apo A-I and Lp A-I. These results suggest that HDL-C, apo A-I and Lp A-I are only in part affected by the same genes and that the measurement of lipids as well as the apo A-I and Lp A-I gives complementary and different information on the metabolic and genetic aspects.


Arterial response to mechanical injury: balloon catheter de-endothelialization. Coronary angioplasty has been used clinically for over a decade. Its initial promise as an alternative to coronary bypass surgery has only partially been fulfilled because of the high rate of post-operative restenosis. A number of animal models have been devised to study this phenomenon and although none is entirely satisfactory, they have, together with recent advances in molecular biology provided an insight into the cellular mechanisms that may contribute to this complication. This knowledge may ultimately lead to a means of therapeutic intervention. This review summarises our present understanding of the pathology of post-angioplasty re-stenosis as revealed by studies using the balloon catheter de-endothelialization model, and discusses some of the intervention strategies that have been attempted.


Age dependency of cystathionine beta-synthase activity in human fibroblasts in homocyst(e)inemia and atherosclerotic vascular disease. In order to clarify whether cystathionine beta-synthase (CBS) could differentiate groups of patients with various vascular diagnosis, CBS was studied in cultured human skin fibroblasts from 99 human subjects diagnosed as homozygotes or heterozygotes for CBS deficiency or suffering from atherosclerotic vascular disease or Down's syndrome (prone to less atherosclerosis). In addition, embryonic human skin fibroblasts and controls were analysed for CBS. We found significant group differences but the overlap in the hetero- and homozygotes for CBS deficiency was too extensive to allow any individual diagnosis based on cell culture studies. CBS activity was significantly lower in the atherosclerotic patients as compared to control subjects. The difference was mostly due to much higher CBS activity in the younger controls. Age dependency was markedly emphasized by very high values from embryonic cells. A strong negative correlation was noted for age and CBS activity in control subjects but not in the atherosclerotic patients. The results are important for the discussion of homocysteine in atherosclerosis and point to the importance of donor age on CBS activity in cultured cells. In addition, diagnosis of hetero-homozygosity for CBS activity is not possible on an individual basis by this method. Further studies in cell culture systems are needed to investigate if young patients (less than 45 years old) with atherosclerotic disease could be identified by low CBS activity in fibroblast cultures as indicated by this study.


Correlation of apolipoprotein(a) isoproteins with Lp(a) density and distribution in fasting plasma. Lp(a) is an LDL-like lipoprotein which contains an additional apolipoprotein called apo(a). Apo(a) exhibits a significant size polymorphism and its size is inversely correlated with plasma Lp(a) levels. We investigated the distribution of different apo(a) isoproteins in lipoprotein density fractions. Fasting plasma samples were subjected to non-equilibrium density gradient ultracentrifugation. After SDS-PAGE and anti-apo(a) immunoblotting, apo(a) concentrations in individual density fractions were evaluated by densitometry. In series I, analysis of selected density fractions from 35 coronary heart disease (CHD) patients demonstrated that although most of the apo(a) was present in the Lp(a) density range, apo(a) was consistently found in both the VLDL and IDL fractions as well. In series II, density fractions from 9 normolipidemic subjects with 6 different apo(a) isoproteins were evaluated. A strong association between the size of the apo(a) isoprotein and the density of the associated Lp(a) particle was established (r = 0.976, P less than 0.001). Lp(a) densities ranged from 1.057 g/ml for the B isoprotein to 1.09 g/ml for the S5 isoprotein. Overall, 75% of the total apo(a) was detected in the Lp(a) density range (d = 1.05-1.12 g/ml), with 9% and 10% in the LDL (d = 1.019-1.05 g/ml) and HDL (d = 1.12-1.21 g/ml) fractions, respectively. VLDL contained an average of 4% of the total apo(a) in fasting normolipidemic plasma. Two hypertriglyceridemic subjects had substantially greater amounts of apo(a) in the fasting triglyceride-rich fraction. The results of this study indicate that the size of the apo(a) isoprotein strongly influences the density of its associated Lp(a) particle and that apo(a) is consistently found in the triglyceride-rich lipoproteins of fasting plasma.


Effects of niceritrol on levels of serum lipids, lipoprotein(a), and fibrinogen in patients with primary hypercholesterolemia. Thirty-three consecutive unselected patients with primary hypercholesterolemia received niceritrol 1.5 g daily for 12 weeks, with the effect of administering divided dose (twice daily (b.i.d.) and three times daily (t.i.d.)) evaluated. The serum concentrations of lipoprotein(a) (Lp(a)), lipids, the major apolipoproteins (apo), cholesteryl ester transfer activity and fibrinogen were determined before and after treatment. The b.i.d. and t.i.d. regimens each significantly reduced the serum levels of total cholesterol and triglyceride. The mean changes in serum lipids and lipoproteins did not differ significantly between the two groups. After 12 weeks of treatment, there was a significant decrease in total plasma cholesterol, triglyceride, low density lipoprotein cholesterol, apo A-II, apo B and fibrinogen and an increase in the high density lipoprotein cholesterol levels. Although the serum level of Lp(a) did not change in every patient, niceritrol significantly reduced the serum Lp(a) level in those with an initially high level of Lp(a) (greater than or equal to 20 mg/dl).


