K+ channel involvement in induction of synaptic enhancement by mast cell degranulating (MCD) peptide. A bee venom, mast cell degranulating peptide (MCD), which induces long-term potentiation (LTP) of synaptic transmission in hippocampal slices, was found to possess multiple functions. They include (1) binding and thereby inhibiting a voltage-dependent K(+)-channel in brain membranes, (2) incorporation in a lipid bilayer to form voltage-dependent and cation-selective channels by itself, and (3) activation of a pertussis toxin (Ptx)-sensitive GTP-binding proteins. In this study, we prepared several derivatives and analogues of MCD and investigated which function is more closely related to the induction of LTP. Another bee venom, apamin, formed ion channels in a lipid bilayer which were indistinguishable from those formed by MCD. D-MCD, an optical isomer of MCD, activated a Ptx-sensitive GTP-binding protein. However, these peptides did not induce LTP in the hippocampal slices. A snake venom, dendrotoxin-I (DTX-I), bound to the same K(+)-channels as MCD and did induce LTP. These results suggest that the most potent aspect of MCD involved in LTP inducibility is its interaction with the voltage-dependent K(+)-channel.


Clinical and cytologic aspects of ocular late-phase reaction in the guinea pig. Conjunctival tissue of 12 guinea pigs (6 experimental animals, 6 controls) was sensitized with IgG1 (PCA titer 1:2,000) antisera to dinitrophenol carrier and challenged topically with hapten. Periorbital swelling, conjunctival edema and conjunctival redness were evaluated clinically at 0.5 h and hourly for 12 h, and again at 24 and 48 h. Tears were collected at each time point for cytology. Results showed an early-phase reaction (EPR) peaking at 0.5 h and a late-phase reaction (LPR) peaking at 5-7 h. Periorbital swelling was minimal 4 h after appearance of the EPR. Conjunctival edema and redness increased in both the early and late phases, but without two distinct peaks. However, individual animals in both groups did show biphasic reactions. Tear cytology showed an abnormal increase in neutrophils and eosinophils in the later time periods and was a significant way to detect ocular anaphylaxis. In conclusion, if the presence of two significant peaks of clinical inflammation after one antigen challenge is taken as the narrowest definition of a late phase in anaphylaxis, our results show an ocular LPR occurring in our animal model.


Clonazepam treatment of myoclonic contractions associated with high-dose opioids: case report. A 30-year-old man with chronic abdominal pain was treated with high doses of hydromorphone intravenously and developed severe and frequent myoclonic contractions. Several medications including lorazepam failed to control the contractions; however, clonazepam in normal doses reduced the myoclonus dramatically.


Effects of intrathecal antibodies to substance P, calcitonin gene-related peptide and galanin on repeated cold stress-induced hyperalgesia: comparison with carrageenan-induced hyperalgesia. Rats exposed to a cold environment (4 degrees C) for 30 min every 1 h during the day and at night show a gradual decrease in the nociceptive threshold for pressure stimulation. Such hyperalgesia, referred to as repeated cold stress (RCS)-induced hyperalgesia, is stable for at least 4 h and maintained for 3 days only by exposing to cold overnight; thus, no adaptation to RCS is apparent. Hyperalgesia gradually returns over 4 days after cold exposure ceases. To determine whether three neuropeptides, substance P (SP), calcitonin gene-related peptide (CGRP) and galanin (GAL), which are present in the superficial dorsal horn including primary afferent terminals, would be responsible for RCS-induced hyperalgesia, we examined the effects of intrathecal injections of their antibodies (used as inhibitors of neuropeptide-mediated synaptic transmission) on the nociceptive threshold of RCS rats, and compared this with the antibody effect on carrageenan-induced hyperalgesia. An intrathecal injection of anti-SP antibody significantly inhibited the hyperalgesia of RCS rats as well as carrageenan-induced hyperalgesia, and slightly increased the nociceptive threshold of non-RCS rats. Anti-CGRP antibody produced an improvement in the hyperalgesia of RCS rats as well as carrageenan-induced hyperalgesia without having an effect on the nociceptive threshold of non-RCS rats. Although anti-GAL antibody significantly inhibited carrageenan-induced hyperalgesia, it did not affect the nociceptive threshold of RCS and non-RCS rats. The present results suggest that enhancement of synaptic transmission mediated by SP and CGRP, but not GAL, in the spinal dorsal horn is, at least in part, involved in RCS-induced hyperalgesia.


[The keratinocyte].

[The keratinocyte]. This review summarizes recent data on the keratinocyte, the major cell type in the epidermis. A two-state model is proposed: in the "resting" state, the keratinocyte is committed to a program of terminal differentiation focusing on production of an efficient barrier, the stratum corneum, and of the epidermal basement membrane; the activated state, first discovered in cell culture studies, occurs in vivo during epidermal wound healing and inflammatory skin diseases.


[Biophysical characterization of the stratum corneum. Relationship between structure and properties]. Studies carried out over the last ten years have determined the structure and chemical composition of the stratum corneum and their significance for biologic function; the stratum corneum (SC) is composed of flat layers linked to each other by desmosome plates. Intercellular spaces are composed mainly of double lipid layers which are responsible for the main part of the barrier function of the SC. This barrier between the body and the environment must be effective despite the deformations and various insults (chemical, physicochemical) to which the SC is subjected. The SC fulfills its barrier function through remarkable physical properties of which most originate in an ability to bind water molecules to protein sites whose presence depends on the humidity of the environment in which the SC was produced. Lastly, the SC exhibits some enzyme activities and consequently can no longer be considered as an inert membrane.


2-5A synthetase expression in mice infected with Trypanosoma cruzi. This study describes the production and action of interferon in mice infected with Colombian and Y strain T. cruzi. The production of interferon was monitored by an in vitro assay of plasma and extract of spleen, lung and heart for interferon activity. The action of interferon in mice was assessed by measuring an interferon-mediated enzyme activity, 2-5A synthetase. Infected mice (strain Balb/c) were sacrificed at different time intervals, and the level of this enzyme was measured in extracts of spleen, lung and heart. Colombian strain infection induced higher levels of interferon than Y strain under the same conditions; consequently, a greater increase in 2-5A synthetase induction was observed in the former of the two strains. These results suggest that interferon produced by T. cruzi infected mice is active, since a variety of organs respond to its presence by producing elevated levels of 2-5A synthetase.


[Interactions between peptides and major histocompatibility complex molecules]. T-cell receptors recognize foreign proteins as peptide fragments associated with major histocompatibility complex (MHC) molecules. Properties of antigenic peptides, methods for detecting peptide-MHC molecule combinations, and the characteristics of the interaction are reviewed. Possible explanations for graft rejection and autoimmune disease are suggested in the light of these data.


Tachycardia sensing failure of an implantable cardioverter defibrillator in a patient with hypertrophic cardiomyopathy. A 17-year-old white male was found to have nonobstructive hypertrophic cardiomyopathy after suffering three severe syncopal episodes. He experienced an episode of sustained polymorphic ventricular tachycardia during exercise tolerance testing that required cardioversion. Electrophysiological studies were able to reproduce sustained polymorphic ventricular tachycardia that was unresponsive to standard pharmacotherapy. An automatic implantable defibrillator was placed. However, during implantation with the rate sensing electrodes on the left ventricle, it was found that the extremely polymorphic nature of the tachycardia caused such rapid fluctuations in the sensed R wave signal that the device could not properly detect the tachycardia. This was felt to be due to the automatic gain control circuit of the Ventak 1550. The problem was solved by moving the rate sensing electrodes to the lateral right ventricle. This case suggests that the unique arrhythmic substrate of hypertrophic cardiomyopathy may present sensing difficulties during automatic implantable defibrillator insertion.


Bedside termination of sustained ventricular tachycardia by transesophageal atrial pacing. Transesophageal atrial pacing was used to terminate hemodynamically stable sustained monomorphic ventricular tachycardia in two patients. The procedure was performed at the bedside, no anesthesia was required, there were no complications, and one of the patients went home after the procedure was performed. This method should be considered prior to using direct current cardioversion in patients with hemodynamically stable sustained monomorphic ventricular tachycardia.


Intraoperative identification of a radiofrequency lesion allowing validation of catheter mapping of ventricular tachycardia with a computerized balloon mapping system. Radiofrequency current was utilized to mark the anatomic location of earliest endocardial activation during catheter mapping of ventricular tachycardia. Intraoperative identification of the radiofrequency lesion allowed validation that the site of earliest endocardial activation determined by a catheter mapping study was the same as assessed by a computerized balloon mapping system. Radiofrequency current may be a useful method of marking areas of endocardium thought to be potential sites for ablative surgery as well as allowing correlation between different mapping techniques.


Ventricular fibrillation produced by stimulation of external transthoracic electrodes--an experimental study. The threshold for ventricular fibrillation was determined in 12 pentobarbital anesthetized dogs using transthoracic electrodes located at the optimal axillary electroventilation sites. Electroventilation is the name used to designate inspiration produced by stimuli applied to body surface electrodes. The optimal stimulation site for electroventilation was first determined using hand-held electrodes. Then electrodes, 4.1 cm in diameter, were sutured bilaterally to the optimal anterior axillary stimulation site. The threshold current for producing ventricular fibrillation was determined using single pulses ranging from 0.1-10 msec in duration delivered during the vulnerable period of the cardiac cycle. Fibrillation was produced in all dogs with the 10- and 5-msec pulse durations, in 11 dogs with 0.3-msec, in 6 dogs with 0.2-msec, and in 1 dog with 0.1-msec pulse duration. In all dogs, the current required to produce ventricular fibrillation increased greatly as the pulse duration was decreased. The current required for fibrillation was much in excess of that required to produce one tidal volume. With the longer duration pulses, the ratio was about 80. With the 8 microseconds duration pulses used for electroventilation the estimated ratio is about 800.


