Automated 'pain drawing' analysis by computer-controlled, patient-interactive neurological stimulation system.

We have developed a new method for the collection and analysis of pain drawings, as part of a computer-controlled, patient-interactive system for use with implanted neurological stimulators. The system has been tested in 44 patients with permanently implanted spinal cord stimulators for the relief of chronic, intractable pain. Patients interact directly with the system, using a graphics tablet, to enter pain drawings and corresponding outlines of their perceptions of stimulation paresthesias, for different stimulating pulse parameters and electrode geometries. Image analysis software has been developed to examine these data quantitatively. This precludes the inter-rater inconsistencies reported for manual pain drawing scoring techniques. A highly significant correlation has been observed between patients' analog ratings of the overlap of pain by paresthesias and the results of our automated analysis of graphic data. This in turn has been found to correlate with clinical observations of pain relief. The contemporary implantable stimulation devices supported by our system permit non-invasive selection of stimulating anodes and cathodes from a linear array of 4 electrodes. The 50 possible electrode combinations have certain geometric features, which we have entered into a multivariate statistical analysis, to determine their relationship with the overlap of pain by paresthesias. One particular configuration (cathode(s) flanked by anode(s) above and below) is significantly better, by this measure, than all the alternatives. This is consistent with prior clinical observations that this configuration is favored by patients whose systems have been adjusted by conventional, manual methods. Pain drawing' entry and analysis by a computerized, patient-interactive system has been useful in this specialized setting and may have broader applications.


The use of methylphenidate in patients with incident cancer pain receiving regular opiates. A preliminary report.

In this open, uncontrolled trial, 15 patients with severe incident cancer pain receiving regular opiates were administered 10 mg oral methylphenidate (MP) at 08.00 h and 15 mg at 12.00 h in order to antagonize opiate-induced sedation. The daily dose of opiate was increased by 30% 24 h after starting MP, followed by a 10% increase twice a day until maximal tolerated dose. In 14 evaluable patients, pain (VAS 0-100 mm), sedation (VAS), and mean equivalent daily dose (MEDD) of morphine were 55 +/- 17, 65 +/- 18 and 248 +/- 150 48 h before MP, versus 38 +/- 12 (P less than 0.01), 42 +/- 12 (P less than 0.01), and 405 +/- 130 (P less than 0.01) 48 h after MP, respectively. After 48 h of treatment, 12 of 14 patients felt better on MP, 2 of 12 patients felt no difference, and no patients felt worse (P less than 0.05). We conclude that the addition of MP allowed for an increase in the MEDD of morphine and increased pain control. Controlled double-blind trials should be performed.


Right atrial diverticulum presenting as Wolff-Parkinson-White syndrome.

A 7-year-old male presenting with Wolff-Parkinson-White syndrome and tachycardia was suspected by echocardiographic and magnetic resonance imaging evaluation to have an associated pericardial cyst anterior to the right atrium and ventricle. Electrophysiological evaluation demonstrated short antegrade and retrograde accessory connection refractory periods, with inducible orthodromic atrioventricular reentrant tachycardia. Surgical observation revealed a rare congenital right atrial diverticulum bridging the anterior right atrioventricular groove, with the functional accessory connection lateralized to the medial aspect of this structure. Endocardial and epicardial incisions and cryolesions placed along the anterior right atrioventricular groove initially appeared successful, but preexcitation recurred within 4 weeks postoperatively.


Coronary air embolism complicating transseptal radiofrequency ablation of left free-wall accessory pathways.

Radiofrequency catheter ablation is an important new technique for curing patients with accessory pathway-mediated tachycardia. Ablation of left free-wall accessory pathways may be accomplished either by a retrograde, transarterial approach or via a transseptal approach using a long vascular sheath. We describe air embolization into the coronary artery as a complication of the transseptal approach, which was temporally associated with catheter exchange. While there were no permanent adverse sequelae, this report emphasizes the need for scrupulous attention to the possible insinuation of air during procedures involving long vascular sheaths across the atrial septum. To prevent air embolism, we recommend slow removal of the ablation catheter along with continuous flushing with heparinized saline during exchanges.


Does dual chamber or atrial pacing prevent atrial fibrillation? The need for a randomized controlled trial.

Partially due to recent reports that cardiac antiarrhythmic therapy may have adverse effects on patient survival, clinicians have become more interested in the nonpharmacological prevention of atrial fibrillation. There is a large body of literature that suggests that the rate of development of atrial fibrillation in paced sick sinus syndrome patients is much lower in those patients who have received an atrial-based system, rather than a VVI system. However, all the published studies to date are retrospective, and fraught with potential bias favoring the AAI or DDD group. The authors strongly believe that the only way to determine if these suggestive but uncertain retrospective analyses are correct is to apply the same scientific rigor to this problem as has been applied to many other problems in cardiovascular medicine and perform a prospective randomized trial. A proposed trial design is discussed.


Significance of ventricular pacing site in manifest entrainment during orthodromic atrioventricular reentrant tachycardia with left-sided accessory pathway.

We examined entrainment by ventricular pacing in six patients during orthodromic atrioventricular reentrant tachycardia (AVRT) utilizing a left-sided lateral accessory pathway. Constant fusion and progressive fusion were demonstrated in all patients by left ventricular pacing during tachycardia, but in none of the patients by right ventricular pacing. When left ventricular pacing was performed during AVRT, the antidromic wave front from the pacing impulse (n) collided with the orthodromic wave front of the previous pacing beat (n - 1) within the ventricle, therefore, constant fusion and progressive fusion were demonstrated in the surface electrocardiographic QRS complexes. On the other hand, when right ventricular pacing was performed during orthodromic AVRT, the antidromic wave front from the pacing impulse (n) collided with the orthodromic wave front of the previous paced beat (n - 1) within the normal atrioventricular pathway, and constant fusion and progressive fusion were therefore not demonstrated. These phenomena were explained by the relationship of the ventricular pacing site and the reentrant circuit. This study demonstrates the importance of the pacing site in manifest entrainment of orthodromic AVRT during ventricular pacing.


Radiofrequency ablation of accessory pathways: characteristics of transiently and permanently effective pulses.

The purpose of this study was to characterize and compare the radiofrequency current applications that produced permanent or transient accessory pathway conduction block. One hundred fifty-two radiofrequency energy applications that induced permanent (permanently effective pulses, n = 48) or transient (transiently effective pulses, n = 104) accessory pathway block in 57 patients with 60 accessory pathways were analyzed. The time from the onset of current application to disappearance of preexcitation or termination of supraventricular tachycardia by permanently effective pulses was 1-15 seconds (mean 3.6 +/- 3.8 sec) compared to 2-29 seconds (mean 11.5 +/- 7.5 sec) by transiently effective pulses (P less than 0.01). After transiently effective pulses that induced block in accessory pathway, conduction resumed within 5 minutes while induced block by permanently effective pulses persisted in 44 of 48 patients (92%) during follow-up of 11 +/- 12 months. The accessory pathway conduction returned in the remaining four patients after ablation 2 weeks to 7 months. After transiently effective pulses, 41 impulses were delivered to the same site using a higher power output (n = 32) and/or longer energy delivery duration (n = 20) without new mapping of accessory pathway location. Thirty-six of these impulses again resulted in transient accessory pathway block, four had no effect, only one impulse induced a permanent block in the accessory pathway. Pulses with higher power outputs tended to induce transient effects more frequently than pulses with lower energy.(ABSTRACT TRUNCATED AT 250 WORDS)


Driving restrictions advised by midwestern cardiologists implanting cardioverter defibrillators: present practices, criteria utilized, and compatibility with existing state laws.

Although some patients remain at risk of losing physical control or collapsing after implantation of a cardioverter defibrillator for sustained ventricular arrhythmias, little is known about restrictions advised by arrhythmia specialists to patients with implanted devices concerning physical activities such as driving. In this study, all of the 58 cardiologists implanting cardioverter defibrillators in three contiguous midwestern states were surveyed to determine present practices and the compatibility of these practices with existing state law. Of the 51 respondents (88%), 27 cardiologists (53%) advised only those implanted patients who had had arrhythmia-induced presyncope or physical collapse to cease driving. Twenty two of the remaining cardiologists (43%) advised all implanted patients to cease driving, whereas two cardiologists (4%) never advised any implanted patient to restrict driving. Permanent driving abstinence was advised by seven of the responding cardiologists (14%), while temporary driving abstinence for periods of 2-12 months (mean 6 +/- 3 months) was recommended by the remaining 42 respondents (82%) who advised against driving. The criteria utilized, driving restrictions advised, and durations advised for driving restrictions were not uniform in any of the 13 surveyed university and nonaffiliated cardiology practices with greater than or equal to 2 implanting cardiologists. Overall, 38 cardiologists (74%) advised against driving and recommended durations that equaled or exceed their state's minimum legal requirements, although only 27 of the 51 cardiologists (53%) based their practice upon knowledge of their state's driving laws. The results of this survey suggest that the majority of cardiologists who implant cardioverter defibrillators advise their patients against driving postoperatively.(ABSTRACT TRUNCATED AT 250 WORDS)


Direct current application: easy induction of ventricular fibrillation for the determination of the defibrillation threshold in patients with implantable cardioverter defibrillators.

For the determination of the defibrillation threshold, the induction of ventricular fibrillation is mandatory. However, in severely damaged hearts it is sometimes difficult to induce ventricular fibrillation by rapid stimulation or alternating current. Only rapid nonclinical ventricular tachycardias may result, and their cardioversion threshold may be different from the defibrillation threshold. Therefore, it was the purpose of this study to test the potential of direct current (DC) application to rapidly induce ventricular fibrillation in patients with an implanted cardioverter defibrillator. The defibrillation threshold had to be determined in 13 patients (9 with coronary heart disease, 4 with dilative cardiomyopathy, ejection fraction 35%) during and 2 weeks after the implantation of a cardioverter defibrillator. DC was applied 37 times by a commercially available 9-V DC battery via a bipolar catheter for about 3 seconds. Ventricular fibrillation was induced 23 times (62%) and rapid nonclinical ventricular tachycardias were induced six times (16%). In one patient clinical ventricular tachycardia was observed. In seven instances (19%) sinus rhythm remained. In 12 of the 13 patients, ventricular fibrillation could be induced by DC. Thus, the induction of ventricular fibrillation by DC application may serve as an additional tool to induce ventricular fibrillation, determining the defibrillation threshold in implantable cardioverter defibrillator patients.


Transtelephonic interrogation of the implantable cardioverter defibrillator.