Effects of the gonadotrophin-releasing hormone agonist 'Zoladex' upon pituitary and gonadal function in hypogonadal (hpg) male mice: a comparison with normal male and testicular feminized (tfm) mice. Hypogonadal (hpg) mutant mice, with a congenital deficiency of hypothalamic gonadotrophin-releasing hormone (GnRH), and testicular feminized (tfm) mice, which lack a functional androgen receptor, were used to study the effects of the potent GnRH agonist 'Zoladex' (ICI 118630; D-Ser (Bu(t))6, Azgly10-GnRH) on pituitary and gonadal function. Zoladex (0.5 mg) in a sustained-release lactide-glycolide copolymer depot was administered subcutaneously under anaesthesia and was left in place for 7 days, after which time the effects of the drug upon pituitary and serum gonadotrophin concentrations, glycoprotein hormone subunit mRNAs and testicular morphology were investigated. At the pituitary level, Zoladex treatment resulted in a substantial reduction in LH content in normal males, and LH content was depressed in hpg mice even below the basal levels normally found in these mutants. Pituitary LH content in the Zoladex-treated animals was depressed in the tfm groups, but not to the same levels as those found in the normal and castrated normal mice. Zoladex treatment at the time of castration prevented the post-operative elevation in serum LH associated with castration alone. In the androgen-deficient tfm mouse, Zoladex did not depress the normally elevated serum LH levels. Serum LH in the hpg animals was, in all cases, below the limit of detection of the assay. Pituitary FSH content was depressed into the hpg range in both the normal and castrated animals, but there was no further depression in the hpg mice. The pituitary content was reduced in the tfm mice, again the effects not being as dramatic as in the normal and castrated animals. Serum FSH content, as measured by radioimmunoassay, was depressed by 50% in normal mice; there was no reduction in the hpg mice, however. With regard to pituitary gonadotrophic hormone gene expression, Zoladex administration to normal mice caused a dramatic reduction in LH beta mRNA content, to a level approximating that found in untreated hpg mice. The drug also depressed LH beta mRNA in the castrated group to the hpg range when given at the time of castration, whereas in untreated castrated mice there was a significant increase in LH beta mRNA. In the tfm mouse, which can be considered as a model for long-term failure of androgen feedback, Zoladex again induced a fall in LH beta mRNA, but not to the same extent as in the normal and normal castrated group. Zoladex had no effect on the already low levels of LH beta mRNA found in hpg mice.(ABSTRACT TRUNCATED AT 400 WORDS)


[Laparoscopic cholecystectomy in pediatrics. A report of the first case in the Mexican literature]. The authors present the case of a 9.8-year-old girl with 5 a month history of epigastric pain which was due to gallstones as demonstrated by ultrasound. After an extensive work-up failed to reveal associated hematologic problems, she was successfully treated by laparoscopic cholecystectomy, resulting in complete recovery, hospitalization of 22 hours and return to school three days after major surgery. We could not find references of laparoscopic cholecystectomy in the pediatric age group in the Mexican literature and only 3 in English language publications. In cases of cholelithiasis without associated diseases in children, the operation of choice should be laparoscopic cholecystectomy.


Atrial and digitalis-like natriuretic hormones in essential hypertension under functional loading. To assess the relation of the two natriuretic hormones, atrial natriuretic peptide (ANP) and digitalis-like natriuretic factor (DLF), to hypertension, levels of ANP and DLF were measured under basal conditions and after salt and water loading in 31 normal subjects and 36 and 57 patients with Stage I or II essential hypertension (EH). DLF levels were higher in normal women than men; in EH-II patients, DLF levels were elevated among men but subnormal in women (P less than .02) and rose with water loading in both genders. In all groups ANP levels tended to be higher in women. Water loading increased ANP levels in EH-I patients (P less than .001) and caused less marked increases of ANP in control and EH-II women and men. ANP also tended to increase with salt loading. Both DLF and ANP were related to blood pressure in the subject groups (r = 0.75 to 0.96 and r = 0.27 to 0.75, respectively) and were also related to each other (r = 0.20 to 0.47). The role of ANP and DLF in hypertension are likely to be compensatory and directed against water-electrolyte metabolism disorders associated with elevated arterial pressure.