Slowing of ventricular tachycardia as a possible endpoint for serial drug testing at electrophysiological study. Certain patients who cannot be rendered noninducible by serial drug testing during electrophysiology study demonstrate significant slowing of their ventricular tachycardia rate with selected agents. We evaluated the characteristics and outcome of 19 such patients to assess whether this slowing could be considered an acceptable endpoint for treatment. This group consisted of 14 males and 5 females (mean age 63 +/- 9) with a mean ejection fraction of 28 +/- 13% and inducible sustained ventricular tachycardia. Sixteen patients had known coronary artery disease and 13 had prior myocardial infarction. The other three patients had idiopathic cardiomyopathy. Serial drug testing during an electrophysiology protocol that used up to three extrastimuli at two or three cycle lengths at two right ventricular sites was used to select a medication regimen that provided optimal ventricular tachycardia slowing without limiting side effects. Five patients were treated with amiodarone, three with Ic agents (all with ejection fraction greater than 30%), and the remainder with Ia and Ib agents alone or in combination. Mean initial ventricular tachycardia rate was 219 +/- 26 beats/min with posttreatment ventricular tachycardia rate 137 +/- 17 (mean initial cycle length 278 +/- 35 msec, posttreatment 443 +/- 53 msec). Two groups were identified, those who had recurrent (although well-tolerated) ventricular tachycardia (group I, n = 8, mean time to recurrence = 15 months), and those who did not (group II, n = 11, mean follow-up 22 months). Overall sudden death rate was 5%, while total mortality was 11% (all mortality in group I).(ABSTRACT TRUNCATED AT 250 WORDS)


Clinical evaluation of the safety of repetitive intraoperative defibrillation threshold testing. One goal of the initial implantation procedure for a cardioverter defibrillator is determination of the configuration and patch location with the lowest defibrillation threshold (DFT). To determine the safety of multiple defibrillation tests, an analysis of the intraoperative defibrillation threshold tests (DFTT) in our patients was performed. In 84 patients, the mean number of DFT trials was 5.27; the mean number of joules received was 275.0. The maximum number of shocks in one implant procedure was 50 for a total of 4,895 joules without complications. Four patients received 30 or more DFT shocks without complication. There were two complications related directly to the DFTT: one patient with severe noninflammatory cardiomyopathy developed electromechanical dissociation and was subsequently resuscitated and survived; the second patient with severe triple vessel coronary artery disease suffered an intraoperative myocardial infarction during testing and eventually died 22 days postoperatively. All patients received an ICD unit; six patients had DFTs of greater than 20 joules. Based on our experience, we followed the clinical status (heart rate, blood pressure, ECG changes, fluid status, total anesthesia time) during the DFTT to determine the extent and duration of our testing protocol. Multiple shocks due to repositioning of the leads in a stable patient should not prohibit extensive testing as adverse consequences do not appear to be cumulative.


Relationship between beat-to-beat changes in hemodynamic state and action potential duration of the left ventricle during rapid ventricular pacing in man. A relationship between beat-to-beat changes in hemodynamic state and action potential duration (APD) of the left ventricle was studied by pacing the right ventricle with a constant cycle length (400 msec) for 3 minutes and recording simultaneously the intraarterial pressure and left ventricular monophasic action potential in 16 patients (mean age 51 +/- 8 years) undergoing routine cardiac catheterization. The APD measured at the point of 90% repolarization (APD-90) shortened gradually from a baseline value of 305 +/- 25 msec to a minimum of 246 +/- 25 msec (P less than 0.001) by 160 +/- 10 seconds after the onset of pacing. After reaching the minimum duration, the APD and blood pressure were measured from 30 consecutive beats. The magnitude of beat-to-beat variation in the APD was directly correlated to variation in the mean arterial blood pressure (r = 0.65, P less than 0.01). Beat-to-beat changes in hemodynamic and electrical state were related in that an increase of at least 10 mmHg in the blood pressure of one beat was associated with an increase in the APD of the concomitant beat by at least 5 msec. In six patients with ventriculoatrial dissociation during the rapid ventricular pacing, the sequential ventriculoatrial pacing decreased the beat-to-beat variation of APD from 2.8% +/- 1.4% to 0.8% +/- 0.7% (P less than 0.01) and variation of blood pressure from 6.4% +/- 3.2% to 1.4% +/- 0.9% (P less than 0.01). The observed association between beat-to-beat changes in hemodynamic state and APD of the left ventricle demonstrates that an immediate force-interval relationship exists in the human left ventricle.


Adaptive rate pacing controlled by the right ventricular preejection interval: clinical experience with a physiological pacing system. In the Precept pacing system, the right ventricular intracardiac impedance waveform is used to evaluate either of two indicators of metabolic demand relative right ventricular stroke volume and preejection interval (PEI). PEI is known to reliably parallel contractility changes, which is reflective of physical and emotional stress. The stability and dynamic behavior of PEI were tested in ten patients with a Precept pacing system under various forms of exercise and during postural changes. Although significant patient-to-patient variability of the sensor values was observed, reflecting individual physiological differences, the chronic stability of PEI was excellent in the total device experience of 147 months. In all patients, PEI shortened significantly during bicycle ergometry from a mean value of 137.7 +/- 17.8 (range 96-162) to a mean value of 103.0 +/- 21.6 (range 92-109) (P less than 0.05). Low level bicycle exercise of short duration resulted in a prompt decrease in PEI and increase in pacing rate in all patients. There were no uniform postural responses overall, although some posture related rate changes were observed in two patients. We conclude that the first generation of a PEI based pacing system holds promise for adaptive rate pacing.


The value of transesophageal atrial pacing in predicting the efficacy of antiarrhythmic drugs in patients with paroxysmal narrow QRS complex tachycardia. Transesophageal atrial pacing (TAP) is used in the diagnosis and treatment of paroxysmal narrow QRS complex tachycardia (NQT). The aim of this study was to assess the value of this technique in predicting the efficacy of antiarrhythmic therapy. The study group consisted of 30 consecutive patients with spontaneous NQT whose clinical tachycardia was inducible by TAP. Baseline TAP was performed off all antiarrhythmic medication and repeated during oral antiarrhythmic drug therapy. The pacing protocol consisted of three stages: a single extrastimulus introduced at progressively shorter coupling intervals during sinus rhythm, pacing at incremental rates to the point of second-degree AV block, and bursts of rapid pacing. On repeat stimulation while on oral antiarrhythmic therapy (37 pacing studies) NQT was still inducible in 12 cases. During the follow-up period ten patients developed a recurrence of NQT:nine cases out of 12 (75%), in whom NQT was inducible while on antiarrhythmic therapy, and one case out of 25 (4%), in whom NQT was not inducible (P less than 0.001). The sensitivity of TAP in predicting the outcome of the patients with NQT was 90%, and the specificity 89%. The negative predictive value of TAP (prediction of no recurrence of NQT) was 96%, and the positive predictive value (prediction of recurrence of NQT) was 75%. We conclude that TAP is a simple and accurate method for predicting the efficacy of antiarrhythmic treatment in patients with NQT.


A double-blind study of submaximal exercise tolerance and variation in paced rate in atrial synchronous compared to activity sensor modulated ventricular pacing. To assess the variation in paced rate during everyday activity and the importance of atrioventricular synchronization (AV synchrony) for submaximal exercise tolerance, atrial synchronous (DDD) and activity rate modulated ventricular (VVI,R) pacing were compared in 17 patients with high degree AV block. The patients were randomly assigned to either mode and evaluated by treadmill exercise to moderate exertion and by 24-hour Holter monitoring after 2 months in the DDD and VVI,R modes, respectively. At the end of the study, the patients were programmed to the pacing mode corresponding to the preferred study period. During the treadmill test, the mean exercise time to submaximal exertion (Borg 5/10), exertion ratings and respiratory rate did not differ between pacing modes despite a significantly lower ventricular rate in the VVI,R mode. The atrial rate during VVI,R pacing was significantly higher than the ventricular rate, but did not differ from the ventricular rate during DDD pacing. There was a diurnal variation in paced rate in both pacing modes. Paced ventricular rate was, however, higher and variation in paced rate greater in DDD compared to VVI,R pacing. Nine patients preferred the DDD mode, three patients preferred the VVI,R mode, while five subjects did not express any preference. The results from this study indicate that the variation in paced rate during activity sensor-driven VVI,R pacing does not match that during DDD pacing neither during everyday activities nor during submaximal treadmill exercise. Nevertheless, no differences in exercise time, Borg ratings, and respiratory rate during submaximal exercise were found. Thus, for most patients with high degree AV block, DDD and VVI,R pacing seem equally satisfactory for submaximal exercise.


TU alternans, long QTU, and torsade de pointes: clinical and experimental observations. T or U wave alternans in association with long QTU and torsade de pointes (TdP) is uncommon and its mechanism(s) is unknown. We studied three patients with TU alternans, long QTU, and TdP: patient 1 was a newborn with congenital long QTU; patient 2 had marked hypokalemia and hypomagnesemia; and patient 3 was receiving procainamide. In the three patients, TU alternans was tachycardia dependent and preceded the onset of TdP. In the patient on procainamide, TU alternans and TdP occurred at long cardiac cycles. In this patient, endocardial monophasic action potential (MAP) recordings showed that TU alternans was associated with alternation of the duration of the plateau. A deflection consistent with early afterdepolarization (EAD) arose at a constant time interval from phase 0 but alternated from high and low levels of phase 3. The first ectopic beat of TdP arose on the descending limb of the EAD. TU alternans was investigated by MAP recordings in six normal dogs, following the administration of anthopleurin-A (AP-A), a drug shown to delay sodium inactivation and to induce bradycardia dependent long QTU, EADs, and TdP. In two dogs TU alternans was associated with 2:1 recordings of EAD and nearly constant plateau duration. In three dogs, TU alternans was associated with EAD that occurred in consecutive beats at constant time intervals from phase 0, but alternated from high and low phase 3 because of alternation of the duration of the plateau. In one dog, alternation of EAD and plateau duration occurred. In 36 separate episodes of TdP that were analyzed in the six dogs, 32 were bradycardia dependent but four developed on abrupt shortening of the cardiac cycle associated with alternation of action potential duration. Our results suggest: (1) TU alternans may be due to 2:1 propagation of an EAD or to alternation of the recovery kinetics of a repolarization current; (2) The constant occurrence of EAD in relation to phase 0 in spite of alternation of plateau duration suggests an ionic mechanism synchronized to depolarization; (3) Tachycardia dependent TdP in clinical and experimental examples of long QTU seems to be characteristically associated with TU alternans. Dispersion of repolarization may underlie the increased ventricular electrical instability in these cases.


Effect of successful coronary angioplasty on the signal-averaged electrocardiogram. The effect of successful coronary artery angioplasty on the signal-averaged electrocardiogram (SAECG) was examined in 50 patients (41 men, 9 women, aged 55 +/- 8 years) with stable (26 patients) or unstable angina (24 patients) and good overall left ventricular function (ejection fraction = 55% +/- 8%). The SAECG was recorded before and within 24-48 hours after the angioplasty and was filtered at 40-250 Hz, with 250 beats averaged. The noise level averaged 0.57 +/- 0.15 microV before and 0.56 +/- 0.17 microV after the procedure. There was no overall significant difference between pre- and postangioplasty SAECGs. Subgroup analysis showed that 14 patients had a significant increase of the root mean square voltage of the last 40 msec of the filtered QRS that was independent of noise level changes, previous myocardial infarction, stable or unstable angina status, positive or negative baseline SAECG, or vessel being dilated. Eleven patients (22%) had late potentials at baseline, of whom four (36%) lost them after angioplasty, while one patient developed them after the procedure, all due to root mean square voltage changes. Thus, successful angioplasty exerted no significant overall effect on the SAECG, suggesting that the substrate of late potentials was not grossly altered by the procedure in our patients. However, there appear to be some patients, constituting approximately one third of this study population, who derive a favorable influence on the SAECG from angioplasty, a subgroup that needs to be further defined in future studies.


