Effect of muscle stretching on the activity of neuromuscular transmission. The purpose of this study was to investigate how muscle stretching affects the activity of neuromuscular transmission. The magnitude of the post-tetanic potentiation (PTP) of miniature end-plate potential (m.e.p.p.) frequency was measured in the rat soleus muscle at resting length and stretched length. The parameters of the magnitude of PTP can be indicators of the kinetics of Ca2+ metabolism in the nerve terminal. One of the parameters, normalized initial post-tetanic frequency (f), was significantly increased by stretching muscle 10% (P less than 0.05) and 20% (P less than 0.01) of its resting length. Another parameter, the time constant of augmentation (tau a), was not significantly changed by muscle stretching. The time constant of potentiation (tau p) was significantly increased by 20% muscle stretching (P less than 0.05). These results indicate that the Ca2+ conductance, especially the voltage-dependent Ca2+ influx, of the nerve terminal could be increased by muscle stretching. Greater Ca2+ conductance of the nerve terminal would increase intracellular free Ca2+. Consequently, the probability of transmitter release would be increased, because Ca2+ is closely related to the activity of the transmitter release mechanism.


Dextran-magnetite particles: contrast-enhanced MRI of blood-brain barrier disruption in a rat model. Dextran-magnetite particles (DMP) were studied for their use as a MR contrast agent to visualize lesions with a blood-brain barrier (bbb) disruption. A freezing injury to the rat cerebral cortex was used as a model of bbb disruption. The biodistribution of iv-injected DMP was studied using atomic absorption spectrophotometry, electron microscopy, and MRI. One hour after injection, focal accumulation of the particles in capillary endothelial cells could be demonstrated in the freezing lesion. Despite the observation that the relaxivity of DMP in vivo appears to be less well pronounced than that in vitro, the MR imaging studies show that DMP can be used to visualize bbb disruption with adequate contrast.


[Studies on stable control of blood sugar by continuous administration of insulin through isolated intestinal loop]. Insulin (INS) has been known to be absorbed through the intestine in small amounts and to decrease blood sugar (BS) levels. However, stable control of BS has not been attained by intestinal intake of INS. To elucidate the possibility of a surgical approach to control hyperglycemic states, Thiry-Vella loops or upper jejunal loop fistulae were constructed in mongrel dogs in hyperglycemic states. Hyperglycemic states (around 400mg/dl) were induced by injecting streptozotocin. Lente insulin was administered every day to control hyperglycemia, except for the days for studies. As we confirmed that co-administration of aprotinin (TR) or nafamostat mesilate (FT) was necessary, to decrease BS by infusing INS into the intestinal loops, safe and effective dose levels were determined by series of preliminary infusion studies in the intestinal loops of diabetic dogs. By continuous infusion of these doses into the jejunal loop fistulae, a long term stable blood sugar control was attained in the diabetic dogs as long as 7 days. These results suggest that continuous infusion of INS with antiproteolytic adjuvants like TR or FT into the isolated small intestinal loop can control BS in hyperglycemic states.


Involvement of cDNA in homologous recombination between Ty elements in Saccharomyces cerevisiae. Strains carrying a marked Ty element (TyUra) in the LYS2 locus were transformed with plasmids bearing a differently marked Ty1 element (Ty1Neo) under the control of the GAL promoter. When these strains were grown in glucose, a low level of gene conversion events involving TyUra was detected. Upon growth on galactose an increase in the rate of gene conversion was seen. This homologous recombination is not the consequence of increased levels of transposition. When an intron-containing fragment was inserted into Ty1Neo, some of the convertants had the intron removed, implying an RNA intermediate. Mutations that affect reverse transcriptase or reverse transcription of Ty1Neo greatly reduce the induction of recombination in galactose. Thus, Ty cDNA is involved in homologous gene conversion with chromosomal copies of Ty elements. Our results have implications about the way families of repeated sequences retain homogeneity throughout evolution.