Third generation implantable cardioverter defibrillators (ICDs) have extensive memory capability to store data about the patient's arrhythmias and the effect of therapies delivered by the ICD. However, this data has so far been accessible only when the patient attends the pacing clinic. Two Medtronic 9421 PCD TeletraceR transmitters have been used to interrogate Medtronic 7216A and 7217B PCD S at distances of up to 300 miles from our hospital and transmit the data to a 9420 PCD TeletraceR receiver. Successful transmission of data has been obtained on 50 occasions with 100% data concordance with repeat transmission. The system can reduce the number of unscheduled clinic visits, reduce delay in making a diagnosis following unexpected delivery of a shock therapy, and reassure patients about to be discharged following ICD implantation. The benefits are magnified where patients reside far away from the implanting center.


Improved chronic epicardial pacing in children: steroid contribution to porous platinized electrodes.

Although new "low threshold" epicardial electrodes combine steroid with a porous, platinized-platinum surface, the actual contribution of steroid elution has not been established. We evaluated this new electrode surface design with and without steroid in 13 children, ages 1-22 years. Both electrodes are unipolar and of similar surface area. The Medtronic Model 4951-P is a barb design for epimyocardial insertion without steroid while the Model 10295A is a steroid eluting, epicardial disk-shaped design. Both electrodes were implanted for atrial and ventricular pacing. At implant, sensed P and R waves, and pacing impedances were comparable between both electrodes. There were no significant differences between initial measured pulse width or calculated energy thresholds for the first 2 months following implant. Strength-duration curves for both electrodes at 1 month were comparable to implant values. After 2 months, the threshold of the nonsteroid electrode peaked and stabilized at a significantly higher (P less than 0.05) level than the more constant steroid eluting electrode. This difference continued for the first year following implant. We conclude that the new porous, platinized-platinum electrode design intrinsically limits initial electrode-tissue interface reactivity in children and improves epicardial pacing with low chronic threshold values. Steroid elution augments these intrinsic qualities by maintaining fibrous capsule stability with more constant low thresholds over time.


Comparison of isoproterenol and exercise tests in asymptomatic subjects with Wolff-Parkinson-White syndrome.

In order to evaluate the effects of increases of sympathetic tone in ventricular response during atrial fibrillation and in the relationship between the accessory pathway effective refractory period (ERP) and ventricular rate during atrial fibrillation, 20 male subjects, aged 19 +/- 6 years, were studied electrophysiologically in basal conditions, after isoproterenol infusion (2-4 micrograms/min) and during submaximal bicycle exercise test, at a constant workload equal to that which increases the sinus rate to the same extent (140 beats/min) induced by isoproterenol infusion. Accessory pathway ERP was evaluated at the same driven rate (150 beats/min) in both instances. In the control study as during both tests atrial fibrillation paroxysms were induced by burst stimulation. In control conditions the rate increase from 100 to 150 beats/min induced a reduction of accessory pathway ERP from 266 +/- 27 msec to 244 +/- 22 msec (P less than 0.005). At the same driven rate of 150 beats/min, isoproterenol infusion and exercise test induced a more marked shortening of accessory pathway ERP to 211 +/- 28 msec (P less than 0.005) and to 214 +/- 29 msec (P less than 0.005), respectively. Atrial fibrillation paroxysms lasting more than 10 seconds were induced in 20/20 cases in the control study, in 15/20 during isoproterenol infusion and in 13/19 cases during exercise test. The shortest cycle length during atrial fibrillation was reduced from a basal value of 253 +/- 72 msec to 204 +/- 27 msec (P less than 0.05) during isoproterenol infusion and to 236 +/- 32 msec (NS) during exercise test.(ABSTRACT TRUNCATED AT 250 WORDS)


Ablation of ventricular tachycardia using multiple sequential transcatheter application of radiofrequency energy.

Multiple sequential radiofrequency energy was applied in the left and right ventricles of 24 dogs to produce large ablated areas limited to endocardial and subendocardial regions. Endocardial ablation was performed in nine dogs with normal ventricles and 15 that had survived remote myocardial infarcts, three with inducible sustained monomorphic ventricular tachycardia. A quadripolar catheter was positioned either at the site of earliest ventricular activation during induced monomorphic ventricular tachycardia or at circumscribed areas of the left ventricle. Radiofrequency energy was delivered between two adjacent poles of the catheter, successively applying radiofrequency energy to the distal, middle, and proximal electrode pairs; this was repeated 9 to 11 times with the catheter in a slightly different position. A cumulative energy of 9,688 +/- 4,191 joules resulted in an ablated endocardial/subendocardial surface area of 4.7 +/- 2.2 cm2 (range 2.4-10 cm2, maximum depth 4 mm). Sustained tachycardia was not inducible by aggressive programmed ventricular stimulation in the dogs with previously inducible tachycardia, indicating successful ablation of the tachycardia foci. Only seven normal dogs were available for electrophysiological studies; three were used in acute and four in chronic studies. Ventricular tachycardia was not induced in the remaining dogs either before or after radiofrequency ablation, indicating the lack of an arrhythmogenic effect of this method. Histologic examination was performed in all nine normal dogs (five were sacrificed for acute pathological examination) as well as in the 15 with myocardial infarction. The late pathological examination of the radiofrequency lesion in these 19 animals showed homogeneous areas of coagulation necrosis and endocardial proliferation. Thus, this modified technique of radiofrequency ablation produced large homogeneous endocardial/subendocardial scars suitable for treating ventricular tachycardia and showed no evidence of an arrhythmogenic influence.


The range of sensors and algorithms used in rate adaptive cardiac pacing.

Implantable sensors play an important role in physiological cardiac pacing. Sensors can be classified according to the technical methods in which sensing is achieved: the sensing of the evoked ventricular response, intrathoracic impedance and body acceleration forces, and the incorporation of special sensors on pacing electrodes. These sensors differ in their relative merits in terms of speed, proportionality, sensitivity, and specificity of rate response. The efficacy of a sensor can be significantly modified by the algorithm used in relating sensor signal to a pacing rate change. The currently available types of sensors and algorithms are summarized and compared in this review article. The relative merits of these sensors and algorithms form the basis for designing a multisensor pacing system.


A case of cardiac foreign bodies associated with four types of tachycardias.

A case of cardiac foreign bodies leading to development of four varieties of automatic tachycardia is reported. A 51-year-old male with aortic regurgitation was admitted to our hospital because of palpitations. An electrocardiogram revealed junctional tachycardia with or without left bundle branch block, and two types of fascicular tachycardias. Computed tomography showed metallic foreign bodies from a fractured guidewire in the membranous portion of the interventricular septum, which was inadvertently retained when he underwent diagnostic cardiac catheterization at the age of 27.


Blood culture results as determinants in the organism identification of bacterial meningitis.

The diagnosis of bacterial meningitis depends on a lumbar puncture (LP). Sometimes, antibiotics are administered before a LP that is delayed owing to prior need for computerized tomography (CT) scan, technical problems, inability to obtain consent, or an unstable patient. We examined the accuracy of blood culture, cerebrospinal fluid (CSF) Gram's stain, and antigen detection by latex for organism identification of meningitis. All patients admitted to the Children's Hospital of Buffalo between January 1, 1984 and December 31, 1989 and having a CSF culture diagnosis of bacterial meningitis had their charts retrospectively reviewed. Patients excluded from the study were those with neural tube defects or CSF catheters, those admitted directly to the Intensive Care Nursery (ICN), those whose positive CSF cultures were determined to be a contaminant, those whose medical records were not found, or those older than 16 years. We analyzed a total of 178 patients with positive CSF cultures and the confirmed diagnosis of bacterial meningitis. Of 169 patients who had a blood culture performed, 86% had the organism responsible for meningitis recovered by this test, with the highest yield of 91% occurring in the 2.5-month to 24-month age group. Blood culture identified the bacteria in 94% of those patients with Haemophilus influenzae meningitis, and this yield increased to 100% when patients who had been pretreated with antibiotics were excluded. The combination of blood culture, CSF Gram's stain, and/or latex agglutination identified the causative bacteria in 92% of patients with meningitis. Blood culture, CSF Gram's stain, and latex agglutination are useful in identifying the organism causing pediatric meningitis.(ABSTRACT TRUNCATED AT 250 WORDS)


Endogenous cholecystokinin, the major factor responsible for dietary protein-induced pancreatic growth.

This study was undertaken to clarify the role of endogenous cholecystokinin (CCK) in induction of pancreatic growth stimulated by a high protein diet. Rats with i.v. jugular cannulae in place and kept in Bollman cages were adapted to 5% casein diet for 9 days and switched to 70% casein for 2 days. MK-329, a CCK receptor antagonist, and SMS 201-995, a somatostatin agonist, were continuously infused at 0.5 mg/kg/h and 5 micrograms/kg/h, respectively, starting at the onset of feeding 70% casein. The 5 and 70% casein control groups were infused with saline. Feeding 70% casein significantly stimulated pancreatic hyperplasia and tissue hypertrophy. MK-329 and SMS 201-995 totally prevented 70% casein-induced increases in pancreatic weight and total RNA and DNA contents. The results indicate that endogenous CCK is the major factor responsible for pancreatic growth induced by a high protein diet.


Trophic effects of unsulfated cholecystokinin on mouse pancreas and human pancreatic cancer.

The effect of unsulfated cholecystokinin on pancreatic growth was evaluated in two experimental models in vivo and in vitro. Mice were injected with sulfated cholecystokinin (CCKs) or unsulfated cholecystokinin (CCKu) (10 or 20 micrograms/kg) or vehicle twice daily for 15 days. Animals were then killed and pancreatic weights, protein, amylase, and DNA content were evaluated. In vitro, growth was evaluated by DNA synthesis and viable cell counts. MIA PaCa-2 and BxPC-3 human pancreatic cancer cells were treated with CCKs or CCKu (10(-12) to 10(-9) M) for 48 or 72 h in the presence of [3H]thymidine to evaluate DNA synthesis. Viable cell counts were performed on both cell lines grown in the presence or absence of unsulfated CCK (10(-12) to 10(-9) M) for 96 h. Pancreatic weight, protein, amylase, and DNA were significantly increased in animals treated with either CCKs or CCKu. However, pancreatic weight, protein, and amylase were significantly higher in mice treated with CCKs compared to CCKu (p less than 0.005). DNA content and index of hyperplasia were the same whether mice were treated with CCKs or CCKu. CCKu was as potent a stimulus for DNA synthesis as CCKs in MIA PaCa-2 and BxPC-3 cells. Finally, CCKu increased cell counts in both pancreatic cancer cell lines. These data suggest that the mechanisms responsible for CCK-induced growth of normal pancreas and pancreatic cancer may differ from those that regulate secretion.


Effects of stress on the development of chronic pancreatitis.