Magnetic resonance imaging of cardiac hypertrophy in malignant arterial hypertension. The major target organs that become damaged as a consequence of long-standing arterial hypertension are the kidneys, heart, and brain. Left ventricular hypertrophy (LVH) cannot be considered only as an adaptive process to elevated blood pressure (BP), and the heart is also a major target organ in malignant arterial hypertension (MH). Magnetic resonance (MR) was used as a method for visualization of the heart in 68 patients with MH including 18 with essential hypertension, 16 with chronic glomerulonephritis, 13 with chronic pyelonephritis, 16 with renovascular hypertension, eight with adrenal tumors, and in 20 healthy volunteers (as a comparison group). Electrocardiogram-gated, double spin-echo magnetic resonance imaging was performed to image the right and left ventricles (RV and LV), interventricular septum, apex and LV posterior wall, left atrium, and aortic root. In all the patients, symmetric LV hypertrophy was registered and in the most severe cases LV wall thickness was more than 20 mm. There was no LV cavity enlargement or local contractility abnormalities. There was close correlation of LVH and diastolic BP. The degree of LVH and diastolic dimensions of the LV differed between etiologies of MH. These findings show that different pathophysiologic mechanisms of development of MH influence the processes of myocardial hypertrophy. The highly informative yield of MR tomography for evaluating structural and functional changes of the heart under MH must be underlined.


Effects of sertraline and citalopram given repeatedly on the responsiveness of 5-HT receptor subpopulations. The effect of repeated treatment (5 and 10 mg/kg, po, twice daily, 14 days) with sertraline and citalopram (antidepressants which selectively inhibit the reuptake of 5-hydroxytryptamine (5-HT)) on the responsiveness of different 5-HT receptors to their agonists, was examined in rats and mice. Sertraline and citalopram (both at a dose 5 and 10 mg/kg) antagonized (the first one more potently) the hypothermia induced in mice by 8-OH-DPAT (a 5-HT1A agonist), but not the behavioural syndrome induced in rats by this substance. The m-chlorophenylpiperazine-induced hypothermia in mice (a 5-HT1B effect) was increased by sertraline and citalopram (only in a dose of 10 mg/kg). Both antidepressants, given repeatedly (as well acutely) attenuated exploratory hypoactivity induced in rats by m-chlorophenylpiperazine (a 5-HT1C effect). L-5-HTP-induced head twitches in mice (5-HT2 effect) were antagonized dose-dependently by both repeated sertraline and citalopram. Both antidepressants (citalopram only in higher dose) reduced the fenfluramine-induced hyperthermia in rats (5-HT2 effect). The results indicate that sertraline and citalopram given repeatedly decrease the responsiveness of 5-HT1A (presynaptic) and 5-HT2 receptors but increase the responsiveness of 5-HT1B receptors to respective agonists.


Complete rescue of photoreceptor dysplasia and degeneration in transgenic retinal degeneration slow (rds) mice. retinal degeneration slow (rds) is a semidominant mutation of mice with the phenotype of abnormal development of rod and cone photoreceptors, followed by their slow degeneration. The rds gene has been putatively cloned and its novel protein product initially characterized biochemically. In the present study we undertook to correct in vivo the retinal phenotype of mice with the rds mutation. We assembled a transgene containing a regulatory segment of the opsin gene positioned upstream of the wild-type rds coding region. Mice from three transgenic lines, homozygous for the rds mutation, were analyzed for expression of the transgene and for their retinal phenotypes. In two high expressing lines, we observed complete reversion to wild-type retinal morphology. In a third, low expressing line, we observed a retinal phenotype intermediate between wild type and rds/rds, suggesting partial rescue of the mutation. These results constitute formal proof that we have cloned the rds gene.


[Significance of plasma atrial natriuretic peptide measurement during the management of hyponatremia in neurosurgical patients]. We have examined the total number of admitted cases to clarify the pathogenesis of hyponatremia during the management of neurosurgical patients. We experienced 32 cases of hyponatremia during the past year by measuring the sodium balance and atrial natriuretic peptide (ANP) level. According to these two factors, we divided the cases into three groups. The first group shows normal ANP levels in spite of hyponatremia. Low administration of the sodium was thought to be the cause in these cases. The second group shows the elevated ANP levels with a positive sodium balance. Elevated circulatory volume due to the inadequate level of antidiuretic hormone and mild heart and/or kidney failures cause these conditions. Water restriction and/or diuresis were effective methods in the management of the cases. The last group shows the elevated ANP levels with a negative sodium balance. There is a statistically significant negative correlation between sodium balance and the ANP level. Marked natriuresis due to the elevated ANP causes the decrement of the circulatory volume in these cases. Pathogenesis of the last group is very important in the management of neurosurgical patients in an acute state, especially in subarachnoid hemorrhage cases. The decrement of the systemic circulatory volume would jeopardize the patient's neurological condition. In this group, water restriction that has been commonly recommended is contraindicated. Satisfactory water and sodium replenishment seems to be the best recommended treatment for this group.