Syncope--brain or heart? A case report.

Syncope--brain or heart? A case report. A 44-year-old man suffered from recurrent episodes of unconsciousness, without any other concomitant manifestations. After routine workup, EEG and CT had proven nondiagnostic, prolonged Holter monitoring revealed a single episode of asystole, lasting 7.6 seconds. A pacemaker was inserted but did not abolish his episodic syncope. Subsequently, long-term EEG recording revealed epileptiform activity with independent foci in both temporal lobes. Antiepileptic treatment relieved the patient of his symptoms. This case illustrates the intimate relationship between the heart and the brain that sometimes lies behind syncope.


The next step in cardiac pacing: the view from 1958. At a 1-day conference sponsored by the Rockefeller Institute in September 1958, physicians and engineers held a confused debate over future directions for the young field of cardiac pacing. Many of the existing impediments to long-term pacing for chronic illness received little attention; the participants focused instead on whether atrial synchrony would be a requirement for long-term pacing. The Rockefeller conference illustrates the observation that cardiac pacing has undergone several major redefinitions. This process is rarely a smooth one but involves uncertainty, intense debate, and explicit choice.


Anti-desmin vs. anti-actin for quantifying the area density of prostate smooth muscle. Anti-desmin and anti-actin are commercially available antibodies that bind to smooth muscle. The present study was designed to compare the staining properties of anti-desmin and anti-actin in the human prostate in order to determine the optimal antibody for quantifying the smooth muscle content of the human prostate. Nineteen male subjects with symptomatic BPH underwent needle biopsy of the prostate. Double-immunoenzymatic staining was performed with peroxidase-anti-peroxidase (PAP) and alkaline phosphatase-anti-alkaline phosphatase (APAAP) techniques. Rabbit anti-desmin:mouse anti-human prostatic acid phosphatase and mouse anti-actin:rabbit anti-human prostatic acid phosphatase were utilized. Computer assisted color image analysis was performed using the Bioquant image analysis system. The percent area density of stroma and epithelium was independent of the antibodies used. The percent area density of smooth muscle in the anti-actin stained tissue sections was twofold greater than the anti-desmin stained tissue sections. A direct relationship was observed for the area density of smooth muscle (r = 0.71; P = 0.0006) and the area density of connective tissue (r = 0.82; P less than 0.001) determined from anti-desmin and anti-actin stained tissue sections. Anti-actin represents the optimal antibody for quantifying the area density of prostate smooth muscle. The reproducibility of the immunoenzymatic staining technique is inferred from the direct relationship observed for area density of epithelium between the different staining techniques.


Inhibition of growth of PC-82 human prostate cancer line xenografts in nude mice by bombesin antagonist RC-3095 or combination of agonist [D-Trp6]-luteinizing hormone-releasing hormone and somatostatin analog RC-160. The effects of treatment with a bombesin receptor antagonist [D-Tpi6, Leu13 psi (CH2NH) Leu14]BN(6-14)(RC-3095) and the combination of an agonist of luteinizing hormone-releasing hormone [D-Trp6]-LH-RH and somatostatin analog D-Phe-Cys-Tyr-D-Trp-Lys-Val- Cys-Trp-NH2 (RC-160) were studied in nude mice bearing xenografts of the hormone-dependent human prostate tumor PC-82. During the 5 weeks of treatment, tumor growth was decreased in all treated groups compared with controls. Bombesin antagonist RC-3095 and the combination of [D-Trp6]-LH-RH and RC-160 caused a greater inhibition of tumor growth than [D-Trp6]-LH-RH or RC-160 alone as based on measurement of tumor volume and percentage change in tumor volume. The largest decrease in tumor weight was also seen in the groups treated with the bombesin antagonist and with the combination of RC-160 and [D-Trp6]-LH-RH. Serum prostatic-specific antigen levels were greatly decreased, and insulin-like growth factor I (IGF-I) as well as growth hormone levels were reduced in all treated groups. Specific binding sites for [D-Trp6]-LH-RH, epidermal growth factor (EGF), IGF-I, and somatostatin (SS-14) were found in the tumor membranes. Receptors for EGF were significantly down-regulated by treatment with the bombesin antagonist or RC-160. Combination of LH-RH agonists with somatostatin analog RC-160 might be considered for improvement of hormonal therapy for prostate cancer. The finding that bombesin antagonist RC-3095 inhibits the growth of PC-82 prostate cancer suggests the merit of further studies to evaluate the possible usefulness of antagonists of bombesin in the management of prostatic carcinoma.


Immunohistochemistry and biochemistry in detection of androgen, progesterone, and estrogen receptors in benign and malignant human prostatic tissue. The relative distribution of androgen (AR), progesterone (PR), and estrogen receptors (ER) was localized and estimated in human prostate tissue by immunohistochemistry in five normal tissue samples, in eight benign hyperplastic (BPH) samples, in nine primary cancers, and in seven prostate cancer metastases. Moreover, three prostatic cancer cell lines (LNCaP, DU 145, and PC 3) were analyzed. A comparison between the results obtained by radioligand binding assays and immunohistochemistry was performed for the AR and PR. Using immunohistochemistry, the AR was exclusively detected in the nuclei of both benign and malignant prostatic epithelial cells. The highest proportion of AR-positive cells was found in BPH and in prostate cancer metastases as compared with normal prostatic tissue. In a majority of the cases, the PR was only present in the nuclei of stromal cells. Benign hyperplastic prostates contained higher proportions of PR-positive cells as compared with primary carcinoma. PR was sparse in epithelial cells. ER-positive stromal cell nuclei were only detected in carcinomatous prostates. A few ER-positive epithelial cell nuclei were found in one sample each of a BPH and normal prostate. All cells from the androgen-dependent, LNCaP, cell line and a majority of the cells from the androgen-independent, DU 145, cell line were AR-positive. In contrast, the cells from the androgen-independent, PC 3, cell line were all AR-negative. All three cell lines were PR- and ER-negative. The radioligand binding technique detected the AR in extracts from both the cytosol and the nucleus. Again BPH contained higher amounts of AR as compared with normal prostatic tissue. The LNCaP cells contained high amounts of cytosolic AR while cells from the DU 145 and PC 3 cell lines lacked detectable AR as estimated by biochemical techniques. There seemed to be a discrepancy between biochemically measured and immunohistochemically estimated receptor content.


Prognostic significance of antigenic heterogeneity, Gleason grade, and ploidy of lymph node metastases in patients with prostate cancer. We retrospectively evaluated 51 prostate cancer patients found to have pelvic lymph node metastases at the time of pelvic lymphadenectomy and 125I implantation. All of them were followed until death or for a minimum of 70 months. Rabbit polyclonal anti-PSA, anti-PAP, anti-PSP-94, and mouse TURP-27 monoclonal antibodies were used in immunohistochemical evaluation of the metastatic lesions. In addition, Gleason grade and ploidy were assessed and correlated. No tumor with a Gleason grade of less than 7 could be found in the metastatic lymph nodes. Time to progression (P = .003), disease-specific survival (P = .009), and overall survival (P = .003) were significantly shorter in patients whose tumors had a primary Gleason pattern of 5 (grade 9 or 10). In the PSA study, patients whose tumors were reactive in more than 75% of cancer cells experienced significantly longer survival than those with less than 75% of cancer cells expressing PSA (P = .0006 log rank test). The means of overall survival +/- SEM were 71.5 +/- 5.0 and 34.9 +/- 5.4 months, respectively. Similar correlations were found with disease-specific survival and time to progression. Patterns of PAP expression and TURP-27 reactivity were not prognostically useful, whereas PSP-94 expression may add some additional information. These data suggest that evaluation of tissue PSA heterogeneity in lymph node metastases may offer additional prognostic information on prostate cancer patients. Better prediction of individual prognosis may be possible with the combined use of Gleason grade, flow cytometry, and PSA expression.


Inhibitory effect of dicumarol on the excitability of the sarcolemma of skeletal muscle. Dicumarol (7.5 x 10(-6) M -7.5 x 10(-5) M) inhibited the twitches and the simultaneously recorded electromyogram of the indirectly stimulated rat diaphragm preparation. The directly stimulated curarized preparation was inhibited in a similar way. Accordingly, the excitability of the sarcolemma was inhibited by dicumarol. The inhibitory effect was irreversible upon washing. Intracellular microelectrode recordings disclosed that the inhibitory effect was accompanied by a depolarization, and the inhibition was potentiated in high K+ (9.5 mM) solution. In addition, the ability of the preparations to produce a depolarization contracture with KCl (100 mM) was inhibited by dicumarol. Therefore, a part of the inhibitory effect might be caused by the depolarization. On the other hand, subthreshold direct stimulation disclosed that the inhibitory effect might be due to an increase of the threshold. However, a lack of synergistic interaction with lidocaine (0.2 mM) showed that this effect of dicumarol was probably not a membrane stabilizing or local anaesthetic effect.


Reversible alterations in myocardial gene expression in a young man with dilated cardiomyopathy and hypothyroidism. Thyroid hormone effects on myocardial gene expression have been well defined in animal models, but their relationship to the pathogenesis of cardiac dysfunction in hypothyroid humans has been uncertain. We evaluated a profoundly hypothyroid young man with dilated cardiomyopathy. Before and during 9 months of thyroxine therapy, serial assessment of myocardial performance documented substantial improvements in the left ventricular ejection fraction (16-37%), left ventricular end-diastolic diameter (7.8-5.9 cm), and cardiac index (1.4-2.7 liters.min-1.m-2). Steady-state levels of mRNAs encoding selected cardiac proteins were measured in biopsy samples obtained before and after thyroxine replacement. In comparison with myocardium from nonfailing control hearts, this patient's pretreatment alpha-myosin heavy-chain mRNA level was substantially lower, the atrial natriuretic factor mRNA level was markedly elevated, and the phospholamban mRNA level was decreased. All of these derangements were reversed 9 months after restoration of euthyroidism. These observations in an unusual patient with profound myxedema and cardiac dilatation permit correlation between the reversible changes in myocardial function and steady-state mRNA levels in a cardiomyopathy. They suggest that alterations in gene expression in the dilated myopathic heart may be correctable when a treatable cause is identified.