In this study, the effects of chronic water immersion stress on the pancreas were investigated in four groups of rats (each group, n = 9): stress + cerulein group, stress group, cerulein group, and control group. Stress + cerulein rats were treated with water immersion stress for 5 h and two intraperitoneal injections of 20 micrograms/kg body wt of cerulein once a week for 16 weeks. In the macroscopic findings of the pancreas, all rats in the stress+cerulein group exhibited moderate or distinctive congestion of blood vessels, gland atrophy, and fatty changes, while some of them showed bleeding. Microscopically, they all exhibited moderate or severe fibrosis, inflammatory cell infiltration, fatty changes, destruction of lobular architecture, and hemosiderin deposits, while some of them also showed bleeding. The stress group without treatment with cerulein injections showed slight fibrosis, hemosiderin deposits, and bleeding. The cerulein group without stress treatment showed fatty changes, but no inflammatory cell infiltration or fibrosis. In the stress + cerulein group only, the contents of digestive enzymes and protein in the pancreas were approximately 55% lower than those of the control group, whereas those in other groups did not show significant reduction. These findings suggest that stress plays some role in the development of chronic pancreatitis, perhaps by causing circulatory disturbance and blood vessel injury.


Evidence for a relationship between recovery from anaesthesia, modified state of consciousness and striatal voltammetric levels of ascorbic acid.

The effects of various procedures which modify consciousness were studied on the extracellular concentration of ascorbic acid (AA), 3,4-dihydroxyphenylacetic acid (DOPAC) and 5-hydroxyindolacetic acid (5HIAA) in rat striatum, as measured by differential pulse voltammetry (DPV) with electrically pretreated carbon fibre micro-electrodes (CFE). Recovery from anaesthesia (produced by 500 mg/kg i.p. chloral hydrate) was accompanied by a six-fold increase in extracellular striatal AA levels, while negligible changes in DOPAC and 5HIAA occurred. Following complete recovery from anaesthesia, the animals were re-injected with the same dose of anaesthetic which specifically reduced AA levels by 90% (DOPAC levels were unchanged and 5HIAA concentrations slightly reduced). In conscious rats, the neuroleptic haloperidol (1 mg/kg i.p., n = 5) and the minor tranquillizer diazepam (10 mg/kg i.p., n = 5), both considered as behaviourally depressant drugs, reduced extracellular AA levels to 50% of controls. The psychomotor stimulant D-amphetamine (1 mg/kg i.p., n = 5) increased AA levels by 90% over controls. Stress activation of animals (handling for 10 min, n = 10) also produced a transient, significant increase (180% of control values) in this striatal parameter. Taken together with previous reports, our results suggest a close relationship between the state of consciousness and extracellular AA levels in the rat striatum and that this relationship appears to be more correlated to AA as no such clear interdependence was noted between the levels of consciousness and extracellular striatal DOPAC or 5HIAA.


Carbonic anhydrase II gene expression in mouse pancreatic duct cells.

Our goal is to create a transgenic mouse model for human pancreatic duct cell adenocarcinoma using the promoter/enhancer region of the carbonic anhydrase (CA) II gene to drive the expression of SV-40 T-antigen in pancreatic duct cells. This requires that the CA II gene be expressed in mouse pancreatic duct cells and not in other pancreatic cells, as has already been shown to be the case in the human and guinea pig pancreas. We have shown with an enzyme histochemical assay that mouse pancreatic duct cells contain CA activity in both intact pancreas and cultured interlobular duct epithelium. In addition, CA activity was detected with a biochemical assay in homogenates of cultured duct epithelium. The specific activity of duct cells was 2.75-fold greater than in whole pancreas, suggesting that a substantial amount of total pancreatic CA activity is contributed by duct cells. At least some of the CA in cultured duct cells was inferred to be CA II by Northern blot analysis of RNA extracted from the cells. The concentration of CA II mRNA in the cultured duct cells was substantially greater than in whole pancreas and would appear to account for the majority, if not all, of the CA II in the mouse pancreas.


Central monoaminergic changes induced by morphine in hypoalgesic and hyperalgesic strains of domestic fowl.

The present research examined morphine dose-response effects on both the formalin test and on CNS monoamine (MA) levels and the metabolites dihydroxyphenylacetic acid (DOPAC) and 5-hydroxyindoleacetic acid (5-HIAA) in hypoalgesic (76) and hyperalgesic (SN) strains of domestic fowl. Morphine produced a significant hypoalgesic response in the 76 strain at 15-45 mg/kg and a significant hyperalgesic response in the SN strain at 5-10 mg/kg. In subsequent experiments, analyses of whole brain (minus tectum), brainstem, and spinal cord MA, DOPAC, and 5-HIAA via high-performance liquid chromatography with electrochemical detection (HPLC-EC) were performed following morphine administration in both the 76 and SN strains. Morphine produced a significant elevation of brain dopamine (DA) and a significant elevation of brain, brainstem, and spinal cord serotonin (5-HT) in both the 76 and SN strains. Morphine elevated brain norepinephrine (NE) in the 76 strain. However, morphine failed to affect brain NE in the SN strain. This distinct morphine effect on brain NE differentiates strain-dependent hypoalgesia and hyperalgesia in domestic fowl.


Rapid histochemical identification of motor and sensory fascicles: preparation of solutions.

We report a rapid and inexpensive staining method for identification of motor and sensory fibers within 1 hour (intraoperatively). This method should be easy to use in any hospital where frozen histologic sectioning is available.


[Determining the time of diabetes mellitus onset by the level of glycosylated keratin in hair].

The levels of glycosylated hemoglobin (HbA1c) and glycosylated keratin were studied along the hair length in 17 patients with newly detected insulin dependent diabetes mellitus (IDDM) aged 7 to 33, in 14 patients with newly detected noninsulin dependent diabetes mellitus (NIDDM) aged 46 to 73, and in 41 healthy subjects. The level of glycosylated keratin in healthy hairs along the entire length varied within 0.094-0.124 mumol of fructosamine per 100 mg of hair. In 10 of 17 patients of the IDDM group measurements of glycosylated keratin levels made it possible to determine the time of appearance of diabetes mellitus, in 13 patients of the NIDDM group these levels along the entire hair length were elevated. The determination of these levels permitted the estimation of the time of appearance of IDDM. Its manifestation followed a long period (1-2 yrs.) of latent derangements of carbohydrate metabolism. In NIDDM patients the time of appearance of disease is difficult to determine because of the limited hair length and a long latent period of disease (over 2-3 yrs.).


Risperidone: clinical safety and efficacy in schizophrenia.

Risperidone represents a unique pharmacology of potent antagonism of both serotonin and dopamine receptors. In a randomized, parallel-group, double-blind trial of risperidone vs. haloperidol and placebo in 36 schizophrenic patients in acute exacerbation, risperidone showed a quicker onset of antipsychotic activity than did haloperidol. Risperidone treatment was statistically superior to placebo, with a trend toward superiority to haloperidol. Risperidone did not differ from placebo on assessment scales of extrapyramidal side effects, but produced significantly less than did haloperidol. There were no major adverse reactions associated with risperidone use, but it was noted to reduce the signs of tardive dyskinesia. This study suggests that risperidone may offer a superior side-effect profile, and possibly greater efficacy, than a standard neuroleptic such as haloperidol.


Neurotransmitter changes in early- and late-onset Alzheimer-type dementia.

Neurotransmitter-related markers were examined in Alzheimer-type dementia (ATD) and studied whether or not there is biochemical difference between the early- and late-onset sub-groups. Postmortem brains were obtained from neuropathologically diagnosed ATD patients and control subjects with no clinico-neuropathological findings indicative of neuropsychiatric diseases. Neurochemical data in the early- and late-onset groups were compared to the age-matched younger and older control groups, respectively, and expressed as a percentage of the mean value in the respective controls. Choline acetyltransferase activity and concentrations of serotonin and noradrenaline were more severely depleted in the early-onset ATD group than in the late-onset ATD group. These findings indicative of heterogeneity of ATD patients were discussed from the pathogenetic and therapeutic viewpoints.


Modulation of NMDA receptors by ACh in the central nervous system.

The effect of acetylcholine (ACh) on N-methyl-D-aspartate (NMDA)-induced responses in neurons acutely dissociated from the rat nucleus basalis of Meynert (nBM) was investigated with a conventional patch-clamp technique. The whole-cell recording revealed that NMDA-induced inward current (INMDA) consisted of transient peak and successive steady-state components. With outside-out recording, these components were shown to have the identical single channel conductance. Pretreating the nBM neurons with the low concentration of ACh suppressed the peak component of INMDA, whereas the high concentration potentiated it. The modulatory action of ACh was observed in nBM neurons dissociated from mature rats. The action was deficient, however, in those from immature animals. These results suggest that ACh is either an inhibitory or an excitatory modulator of glutamatergic transmission in the central nervous system.


Subcellular localization of transferrin and pyrophosphate in liver cells after perfusion in situ or incubation in vitro.

Rat livers were perfused in situ with [125I]-transferrin, FITC-dextran and [32P]-inorganic pyrophosphate (PP). In a parallel set of experiments rat liver homogenates was incubated with [125I]-transferrin and [32P]-PPi. In both sets of experiments cellular organelles were prepared by density gradient centrifugation in metrizamide. In the perfusion experiments [125I]-transferrin and FITC-dextran accumulated in fractions rich in plasma membrane enzymes, whereas in the homogenate experiments [125I]-transferrin remained in soluble fractions. [32P]-PPi remained in soluble fractions in both sets of experiments, i.e. PPi did not co-localize with transferrin taken up by intact cells. In another set of experiments fractions rich in endosomes were tested for their ability to synthesize PPi from ATP. No PPi was found. The results argue against a role for PPi in transferrin-iron release within endosomes.


Serum, disc and egg ELISA for the serodiagnosis of Salmonella gallinarum and S. enteritidis infections in chickens.

An ELISA was evaluated for the serodiagnosis of fowl typhoid and paratyphoid due to Salmonella enteritidis in chickens. The hot phenol: water lipopolysaccharide (LPS) extract of Salmonella was used as the antigen. Chicken serum, eggs and discs impregnated with chicken blood were tested for the presence of antibodies against Salmonella factor 'O' 9 antigen. The substrate and chromogen used were hydrogen peroxide and orthophenylenediamine respectively. Serological results from the experimentally and naturally infected chickens showed close agreement between the conventional Serum Tube Agglutination Test (SAT) and serum ELISA while serum ELISA results were in close agreement with the egg and disc ELISA results. It was noted that ELISA was highly sensitive, convenient and versatile. It is concluded that ELISA, especially disc ELISA, ought to replace SAT for seroscreening chickens against S. gallinarum and other Salmonella Group D infections.