Inhibition of inhaled metabisulphite-induced bronchoconstriction by inhaled frusemide and ipratropium bromide. 1. The effect of inhaled frusemide and high dose inhaled ipratropium bromide on bronchoconstriction induced by inhaled metabisulphite was studied in 10 atopic volunteers. 2. Frusemide (40 mg), ipratropium bromide (0.5 mg) or saline placebo were administered by nebuliser in a double-blind fashion, prior to construction of a dose-response curve to metabisulphite (2.5-100 mg ml-1). 3. Geometric mean of the provocative dose of metabisulphite that caused a 35% fall in specific airways resistance (sGaw) after placebo was 13 (95% confidence intervals CI 4-36 mumol) compared with 36 (16-78) mumol after ipratropium bromide and 45 (22-94) mumol after frusemide. 4. Mean maximum fall in sGaw was 49 (40-57)% after placebo, 11 (0-22)% after frusemide and -1 (-25-22)% after ipratropium bromide. 5. Frusemide significantly protected against metabisulphite induced bronchoconstriction (P less than 0.005). The protection from high dose ipratropium bromide was also significant (P less than 0.05), but the response was more variable between subjects.


Agreement and reproducibility of the estimates of cardiovascular function by impedance cardiography and M-mode echocardiography in healthy subjects. The reproducibility and agreement of the estimates of stroke volume (SV), cardiac output (CO) and total peripheral resistance (TPR) by transthoracic impedance cardiography (ZCG) and M-mode echocardiography (ECHO) were analyzed before and after the placebo-controlled administration of ascending doses of isosorbide dinitrate and nicorandil in 12 healthy subjects. There was no biostatistical agreement between the two methods in estimating cardiovascular function either before or after dosing (ZCG estimated substantially larger SV, CO and lower TPR). But, ZCG and ECHO estimated about similar overall treatment related changes (across treatments and periods) and reached substantially better agreement when the values were expressed as ratio of the baseline before dosing. Such improvement did not occur when the data were expressed as arithmetic difference from baseline. In spite of the improvement of agreement by expressing the data as ratio of baseline, the coefficients of reproducibility between the two methods (circa 25% of baseline) remained too large to judge the methods interchangeable.


A meta-analysis of randomized trials of fish oil in prevention of restenosis following coronary angioplasty. The efficacy of fish oil in decreasing restenosis following percutaneous transluminal coronary angioplasty (PTCA) remains controversial despite seven published reports of randomized trials involving 951 patients. We performed a meta-analysis to determine whether these trials, viewed in aggregate, demonstrate a significant benefit. We evaluated rates of restenosis two to 12 months after PTCA and calculated an estimate of the overall effect and 95% confidence interval (CI). The typical odds ratio (treatment versus control) was 0.71 (95% CI 0.54, 0.94), P = 0.016 (two-tailed). The data show a strong and highly significant (P less than .0001) relationship between daily fish oil dose and gastrointestinal side effects. While compatible with a small to moderate benefit of fish oil on rates of restenosis, these results require confirmation in a randomized clinical trial large enough to distinguish reliably between a clinically meaningful benefit and a null result.


Annexin I-mediated vesicular aggregation: mechanism and role in human neutrophils. Whole cytosol isolated from human neutrophils was found to accelerate the Ca(2+)-dependent fusion of phospholipid vesicles with neutrophil plasma membranes as measured by several fluorescence resonance energy transfer lipid dilution assays or by the fate of an encapsulated aqueous soluble fluorophore. The Ca2+ (threshold of 2-10 microM) and protein concentration dependencies for fusion mediated by purified human neutrophil annexin I (lipocortin I), recombinant annexin I and des(1-9)annexin I showed behavior similar to that of whole cytosol. A monoclonal antibody against the N-terminal region of annexin I strongly inhibited the action of isolated annexins as well as whole cytosol, indicating that annexin I is the major activity of this type in whole neutrophil cytosol and that it functions even in this complex mixture of proteins. Residual Ca(2+)-dependent fusion activity in the absence of cytosol or annexin I was not inhibited by several antibodies against annexin I, implicating an as yet unknown protein. Kinetic analysis of liposomal fusion showed that annexin I, as in the case of synexin, accelerates aggregation of vesicles but not the actual fusion event per se. The disposition of annexin I in liposomal aggregates was studied by monitoring binding of the protein with a pyrene-phospholipid and by simultaneously monitoring vesicular aggregation by turbidity. An antibody to the N-terminus of annexin I inhibited vesicular aggregation but not binding, suggesting that initial binding of annexin I is similar to that of annexin V. A relatively small proportion of the bound annexin was involved in intervesicular linkage, and no exchange of bound annexin to subsequently added vesicles was observed. The lack of extensive contact between lipids of aggregated vesicles was supported by a lack of energy transfer between phospholipid probes on separate aggregating vesicles. Covalent linkage of maleimidyl or photoaffinity phospholipid derivatives with annexin I in vesicular aggregates did not allow complete disaggregation of vesicles by EDTA, suggesting that monomers of annexin I can contact two membranes simultaneously at the point of intervesicular linkage. These data are discussed in terms of possible models for the structure of this site.