Parathyroid hormone-related protein in the rat urinary bladder: a smooth muscle relaxant produced locally in response to mechanical stretch. Parathyroid hormone-related protein (PTHrP) gene expression in the pregnant rat uterus has been shown to be dependent on occupancy of the uterus by the fetus. To further test the hypothesis that the synthesis of PTHrP in smooth muscle tissue is regulated by mechanical stretch, we conducted experiments using the rat urinary bladder as a model of an expansible hollow organ. The results indicate that PTHrP mRNA levels do change in response to the stretch of the bladder wall. Under normal conditions PTHrP mRNA levels in the bladder correlated with the urine volume-namely, the extent of bladder distension. When bladders were maintained empty in vivo, PTHrP mRNA levels decreased gradually. Conversely, when bladders were distended by the accumulation of urine, levels of PTHrP mRNA increased dramatically with time. When distension was limited to one-half of the bladder, the increase in PTHrP mRNA was observed only in the distended portion. Histochemical studies performed on distended bladder tissue indicated the presence of PTHrP immunoreactivity in smooth muscle cells. Isolated organ bath studies were used to examine the possible physiological role of PTHrP in smooth muscle tonicity. In vitro responsiveness of bladder muscle strips to exogenous PTHrP was dependent on the in vivo condition of the bladder. In muscle strips obtained from bladders kept empty in vivo, PTHrP-(1-34)-NH2 relaxed carbachol-induced contraction in a dose-dependent manner but failed to relax the contraction in muscle strips from distended bladders that had high endogenous PTHrP expression. These results and the previous findings in the rat uterus suggest a physiological role of PTHrP in bladder smooth muscle function.


Purification of a distinctive form of endotoxin-induced nitric oxide synthase from rat liver. An endotoxin-induced form of nitric oxide synthase (EC 1.14.23) was purified to homogeneity from rat liver by sequential anion-exchange chromatography and affinity chromatography using 2',5'-ADP-Sepharose. The enzyme has a subunit molecular mass of 135 kDa as determined by SDS/PAGE, a maximum specific activity of 462 nmol of citrulline formed from arginine per min per mg, and a Km for arginine of 11 microM. The enzyme was strongly stimulated by the addition of calmodulin with an EC50 of 2 nM, but removal of free calcium from the assay medium only reduced activity by 15%. Calmodulin inhibitors significantly reduced the enzyme activity. Tetrahydrobiopterin, FAD, and FMN were all required for full enzyme activity. This form of endotoxin-induced nitric oxide synthase from liver differs from the inducible enzyme found in macrophages and is unusual in that it is stimulated by calmodulin with little dependence on the calcium ion concentration.


Phosphorylation-dependent epitopes of neurofilament antibodies on tau protein and relationship with Alzheimer tau. We have studied the phosphorylation of tau protein from Alzheimer paired helical filaments, of tau from normal human brain, and of recombinant tau isoforms. As a tool we used monoclonal antibodies against neurofilament protein [Sternberger, N., Sternberger, L. & Ulrich, J. (1985) Proc. Natl. Acad. Sci. USA 82, 4274-4276] that crossreact with tau in a phosphorylation-dependent manner. This allowed us to deduce the state of phosphorylation in normal and pathological tau, as well as antibody epitopes. The epitope of antibody SMI33 is at the first Lys-Ser-Pro sequence motif (residues 234-236) and requires an unphosphorylated Ser-235. Antibody SMI31 binds between Ser-396 (in the second Lys-Ser-Pro motif) and Ser-404, both of which must be phosphorylated. SMI34 has a conformational epitope that depends on the interaction between regions on either side of the microtubule-binding region; it also requires phosphorylation. The phosphorylatable serines detected by the SMI antibodies are part of Ser-Pro motifs and can be phosphorylated by a protein kinase activity that can be used to induce a paired helical filament-like state in human brain tau in vitro. The phosphates are incorporated in several stages that can be identified by antibody reactivity and gel shift. This suggests a role for the phosphorylation sites in Alzheimer disease, as well as the involvement of a Ser-Pro-directed protein kinase.


A family of concanavalin A-binding peptides from a hexapeptide epitope library. The lectin concanavalin A (Con A) binds methyl alpha-D-mannopyranoside (Me alpha Man) as well as alpha-D-mannosyl groups at the nonreducing terminus of oligosaccharides. Ligand peptides that mimic the binding of Me alpha Man to Con A were identified from screening an epitope library composed of filamentous phage displaying random hexapeptides. A consensus sequence was identified among affinity-purified phage; Con A binds phage bearing this sequence and is inhibited from doing so by Me alpha Man. When tested for binding against a panel of lectins, phage bearing this sequence bind only weakly to a closely related D-mannose-binding lectin, indicating that binding to Con A is highly selective. A synthetic peptide bearing the consensus sequence blocks the precipitation of Con A by dextran with an inhibition strength equivalent to that of methyl alpha-D-glucopyranoside. These results demonstrate that the specificity of Con A is not limited to carbohydrates and that highly selective sugar-mimics for lectins of plant, animal, or bacterial origin may be identified from epitope libraries.


Brief visual experience induces immediate early gene expression in the cat visual cortex. Brief visual experience causes rapid physiological changes in the visual cortex during early postnatal development. A possible mediator of these effects is the immediate early genes whose protein products are involved in the rapid response of neurons to transsynaptic stimulation. Here we report evidence that the levels of immediate early gene mRNAs in the visual cortex can be altered by manipulating the visual environment. Specifically, we find that brief (1 h) visual experience in dark-reared cats causes dramatic transient inductions of egr1, c-fos, and junB mRNAs in the visual cortex but not in the frontal cortex. Levels of c-jun and c-myc mRNAs are unaffected. These results suggest that select combinatorial interactions of immediate early gene proteins are an important step in the cascade of events through which visually elicited activity controls visual cortical development.


Random PCR mutagenesis screening of secreted proteins by direct expression in mammalian cells. We have developed a general method for screening randomly mutagenized expression libraries in mammalian cells by using fluorescence-activated cell sorting (FACS). The cDNA sequence of a secreted protein is randomly mutagenized by PCR under conditions of reduced Taq polymerase fidelity. The mutated DNA is inserted into an expression vector encoding the membrane glycophospholipid anchor sequence of decay-accelerating factor (DAF) fused to the C terminus of the secreted protein. This results in expression of the protein on the cell surface in transiently transfected mammalian cells, which can then be screened by FACS. This method was used to isolate mutants in the kringle 1 (K1) domain of tissue plasminogen activator (t-PA) that would no longer be recognized by a specific monoclonal antibody (mAb387) that inhibits binding of t-PA to its clearance receptor. DNA sequence analysis of the mutants and localization of the mutated residues on a three-dimensional model of the K1 domain identified three key discontinuous amino acid residues that are essential for mAb387 binding. Mutants with changes in any of these three residues were found to have reduced binding to the t-PA receptor on human hepatoma HepG2 cells but to retain full clot lysis activity.


Immunoelectron microscopic localization of the ubiquitin-activating enzyme E1 in HepG2 cells. As the first enzyme in the ubiquitin system the ubiquitin-activating enzyme E1 plays a pivotal role in all pathways of protein ubiquitination. In an effort to learn more about the cell biology of this pathway, we have purified the 110-kDa enzyme to homogeneity and generated a panel of distinct monoclonal antibodies to it. Using quantitative electron microscopic immunolocalization with these anti-E1 monoclonal antibodies, we find that E1 is abundant both within the cytoplasm and nucleus. Within the cytoplasm, E1 was found throughout the cytoplasmic volume as well as enriched along the cytoplasmic face of the rough endoplasmic reticulum and associated with the dense material along the desmosomal junctions. E1 was also found associated with the cytoplasmic surface of endosomal/lysosomal vacuoles. Interestingly, E1 was also found within the mitochondria. The lumen of rough endoplasmic reticulum, Golgi complex, endosomes, and lysosomes were negative. The specific localization of E1 to distinct subcellular organelles suggests that E1 may play multiple physiological roles within the cell.


A1 adenosine-receptor antagonists activate chloride efflux from cystic fibrosis cells. A1 adenosine-receptor-antagonist drugs such as 8-cyclopentyl-1,3-dipropylxanthine (CPX) and xanthine amine congener (XAC) are found to activate the efflux of 36Cl- from CFPAC cells. These cells are a pancreatic adenocarcinoma cell line derived from a cystic fibrosis (CF) patient homozygous for the common mutation, deletion of Phe-508. The active concentrations for these compounds are in the low nanomolar range, consistent with action on A1 adenosine receptors. In addition, drug action can be blocked by exogenous agonists such as 2-chloroadenosine and also can be antagonized by removal of endogenous agonists by treatment with adenosine deaminase. Cells lacking the CF genotype and phenotype, such as HT-29 and T84 colon carcinoma cell lines, appear to be resistant to activation of chloride efflux by either drug. CFPAC cells transfected with the CF transmembrane regulator gene, CFTR, are also resistant to activation by CPX. We conclude that, since these antagonists are of relatively low toxicity and appear to act somewhat selectively, they might be considered as promising therapeutic candidates for CF.


Cloning of a rat kidney cDNA that stimulates dibasic and neutral amino acid transport and has sequence similarity to glucosidases. The transport of amino acids across cell membranes is believed to be mediated by integral membrane proteins with distinct substrate specificities. Using expression cloning in Xenopus oocytes and assaying for the uptake of 14C-labeled cystine, we isolated a 2.3-kilobase cDNA (D2) from a rat kidney library. D2 is expressed specifically in kidney and intestine and induces the transport of both neutral and cationic amino acids. The deduced amino acid sequence predicts a 78-kDa protein with a single transmembrane domain, a structure not typical of the known membrane transport proteins, which generally have multiple membrane-spanning regions. The putative extracellular region is highly similar to the 4F2 heavy-chain cell surface antigen and to a family of alpha-glucosidases, which raises the possibility that D2 encodes a transport activator or regulatory subunit.


Expression cloning of a cDNA from rabbit kidney cortex that induces a single transport system for cystine and dibasic and neutral amino acids. We have isolated a cDNA clone by screening a rabbit kidney cortex cDNA library for expression of sodium-independent transport of L-arginine and L-alanine in Xenopus laevis oocytes. Expressed uptake relates to a single component of sodium-independent transport for dibasic and neutral amino acids. This transport activity resembles the functionally defined system b0,+ and carries cystine and dibasic amino acids with high affinity. The rBAT (b0,+ amino acid transporter-related) mRNA is found mainly in kidney and intestinal mucosa. It encodes a predicted 77.8-kDa protein with only one putative transmembrane domain and seven potential N-glycosylation sites. This protein could either be a constitutive element or a specific activator of system b0,+.