Enhanced spontaneous and induced LAK function in baboons receiving total lymphoid irradiation (TLI).

TLI has potent immunosuppressive effects, but there is concern over its possible carcinogenic properties. The aim of the study was to assess to what degree TLI may compromise natural killer (NK) and lymphokine activated killer (LAK) cell function. There was a significant increase (P less than 0.0001) in both NK-cell function (measured against K562 targets) and spontaneous LAK-cell function (measured against DAUDI targets) in fresh blood lymphocytes throughout a course of 8 x 100 cGy fractionated TLI. This may be related to a three- and sevenfold increase, respectively, in CD16+ CD8- and CD56+ CD2- cell frequencies over the same period. Mitogen-induced interleukin 2 (IL-2) synthesis from blood lymphocytes was inhibited by up to 75% with as little as 100 cGy of TLI. Expression of IL-2 receptors on fresh lymphocytes did not vary and remained low. Therefore spontaneous LAK occurrence appeared to be triggered through an IL-2-independent pathway. The in vitro addition of IL-2 verified that cells retained their ability to respond to this lymphokine resulting in greatly enhanced induced LAK function. This was most probably mediated by CD56+ cells which were found to readily express IL-2 receptors upon mitogen stimulation. In conclusion, fractionated low-dose TLI appears to enhance MHC unrestricted immune surveillance in a manner independent of IL-2 production.


Production, immunoanalysis and selection of monoclonal antibodies for studies of polymorphism of bovine MHC class I antigens.

Two monoclonal antibodies (MoAbs) were raised against bovine lymphocyte antigens of the major histocompatibility complex (MHC) for studies of the polymorphism of MHC class I antigens of African cattle. Immunoanalysis of lymphoid tissues and molecular weight data were used to characterize the epitopes seen by the MoAbs. Competitive binding assays indicated specificity of the MoAbs for two distinct epitopes. Application of the MoAbs for MHC typing of a cattle population showed that the epitopes were co-expressed on the target MHC antigen but occurred independently on non-target MHC antigens. It is concluded that, to obtain MoAbs for studies of MHC polymorphism, it is important to conduct large-scale analysis using immunoassays and population studies.


Development of a malaria T-cell vaccine for blood stage immunity.

We have defined a strategy for the development of a T-cell vaccine for blood stage immunity, taking into consideration the central role of T cells and MHC restriction in malaria immune responses. We have used the AMPHI computer algorithm to identify putative T-cell epitopes from conserved regions of 11 Plasmodium falciparum asexual stage proteins. Ten of the eleven proteins are currently candidates for vaccine development. Using this algorithm we selected 22 putative T-cell epitope peptides and 8 control peptides. These peptides were used to test the T-cell responses of three defined populations of Caucasians who have (1) recovered from P. falciparum malaria, (2) been exposed, but never clinically infected, (3) never been exposed or infected. Preliminary analysis of our data shows population differences in T-cell responses to putative T-cell epitope peptides. Ultimately, these studies will help to identify those T epitopes that can be incorporated into a T-cell vaccine for protective immunity.


Coagulation dynamics after hemodilution with polyhemoglobin.

The effect of polyhemoglobin solution (PHS), a candidate erythrocyte cell substitute, on blood coagulation, was investigated. Whole blood samples from six male Sprague-Dawley rats were diluted in vitro 3:1, 1:1 and 1:3 with normal saline solution, stroma-free hemoglobin (SFH) (7 grams of hemoglobin per deciliter), PHS (14 grams of hemoglobin per deciliter) or 5 percent bovine albumin (ALB) and tested with a thrombelastograph (TEG) for effects of hemodilution on coagulation. For in vivo tests, 24 rats were randomly divided into four groups of six each and 50 percent of the estimated blood volume was replaced with SFH, PHS, ALB or 6 percent hydroxyethyl starch. Blood samples collected before and after the hemodilution were tested with a TEG. The TEG parameters (r, k, Ma, and fi) were determined and analyzed statistically. At 50 percent in vitro or in vivo hemodilution, PHS did not cause significant alteration (p greater than 0.05) in initial coagulation mechanism, while SFH slightly, but significantly (p less than 0.05), accelerated coagulation. Tensile strength of formed clot (Ma) did decrease after hemodilution with PHS (p less than 0.05), but the effect was attributable to dilutional effect. At moderate hemodilution (50 percent), PHS does not seem to cause undue coagulopathy in this rat model.


Santiago Ramón y Cajal and methods in neurohistology.

Controversy, misunderstanding or uninformed opinion abound over the extent to which the great Spanish neurohistologist, Santiago Ramón y Cajal, specified his staining methods in his analytical papers, the methods by which he analysed and presented his data, and the microscopes available to him. In this paper, we have attempted to outline the information on these points that we have been able to obtain from a detailed examination of his writings and a study of the evidence remaining in the Cajal Museum in Madrid.


Three-dimensional images of Ramón y Cajal's original preparations, as viewed by confocal microscopy.

The drawings prepared by Santiago Ramón y Cajal have an inherent beauty as well as being accurate and arose partly as a result of the difficulty contemporary photographic methods had in reproducing informative images of the thick sections he used. Modern methods are better and in this brief article Alan Boyde presents some three-dimensional images of these historical preparations, generated by confocal microscopy.


Mechanoelectrical transduction by hair cells.

Hair cells of the inner ear are one of nature's great success stories, appearing early in vertebrate evolution and having a similar form in all vertebrate classes. They are specialized columnar epithelial cells, with an array of modified microvilli or stereocilia on their apical surface, interconnected by a series of linkages. The mechanical stimulus causes deflection of the stereocilia, stretching linkages between them, and opening the mechanotransducer channels. On a slower timescale, hair cells adapt in order to maintain optimum sensitivity, with an adaptation motor within the stereocilia acting to keep the resting tension on channels constant.


Changes in Ca(2+)-binding proteins in human neurodegenerative disorders.

The cellular distribution of Ca(2+)-binding proteins has been extensively studied during the past decade. These proteins have proved to be useful neuronal markers for a variety of functional brain systems and their circuitries. Their major roles are assumed to be Ca2+ buffering and transport, and regulation of various enzyme systems. Since cellular degeneration is accompanied by impaired Ca2+ homeostasis, a protective role for Ca(2+)-binding proteins in certain neuron populations has been postulated. As massive neuronal degeneration takes place in several brain diseases of humans, such as Alzheimer's disease, Parkinson's disease and epilepsy, changes in the expression of Ca(2+)-binding proteins have therefore been studied during the course of these diseases. Although the data from these studies are inconsistent, the detection and quantification of Ca(2+)-binding proteins and the neuron populations in which they occur may nevertheless be useful to estimate, for example, the location and extent of brain damage in the various neurological disorders. If future studies advance our knowledge about the physiological functions of these proteins, the neuronal systems in which they are expressed may become important therapeutical targets for preventing neuronal death in an array of neurodegenerative diseases.


Neurotransmitter transporters (plus): a promising new gene family.

Neurotransmitter transporter genes encode proteins with 12 putative transmembrane regions, which mediate Na(+)-dependent reaccumulation of released neurotransmitters into presynaptic terminals and are the sites of action of important abused and therapeutic drugs. Studies of these genes show promise for improving our understanding of transport mechanisms and modes of drug action, and may even uncover previously unanticipated neurotransmitters.


A rapid method for detection of Yersinia enterocolitica serotype O:3 in pig feces using monoclonal antibodies.

A simple colony immunoblotting method using monoclonal antibodies (MAbs) was developed to detect Y. enterocolitica serotype O:3 in pig feces. One of the MAbs studied was able to detect single colonies of the organism in the presence of calculated 3.1 x 10(8) heterologous organisms in pig feces. The MAb was found to be specific for the lipopolysaccharide (LPS) O-antigens of Y. enterocolitica serotype O:3. No significant cross-reactivity was found against a variety of closely related serotypes and Gram-negative organisms. The MAb could also be used in a slide agglutination test and an indirect fluorescence antibody assay for rapid identification of Y. enterocolitica serotype O:3.


Purification and adhesion receptor phenotype of ovine bone marrow-derived haemopoietic colony-forming cells.

Ovine haemopoietic progenitor cells that form colonies (CFC) in soft agar cultures were compared to more mature bone marrow cells for their level of expression of the adhesion receptor molecules ovine (ov) CD44, ov CD11a (LFA-1) and ov CD58 (LFA-3) as well as the 175-antigen using specific monoclonal antibodies. Ov CD44, ov CD11a and ov CD58 were expressed on all CFC of the myeloid (non-erythroid) series, whereas ov CD44 and ov CD11a expression was very low or absent from a small number of blast and erythroid series CFC. Within the mature non-erythroid population of myeloid cells, neutrophils retained a low level of expression of ov CD11a. Most CFC representing all lineages strongly expressed the ov CD44 antigen. In contrast, the majority of CFC lacked the 175-antigen, as did bone marrow lymphocytes, basophils and mast cells. This property of CFC was exploited in a negative selection technique using panning and immunomagnetic beads to select CFC from other bone marrow cells with a 116-125-fold enrichment, 12-14% purity and 29-40% yield. These results demonstrate that ovine CFC express some of the molecules necessary to allow adhesion to haemapoietic stromal cells and vascular endothelium in the tissues. Future studies will concentrate on the function of the adhesion receptor molecules in medullary and extra-medullary haemopoiesis and inflammatory cell development in sheep.


Changes in cytokeratin expression accompany squamous metaplasia of the human respiratory epithelium.

To determine the characteristics of metaplastic changes of the nasal respiratory epithelium, the distribution of individual cytokeratins (CKs) was studied immunohistochemically and by two-dimensional gel electrophoresis. The authors define four types of changes of the normal pseudostratified columnar epithelium: (1) transitional pseudostratified epithelium (first unusual CK.: no. 13); (2) stratified columnar epithelium (increased expression of CKs 4 and 13; CKs 7, 8, 18 and 19 reduced); (3) stratified squamous epithelium, nonkeratinized (appearance of CK 16); and (4) stratified squamous epithelium, keratinized (expression of CKs 1 and 10, variable CK5 and 14 patterns in basal cells). These phenotypes were found simultaneously within single specimens, resulting in apparent overall variability in the immunohistochemical staining patterns. Spatially, changes in CK expression towards "normal" parts were not abrupt but rather gradual. Biochemical data confirmed the immunohistochemical findings and added CK 6 to the pattern of altered nasal mucosa. The findings of this study suggest a stem cell metaplasia in the nasal epithelium which is based on its inherent bimodal developmental programme. A gradual loss of normal respiratory epithelial differentiation, as seen by the loss of CKs 7, 8, and 18, was paralleled by the appearance of squamous epithelial type CKs, e.g. the expression of CKs 1, 10 and 13. Basal cell types CKs 5, 14, 17 and 19 were maintained during this process. Implications of these results for general concepts of CK expression in the metaplastic process are discussed.