Zero-length crosslinking between subunits delta and I of the H(+)-translocating ATPase of chloroplasts. Treatment of spinach thylakoids with 1-ethyl-3-(dimethylaminopropyl)-carbodiimide (EDC)/N-hydroxysulfosuccinimide (sulfo-NHS) induced formation of a zero-length crosslink of an apparent molecular mass of 38 kDa. This product was shown, by immunodetection, to consist of subunit delta of CF1 and subunit I of CF0. The crosslink was isolated by preparative SDS gel electrophoresis and subjected to cyanogen bromide cleavage. Electrophoretic and immunological analysis of the resulting peptides suggested that the crosslink was formed between a glutamyl or aspartyl residue at the C-terminal end of subunit I and a basic amino acid of subunit delta in the range between Val-1 to Met-165. Treatment of thylakoids with EDC/Sulfo-NHS resulted in inhibition of photophosphorylation and CF0CF1-catalyzed ATP hydrolysis without affecting formation of a proton gradient related to phenazine methosulfate-mediated cyclic electron transport. Inhibition of H+ transport-coupled ATP hydrolysis was more pronounced than non-coupled methanol-stimulated ATP hydrolysis. The results suggest that subunits delta and I form a connection between the partial complexes CF1 and CF0 in situ. Crosslinking of the two subunits may impede the translocation of protons through CF0CF1.


ATP synthase: structure-function relationships. Recent work has focused on obtaining a better understanding of the three-dimensional structural relationships between the alpha and beta subunits of the F1 moiety and the location of nucleotide binding domains within these subunits. Four types of approach are currently being pursued: X-ray crystallographic, chemical, molecular biological and biochemical. Here we briefly review some of the major conclusions of these studies, and point out some of the problems that must be resolved before an adequate model that relates structure to function in the ATP synthase molecule can be formulated.


F-type or V-type? The chimeric nature of the archaebacterial ATP synthase. Archaebacterial plasma membranes contain an ATPase acting in vivo as a delta mu H(+)-driven ATP synthase. While functional features and their general structural design are resembling F-type ATPases, primary sequences of the two large polypeptides from the catalytic part are closely related to V-type ATPases from eucaryotic vacuolar membranes. The chimeric nature of archaebacterial ATPase from Sulfolobus was investigated in terms of nucleotide interactions and related to specific sequence parameters in a comparison to well known F- and V-type ATPases. The study disclosed a general difference of F- and V-type ATPases at one class of the nucleotide binding sites.


Humoral immunity in Down's syndrome.

Humoral immunity in Down's syndrome. Carried out on 57 outpatients, the study was aimed to prove the existence of a disgammaglobulinemia in Down's syndrome. The event could not be attributed to hospitalization. Data indicate increased A and G immunoglobulinemias. In agreement with other authors Ig A level was found to be very high especially after the age of 5. Ig M titre in patients was reduced in comparison with that of the controls, regardless of the age group. Increase timing of IgA and Ig G concentrations tends to be more marked for the Ig A fraction. In patients ranging from 0 to 5 years the Ig A serum level is deficient.


The ATPases of Propionigenium modestum and Bacillus alcalophilus. Strategies for ATP synthesis under low energy conditions. In Propionigenium modestum, ATP synthesis is coupled via delta mu Na+ to the decarboxylation of (S)-methylmalonyl-CoA. The low energy yield of this reaction implies that approx. 4 decarboxylation cycles are necessary to synthesize 1 molecule of ATP. Theoretical considerations in accord with experimental results suggest ATP synthesis in P. modestum at delta mu Na+ = -110 mV. Other anaerobic bacteria synthesize ATP at a delta mu H+ of similar size and alkaliphilic bacteria at pH 10.3 have a delta mu H+ of only -103 mV. In these cases, the H+(Na+) to ATP stoichiometry must be at least 4.


Chemical modification of cyclomaltodextrin glucanotransferase from Bacillus circulans var. alkalophilus. Counting of integral numbers of cysteine residues of the reduced and denaturated form of cyclomaltodextrin glucanotransferase (CGTase) from Bacillus circulans var. alkalophilus (ATCC 21783) showed two cysteine residues per enzyme molecule. Titrations of the enzyme with 5,5'-dithiobis-(2-nitrobenzoic acid) led to the same result. No free SH-group was detected in denatured form of CGTase, indicating that the two cysteine residues are linked by one disulfide bridge. Cyclizing activity of the GdmCl-denaturated and reduced enzyme was 13% of that of the native one. Incubation of CGTase with diethylpyrocarbonate (DEP) showed a pseudo-first-order inhibition with second-order rate constant of 3.2 M-1 s-1. Reaction with hydroxylamine and spectroscopic studies implied that inactivation of CGTase by DEP is due to modification of one histidine residue concomitantly with a 50% decrease in the cyclizing activity (t1/2 = 10.8 min). The inhibition was partially reversible. CGTase was protected against inactivation by alpha- and beta-cyclodextrins suggesting that the modified histidine residue is at or near the active site. Conversion of starch with DEP-modified enzyme resulted in a decreased formation of cyclodextrins while the relative amount of reducing sugars increased. Preliminary results on modification of CGTase with other reagents, e.g., Woodward's reagent K, 2,3-butanedione and carbodiimide are included.