A collection of mRNA species that are inducible in the RAW 264.7 mouse macrophage cell line by gamma interferon and other agents. To identify genes induced during macrophage activation, a cDNA library was prepared from cultures of the RAW 264.7 mouse macrophage cell line that had been treated with conditioned medium from mitogen-stimulated spleen cells, and the cDNA library was screened by differential plaque hybridization. Eleven cDNA clones, designated CRG-1 through CRG-11, corresponding to mRNA species inducible in RAW 264.7 cells by the spleen cell conditioned medium, were isolated. Inductions were not blocked by cycloheximide. All of the mRNAs were inducible by gamma interferon, and some were also inducible by alpha and beta interferons, by lipopolysaccharide, by phorbol 12-myristate 13-acetate, and by the calcium ionophore A23187. Sequencing of the cDNAs revealed that CRG-1, CRG-3, and CRG-5 are cDNAs of recently identified transcription factors IRF-1, zif/268, and LRF-1 respectively. As previously reported, CRG-2 and CRG-10 (MIG) encode new members of the platelet factor 4 family of cytokines. CRG-6 corresponds to a new member of a family of interferon-inducible genes clustered on mouse chromosome 1, CRG-9 corresponds to a prostaglandin synthase homolog, CRG-8 corresponds to beta 2-microglobulin, and CRG-4 corresponds to metallothionein II. CRG-11 contains sequences of a truncated L1Md repetitive element as well as nonrepetitive sequences. The nonrepetitive sequence of CRG-11 as well as the sequences of CRG-7 are not closely related to published sequences. The CRG genes and proteins are of interest because of their involvement in macrophage activation, because of their roles as mediators of the effects of gamma interferon and other pleiotropic agents, and because of their usefulness as tools for studying the signal pathways through which gamma interferon and other inducers exert their effects on gene and protein expression.


A lymphoid cell-specific nuclear factor containing c-Rel-like proteins preferentially interacts with interleukin-6 kappa B-related motifs whose activities are repressed in lymphoid cells. The proto-oncoprotein c-Rel is a member of the nuclear factor kappa B transcription factor family, which includes the p50 and p65 subunits of nuclear factor kappa B. We show here that c-Rel binds to kappa B sites as homodimers as well as heterodimers with p50. These homodimers and heterodimers show distinct DNA-binding specificities and affinities for various kappa B motifs. In particular, the c-Rel homodimer has a high affinity for interleukin-6 (IL-6) and beta interferon kappa B sites. In spite of its association with p50 in vitro, however, we found a lymphoid cell-specific nuclear factor in vivo that contains c-Rel but not p50 epitopes; this factor, termed IL-6 kappa B binding factor II, appears to contain the c-Rel homodimer and preferentially recognizes several IL-6 kappa B-related kappa B motifs. Although it has been previously shown that the IL-6 kappa B motif functions as a potent IL-1/tumor necrosis factor-responsive element in nonlymphoid cells, its activity was found to be repressed in lymphoid cells such as a Jurkat T-cell line. We also present evidence that IL-6 kappa B binding factor II functions as a repressor specific for IL-6 kappa B-related kappa B motifs in lymphoid cells.


The neuromuscular basis of hereditary kyphoscoliosis in the mouse. We describe a new neuromuscular disorder in the kyphoscoliotic mouse mutant (ky). Mice were killed at ages from birth to 210 days, and tissues were taken for standard light microscopy, histochemistry, nerve ending studies, and electron microscopy. At birth a few myofibers showed phagocytosis ultrastructurally. Between 6 and 25 days there was prominent necrosis and regeneration in soleus, gracilis, paraspinal, and back muscles. At 47 days, these muscles were atrophic and necrosis and regeneration were rare. At 136 days, all muscle groups, including head muscles, showed some degree of myofiber atrophy and gracilis was fibrotic. Prominent intramuscular axonal sprouting was present from 31 days. Peripheral nerves and anterior horn cells were normal. The findings indicate a neuromuscular basis of hereditary kyphoscoliosis in the mouse. The animal may be useful as a model of human muscle disease and scoliosis.


Down-regulation of cystic fibrosis transmembrane conductance regulator gene expression by agents that modulate intracellular divalent cations. In cystic fibrosis (CF), epithelial cells are unable to normally up-regulate apical membrane Cl- secretion in response to agents which increase cyclic AMP, but they do increase Cl- secretion in response to increases in intracellular Ca2+. Since intracellular divalent cations regulate the expression of many genes, we hypothesized that mobilization of intracellular Ca2+ and/or other divalent cations might modulate not only Ca(2+)-dependent Cl- channels but also cystic fibrosis transmembrane conductance regulator (CFTR) gene expression. To evaluate this concept, HT-29 human colon carcinoma cells were cultured under various conditions designed to manipulate intracellular divalent cation concentrations and CFTR gene expression was quantified at the levels of transcription, mRNA accumulation, mRNA half-life, and protein. Exposure to the divalent cation ionophores A23187 and ionomycin (agents which increase intracellular divalent cation concentrations) caused dose- and time-dependent reductions of CFTR mRNA levels, which could be blocked by the use of Ca(2+)- and Mg(2+)-free media. Ionophore-induced CFTR gene modulation was also observed with T84 human colon carcinoma cells and freshly isolated normal human bronchial epithelial cells. Incubation of HT-29 cells with thapsigargin, an agent that releases Ca2+ from intracellular stores, or in medium containing increased extracellular concentrations of Ca2+ or Mg2+ also caused down-regulation of CFTR mRNA levels. Transcription run-on analysis showed that, parallel with the decrease in CFTR mRNA levels, A23187 reduced the rate of transcription of the CFTR gene, while CFTR mRNA transcript half-life was unaffected. Consistent with the down-regulation of CFTR gene expression, CFTR protein levels also decreased after exposure to A23187. Thus, despite the independence of Ca(2+)-dependent Cl- channels and cyclic AMP-dependent CFTR-related Cl- channels in epithelial cells, increases in intracellular divalent cation concentrations down-regulate the expression of the CFTR gene at the transcriptional level, with consequent decreases in CFTR mRNA and protein.