Keratin profiles in normal/hyperplastic prostates and prostate carcinoma.

Immunoreactivities in 25 cases of prostatic adenocarcinoma and 10 normal/hyperplastic prostates were investigated in methacarn-fixed, paraffin-embedded serial sections using a panel of nine anti-keratin monoclonal antibodies (mAbs); 34 beta E12, CK8.12, 312C8-1, CK4.62, RPN1165, RPN1162, 35 beta H11, CK5, M20, and one of anti-actin mAb, HHF35. In normal/hyperplastic prostates, RPN1162, 35 beta H11, CK5 and M20 stained luminal cells without staining basal cells, and 34 beta E12, CK8.12 and 312C8-1 stained basal cells but not luminal cells. Other mAbs, CK4.62 and RPN1165, stained basal cells as well as luminal cells. All of the mAbs labelling luminal cells stained cancer cells with variable frequencies in a manner unrelated to the grade of tumour differentiation. Of the prostate cancer cases 92% were scored positive with M20, 84% with 35 beta H11, 80% with CK5, 68% with CK4.62, 60% with RPN1165 and 4% with RPN1162. However, basal cell-specific keratins labelled with 34 beta E12, CK8.12 and 312C8-1 were totally negative in the cancer cells. HHF35 showed no labelling in normal, hyperplastic or neoplastic epithelial cells of the prostate. Our findings indicate that the major part of the cells of prostatic adenocarcinomas have keratin phenotypes similar to luminal cells but not basal cells, and that no myoepithelial differentiation can be detected in epithelial cell of the prostate. Thus, mAbs for keratins facilitate the identification of epithelial cell phenotypes in normal, benign and malignant conditions of the prostate.


Testicular sex cord-stromal lesions: immunohistochemical analysis of cytokeratin, vimentin and steroidogenic enzymes.

We have studied immunolocalization of all steroidogenic enzyme involved in sex steroids biosynthesis, P-450 side chain cleavage (P-450scc), 3 beta hydroxy steroid dehydrogenase (3 beta-HSD), P-450 17 alpha hydroxylase (P-450(17 alpha)) and P-450 aromatase (P-450arom) and that of vimentin and cytokeratin in 14 cases of testicular sex cord-stromal tumours (6 Leydig cell tumours, 5 Sertoli cell tumours, 2 fibromas and 1 granulosa cell tumour) as well as 4 cases of hyperplasia (2 Leydig and 2 Sertoli). Leydig cell tumour expressed all four steroidogenic enzymes examined, indicating that this tumour can synthesize oestrogen from cholesterol. In 2 cases of Sertoli cell tumour, the tumour cells with clear cytoplasm and without Reinke's crystals expressed P-450ssc, 3 beta-HSD and P-450(17 alpha), suggesting the capability of androgen production in these tumour cells. Fibromas and granulosa cell tumour were negative for the enzymes examined. In immunohistochemistry of intermediate filaments, Leydig cell tumours demonstrated only vimentin. Sertoli cells in hyperplasia and non-neoplastic testis expressed only vimentin but Sertoli cell tumours expressed both cytokeratin and vimentin. Cytokeratin immunoreactivity was correlated with morphological epithelial differentiation in Sertoli cell tumour. These findings in testicular Sertoli cell tumour are considered to represent the multiple differentiation capacity of this neoplasm. Immunohistochemical study of steroidogenic enzymes and intermediate filaments provided new insight into neoplastic steroidogenesis and the differentiation capacity of testicular sex cord-stromal neoplasms.


Establishment and characterization of a human papillary thyroid carcinoma cell line with oxyphilic differentiation (ONCO-DG 1).

In the present study the establishment and characterization of a new oxyphilic papillary thyroid carcinoma cell line--ONCO-DG1- is given. With immunohistological, histochemical and flow cytometric methods, ONCO-DG 1 cells revealed features of epithelial differentiation. Furthermore the cells formed von Kossa-positive deposits resembling psammoma bodies in monolayer and spheroid culture until late passages. The tumor cell line is now in the 40th subculture. Because of the ability to form multicellular tumor spheroids (MCTS), this cell line is a good model for examining the interaction between thyroid tumor cells and confluent human endothelial cells on extracellular matrix in vitro. It is also suitable for xenotransplantation studies, because it is tumorigenic in NMRI nude mice in vivo.


Pancreatic islet transplantation: isolation, separation, and microvascularization.

Twenty years ago Ballinger and Lacy were the first to report cure of experimental diabetes in rats after free transplantation of isolated pancreatic islets. In a remarkable number of experimental studies on transplantation of isolated islets of Langerhans reversal of diabetes and restoration of normal glucose metabolism has been achieved in rodents. However, studies in larger animals and clinical attempts have been less successful. Beside graft rejection, the isolation and purification of a sufficient mass of islets have been identified as major obstacles. In addition, insufficient revascularization is thought to be, in part, responsible for early graft failure. Recent developments allow for consistently high yields of isolated islets using an automated method, in particular for isolation of human pancreatic islets. Furthermore, a variety of techniques have been implemented during the past years improving purification, including gradient separation, fluorescence-activated sorting, magnetic microsphere extraction and cryopreservation. Revascularization of the islet grafts may be enhanced by selection of adequate sites for transplantation and/or by supportive application of angiogenic peptides. Progress achieved during the past years allow for new hope for successful clinical islet transplantation.


[Value of 24-hour blood pressure measurement in patients with arterial hypertension, left heart enlargement and increased cardiac risk].

Non-invasive 24-h blood pressure and a 24-h electrocardiogram were recorded in 45 normotensive and 97 matched, untreated, hypertensive subjects, with and without echocardiographic signs of left-ventricular hypertrophy and without signs of coronary artery disease. Forearm vascular resistance was calculated from mean blood pressure and postischemic blood flow, which was measured by venous plethysmography. Systolic ambulant 24-h blood pressure exhibited the closest correlation with left-ventricular mass index in hypertensives (r = 0.48; p less than 0.001). No relation could be found between blood-pressure fall overnight and left-ventricular hypertrophy. Blood-pressure variability was significantly higher in hypertensive subjects. Furthermore, systolic 24-h blood pressure significantly correlated with the rate of ventricular arrhythmias in hypertensives. Casual blood pressure exhibited no comparable correlations. A significant close correlation between 24-h blood pressure and vascular resistance was identified in hypertensives. Furthermore, left-ventricular mass index and vascular resistance were correlated (in hypertensives: r = 0.32; p less than 0.01). This finding speaks for a parallel development of total peripheral resistance and left-ventricular hypertrophy in essential hypertension. The close relationships between ambulatory 24-h blood pressure, left-ventricular hypertrophy, ventricular arrhythmias, and vascular resistance demonstrates the high diagnostic and, possibly, the prognostic value of long-term blood-pressure measurement.


Intrinsic organization of the paramedian pontine reticular formation of the cat.

A morphoquantitative analysis was carried out to clarify the cytoarchitectural organization of the paramedian pontine reticular formation (PPRF) which is considered to be an important site in the control of eye movements. The study was carried out on the cat, using the Golgi staining method. The topographic position and detailed structure of the neurons were demonstrated using morphoquantitative methods. On the basis of their neuronal arborization, fusiform neurons and two types of multipolar cells were identified. Fusiform neurons show dendrites which are given off from the two poles of the small- to medium-sized cell body. The arborization generally runs caudorostrally, ending inside the PPRF. These neurons are ubiquitous. Type 1 multipolar neurons, the most frequent elements of the neuronal population (60%), have a small- to large-sized cell body from which 2 or 3 primary spiny dendrites and the axon emerge. Their dendritic field is oval and generally oriented in the vertical plane. These neurons are scattered everywhere in the PPRF. Type 2 multipolar cells are large neurons endowed with numerous primary spiny dendrites constituting a wide round dendritic field and with a thick axon. They are located almost exclusively at the boundaries of the PPRF and preferentially in the caudal region. The characteristics of the neurons suggest that the fusiform cells may play an interneuronal role, while the multipolar neurons could have both a projective function and an important receptive role for the afferent fibers to the PPRF. The lack of homogeneity found among the multipolar neurons is in agreement with the variety of projective elements shown by functional investigations.(ABSTRACT TRUNCATED AT 250 WORDS)


[Morphometric analysis in the rat spinal cord following peripheral nerve transection with the use of MAP2 immunohistochemistry].

The physiological function of the spinal cord is regulated by two different kinds of information, namely, afferent inputs from the primary sensory fibers and descending inputs from the brain. The former conveys the sensory information from the peripheral region via the dorsal root ganglia to spinal interneurons or directly to motor neurons and brings about the voluntary and/or involuntary muscle movement. In the present study, the time-related changes in the dendrocytoarchitecture of the rat spinal cord following the transection of afferent and efferent connections from/to the peripheral organs were observed by using immunohistochemistry for microtubule-associated protein 2 (MAP2) and Nissl staining. To investigate the changes quantitatively, morphometric analyses were performed on the section of gray matter of tissue samples stained with MAP2-antisera. One to 24 weeks after hemitransection of the dorsal and ventral roots, a remarkable reduction in the area and progressive changes in the shape of the gray matter were seen in both the anterior and posterior horns on the lesioned side. These changes were found to be due to the reduction of the width of the anterior and posterior horns on the lesioned side rather than of the height. At the later postoperative stage, atrophy of the Fasciculus gracilis and disappearance of motoneurons on the lesioned side were recognized in Nissl stained tissue samples.


Cross-reactivity of a commercial chemiluminescence immunoassay for human chorionic gonadotropin with the free beta-subunit.

An enhanced chemiluminescence immunoassay for the determination of serum human chorionic gonadotropin (HCG) in specimens from oncology patients has been assessed with respect to its cross-reactivity with the free HCG beta-subunit (HCG-beta). The assay, standardized against the First International Reference Preparation 75/537, had a cross-reactivity with the free beta-subunit of 625% (molar basis). Therefore this assay achieves high sensitivity for the detection of either intact HCG or free HCG-beta in serum of patients with seminomatous or nonseminomatous testicular cancers. Results of both assays, the in-house immunoradiometric assay (+HCG-beta) and the Amerlite HCG-60 assay, showed a close correlation (R = 0.854-0.960) when serum samples from tumour patients were analyzed. Moreover, the content of free beta-subunit determined in a specific HCG-beta assay, could be quantitatively measured in the enhanced chemiluminescence immunoassay. Thus, this assay is suitable for oncology use, but also highlights the limitations of measuring HCG in serum samples.