Surface-immobilized polyethylene oxide for bacterial repellence. Polyethylene terephthalate films were surface-modified with polyethylene oxide (18,500 g/mol) using a solution technique described previously. These films were investigated for their resistance to bacterial adhesion. Three bacterial strains most commonly associated with implant infections, Staphylococcus epidermidis, Staphylococcus aureus and Pseudomonas aeruginosa, were cultured in tryptic soya broth, human plasma and human serum on the polymeric substrates. Significant reductions (between 70 and 95%) in adherent bacteria were observed on the polyethylene oxide-modified substrates compared to the untreated control polyethylene terephthalate. Surface modification with polyethylene oxide may reduce the risk of implant-associated infections. Plasma fibrinogen was observed to play an important role in the adhesion of all three of these species on both the polyethylene oxide-modified and control polyethylene terephthalate materials.


Tissue response to intraperitoneal implants of polyethylene oxide-modified polyethylene terephthalate. Polyethylene terephthalate films surface modified with polyethylene oxide of mol wt 18,500 g/mol (18.5 k) by a previously described technique, were implanted in the peritoneal cavity of mice, along with their respective untreated controls, for periods of 1-28 d. The implants were retrieved and examined for tissue reactivity and cellular adherence. The control polyethylene terephthalate surfaces showed an initial inflammatory reaction followed by an extensive fibrotic response with a mean thickness of 60 microns at 28 d. By contrast, polyethylene oxide-modified polyethylene terephthalate showed only a mild inflammatory response and no fibrotic encapsulation throughout the implantation period: at 28 d a cellular monolayer was observed. Apparently either the polyethylene oxide-modified surface was stimulating less inflammation, which was in turn stimulating less fibroblastic overgrowth, or the cellular adhesion to the polyethylene oxide-modified surface was too weak to support cellular multilayers.


Acute upregulation of interleukin-1 receptor by ligand. In this study we have investigated the effect that interleukin 1 (IL-1) has on cell surface IL-1 receptor expression in the murine thymoma cell line, EL4 6.1. These cells express IL-1 receptors with both high affinity (Kd = 65 pM, 986 receptors/cell) and low affinity (Kd = 14.5 nM, 10,417 receptors/cell). The high- and low-affinity receptors are indistinguishable by crosslinking studies performed at both high and low ligand concentrations. However, the two affinity states could be functionally distinguished on the basis of their internalization of ligand. Receptor-mediated endocytosis was dependent upon the concentration of ligand bound to the cells. In the presence of low IL-1 concentrations receptor-mediated endocytosis was slow, whereas at high IL-1 concentrations, endocytosis was more rapid. Furthermore, receptor-mediated endocytosis of IL-1 did not result in downregulation of surface IL-1 receptors. Indeed, both kinetic and equilibrium binding studies revealed that pre-incubation of cells with IL-1 alpha resulted in an acute upregulation of 125IL-1 alpha binding to high affinity surface receptors in a time and energy dependent manner. Examination of the association kinetics suggested that increased binding was not attributable to positive co-operativity of the high affinity IL-1 receptor, but was due to increasing IL-1 receptor number. This observation was confirmed by equilibrium binding studies. Moreover, receptor numbers were not enhanced by de novo synthesis, nor release of receptors from an intracellular pool. The observed increases in surface ligand binding were most probably due to conversion of the surface pool of low affinity receptors into high affinity receptors.


Effects of human anti-IL-1 alpha autoantibodies on receptor binding and biological activities of IL-1. Immunoglobulin G (IgG) antibodies to interleukin 1 alpha (IL-1 alpha) are frequently found in the sera of healthy human individuals. The effects of these autoantibodies on receptor binding and biological activities of human IL-1 were tested. Using the murine T-lymphocyte line NOB-1, human thyrocytes and human foreskin fibroblasts, the antibodies competitively inhibited the biological activity of human recombinant IL-1 alpha (rIL-1 alpha). The degree of inhibition correlated with 125I-rIL-1 alpha binding to IgG in different immunoglobulin preparations and in individual sera. These antibodies also neutralized the IL-1 activity of isolated membrane fragments and lysates of human blood monocytes activated by lipopolysaccharide. In contrast, the supernatant IL-1 activity was not affected. Stronger inhibition of biological activity and cell binding of 125I-rIL-1 alpha was obtained with NOB-1 cells than with human thyrocytes. The antibodies failed to interfere with the biological activity of rIL-1 beta. It is concluded that IgG autoantibodies of IL-1 alpha in the sera of healthy humans selectively inhibit the biological activity of the soluble and membrane-associated forms of IL-1 alpha in vitro, and that the degree of biological inhibition afforded by these antibodies depends upon the target cell.