Stimulation of system y(+)-like amino acid transport by the heavy chain of human 4F2 surface antigen in Xenopus laevis oocytes. A kidney cortex cDNA clone (rBAT) has recently been isolated, which upon in vitro transcription and capping complementary RNA (cRNA) and injection into Xenopus laevis oocytes induces a system b0,(+)-like amino acid transport activity. This cDNA encodes a type II membrane glycoprotein that shows significant homology to another type II membrane glycoprotein, the heavy chain of the human and mouse 4F2 surface antigen (4F2hc). Here we demonstrate that injection of human 4F2hc cRNA into oocytes results in the activation of a cation-preferring amino acid transport system that appears to be identical to the y(+)-like transport already present in the oocyte. This is based on the following results: (i) Injection of in vitro transcripts from 4F2hc cDNA (4F2hc cRNA) into oocytes stimulates up to 10-fold the sodium-independent uptake of L-arginine and up to 4.1-fold the sodium-dependent uptake of L-leucine. In contrast, 4F2hc cRNA does not increase the basal sodium-independent uptake of L-leucine. (ii) Basal and 4F2hc cRNA-stimulated sodium-independent uptake of L-arginine is completely inhibited by L-leucine in the presence of sodium. Similarly, the basal and 4F2hc cRNA-stimulated sodium-dependent uptake of L-leucine is entirely inhibited by L-arginine. (iii) The stimulation of sodium-independent uptake of L-arginine and the stimulation of sodium-dependent uptake of L-leucine induced by injection of 4F2hc cRNA are both completely inhibited by dibasic L amino acids and to a lesser extent by D-ornithine. (iv) Both basal and 4F2hc cRNA-stimulated sodium-independent uptake of L-arginine show two additional characteristics of the system y+ transport activity: inhibition of L-arginine uptake by L-homoserine only in the presence of sodium and an increase in the inhibition exerted by L-histidine as the extracellular pH decreased. Our results allow us to propose that an additional family of type II membrane glycoproteins (composed by rBAT and 4F2hc) is involved in amino acid transport, either as specific activators or as components of amino acid transport systems.


Sequence and molecular characterization of human monocyte/neutrophil elastase inhibitor. cDNA encoding human monocyte/neutrophil elastase inhibitor (EI), a M(r) approximately 42,000 protein with serpin-like functional properties, has been sequenced. The 1316-base-pair sequence was obtained from overlapping clones and amplified DNA from libraries of monocyte-like and neutrophil-like cells. Hybridization with EI cDNA identified three EI mRNA species of 1.5, 1.9, and 2.6 kilobases in U937 monocyte-like cells and no hybridizing mRNA in lymphoblastoid cells lacking detectable EI. The cDNA open reading frame encodes a 379-amino acid protein, of which 167 residues were confirmed by tryptic peptides. Although EI may function extracellularly as well as intracellularly, its deduced sequence lacks a typical cleavable N-terminal signal sequence. Sequence analysis established that EI is a member of the serpin superfamily. EI has greatest homology (50.1% identity of amino acids) with plasminogen activator inhibitor 2, also a monocyte protein, and ovalbumin and gene Y, which were previously grouped as an ancient branch of the serpin superfamily. The extent of EI identity with the functionally related serpin alpha 1 antitrypsin is only 30.1%. Sequence alignment indicates that the reactive center P1 residue is Cys-344, consistent with abrogation of elastase inhibitory activity by iodoacetamide and making EI a naturally occurring Cys-serpin. The cleavable bond, Cys-Met, suggests an oxidation-sensitive molecule capable of inhibiting more than one serine protease. Oxidation sensitivity would limit the place of action of EI to the immediate vicinity of carrier cells. The molecular structure will help clarify the likely role of EI in regulating protease action and preventing tissue damage by phagocytic cells.


Predominant expression and activation-induced tyrosine phosphorylation of phospholipase C-gamma 2 in B lymphocytes. The triggering of T- or B-cell antigen-specific receptors is accompanied by rapid tyrosine phosphorylation of distinct cellular substrates, one of which is the gamma 1 isoform of inositol phospholipid-specific phospholipase C (PLC-gamma 1). This phosphorylation event, mediated by a putative protein tyrosine kinase coupled to the antigen receptor, probably stimulates the enzymatic activity of PLC-gamma 1, thereby promoting inositol phospholipid hydrolysis and other downstream signal transduction events. Recently, another ubiquitously expressed PLC isoform, PLC-gamma 2 (which shares 50.2% amino acid homology with PLC-gamma 1), has been identified. PLC-gamma 2-specific antibodies were used to evaluate the distribution and potential signaling role of this isoform in lymphocytes. Here, we report that, in contrast to T lymphocytes that express predominantly PLC-gamma 1, the major isoform expressed in murine and human resting B cells is PLC-gamma 2. Among B-cell tumor lines, all five murine B-lymphoma lines tested and one of six human B-lymphoblastoid cell lines also expressed predominantly PLC-gamma 2. However, three other human lines preferentially expressed PLC-gamma 1, and two others displayed similar levels of the two PLC-gamma isoforms. Furthermore, the triggering of B-cell surface immunoglobulin by anti-receptor antibodies was accompanied by a rapid tyrosine phosphorylation of PLC-gamma 2, which peaked after 5 min of stimulation. Conversely, and in agreement with recent reports, triggering of the T-cell antigen receptor complex led to the predominant phosphorylation of PLC-gamma 1 on tyrosine. These findings identify PLC-gamma 2 as a substrate for a B-cell putative protein tyrosine kinase coupled to the antigen receptor and suggest that its tyrosine phosphorylation constitutes a critical and early event in B-cell activation and, furthermore, that PLC-gamma 1 and PLC-gamma 2 may participate in similar but distinct signal transduction pathways in lymphocytes.


Expression cloning of the murine and human interleukin 9 receptor cDNAs. Interleukin 9 (IL-9) is a T-cell-derived lymphokine that induces the proliferation of various lymphoid and hemopoietic cells. A cDNA clone encoding the murine IL-9 receptor was isolated by expression cloning in COS cells and screening with 125I-labeled IL-9. Transient expression of this cDNA produced high-affinity binding sites for IL-9. The predicted 52-kDa protein contains a putative signal peptide and a typical transmembrane domain. A cDNA for the human homologue was isolated by cross-hybridization. Transfection of this cDNA in a murine T-cell clone conferred responsiveness to human IL-9. Sequence analysis revealed that the IL-9 receptor belongs to the recently described hematopoietin receptor super-family and is expressed in membrane-bound and soluble forms.


Presenting a routine screening test in antenatal care: practice observed. People's knowledge of screening tests for which they are eligible and which they may have undergone is frequently low. The aim of the current study is to determine the extent to which this is due to how a test is offered and explained. Routine consultations (n = 102) between midwives, obstetricians and pregnant women were tape-recorded to determine how a routine screening test for fetal abnormalities (maternal serum alpha-fetoprotein) is presented. The test was presented in the vast majority of consultations. Overall, little information was provided about the test, the conditions screened for, and the meaning of either a positive or a negative result. Screening was presented in such a way as to encourage women to undergo the test. The way in which routine prenatal screening is presented is unlikely to maximise informed decisions about whether to participate in this screening programme. Factors likely to be influencing test presentation include knowledge, attitudes and skills of staff, as well as the attitudes of pregnant women. The results of this study highlight a need to train the heath professionals implementing screening programmes in how to inform people fully about low probability but serious events without alarming them unduly, or reassuring them falsely.


Percutaneous drainage of 335 consecutive abscesses: results of primary drainage with 1-year follow-up. Retrospective review of percutaneous abscess drainage (PAD) of 335 abscesses in 323 consecutive patients was undertaken. Particular attention was directed to body location, associated organ system, communications and fistulae, and to the underlying immunologic status of the patient. One-year follow-up was available in all patients. Overall, the cure rate was 62.4% (209 of 335 abscesses), with a failure rate of 8.95% (30 of 335 abscesses). There were 14.2% (46 of 323 patients) deaths in the follow-up period, of which 4.6% (15 of 323 patients) were believed attributable to sepsis or septic complications. The overall complication rate was 9.8% (33 of 335 abscesses), most of which were minor in nature. For the patient exhibiting immunocompromise, representing 53.1% (172 of 323 patients) of the patient population, the cure rate was 53.4% (95 of 178 abscesses), which was significantly lower than the cure rate of 72.6% (114 of 157 abscesses) for the immunocompetent patient population (n = 151) (P less than .001). The recurrence rate was 2.1% (seven of 335 abscesses), with all recurrences within 3 months of initial drainage. PAD is effective and permanent treatment for both immunocompromised and immunocompetent patients.


Localized prostate cancer: use of serial prostate-specific antigen measurements during radiation therapy. The serum half-life of prostate-specific antigen (PSA) after radical prostatectomy is about 3 days; to the authors' knowledge, the PSA half-life during radiation therapy (RT) has not been investigated with weekly serial measurements. To determine the rate of decline and the half-life of PSA, serial measurements were obtained during 6-8 weeks of external-beam RT for localized prostate cancer. PSA values were determined immediately before and approximately 24 hours after the first dose of RT; thereafter, weekly measurements were made. There was a downward trend in PSA levels in 19 patients, with a median half-life of 58.5 days; the mean decline was 1.6% per day. However, in four patients, PSA levels either rose and fell to pre-RT values or increased steadily. The effect of digital rectal examination (DRE) on PSA levels was also analyzed. When the dates of DRE and subsequent PSA levels were inspected, no increase in PSA levels subsequent to DRE was found, although three of the four patients in whom PSA levels did not decrease underwent multiple DREs. The authors found a statistically significant (P = .023) transient elevation in the mean PSA values after the first fraction of RT (2 Gy) was administered; the mechanism and importance of which are not known.


Relative abundance of human placental phospholipase A2 messenger RNA in late pregnancy. The aim of this study was to determine messenger RNA (mRNA) levels for a specific phospholipase A2 (PLA2) in human placenta during late gestation prior to the onset of labour. The relative abundance of human placental mRNA for a nonpancreatic Group II PLA2 was determined using cDNA clone specific for this PLA2. Twenty seven placentae were collected from women undergoing elective (i.e. before the onset of labour) Caesarean section between 37 and 41 completed weeks gestation. The relative amount of human placental PLA2 mRNA did not change significantly over this period of late pregnancy (p greater than 0.8). Previously, we have demonstrated that placental PLA2 messenger RNA levels are increased in association with spontaneous onset labour at term in the human. The data obtained in this current study are consistent with the conclusion that the regulation of this human placental PLA2 gene is tightly controlled around the time of parturition and that its expression is increased acutely in association with labour.