Insulin is a prominent modulator of the cytokine-stimulated expression of acute-phase plasma protein genes. Several endocrine hormones which influence liver metabolism are known to increase in activity during the acute phase of injury or inflammation. We determined whether these hormones have the potential to influence acute-phase protein production in human and rat hepatoma cells. Catecholamines, glucagon, growth hormone, triiodothyronine, and cyclic nucleotides individually or in combination did not modulate the basal or the interleukin-1 (IL-1)-, IL-6-, and dexamethasone-stimulated levels of acute-phase plasma proteins. Insulin, however, was found to be a rapid, nonspecific, and dose-dependent inhibitor of the cytokine and glucocorticoid stimulation of acute-phase protein gene expression and to exert its effect at the transcriptional level. The insulin inhibition applied to all cytokines tested but to various degrees, depending upon the particular acute-phase gene. Insulin resulted in an early and prominent increase in the transcription of genes encoding the AP-1 components of JunA, JunB, and c-Fos, as has been observed for other growth factors. However, the effect of insulin on C/EBP beta was unexpected and paradoxical: while insulin completely inhibited the transcriptional activation of the C/EBP beta gene in cytokine- and dexamethasone-treated cells, the level of cytoplasmic C/EBP beta RNA was elevated. Quantitation of C/EBP beta mRNA by Northern (RNA) blot analysis and of C/EBP beta DNA binding activity by Southwestern (DNA-protein) blot analysis showed that insulin, when combined with cytokines and dexamethasone, stimulated both the mRNA and DNA binding activity by a factor of 1.6 compared with that of cells treated with cytokines and dexamethasone alone. Transient transfection of H-35 and HepG2 cells with a chloramphenicol acetyltransferase (CAT) gene expression vector containing the C/EBP beta response element also resulted in a 1.5-fold increase of C/EBP beta-mediated transcription in insulin-treated cells. Transfection of CAT gene constructs containing increasing lengths of heptaglobin gene 5' flanking sequences indicated that insulin inhibition of IL-6 stimulation required the presence of the region from -4100 to -1030. These results suggest that insulin has the potential to control the transcription of acute-phase genes by at least two separate mechanisms.


Lymphoproliferation disorder in mice explained by defects in Fas antigen that mediates apoptosis. Fas antigen is a cell-surface protein that mediates apoptosis. It is expressed in various tissues including the thymus and has structural homology with a number of cell-surface receptors, including tumour necrosis factor receptor and nerve growth factor receptor. Mice carrying the lymphoproliferation (lpr) mutation have defects in the Fas antigen gene. The lpr mice develop lymphadenopathy and suffer from a systemic lupus erythematosus-like autoimmune disease, indicating an important role for Fas antigen in the negative selection of autoreactive T cells in the thymus.


C. elegans cell-signalling gene sem-5 encodes a protein with SH2 and SH3 domains. The induction of the hermaphrodite vulva and the migration of the sex myoblasts in the nematode Caenorhabditis elegans are both controlled by intercellular signalling. The gonadal anchor cell induces formation of the vulva from nearby hypodermal cells, and a set of somatic gonadal cells attract the migrating sex myoblasts to their final positions. Many genes required for vulval induction have been identified, including the let-23 receptor tyrosine kinase gene and the let-60 ras gene. We report here the identification and characterization of a new gene, sem-5 (sem, sex muscle abnormal), that acts both in vulval induction and in sex myoblast migration. On the basis of its DNA sequence, sem-5 encodes a novel 228-amino-acid protein which consists almost entirely of one SH2 (SH, src homology region) and two SH3 domains. SH2 and SH3 domains are present in many signalling proteins regulated by receptor and non-receptor tyrosine kinases. Mutations that impair sem-5 activity alter residues that are highly conserved among different SH2 and SH3 domains. Our results indicate that the sem-5 gene encodes a novel protein that functions in at least two distinct cell-signalling processes.