Cloning and chromosome mapping of the human interleukin-1 receptor antagonist gene. By screening a human genomic library with an interleukin-1 receptor antagonist (IL-1ra) cDNA probe, we have isolated a 15 kb clone which contains the entire coding region of the gene as expressed in monocytes, and includes 6 kb of 5'-upstream sequence. The gene contains four exons which code for the secreted form of the IL-1ra, however, our clone does not contain the alternative first exon used to generate an intracellular form of the protein as the protein as found in epithelial cells. Analysis of the sequence reveals a consensus TATA box, and three Alu repeats, two of which are in the upstream region and one in intron 3. The sequence also reveals an 86 bp motif tandomly repeated four times within intron 2, and may reflect the polymorphism known to exist in this region of the gene. By in-situ fluorescence hybridization we have shown that the IL-1ra gene is found on the long arm of chromosome 2 and maps to 2q13-14.1. Previous studies have revealed that IL-1 alpha, and IL-1 beta and both type I and type II forms of the IL-1 receptor all map close to this region of chromosome 2.


Molecular characterization of the interleukin-1 receptor (IL-1R) on monocytes and polymorphonuclear cells. Primary human monocytes and monocytic cells express an interleukin 1 receptor (IL-1R) which is similar in molecular weight and IL-1 binding characteristics to the IL-1R expressed on B lymphocytes (type II). Northern blot analysis of monocytic cells using a cDNA probe from the recently isolated type II IL-1R indicates that this mRNA is detectable by 4 h and accumulates for at least 24 h following treatment with IL-1R inducing drugs. The time course of induction of this mRNA is slower than that of the type I IL-1R mRNA which is also transcribed in monocytic cells but does not appear to be translated. Sequence analysis of a monocyte-derived cDNA corresponding to the type II IL-1R mRNA shows that the monocyte and B-cell mRNAs are identical. Comparison of monocyte IL-1R peptide maps with those of the type II IL-1R suggests that the two surface IL-1R are identical. This was confirmed serologically using a polyclonal antiserum raised against the type II IL-1R. Data are presented which indicate that primary human neutrophils can also be induced to express abundant type II IL-1R.


Bacteriophage cloning and Escherichia coli expression of a human IgM Fab. We have combined the molecular biology methods of the polymerase chain reaction and recombinant DNA cloning in bacteriophage lambda to express a human IgM Fab in Escherichia coli using genes derived from an Epstein-Barr virus transformed cell line. This method comprises three cDNA amplifications and a single cloning step, culminating in the stable overexpression of mammalian heterodimeric recombinant protein in a prokaryotic host.


Both CD8+ and CD16+ human T cell clones can provide B cell help for immunoglobulin production. The functional properties of two CD4+CD8-CD16-, five CD4-CD8+CD16- and three CD4-CD8-CD16+ human T cell clones were compared. All CD4- T cell clones displayed strong cytolytic activity in the lectin-dependent lytic assay against the P815 murine mastocytoma cell line, but only the CD4-CD8-CD16+ T cell clones exhibited lytic activity against the natural killer-sensitive K562 cell line. Upon activation with anti-CD3 monoclonal antibody, all T cell clones were able to support IgM and IgA synthesis in autologous B cells. Both CD4+ and CD4- T cell clones required cell-to-cell interaction with the B cells in order to exert their helper activity for immunoglobulin production. However, unlike CD4+, CD4-CD8+CD16- and CD4-CD8-CD16+ T cell clones provided helper function for immunoglobulin synthesis only when low T/B cell ratios were used in culture. At higher T/B cell ratios, there was a decline in the B cell helper activity of CD4- T cell clones that was probably related to the expression of cytolytic capacity against the antigen-presenting B cell. These data support the notion that under certain experimental conditions even cytotoxic T lymphocytes and natural killer cells may provide B cell helper function.


DNA sequence characterization of the G gene region of bacteriophage Mu. The nucleotide sequence of a 1.2 kb region of bacteriophage Mu DNA was determined. This region contains the 3' end of the F gene, the complete G gene, and the 5' end of the I gene, all late genes involved in Mu virion morphogenesis. Identity of the G gene open reading frame was confirmed by sequencing four Gam mutations. The G open reading frame is predicted to encode proteins of 16.7 or 17.2 kDa, depending on which of two possible start codons are used to initiate translation. Four new nuB mutations in the DNA gyrase-binding site between the G and I genes were also sequenced and found to be identical to the nuB103 mutation sequenced previously.


High energy collision-induced dissociation of alkali-metal ion adducts of crown ethers and acyclic analogs. High energy collision-induced dissociation (CID) techniques were applied for structural elucidation of alkali-metal ion adducts of crown ethers. The CID of alkali-metal adducts of tetraglyme and hexaethylene glycol were also evaluated to contrast the fragmentation pathways of the cyclic ethers with those of acyclic analogs. A common fragmentation channel for alkali-metal ion adducts of all the ethers, which results in distonic radical cations, is the homolytic cleavage of carbon-carbon bonds. Additionally, dissociation by carbon-oxygen bond cleavages occurs, and these processes are analogous to the fragmentation pathways observed for simple protonated ethers. The proposed fragmentation pathways for alkali-metal ion adducts of crown ethers result mostly in odd-electron, acyclic product ions. Dissociation of the alkali-metal ion adducts of the acyclic ethers is dominated by losses of various neutral species after an initial hydride or proton transfer. The CID processes for all ethers are independent of the alkali-metal ion sizes; however, the extent of dissociation of the complexes to bare alkali-metal ions increases with the size of the metal.