Serum prostate-specific antigen in monitoring the response of carcinoma of the prostate to radiation therapy. In order to assess the value of serum prostate-specific antigen (PSA) levels in the monitoring of patients with localized prostatic carcinoma undergoing radical radiation therapy, 146 previously untreated patients were entered into this study. Sixty to 70 Gy were administered to the prostate over 8 to 9 weeks. Serum PSA levels were measured prior to radiotherapy, every 3 months during the first year and every 6 months thereafter. Median follow-up was 28 months. Pretreatment PSA values exceeded 10 ng/ml in 62% (91/146). Initial PSA values were correlated with tumor size and Gleason score. Six months after completion of radiation therapy, PSA levels decreased as compared to initial value in 88.3% of the patients. It had fallen to 10 ng/ml or less in 54 (59%) out of the 91 patients with initial abnormal PSA levels. Patients whose initial PSA exceeded 50 ng/ml attained levels of 10 ng/ml or less in only 19% of the cases (6/32). Only 3 of the 55 patients (5.5%) with both initial and 6-month PSA values less than or equal to 10 ng/ml developed metastasis. Out of the 91 patients with initial PSA values over 10 ng/ml, 54 (59.6%) had a 6-month PSA level of 10 ng/ml or less, and only 4/54 (7.4%) relapsed. By contrast, 13 of the 37 patients (35.1%) with a 6-month PSA value persistently above 10 ng/ml relapsed. The 3-year relapse-free survival is 85.1% for patients with a 6-month PSA level less than or equal to 10 ng/ml, and 50.2% for patients with persistently elevated PSA values.(ABSTRACT TRUNCATED AT 250 WORDS)


[Role of interferon in the treatment of renal cancer in adults]. Alpha interferon is effective in the treatment of metastatic renal cell cancer. To obtain a maximal efficacy, alpha interferon should be used at a dose of 10 MU/m2, or 18 MU total dose, given daily or three times a week. At such doses interferon produces flu-like symptoms and severe asthenia and thus should be proposed only to patients who have a good performance status. The response rate is low, in the order of 15-20%, and the median remission duration is short (6-10 months). However a few patients experience very durable remissions, sometimes for more than 5 years. The response rate may possibly be improved to 20% to 35% by combining interferon alpha with antineoplastic agents or to other cytokines, namely interleukin 2. Finally because of its supposed mechanisms of action-direct tumor growth inhibition and enhancement of the host's immune defenses--interferon may be of benefit in the adjuvant setting.


[Stress and nursing. Comparison of the levels of burnout between surgical departments and intensive care units]. Burnout is a state of workers of helping professions that results from a situation of work stress and is characterised by emotional exhaustion, depersonalization and low personal accomplishment. Burnout levels were measured in three nursing staffs: two of them working in intensive care units and one in a surgical department. The results show significant statistical differences between the two staffs of intensive care units. Such differences seem to be connected with the psychological climate evaluated in the units.


Incidence of atypical bronchioloalveolar cell hyperplasia of the lung: relation to histological subtypes of lung cancer. The incidence of atypical bronchioloalveolar cell hyperplasia (ABH) of the lung was investigated to evaluate the possibility of this lesion being a precancerous stage in the histogenesis of adneocarcinoma. Lobectomy and pneumonectomy specimens of 165 primary and 45 metastatic tumour cases were step-sectioned horizontally and examined histologically. An average of 51 blocks were taken in each case. Sixty-seven ABHs up to 10 mm in diameter were detected, only 2 lesions being associated with scar tissue. Age was one factor apparently related to ABH development, although not the major one. There was no correlation between smoking index and ABH occurrence. In males, the incidence was highest in association with adenocarcinoma (25.5% of cases, 0.8% of sections), followed by large cell carcinoma (25.0% of cases), squamous cell carcinoma (10.5% of cases) and metastatic tumours from other sites (4.8% of cases). In females, ABH was also more common together with adenocarcinoma (8.3% of cases) than with metastatic tumours (4.0% of cases). The differences in male incidences by case and by section between the adenocarcinoma and metastatic tumour categories were statistically significant (P less than 0.05, P less than 0.01 respectively) indicating that ABH may be a precancerous lesion capable of transformation of adenocarcinoma.


The distribution of factor XIIIa-positive cells in the human fetus and placenta. Immunohistochemical staining for factor XIIIa, a transglutaminase, revealed a variety of positively stained cells in human fetal tissues. Factor XIIIa-positive cells were most numerous in the dermis and connective tissues. Numerous large, stellate cells in placental villi, decidua, and chorionic membranes also expressed factor XIIIa at 7-9 weeks gestational age, before the onset of fetal hematopoiesis. There was heterogeneity in the staining for factor XIIIa in the early and late fetal tissues, in both rounded and in dendritic cells. In preparations of consecutive sections and in double-labelling experiments, some cells expressed both factor XIIIa and certain monocyte markers and were identified in close association with blood vessels and lymphoid organs in the late fetus and in the placental villi at the end of gestation. Other rounded and dendritic cells expressed factor XIIIa but not monocyte markers, and were found in adult and fetal connective tissues at all gestational ages. These results suggest that there are two factor XIIIa-positive cell populations. One population is present at all developmental stages, does not express monocyte markers, and probably differentiates in situ from primitive mesenchyme. The other population appears mainly after the onset of fetal hematopoiesis, coexpresses some monocyte markers, is HLA-DR positive and may be capable of antigen presentation.


Electron microscopic study of paired helical filaments and cerebral amyloid using a novel en bloc silver staining method. A one step en bloc silver staining method which was originally established to study nucleolar organizer regions has been applied for the demonstration of both paired helical filaments (PHF) and extracellular cerebral amyloids in semi-thin sections and at the electron microscopic level. The three forms of PHF can be visualized: (1) neurofibrillary tangles are shown in all stages from first appearance in form of intracellular patches of PHF to severely degenerated shadow-like "ghost" tangles; (2) neuropil threads are distinctly stained in great numbers; and (3) PHF are easily detected as neuritic components in amyloid plaques. All forms of fibrillar extracellular amyloid structures, i.e. "diffuse", "classical" and "burnt out" plaques, are well demonstrated; congophilic angiopathy reveals amyloid preferentially in arteries and arterioles of the leptomeninges and cortex ranging from small circumscribed patches to large circumferential amounts with occasional plaque-like condensations or broad loose accumulations of amyloid; perivascular cuffs and laminar subpial deposits of amyloid are stained as well. At the electron microscopic level all lesions are clearly visible in non uranyl/lead-stained specimens, characterized by varying numbers of silver grains on a pale background. The detailed demonstration of structures in archival material, which had been stored in paraffin and re-embedded for electron microscopy, is due to the demonstration of argyrophilic structures by the protective colloidal developer of gelatin and formic acid and to the proteolytic resistance of insoluble PHF and extracellular amyloids in plaques and congophilic angiopathy.


Paraffin section immunohistochemistry in the diagnosis of Hodgkin's disease and anaplastic large cell (CD30+) lymphomas. Morphological and immunohistological studies were carried out on a series of 137 lymphomas including CD30+ anaplastic large cell (ALC) lymphomas (48 cases) and non-lymphocyte predominant Hodgkin's disease (HD) (89 cases), with the aim of assessing in situ expression of a combination of antibodies including anti-CD30/BerH2, epithelial membrane antigen (EMA), CD15 and CD45, in addition to other monoclonal antibodies suitable for paraffin tissues. A greater proportion of cases of ALC lymphomas than of HD exhibited positivity for CD45 (91.7% vs 17.6%), EMA (56.2% vs 4.5%), CD43 (53.6% vs 13.1%) and CD45RO (39.5% vs 3.5%), whereas Reed-Sternberg (RS) cells in HD most frequently expressed CD15 (93.2% vs 20.8%) antigen. Moreover, in 35 of 48 (72.9%) ALC lymphomas tumour cells expressed the CD30+, CD45+, CD15-, EMA- or + phenotypic profile, while in the same percentage (62/85) of HD cases RS cells were found to express the CD30+, CD45-, CD15+, EMA- profile. This study suggests that the differential expression of CD45, EMA, and CD15 may be used in the separation of ALC lymphomas and HD. However, co-expression of CD30, CD45 and CD15 antigens by RS cells in HD (14/85 cases, 16.5% in this series) and by tumour cells in ALC lymphomas (9/48 cases, 18.7% in this series) may be encountered in a non-negligible fraction of cases.


Estimates from two survey designs: national hospital discharge survey. The methodology for the National Hospital Discharge Survey (NHDS) has been revised in several ways. These revisions, which were implemented for the 1988 NHDS, included adoption of a different hospital sampling frame, changes in the sampling design (in particular the implementation of a three-stage design), increased use of data purchased from abstracting service organizations, and adjustments to the estimation procedures used to derive the national estimates. To investigate the effects of these revisions on the estimates of hospital use from the NHDS, data were collected from January through March of 1988 using both the old and the new survey methods. This study compared estimates based on the old and the new survey methods for a variety of hospital and patient characteristics. Although few estimates were identical across survey methodologies, most of the variations could be attributed to sampling error. Estimates from two different samples of the same population would be expected to vary by chance even if precisely the same methods were used to collect and process the data. Because probability samples were used for the old and new survey methodologies, sampling error could be measured. Approximate relative standard errors were calculated for the estimates using the old and new survey methods. Taking these errors into account, less than 10 percent of the estimates were found to differ across survey methodologies at the 0.05 level of significance. Because a large number of comparisons were made, 5 percent of the estimates could have been found to be significantly different by chance alone. When there were statistically significant differences in nonmedical data, the new methods appeared to produce more accurate estimates than the old methods did. Race was more likely to be reported using the new methods. "New" estimates for hospitals in the West Region and government-owned hospitals were more similar than the corresponding "old" estimates to data from the census of hospitals conducted by the American Hospital Association. The numerous significant differences in estimates for bed size categories between the two survey methodologies reflected the change in the universe and definition of beds for the new survey. Few statistically significant differences were found in the medical data using the old and the new survey methods. Two main differences, in estimates for cataract and alcohol dependence syndrome, may have resulted from problems with the new survey. A measurement error, reporting outpatients to the NHDS, is one possible explanation of the higher estimates for diagnosis of cataract using the new survey methods.(ABSTRACT TRUNCATED AT 400 WORDS)


National hospital discharge survey.

National hospital discharge survey. This report presents statistics on the utilization of non-Federal short-stay hospitals based on data collected through the National Hospital Discharge Survey from a national sample of the hospital records of discharged inpatients. Estimates are provided by the demographic characteristics of patients discharged, geographic region of hospitals, conditions diagnosed, and surgical and nonsurgical procedures performed. Measurements of hospital use include frequency, rate and percent of discharges and days of care, and average length of stay.