Effects of dorsal root entry zone lesions on CSF and plasma neuropeptides and catecholamines.

Effects of dorsal root entry zone lesions (DREZLs) on cerebrospinal fluid (CSF) and plasma concentrations of neuropeptides, catecholamines, and cyclic nucleotides were studied in 9 patients with intractable chronic pain. Contents of beta-endorphin-like-material in CSF decreased in all patients 12-17 days following DREZLs during which complete to good pain relief was achieved. Contents of beta-endorphin-like-material in CSF increased again about one month after DREZLs in two and remained unchanged in one of three patients tested, who complained of partial reappearance of pain. Contents of beta-endorphin-like-materials in plasma showed no significant changes after DREZLs. Substance P, noradrenaline, adrenaline, and cyclic nucleotide levels in both CSF and plasma were variable among the subjects and did not change significantly following the operations. Thus, the results suggest that production of beta-endorphin-like-material in the central nervous system is decreased by DREZL, though the increase in its turn-over might not be neglected. The mechanisms of the decrease in contents of beta-endorphin-like-material in CSF after DREZLs were discussed in terms of our current knowledge of pain and pain inhibitory systems.


Cytokeratins in intracranial and intraspinal tissues.

The intermediate filament distribution pattern in cells and tissues of vertebrates reflects their differentiation or functional specialization state, their histogenesis, and their malignant transformation. In the case of cytokeratins, the characteristic epithelial intermediate filaments representing a complex group of about 30 polypeptides, extensive attention has been given to their expression in diverse epithelial and epithelioid cells. However, little is known about their distribution during fetal development and in neuroectodermal cells. This review specifically focuses on the data concerning cytokeratin expression in intracranial and intraspinal tissues, as expressed alone or as coexpressed with other intermediate filament proteins. Furthermore, the expression pattern of individual cytokeratin polypeptides was investigated by immunocytochemistry in diverse human and animal tissues using a broad panel of monoclonal antibodies. Only the cytokeratins typical of simple epithelia with the primary keratin pair 8/18 as a significant component have been detected in neuroectodermal tissues such as the choroid plexus and ciliary body epithelia, the retinal pigment epithelium, the subcommissural organ, and the ependymal cell clusters in fetal pineal gland (only in humans) as well as in various "unspecialized" ependymal cells of brain ventricles and spinal cord ependyma. Focal cytokeratin 19 expression in rat ciliary body and ventricle ependyma represents a rare exception. In addition, a group of intracranial and intraspinal tissues with controversial histogenesis express solely the cytokeratins 8 and 18: endocrine pituitary cells, arachnoid cells, and corneal endothelium. In most cases of tissues with neuroectodermal derivation, coexpression of cytokeratins and vimentin, or triple expression of cytokeratin, vimentin, and GFAP (fetal and neonatal choroid plexus of humans, rat and guinea pig ependymal cells in the neighborhood of the subcomissural, folliculostellate cells of human and guinea pig pituitary) is detectable. The coexpressions are discussed in the light of several hypotheses based on morphological and functional data concerning intermediate filament protein expression. Both the occurrence of more than one intermediate filament protein and the individual cytokeratin composition in the corresponding tumors of neuroectodermal origin reflect, in principle, the patterns found in their normal tissues. In the fetal neuroectodermal tissues studied, the cytokeratin pair 8/18 is the first one to be expressed during embryonic development.(ABSTRACT TRUNCATED AT 400 WORDS)


Improved detection of antibodies to hepatitis C virus using a second generation ELISA.

A screening assay for the detection of antibodies to hepatitis C virus (HCV); ORTHO HCV ELISA Test System, Second Generation, was compared with the currently licensed c100-3 based test (ORTHO HCV ELISA Test System). The second generation ELISA differs from the c100-3 based assay in that it detects circulating antibodies to both structural (nucleocapsid) and non-structural (NS3/NS4) HCV proteins. Specimens tested consisted of a cohort of 35 patients diagnosed with non-A, non-B hepatitis (NANBH) and 3971 presumably healthy volunteer blood donors. Second generation ELISA demonstrated significantly greater clinical sensitivity in patients with acute phase NANBH (80% vs. 60%) as well as chronic disease (88% vs. 72%). Additional specimens reactive only in second generation ELISA, demonstrated reactivity to HCV antigens c33c and/or c22-3 in supplemental testing by the Chiron HCV RIBA Assay System. The second generation ELISA also detected additional RIBA reactive volunteer blood donors (0.18% of the population tested) that were nonreactive in first generation ELISA. This data indicated that second generation ELISA would detect approximately 2 additional anti-HCV reactive donors per 1,000 screened. Specificities obtained with this low risk population were 99.6% for first generation and 99.7% for second generation ELISA.


Transient coronary stenosis associated with arrhythmia and ischemia 24 hours after reperfusion.

We examined the electrophysiologic sequelae of transient reductions in coronary blood flow (CBF) in conscious dogs. Animals were instrumented to measure arterial pressure, heart rate, repetitive extrasystole threshold (RET), and CBF before coronary stenosis, during 90 minutes of a 50% reduction in CBF, and at 1, 24, 48, and 72 hours after reperfusion. RET was decreased at 24 and 48 hours but returned to baseline by 72 hours after reperfusion. Frequent ventricular ectopic activity (VEA), absent during the control period, was noted during stenosis and at 1, 24, and 48 hours after reperfusion. beta-Adrenergic blockade (metoprolol) normalized the decreased RET at 24 hours. Addition of alpha 1-adrenergic blockade (prazosin) abolished VEA. Endocardial blood flow in the posterior papillary muscle of the left ventricle was reduced by 52 +/- 9% during stenosis, was similar to baseline levels by 1 hour after reperfusion, and decreased by 39 +/- 5% at 24 hours. We conclude that episodes of modest coronary artery stenosis may be followed by electrophysiologic changes indicative of increased vulnerability to malignant ventricular arrhythmias.


Combination chemotherapy and radiation for palliation of hepatocellular carcinoma.

Twenty seven patients with hepatocellular carcinoma were treated by sequential methotrexate (75 mg/m2) and 5-fluorouracil (5-FU) (750 mg/m2) on day 1 followed on days 8-36 by external beam radiotherapy (total dose 30 cGy). The response was assessed by liver size on clinical examination. One patient had complete response, and six patients had partial response. The overall response to the treatment was 25.9%. More than a 50% reduction in serum alfa-fetoprotein level was noted in 66.6% patients. Seventy-one percent of patients had palliation of pain following therapy. The median survival of responders was 11 months and of nonresponders, 2 months. Radiation was discontinued in two patients who developed radiation hepatitis. Additional trials with different dosages and schedules are needed to fully evaluate this form of therapy.


Extragonadal abdominal germ cell cancers.

The charts of eleven patients with abdominal germ cell tumors were reviewed; one had a seminoma. They all had normal testes by physical examination. Therapy consisted of cisplatin-based chemotherapy and, in some cases, surgical debulking. A complete clinical response occurred in seven patients (63%). Two patients relapsed after achieving pathology complete responses and died of progressive disease despite second-line chemotherapy. All patients that failed to achieve a complete clinical response died of progressive disease. Five patients (45%) are long-term disease-free survivors, having no recurrence 4-10 years from the time of the diagnosis (median 6 years). The outcome for this group of patients did not differ significantly from that for patients with mediastinal germ cell tumors in this institution. They do not fare as well as patients with testicular cancer.


Radiation therapy in a case of systemic mastocytosis: evaluation of histamine levels and mucosal effects.

Plasma histamine levels were obtained during palliative radiation therapy of the spine involved with systemic mastocytosis in a 68-year-old woman. Baseline plasma histamine levels were obtained before irradiation and compared to levels obtained on the third, fifth, eighth, and tenth treatment days. Despite concerns regarding histamine release with mast cell destruction following irradiation, plasma histamine levels remained within normal limits. No change in dermatologic or other systemic manifestations were observed. No untoward systemic or localized sequelae associated with mast cell degranulation in response to the administered large field of radiation was observed. Effective palliation was accomplished, and it was concluded that radiation therapy can effectively be applied in treatment of systemic mast cell disease without significant morbidity.


CFTR!

Cystic fibrosis (CF) is a fatal genetic disease primarily affecting Caucasians, although cases have been reported from other ethnic groups. CF has a complex etiology, but it is chiefly a disease of electrolyte transport and is characterized by defects in fluid secretion by several epithelia, including the sweat duct, exocrine pancreas, and the pulmonary airways. The link between CF and a defect in cAMP-mediated Cl- transport in secretory epithelia was established in the early 1980s. Since then, numerous electrophysiological studies have focused on the characterization and regulation of individual Cl- channels underlying the macroscopic Cl- currents of secretory epithelia in the airways, sweat ducts, and gut. In this review the results of these studies in the light of current knowledge of the function of the CF gene product, the CF transmembrane conductance regulator (CFTR) protein, will be analyzed. The CFTR protein is a member of a family of ATP-binding proteins that act as unidirectional solute pumps. These proteins are membrane spanning, are found in both prokaryotic and eukaryotic cells, and have two ATP-binding domains. The family includes the p-glycoproteins that are involved with the expression of multidrug resistance in certain tumor cells. The majority of CF chromosomes (70%) have a single codon deletion that translates to a missing phenylalanine residue at position 508 (delta F508) of the protein. Unique for this family of proteins, the CFTR protein possesses an additional highly charged domain (the R domain) containing several consensus polypeptide sequences for kinase phosphorylation. Although CFTR bears structural resemblance to this family of ATP-dependent pumps, overexpression of the protein in a variety of different cell types is associated with the appearence of a cAMP-sensitive Cl- channel. We critically examine current information concerning the structure-function relationships of the CFTR protein obtained from both electrophysiological and biochemical approaches. We also summarize recent evidence suggesting that the CFTR protein may act as a pump and a channel, a hypothesis in keeping with the multifaceted nature of the disease.


Oncotic pressure regulates gene transcriptions of albumin and apolipoprotein B in cultured rat hepatoma cells.