Leukemic phase of intermediate non-Hodgkin's lymphoma with cells showing different matured stages in invaded various organs. A 56-year-old male was admitted to our hospital with lymphocytosis (16.4 x 10(9)/l; 79% lymphocytes including 50% small lymphocytes), generalized lymphadenopathy, massive splenomegaly, and heavily infiltrated bone marrow. Immunophenotype analysis of the neoplastic cells in the bone marrow revealed that they were B cells (CD20 + CD19 + Ia1 + sIgM+) positive for CD10. By contrast, the cells in the lymph node were CD20 + CD19 + Ia1 + sIgM+ but negative for CD10. The patient was tentatively diagnosed as having lymphosarcoma cell lymphoma, however, the final diagnosis was leukemic phase of intermediate lymphocytic lymphoma. We concluded that CD10+ neoplastic cells in the bone marrow and peripheral blood had differentiated to CD10- cells.


Relationship of MMPI cluster type, pain coping strategy, and treatment outcome. The purposes of the present study of chronic pain patients were to (a) assess whether cognitive and behavioral coping style is related to personality factors, (b) assess how coping styles differ across personality types, and (c) assess how outpatient interdisciplinary intervention affects the coping styles of various personality types. Four MMPI clusters (Depression/Pathological, V-type, Marginal Depression, and Marginal V-type) were derived using a hierarchical clustering procedure. Seventy subjects also completed the Coping Strategies Questionnaire before and after a 3-week outpatient pain management program. Pretreatment analyses indicated the Depression/Pathological and Marginal Depression groups used diverting attention less than either V-type group. The V-type group reported using praying/hoping significantly more than either of the marginal groups. At posttreatment the Depression/Pathological group used catastrophizing significantly more than either of the marginal groups. Results of pre-post analyses indicated that the Depression/Pathological group increased their use of diverting attention, reinterpreting pain sensations, and ignoring pain sensations, while decreasing catastrophizing. The V-type group increased their use of reinterpreting pain sensations, while decreasing praying/hoping and catastrophizing. Neither of the Marginal subtypes showed significant pre-post changes in coping strategies. These results suggest that different personality types use different pain coping strategies prior to multidisciplinary treatment. Groups showing more severe psychological distress, perhaps related to an underlying personality disorder, displayed greater changes in coping strategies with treatment, but remained more dysfunctional after treatment. These findings suggest that the alteration of coping strategies may be an important treatment effect needing more individualization to maximize treatment response.(ABSTRACT TRUNCATED AT 250 WORDS)


Plasma beta-endorphin is not affected by treatment with imipramine or paroxetine in patients with diabetic neuropathy symptoms. To determine the possible role of endogenous opioid peptides in the action of imipramine and paroxetine in painful diabetic neuropathy, beta-endorphin concentrations in plasma were measured in 20 patients during a double-blind, placebo-controlled randomized three-way crossover trial. Despite a significant reduction in neuropathy symptoms during both imipramine and paroxetine treatment, the beta-endorphin level was unaltered throughout the study. The plasma concentration of beta-endorphin was not related to plasma drug concentrations. Thus, this study does not provide evidence of a role of endogenous opioid peptides in the mechanism of action of imipramine and paroxetine in painful diabetic neuropathy.


Early detection measures and triage procedures for suicide ideation in chronic pain patients. There is a dearth of writings about early detection of potential suicide patients in chronic pain centers. Early detection measures used at the Vanderbilt Pain Control Center include a Symptom Checklist-90, with questions about depressive symptomatology and "Thoughts of Ending Your Life"; medical and psychological interviews; monitoring of changes in emotional disturbance; and, if warranted, administration of the Scale of Suicidal Ideation. Three case studies are presented that indicate that the results of an assessment measure should be tempered with clinical judgment. Suicidal behavior, including suicidal ideation, is a medical emergency; therefore, there is great need for early detection and triage measures.


Cell cycle control in normal and neoplastic cells. Recent studies of cell cycle control suggest that cyclin-dependent protein kinases play a central role in the cell's commitment to a new division cycle in late G1. The regulation of these kinases in normal and neoplastic growth is becoming clear.


Ephedrine and amphetamine induced pecking in chickens: possible indirect D-1/D-2 dopaminergic mechanism. The involvement of dopaminergic mechanism in pecking behaviour has been suggested. In the present work, the involvement of D-1/D-2 dopamine receptors in pecking induced by ephedrine and amphetamine in chickens has been studied. Both ephedrine and amphetamine induced pecking in a dose-dependent manner. Pretreatment of animals with D-1 antagonist SCH 23390, D-2 antagonist sulpiride or reserpine decreased the pecking response induced by both drugs. Phenoxybenzamine or propranolol (alpha- or beta-adrenergic blockers, respectively) and the antimuscarinic drug atropine did not decrease the response of these drugs. The results may indicate that the pecking induced by these drugs in chickens are mediated indirectly through D-1/D-2 dopaminergic mechanisms.