Influence of the cryoprotective agents glycerol and hydroxyethyl starch on red blood cell ATP and 2,3-diphosphoglyceric acid levels. Because hydroxyethyl starch (HAES) is used for volume replacement therapy and as a cryoprotectant for frozen red blood cells (RBCs), this compound, in contrast to glycerol, does not require labor-intensive removal from thawed cells prior to transfusion. We here report the effect of both glycerol and HAES on the RBC organic phosphates ATP and 2,3-diphosphoglyceric acid (2,3-DPG). The CPD-A1-stabilized RBCs of 20 healthy donors (3 females, 17 males) were separately frozen in either 40% glycerol or 6% HAES, of molecular weight 200,000. ATP and 2,3-DPG concentrations were determined in CPD-A1 RBCs before addition of cryoprotectant and in cryopreserved thawed RBCs after 24 h storage at -80 degrees C (glycerol) and -196 degrees C (HAES). It appears that HAES, but not glycerol, significantly reduces ATP concentrations whereas both lead to a reduction of 2,3-DPG concentrations; this reduction was more pronounced with glycerol than with HAES. Experiments with the blood of 6 donors demonstrated that HAES affects autohemolysis by 16%, in contrast to glycerol, after which cryoprotectant autohemolysis was affected by 3.1% only. RBC recoveries were comparable using glycerol or HAES as cryoprotectants. A distinct pattern of reduction of 2,3-DPG levels by glycerol and less by HAES, and of ATP levels by HAES but not by glycerol, emerges. Our findings may be of importance if HAES is to be introduced as a convenient cryoprotectant.


Evaluation of four different methods for platelet freezing. In vitro and in vivo studies. This report describes our experience with various techniques for the freezing of platelet-rich plasma, removed from the final product after leukapheresis procedures performed on 14 hematological patients. A total of 194 platelet units were frozen for subsequent autologous transfusion, by the following four methods: (1) 6% dimethyl sulfoxide (DMSO); (2) a combination of 5% DMSO/6% hydroxyethyl starch; (3) 3% glycerol; (4) 5% glycerol/4% glucose. Each technique was evaluated by measuring the percentage of platelet recovery, malondialdehyde (MDA) production, and lactate dehydrogenase release. To investigate the safety and therapeutic effectiveness of the previously frozen platelets, in vivo comparison of four platelet freezing methods was made in 8 thrombocytopenic patients, using corrected platelet increment (CCI), determined at 24 h. Our in vitro results indicate that the cryopreservation with 6% DMSO, without controlled cooling rate, provides significantly (p less than 0.05) greater platelet recovery (75%) as compared to other systems. The decrease of MDA production and the increase in plasma lactate measured after the thawing process was less in the DMSO-frozen units than in the other platelet units. When platelets, cryopreserved by this method, were subsequently transfused into patients, a significantly better CCI (greater than 5,000/microliters) was obtained. In our series, 6 patients were entirely supported with frozen autologous platelets. It appears from this study that a better understanding of the physical and biochemical events occurring during the freezing process will improve platelet cryopreservation, allowing a more systematic use of frozen platelets in the support of thrombocytopenic patients.


[Tubal rupture after prostaglandin instillation despite decreasing beta-HCG values]. Over the past years laparoscopic surgery has become widely accepted in the treatment of tubal pregnancy and instillation of prostaglandin is well established. However, the failure rate is around 20%. This report describes a case of tubal pregnancy treated according to this procedure where the therapy initially seemed to be successful. Instillation of prostaglandin F2 alpha was followed by decreasing beta-HCG values, which continued to decrease after discharge. Thus, rupture of the operated tube on day 17 after surgery was completely unexpected.


[Diagnosis of chronic pancreatitis. Studies of duodenal juice after stimulation with the secretin-ceruletide test. Decision limits and evaluation of various parameters]. 187 patients were checked up over 4 years by the secretin-ceruletide test. Independently of the test results they were assigned to various disease groups on the basis of clinical assessment. 131 subjects were divided in a pilot investigation into: subjects with a healthy pancreas (n = 55); subjects with chronic pancreatitis (n = 50); subjects whose pancreatic condition could not be classified clearly (n = 26). 8 parameters were compared by univariate and multivariate statistical procedures in order to confirm or rule out the presence of chronic pancreatitis. The discriminatory power of the following parameters in duodenal fluid proved to be sufficiently high, with less than 15% frequency of misclassification: chymotrypsin (activity) and/or; lipase (activity) and/or; amylase (activity); viscosity. Under routine conditions measurement of the activity of two of these enzymes is sufficient. Their contribution to discrimination proved to be approximately equal. The diagnostic sensitivity and specificity of the parameters bicarbonate, lipase (concentration), trypsin (activity) and volume of duodenal fluid are lower. The classification rules derived from the above pilot group were confirmed by a diagnostic study under routine condition in a test group of 38 patients. Limitation to examining only volume and a maximum of 3 parameters which proved best in distinguishing between patients with chronic pancreatitis and healthy subjects, together with the omission of the first-hour samples after a secretin bolus, considerably reduced laboratory workload without altering the discriminatory power of the secretin-ceruletide test.


Protection from herpes simplex virus type 2 is associated with T cells involved in delayed type hypersensitivity that recognize glycosylation-related epitopes on glycoprotein D. Immunization of mice with a vaccinia recombinant (VP176) that expresses a fully glycosylated herpes simplex virus (HSV) glycoprotein D (gD) induces long-term (greater than or equal to 50 days) HSV-specific lymphoproliferation and delayed type hypersensitivity (DTH) responses, the ability to eliminate a high challenge dose of HSV-2 from the epidermis and protection from fatal disease due to HSV replication in the nervous system. Adoptive transfer studies indicate that protection is mediated by the DTH functions of L3T4+ cells and requires the contribution of a non-specific irradiation-sensitive cell. Long-term protection (defined as that seen at greater than or equal to 50 days after immunization) from fatal HSV-2 challenge, virus clearance from the epidermis, and HSV-specific T-cell responses are not induced by a partially glycosylated gD expressed by a vaccinia recombinant (VP254) in which gD is controlled by a late vaccinia virus promoter. However, mice immunized with VP254 are protected from HSV-2 challenge early (day 10) after immunization. The VP254-induced protection is HSV-specific, but it is not mediated by L3T4+ and Lyt2+ cells. The findings are discussed within the context of future developments of anti-HSV vaccines.


Heterotopic pancreas of the stomach. Histogenesis and immunohistochemistry. Ordinary histological investigation has suggested that heterotopic pancreas of the stomach may have two types of histogenesis; one is development from immigrated fetal pancreas tissue, and the other is development from primitive gastric mucosal epithelium following penetration into the submucosa with subsequent erroneous differentiation into pancreas tissue. It is suspected that type-I lesions include the majority of cases caused by immigration from fetal pancreas, and that some type-II cases arise through erroneous differentiation of primitive gastric mucosal epithelium. With regard to immunohistochemical findings, cells positive for pancreatic polypeptide and amylase were much more numerous in the acini of type-I cases compared with type-II cases. Positive cells were found not infrequently in the acini of type-II cases after staining for pancreatic polypeptide, insulin, glucagon, somatostatin, serotonin, and gastrin. On the other hand, a small number of cells in islets were not infrequently positive for alpha 1-antitrypsin, alpha 1-antichymotrypsin, and amylase. It is considered that in the heterotopic pancreas, ductal cells have the potential to differentiate into acinar cells and islet cells, as is the cases in the orthotopic pancreas.


Establishment of a human hepatoblastoma model in athymic nude mice. Producibility of fatty acid-binding protein, alpha-fetoprotein, and alpha 1-acid glycoprotein. An athymic nude mouse model of human hepatoblastoma, designated HBL-2, was established. HBL-2 produced alpha-fetoprotein (AFP) and the serum level in tumor-bearing mouse was roughly proportional to the tumor weight. Reactivity of AFP to Concanavalin A and Lens culinaris lectins indicated that it contained predominantly fucosylated, non-bisecting N-acetylglucosamine as a sugar moiety. Alpha 1-acid glycoprotein (AAG) was also found in both HBL-2 tumor extract and sera of tumor-bearing mice. The serum level, however, was not proportional to either the tumor weight or the value of the tumor extract. In addition, fatty acid-binding protein (FABP) was detected in approximately 40% of tumor cells and in tumor extract with polyclonal antibody against liver FABP. Thus, HBL-2 is an unique model of human hepatoblastoma, in that it produces FABP in addition to AAG and AFP.


Some observations on the localization of hyaluronic acid in adult, newborn and embryonal rat brain. Hyaluronic acid was localized in acetone-fixed cryostat sections of brain and spinal cord obtained from adult, newborn and embryonal rat. The sections were incubated with glial hyaluronate-binding protein (GHAP) of human origin and the protein was visualized by indirect immunofluorescence with monoclonal antibodies raised to human GHAP and not staining rat brain by immunofluorescence. GHAP is a brain extracellular matrix (ECM) glycoprotein, approximately 60,000 molecular weight, which is structurally related to the HA-binding region of cartilage ECM proteins. The distribution of hyaluronate in adult brain white matter and cerebellar cortex was similar to that previously reported for GHAP. In both cases, the reaction product formed a mesh surrounding myelinated axons and granule cells. Hyaluronate was also found in parts of the brain that did not contain GHAP. A finely reticulated mesh was observed in the neuropil between cell bodies in cerebral cortex and basal ganglia. Scattered cortical neurons were surrounded by a rim of reactive material. Perineural staining was the rule rather than the exception in spinal cord anterior horn motoneurons, inferior olivary nucleus, large bulbar reticular neurons and dentate nucleus of cerebellum. The only part of the brain which appeared relatively free of hyaluronate was the molecular layer of the cerebellum. In newborn and embryonal rat, the densely packed cell bodies in cerebral gray matter, periventricular germinal layer and external granular layer of cerebellum were surrounded by hyaluronate. Small droplets of hyaluronate were observed in between the cylindrical epithelial cells lining the neural tube in 11 day embryos. Non-myelinated fiber tracts and the molecular layer of the developing cerebellum were relatively unstained. No hyaluronate was detected in the ependyma lining the cerebral ventricles and the central canal of the spinal cord.


The structural and neurochemical development of the fetal guinea pig retina and optic nerve in experimental growth retardation. In this study we have examined structural and neurochemical aspects of retinal and optic nerve development in experimentally growth-retarded fetal guinea pigs following maternal unilateral artery ligation. Eye weight (n = 4) and total retinal area (n = 6) at 62 days gestation (term approximately 66 days) were both relatively spared when expressed as a percentage of body weight but in absolute terms were significantly reduced by 18% (P less than 0.001) and 13% (P less than 0.05) respectively when compared with age-matched controls. The numerical density of neurons in the ganglion cell layer was significantly higher at both 52 days (n = 4) and 62 days (n = 4) in growth-retarded fetuses compared with controls. However, there was no difference between the groups in the total number of neurons in this retinal layer at either age, since retinal areas are reduced in growth retardation. The area of neuronal somata in the ganglion and inner nuclear layers was significantly reduced in growth-retarded fetuses compared with controls. There was a concomitant reduction in the width of the cellular layers in the retina and also in the plexiform (synaptic) and photoreceptor layers. The growth of the outer segments of the photoreceptor layer was particularly affected in peripheral retina. The higher packing density of cells and the reduced growth of the plexiform layers suggests a reduction in the growth of the neuropile in growth-retarded fetuses compared with controls. The radial bundling of ganglion cell axons coursing across the retina to enter the optic nerve head was poorly defined in growth retardation. In addition myelination was delayed in the optic nerve with the numerical density of myelinated axons being significantly reduced (P less than 0.005) in growth-retarded fetuses compared with controls. There was a significant reduction (P less than 0.01) in the number of amacrine cells in the inner plexiform layer expressing Substance P-like immunoreactivity in growth-retarded fetuses compared with controls. Glutamate-like immunoreactivity was most intense in the five laminae of the inner plexiform layer and in the outer plexiform layer and less pronounced in photoreceptors, ganglion cells and their axons. There was no qualitative difference in glutamate immunoreactivity between control and growth-retarded fetuses in any of these structures. Thus we have shown that intrauterine growth retardation has specific effects on the development of the fetal guinea pig retina, reducing the growth of several types of neurons and their processes and affecting the expression of the neuropeptide substance-P in amacrine cells.