The mechanism of the accelerated syntheses of albumin and apolipoprotein B (apo B) in response to decreased oncotic pressure was investigated in cultured rat hepatoma H4-II-E cells. Addition of dextran (mol wt 6-9 x 10(4)) to the culture medium decreased the levels of albumin and apo B mRNAs in an oncotic pressure-dependent manner. The reductions of both mRNAs were attenuated with increase in the molecular weight of dextran, which resulted in a decrease in oncotic pressure. Addition of macromolecule increased the viscosity in medium; however, alteration of viscosity appeared not to correlate with albumin and apo B mRNA levels. Transcriptional run-on assays with isolated nuclei from dextran-treated vs. untreated hepatoma cells indicated that the changes in steady-state mRNA levels were mainly controlled at the transcriptional step. Treatment with cycloheximide increased albumin mRNA to the basal level, which was effectively suppressed by dextran, and resulted in superinduction of apo B mRNA. These changes occurred primarily at the transcriptional step. These results suggest that regulations of the expressions of the albumin and apo B genes for adaptive increases in the mRNAs may require the continued synthesis of a labile protein(s) or a limiting transcription factor(s). We conclude that oncotic pressure plays an important role in regulation of expression of the albumin and apo B genes at the transcriptional step.


Low K+ increases Na(+)-K(+)-ATPase alpha- and beta-subunit mRNA and protein abundance in cultured renal proximal tubule cells.

Studies from this laboratory demonstrate that LLC-PK1/Cl4 cells, a cultured renal cell line, respond to incubation in low-K+ medium by coordinately increasing abundance of both alpha- and beta-subunits of Na(+)-K(+)-ATPase but increase only beta- and not alpha-mRNA levels (Lescale-Matys et al. J. Biol. Chem. 265: 17935-17940, 1990) and that alpha-abundance is likely increased as a result of increased efficiency of alpha-mRNA translation (L. Lescale-Matys and A. A. McDonough. J. Cell Biol. 111: 311A, 1990). The aim of this report was to determine if nontransformed kidney cells would respond to low K+ in a similar manner. We incubated primary cultures of rat proximal tubule cells in low K+ (0.25 mM) for up to 24 h to address this aim. Na(+)-K(+)-ATPase activity, measured enzymatically, and abundance of alpha- and beta-subunits, measured by immunoblot, were increased significantly and coordinately by 8 h of low K+, and, by 24 h of low K+, these parameters were increased to 2.17 +/- 0.34 (activity), 2.03 +/- 0.21 (alpha), and 2.39 +/- 0.48 (beta)-fold over control. Pretranslationally, beta-mRNA, measured by Northern blot analysis, increased to 1.76 +/- 0.35 after 3 h of low K+ and to 3.4 +/- 0.75-fold over control after 24 h of low K+. The increase in alpha-mRNA was smaller and delayed compared with the beta-mRNA response, but it was sufficient to account for the observed increase in alpha-protein and Na(+)-K(+)-ATPase activity at steady state: alpha-mRNA increased to 1.27 +/- 0.09 after 6 h and to 1.91 +/- 0.41-fold over control after 24 h in low K+. We conclude that the accumulation of sodium pumps in cultured renal proximal tubule cells, unlike LLC-PK1 cells, can be accounted for by increases in both alpha- and beta-subunit mRNA levels.


Right-angle light scattering to assay basal and regulated plasma membrane Cl- conductances.

We describe a simple and rapid technique for assaying both constitutive and regulated plasma membrane Cl- conductances. The method uses right-angle light scattering to measure the rate of swelling of cells in suspension, in which the anion conductance is rate limiting for swelling, due to introduction of high plasma membrane cation conductance using gramicidin. The technique was verified using Chinese hamster ovary cells and mouse L cells, both stably transfected with the cystic fibrosis transmembrane conductance regulator (CFTR), to confer a specific cAMP-activated Cl- conductance not normally present in these cell types. In agreement with results obtained using other methods for assaying Cl- permeability in these cells, forskolin stimulated a significant increase in plasma membrane Cl- conductance in CFTR-expressing cells, as indicated by an increase in light scattering. That the enhanced light scattering by the cells was the result of cell swelling due to NaCl influx was shown by ion substitution experiments, in which no forskolin-induced increase in light scatter occurred in N-methyl-D-glucamine Cl- or Na+ gluconate medium. Enhanced light scattering was also observed in both CFTR-expressing and control cells stimulated with the Ca2+ ionophore, ionomycin. Extracellular anion substitution, to exploit the inwardly directed halide gradient utilized in this protocol, enabled determination of the anion selectivities of both the cAMP- and Ca(2+)-activated Cl- channels. Thus this technique provides a simple optical method for rapidly assaying not only constitutive and regulated Cl- conductance pathways but also their anion selectivities.


Stretch-dependent regulation of atrial peptide synthesis and secretion in cultured atrial cardiocytes.

We have developed a novel system to study stretch-dependent secretion of atrial natriuretic peptide (ANP) using cultured neonatal rat atriocytes in vitro. Application of tension (i.e., 2 sequential stretches) to cells grown on a flexible culture surface effected a dose-dependent increase in immunoreactive (ir) ANP release into the medium. Analysis of atriocyte cytoplasmic RNA 24 h poststretch revealed an increase in ANP mRNA levels of about ninefold relative to the unstretched controls. Medium ATP levels, measured as an index of cellular damage, were similar in control and stretched cells. Furthermore, cooling the cultures to 0 degrees C suppressed both basal as well as stretch-stimulated release. These findings argue against cellular damage and nonspecific release of irANP as an explanation for the increase in medium immunoreactivity. Stretch was incapable of amplifying the secretory response to prostaglandin F2 alpha, suggesting possible overlap in the pathways whereby these stimuli effect release of the peptide. The calcium channel blocker verapamil had no effect on stretch-dependent irANP release, whereas calmidzolium, a calmodulin inhibitor, suppressed basal as well as stretch-dependent secretion, implying a potentially important relationship between intracellular calcium metabolism and irANP release.


NADPH diaphorase and nitric oxide synthase colocalization in enteric neurons of canine proximal colon.

Considerable evidence has recently been presented that suggests that nitric oxide (NO) is a nonadrenergic noncholinergic (NANC) neurotransmitter in gastrointestinal tissues. One of the criteria that must be satisfied before this hypothesis can be accepted is that enteric neurons must be shown to contain the enzymatic apparatus necessary to synthesize NO. Specific antibodies have been developed for NO synthase (NOS) isolated from rat cerebellum, and studies have shown that NOS copurifies and colocalizes with NADPH diaphorase activity, a commonly used neural marker. We used antibodies raised against the cerebellar NOS to determine the distribution of NOS-like immunoreactivity (NOS-LI) in enteric neurons of the canine proximal colon. We also tested whether NADPH diaphorase staining would label the population of neurons containing NOS-LI in this species. A subpopulation of neurons in myenteric and submucosal ganglia displayed NOS-LI and were colabeled with NADPH diaphorase. Labeled neurons had morphological characteristics similar to the Dogiel type I morphology. Cryostat sections showed NOS-positive nerve trunks throughout the circular and longitudinal muscle layers, but a high density of NOS-LI was observed within the submucosal pacemaker region, as predicted from physiological studies. These studies provide the first morphological support for the hypothesis that NO serves as a NANC neurotransmitter in the canine colon. The study also shows that the NADPH diaphorase reaction provides a useful method to label cells with NOS-LI.


Bronchial epithelial cells release neutrophil chemotactic activity in response to tachykinins.

The purpose of this study was to determine whether substance P (SP), neurokinin A (NKA), and neurokinin B (NKB) induce the release of neutrophil chemotactic activity (NCA) from bovine bronchial epithelial cells (BBEC) and whether neutral endopeptidase (NEP), a membrane-bound metalloenzyme that hydrolyzes tachykinins, modulates these effects. BBEC monolayers were exposed to SP, NKA, and NKB in the absence or presence of phosphoramidon (10(-6) M), a selective NEP inhibitor, for 72 h. Using a modified blind-well in vitro neutrophil chemotaxis assay, we found that tachykinin-exposed BBEC culture supernatant fluids induced significant neutrophil chemotaxis compared with supernatants obtained from unstimulated BBEC. Maximal effect was observed after 48 h of incubation and at SP concentration of 10(-13) M [92 +/- 3 (SP) vs. 64 +/- 2 (media) cells/high-power field (HPF), mean +/- SE, n = 7, P less than 0.05]. Release of NCA was mediated by the COOH-terminal of the SP molecule. The rank order of potency of tachykinins in inducing release of NCA was SP greater than NKA = NKB. SP-induced response was significantly potentiated by phosphoramidon (109 +/- 3 vs. 92 +/- 3 cells/HPF, n = 7, P less than 0.05), whereas other proteinase inhibitors had no effect. The released NCA was composed of protein and lipid-soluble components. These data indicate that mammalian tachykinins induce the release of NCA from BBEC and that NEP modulates these effects. We suggest that tachykinins regulate neutrophil recruitment into the lower respiratory tract, in part, by inducing the release of NCA from airway epithelial cells.


Cat tracheal gland cells in primary culture.

Conditions for the primary culture of isolated cat tracheal gland cells were established. Cells plated onto glutaraldehyde-fixed gels of rat tail collagen grew to confluency after 8 days of culture forming a monolayer of cuboidal cells with ultrastructural characteristics of epithelial cells and immunoreactivity to antikeratins. Cultured cells synthesized and released radiolabeled high-molecular-weight glycoconjugates. Glycoconjugate secretion was increased approximately 10% in response to the cholinergic agonist, carbachol. Secretion of glycoconjugates was unrelated to regulated exocrine secretion, since these cells were devoid of secretion granules as assessed by light and electron microscopy. Confluent cultures also generated a spontaneous potential difference and short-circuit current, which were both inhibited by ouabain and increased by carbachol. This suggested gland cells contribute to fluid secretion by active ion-transport mechanisms. We also plated cells onto unfixed collagen gels that were released from the culture dish at confluency. Cells were columnar with apically oriented secretion granules that stained with alcian blue and for blood group A immunoreactivity. Secretion of radiolabeled high-molecular-weight glycoconjugates was increased 27% by carbachol. These cell culture systems may serve as models to investigate glandular secretory mechanisms and their regulation.


Purification of plasma membranes by aqueous two-phase affinity partitioning.

A rapid method for purifying rat liver plasma membranes of high purity and yield is described. Squashed liver was homogenized in an aqueous polyethylene glycol-dextran two-phase system. After phase separation and reextraction of the bottom phase with fresh top phase, the combined polyethylene glycol-rich top phases were affinity partitioned in the presence of borate buffer with new bottom phase containing dextran-linked wheat-germ agglutinin. Under these conditions the lectin selectively pulled plasma membranes into the dextran-rich bottom phase, while other membranes preferentially distributed in the top phase. The lectin-containing bottom phase was reextracted with fresh top phase before collecting the purified plasma membranes by centrifugation. This protocol resulted in a preparation that was 30- to 40-fold enriched compared to the homogenate in plasma membrane markers for both the apical and basolateral domains and had yields of 55-70%. The contamination by other membranes was low. The entire procedure was completed within 90 min. The method should be useful for purifying plasma membranes also from other sources.