Identification of a functional mononuclear precursor of the osteoclast in chicken medullary bone marrow cultures. Mononuclear cells were isolated from the peritrabecular bone marrow from the medullary bone of laying hens maintained on a calcium-deficient diet for 1 week. These cells were cultured for up to 7 days on devitalized bovine bone slices after removing the nonadherent fraction. The mononuclear precursors of the osteoclast that are present in such cultures adhere to bone matrix. These cells are TRAP+, express the vitronectin receptor at high levels, and also express high levels of sodium pumps and of carbonic anhydrase, enzymes that are characteristically enriched in the mature osteoclast. Finally, the most mature mononuclear precursors were found to be capable of resorbing the extracellular bone matrix before forming multinucleated osteoclasts.


Dietary restriction of calcium, phosphorus, and vitamin D elicits differential regulation of the mRNAs for avian intestinal calbindin-D28k and the 1,25-dihydroxyvitamin D3 receptor. We investigated the regulation of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]-induced calbindin-D28k (CaBP) and of the vitamin D receptor (VDR) by evaluating CaBP protein, CaBP mRNA, and VDR mRNA under conditions of altered intake of vitamin D, calcium, or phosphorus. Chickens were maintained for 10 days on one of four diets: vitamin D-deficient, normal (1.0% Ca and 1.1% P), low calcium (0.1% Ca and 1.2% P), and low phosphorus (1.1% Ca and 0.3% P). CaBP was undetectable in D-deficient duodena and was elevated above normal values by low-calcium (3.1-fold) and low-phosphorus (2.3-fold) intake. Contradictory to published data, we observed a correlation between CaBP protein and mRNA levels in that the CaBP mRNA was absent in D-deficient intestine and augmented threefold and twofold in low-calcium and low-phosphate duodena, respectively. In contrast, VDR mRNA concentrations were identical in vitamin D-deficient and normal duodena, implying that intestinal VDR is not dependent upon 1,25-(OH)2D3 for basal expression. Chickens fed a low-phosphorus diet displayed a twofold increase in VDR mRNA, but those fed a low-calcium diet exhibited a dramatic decrease in VDR mRNA. These data show that CaBP mRNA and protein levels are modulated in a tightly coupled fashion, and they are consistent with previous conclusions that augmented circulating 1,25-(OH)2D3 stimulates CaBP expression when dietary calcium or phosphorus is limiting. However, a more complex regulation of VDR expression occurs in that low-phosphorus restriction enhances VDR mRNA levels, possibly via increased circulating 1,25-(OH)2D3. Conversely, reduced dietary calcium diminishes VDR mRNA despite increased circulating 1,25-(OH)2D3, indicating that another factor, such as parathyroid hormone, is a predominant downregulator of VDR.


The role of histamine on the control of thyrotropin secretion in rats. The presence of histamine (HA) in the hypothalamus, especially in its median eminence, as well as results of previous studies from this laboratory, suggest a participation of this amine in the regulation of TSH (thyroid stimulating hormone, thyrotropin) secretion. In the present investigation, rats were treated with a single injection of cimetidine (CIM - 100 mg/kg ip), an H2-HA receptor antagonist. In vitro release of TSH in response to hypothalamic extracts from control animals, was approximately 20% smaller when evoked from pituitaries obtained from CIM-treated, rather than control animals. The addition of hypothalamic extracts from CIM-treated rats to incubates of pituitaries from either control or CIM treated rats did not significantly change basal TSH secretion, suggesting that TRH (thyrotropin releasing hormone) content was decreased by the anti-histamine treatment. These results point towards a facilitatory role of HA on TSH secretion both at the hypothalamic level where it may interfere with TRH synthesis, as well as at the pituitary level where it may modify TSH response to TRH.


[The evaluation of the effects of bunazosin hydrochloride in the treatment of prostatic hyperplasia]. Thirty-five patients with prostatic hyperplasia were entered into a randomized controlled study using bunazosin hydrochloride(alpha 1-adrenergic blocker). Eighteen patients were allocated to be treated with an initial dose of 1.5 mg/day (group 1) and 17 patients received an initial dose of 3.0 mg/day (group 2). Subjective symptoms and parameters of urodynamic studies were evaluated after 4 weeks and 12 weeks. There were 5 withdrawals or exclusions within 4 weeks and 6 within 12 weeks. Therefore, 15 patients in group 1 and 15 patients in group 2 were eligible for evaluation at 4 weeks, 12 patients in group 1 and 11 patients in group 2 were eligible for evaluation at 12 weeks. The rates of the improvement in subjective symptoms such as retarded urination, protracted urination, weakened urinary stream, abdominal straining on voiding, and sense of residual urine were 40%, 25% in group 1 and 46.7%, 36.4% in group 2, at 4 weeks and at 12 weeks, respectively. The rate of improvement in objective parameters of the urodynamic study such as voiding volume, residual volume, maximum flow rate and average flow rate were 26.7%, 16.7% in group 1 and 26.7%, 20.0% in group 2, at 4 weeks and at 12 weeks, respectively. Thus, these improvements were not correlated with the dosage of drug. Adverse effects such as diarrhea, mild headache, palpitation and gastric disturbance were observed in 2 patients in group 1, and 3 patients in group 2. In the 24 patients treated with the drug over 12 weeks, there were no specific adverse effects due to prolonged administration. These findings showed that bunazosin hydrochloride is beneficial for the treatment of prostatic hyperplasia, and could be safety administered for a prolonged period of time up to 12 weeks.


Genetic epidemiology of beta-thalassemia in Sicily: do sequences 5' to the G gamma gene and 5' to the beta gene interact to enhance HbF expression in beta-thalassemia? The present epidemiological study of the molecular characteristics of beta-thalassemia in Sicily was prompted by the disparate phenotypic expression (in clinical status and absolute HbF level) observed in two beta-thalassemic homozygotes who were also homozygous for the beta-like globin gene cluster haplotype III. We suspected that polymorphisms within haplotype III could be the cause for the discrepancy. Based on the association of particular conformations of the (AT)xT(y) motif (-540 5' to the beta gene) with milder forms of thalassemia and sickle cell anemia, 38 homozygous beta-thalassemia patients were studied to define their haplotypes, the -158 site 5' to the G gamma gene (linked to haplotype III) and the structure of the (AT)xT(y) motif. We found that the patient who was phenotypically mild and homozygous for beta-thalassemia, haplotype III, and the -158 C----T mutation was homozygous for the rare (AT)9T5 motif. In contrast, the patient homozygous for beta-thalassemia, haplotype III, and the -158 mutation, but exhibiting a severe clinical course, was homozygous for the (AT)7T7 configuration. Others have suggested that (AT)9T5 is a negative regulatory protein binding sequence, and it is a silent carrier state for beta-thalassemia. The usual configuration (AT)7T7, has considerably less affinity for regulatory protein binding, and it is the most common configuration in Sicilian beta-thalassemics (67 of the 78 chromosomes studied). Within the 38 patients studied, seven were informative because they had various combinations of the (AT)9T5 and (AT)7T7 motif, and the -158 C----T mutation. The results in these patients suggest that only the co-presence of the (AT)9T5 configuration and a C----T change at -158 5' to the G gamma gene is associated with high HbF expression and a mild clinical phenotype. We postulate that these two regions of the beta-like globin gene cluster interact, when endowed with the proper sequences, to enhance the expression of HbF secondary to anemia.


Acute undifferentiated leukemia (AUL): a case report and a proposed system of classification. This report describes a case of HLADR+, CD34- acute undifferentiated leukemia (AUL) diagnosed in an 18-year-old male. A definition of AUL and a system for its classification are proposed on the basis of the current state of knowledge about phenotypic features of AUL cells and their clonal counterparts that exist during early stages of normal hematopoiesis.


Substitution of arginine for glycine 325 in the collagen alpha 5 (IV) chain associated with X-linked Alport syndrome: characterization of the mutation by direct sequencing of PCR-amplified lymphoblast cDNA fragments. A large kindred with adult-type X-linked Alport syndrome was studied with regard to a defect in the recently described COL4A5 collagen gene. Southern blot analysis with COL4A5 cDNA probes showed loss of a MspI restriction site. Direct sequencing of cDNA amplified from lymphoblast mRNA demonstrated a single-base substitution converting a glycine codon to arginine at position 325 in the alpha 5 chain of type IV collagen. The triple-helical collagenous domain of alpha 5(IV), characterized by a Gly-X-Y repeat sequence, is interrupted 22 times by noncollagenous sequences. The mutation creates an additional interruption in the Gly-X-Y repeat motif, between interruptions 4 and 5. It is interesting that such glycine substitutions inside the COL1A1 or COL1A2 genes have been associated with many cases of osteogenesis imperfecta. This gly325-to-arg substitution presumably alters the triple-helix formation, and, in turn, modifies the ultrastructural and functional characteristics of the type IV collagen network inside the glomerular basement membrane.


Molecular diagnostics for myelin proteolipid protein gene mutations in Pelizaeus-Merzbacher disease. Pelizaeus-Merzbacher disease (PMD) is a clinically heterogeneous, slowly progressive leukodystrophy. The recent detection of mutations in the myelin proteolipid protein (PLP) gene in several PMD patients offers the opportunity both to design DNA-based tests that would be useful in diagnosing a proportion of PMD cases and, in particular, to evaluate the diagnostic utility of single-strand conformation polymorphism (SSCP) analysis for this disease. A combination of SSCP analysis and direct sequencing of PCR-amplified DNA was used to screen for PLP mutations in 24 patients affected with leukodystrophies of unknown etiology. Two heretofore undescribed mutations in the PLP gene were identified, Asp202His in exon 4 and Gly73Arg in exon 3. The ease and efficiency of SSCP analysis in detecting new mutations support the utilization of this technique in screening for PLP mutations in patients with unexplained leukodystrophies.