Development of gel staining techniques for detecting the secretion of procathepsin D (52-kDa protein) in MCF-7 human breast cancer cells.

17 beta-Estradiol stimulates the secretion of the 34- and 52-kDa protein (i.e., cathepsin D and procathepsin D, respectively) from MCF-7 human breast cancer cells and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) inhibits this estrogen-stimulated response. A comparison of the effects of 17 beta-estradiol, TCDD, and their combinations on the secretion of these two proteins was determined using four different assay procedures, namely autoradiographic analysis of the 35S-labeled proteins (from [35S]methionine) separated by polyacrylamide gel electrophoresis (PAGE), densitometric analysis of the silver- and double-stained proteins separated by PAGE, and radioimmunoassay of the proteins using commercially available antibodies to the 52-kDa protein. The results showed that the autoradiographic, staining, and radioimmunoassay procedures gave comparable results with only a few minor differences in the relative amounts of the 52-kDa detected in the various treatment groups. In the medium obtained from 17 beta-estradiol-treated cells that was serially diluted, there was an excellent linear correlation for the relative concentrations of the 52-kDa protein using the double-staining/densitometric procedure and the radioimmunoassay. These results indicate that the double- or silver-staining method may be a useful and rapid method for screening new compounds as antiestrogens in MCF-7 cells.


Determination of amino sugars and amino acids in glycoconjugates using precolumn derivatization with o-phthalaldehyde.

This report examines the RP-HPLC separation of o-phthalaldehyde derivatives of amino acids, amino sugars, and amino sugar alcohols using either 2-mercaptoethanol or 3-mercaptopropionic acid. A method with pmol sensitivity for the analysis of N-acetylamino sugars of glycoconjugates was elaborated. Upon hydrolysis, amino sugars are reduced with borohydride. Automated precolumn derivatization and chromatographic conditions for the resulting hexosaminitols are the same as those used for the analysis of amino acids. The method has been tested with as little as 2 micrograms of bovine fetuin, with a glycopeptide from bromelain and with an oligosaccharide after periodate oxidation.


Application of 3-[3-(3-(trifluoromethyl)diazirin-3-yl)phenyl]-2,3- dihydroxypropionic acid, carbene-generating, cleavable cross-linking reagent for photoaffinity labeling.

N-Hydroxysuccinimide ester of 3-[3-(3-(trifluoromethyl)diazirin-3-yl)phenyl]-2,3-dihydroxypro pionic acid was successfully tested in a ribosomal tRNA binding system. It is an originally designed trifluoromethyl-diazirine-based cleavable cross-linking reagent with a very short distance between the active points (about 8.5 A). The reagent was coupled to the amino acid amino group of Phe-tRNAPhe to obtain a photoactivatable analog of peptidyl-tRNA. This analog was bound to ribosomes and the complex was irradiated with uv light. After isolation, the cross-linked product was cleft by periodate treatment to reveal the properties of the new reagent.


The development of substance P-like immunoreactivity in the goldfish brain.

The development of substance P-like immunoreactivity (SPLI) in the goldfish brain was studied by means of the indirect peroxidase-antiperoxidase technique and an antibody to substance P. By 80 h after fertilization, the first SPLI-cell bodies appear in the ventricular zone of the future diencephalon and the first SPLI-fibers appear in the olfactory bulbs. Two days after hatching (which occurs at 100 h after fertilization), SPLI fibers connecting the olfactory bulbs and hypothalamus are seen. In the optic tectum SPLI-fibers appear for the first time 5 days after hatching. In the brain stem, SPLI-cell bodies appear in juvenile animals 40 days after hatching. The highest number and intensity of SPLI-cell bodies and fibers are found in the area postrema. SPLI-cell bodies are also seen in the gustatory nucleus, nucleus ambiguous, reticular formation of the medulla, dorsal motor nucleus of the vagus and commissural nucleus of Cajal. The significant information gained from the present study is: 1. The rostro-caudal sequence in which the SPLI appears in the developing nuclei of the goldfish brain 2. The reduction of SPLI-cell bodies in some nuclei with age Thus, in the brain stem, SPLI-cell bodies that were labeled in juvenile goldfish were not seen in adults. This might be due to changes in the rate of axonal transport, changes of the SP phenotype during development or cell death. The developmental sequence and relative timing in which SPLI-cell bodies appear in the goldfish, rat and mice are similar.


Use of hypertonic saline/dextran versus lactated Ringer's solution as a resuscitation fluid after uncontrolled aortic hemorrhage in anesthetized swine.

We tested the hypothesis that following aortotomy, administration of hypertonic saline/dextran increases hemorrhage and mortality. We also compared hypertonic saline/dextran with the standard therapy of attempting to replace three times the amount of lost blood with lactated Ringer's solution. In this model of uncontrolled arterial hemorrhage resulting from aortotomy, 24 anesthetized Yorkshire swine underwent splenectomy, stainless steel wire placement in the infrarenal aorta, and instrumentation with Swan-Ganz and carotid artery catheters. The wire was pulled, producing a 5-mm aortotomy and spontaneous intraperitoneal hemorrhage. The animals were randomly assigned to one of three study groups: control; hypertonic saline/dextran group in which six minutes after aortotomy a 4-mL/kg mixture of IV 7.5% NaCl and 6% Dextran-70 was given over one minute; or lactated Ringer's group in which six minutes after aortotomy 80 mL/kg IV lactated Ringer's was given over nine minutes. The volume of hemorrhage and the mortality rate in hypertonic saline/dextran-treated animals were significantly greater than in the nonresuscitated controls (1,340 +/- 230 mL versus 783 +/- 85 mL and five of eight versus zero of eight, respectively; P less than .05). Although the mortality rate in the lactated Ringer's group was not significantly different from the hypertonic saline/dextran group, survival time was significantly shorter than in the hypertonic saline/dextran group. In this model of uncontrolled hemorrhage, immediate IV administration of hypertonic saline/dextran significantly increased hemorrhage volume and mortality. However, the accentuation of hemorrhage and reduction in survival were not as great as that produced by the standard practice of attempting to replace the lost blood with three times that volume of lactated Ringer's.


Carotid artery resection for head and neck cancer.

Occasionally, the head and neck surgeon encounters a patient whose malignancy involves the carotid artery. In these patients, curative or palliative surgery may require excision of the common or the internal carotid artery. However, the high complication and death rates dissuade many surgeons from undertaking carotid artery resection. This study reviews the outcomes in 20 patients treated between 1979 and 1985 at the Department of Otolaryngology-Head and Neck Surgery, The University of Iowa Hospitals and Clinics, with resection of the carotid artery for head and neck cancer. The carotid artery was electively resected in 16 patients, while 4 patients underwent emergent carotid artery ligation. In the group of patients studied the stroke rate was 25%, the death rate 20%, and the combined stroke and death rate 30%. Of the patients who survived the procedure, all but 1 died of complications caused by tumor recurrence. These results are discussed, and compared with results from other studies.


[Current therapies of benign prostatic hypertrophy. Can the new medical treatments replace endoscopic resection?].

There has been a renewed interest in benign prostatic hypertrophy over the last two years due to the development of new therapeutic modalities. A recent consensus conference was organized by the World Health Organization (WHO) in Paris in July 1991, at which conventional treatment by transurethral resection was compared with new methods. An updated review is made of the basic concepts concerning BPH and the indications, technique and results of the various treatments: drug treatment by alpha-blockers, hormones, other instrumental treatments: cryotherapy, bladder neck incision, metal stent, hyperthermia. Transurethral resection will be studied in detail: technique, results. The respective indications for these various modalities are discussed.


[Long-term results of treatment of benign prostatic hypertrophy by trans-rectal prostatic hyperthermia].

From November 1987, 250 patients with symptomatic benign prostatic hyperplasia underwent transrectal prostatic hyperthermia. Forty-six patients had an indwelling catheter while 204 patients were obstructed but could still void spontaneously. Hyperthermia was administered in 5 or 10 sixty-minute sessions, with a calculated intraprostatic temperature of 42.5 or 43 degrees C. At the two-year follow-up, residual urine volume was significantly decreased while peak flow rate, maximum flow nomogram and subjective symptoms were only slightly improved, i.e. patients were still obstructed postoperatively. Transrectal prostatic hyperthermia cannot be considered as a first choice therapy for symptomatic benign hyperplasia but it can be offered only to carefully selected patients who cannot undergo any alternative therapeutic procedure.


[The alpha blockers].

Because the bladder neck contains a large number of alpha-adrenergic receptors, use of alpha-blockers to facilitate micturition in early benign prostatic hypertrophy has been advocated. Effects on micturition and urinary flow were fairly favorable. No adverse effects were recorded. The effect of therapy was transient and slightly greater than the effect of the placebo.


[Current role of cryosurgery in benign prostatic hypertrophy compared to recent minimally invasive methods].

The authors review results of 20 years of cryosurgery for benign prostatic hypertrophy based on 992 cases treated by this method. In particular, they compare the 500 more recent cases of cryocauterisation with the new minimally invasive techniques proposed:balloon dilatation, prostatic stent, thermotherapy, hyperthermia, medical treatment with a 5-alpha-reductase. A study of the cost of these treatments with similar performances places cryosurgery in a good position. It covers the largest range of therapeutic indications with the fewest contraindications in high-risk patients. Good long-term objective results were observed. In their current state of application, thermotherapy and hyperthermia are essentially palliative modalities.


Shared T-cell receptor gene usage in experimental allergic neuritis and encephalomyelitis.

Experimental allergic neuritis, an autoimmune disease of the peripheral nervous system, is a model for human Guillain-Barré syndrome. Experimental allergic neuritis is mediated by CD4+ T cells reactive with myelin P2 protein. We demonstrate that these T cells use the same members of T-cell receptor V gene families for both their alpha (V alpha 2) and beta (V beta 8) chains as T cells that cause experimental allergic encephalomyelitis, an autoimmune disease of the central nervous system. Furthermore, these T cells appear to be idiotypically related. Therefore, completely different T-cell lines with different antigen specificities, producing entirely different diseases, share common T-cell receptors.


Sensory neuropathy associated with monoclonal immunoglobulin M to GD1b ganglioside.

A 67-year-old woman with a sensory polyneuropathy was shown to have a serum monoclonal immunoglobulin M lambda antibody with a titer of 1:10,000 toward GD1b ganglioside. The immunoglobulin M also reacted with some other gangliosides containing disialosyl groups such as GD2, GD3, and GQ1b, but it did not react with GM1, LM1, or GD1a. The principal reactive ganglioside in human cauda equina was GD1b.