Intraventricular interferon and oral inosiplex in the treatment of subacute sclerosing panencephalitis. We treated 22 patients with subacute sclerosing panencephalitis (SSPE) with intraventricular alpha-interferon (IFN) and inosiplex PO and followed them for 2 to 54 months. Three deaths occurred. Clinical improvement, demonstrated by decreasing scores on the Neurological Disability Index, occurred in 11/22 (50%); five patients became stable, and the progression rate of the disease decreased in three. The remission rate was significantly higher than untreated controls from the same institution. Patients who had a slowly progressive disease responded best to treatment. Serious side effects were rare. We recommend intraventricular IFN, combined with oral inosiplex, in the treatment of SSPE.


Conjugal multiple sclerosis: immunogenetic characterization and analysis of T- and B-cell reactivity to myelin proteins. We describe two families with conjugal multiple sclerosis. Onset of symptoms in the husbands occurred 11 and 17 years after onset of relapsing/remitting symptoms in their wives. There were no similarities regarding clinical manifestations of MS within each family. Evaluation of T-cell repertoire by enumeration of cells secreting interferon-gamma in response to proteolipid protein (PLP), myelin basic protein (MBP), and to various synthetic MBP peptides revealed similar patterns of T-cell reactivity within the families both in MS-affected parents and unaffected children. Genomic HLA-DR-DQ typing showed that T-cell reactivity was independent of HLA class II phenotype. Analysis of B-cell responses in blood showed low numbers of cells secreting IgG, IgA, or IgM antibodies against MBP, PLP, myelin-associated glycoprotein, and myelin-oligodendrocyte glycoprotein both in MS-affected and unaffected family members. In conclusion, our study of two families with conjugal MS has shown a dominant T-cell response against the same MBP peptide within the family both in MS-affected parents and unaffected children, and this T-cell response seems to be independent of the HLA class II phenotypes of the family members.


Head and neck malignancies associated with HIV infection. Immunosuppressed persons are at greater risk of developing malignancies. In human immunodeficiency virus (HIV) immunosuppression the most common oral cancers are Kaposi's sarcoma and non-Hodgkin's lymphoma. Squamous cell carcinoma has also been reported to be associated with HIV disease. Kaposi's sarcoma is the most frequent neoplastic disease in acquired immunodeficiency syndrome and is by far the most common in the head and neck area. This article reviews the prevalence, clinical features, and management of these diseases in HIV infection.


The amino-terminal half of pp59c-fyn contains sequences necessary for formation of a 75 kDa form and also repressive elements absent in pp60c-src. Members of the src family of tyrosine kinases are composed of amino acid sequences which can be divided into three regions: the unique amino-terminal 80 residues; the next 180 residues of conserved but non-catalytic sequence; and the catalytic carboxy-terminal half of the molecule. To characterize the elements that regulate the catalytic and transforming activities of two members of this family, pp59c-fyn and pp60c-src, we generated six chimeric proteins by interchanging the three regions of the 531F mutant of pp59c-fyn and of the 527F mutant of pp60c-src. Our data indicate that substituting all or part of the amino-terminal 263 residues of pp59c-fyn for those of 527F inhibited the kinase and transforming activities of 527F. Conversely, substituting the amino-terminal half of pp60c-src for that of 531F resulted in an increase in the transforming potential of 531F. These results suggest that the amino-terminal half of pp59c-fyn contains elements which act to suppress the catalytic and transforming activities of the enzyme and that these suppressive elements are either absent or inactive in pp60c-src. These differences argue that the src family of tyrosine kinases are regulated differently in the cell. In vitro translation of some of the chimeras in rabbit reticulocyte lysates generated a 75 kDa protein in addition to the expected 59 kDa product. This 75 kDa species is analogous to the p75 protein previously detected in wild-type pp59c-fyn translation products. Interestingly, formation of p75 required the presence of DNA sequences encoding the unique amino-terminal residues of pp59c-fyn.


Tyrosine phosphorylation of a 120,000 dalton membrane-associated protein by the neural form of pp60c-src, pp60c-src+. The c-src proto-oncogene encodes a 60,000 dalton tyrosine kinase, pp60c-src, which is the prototype member of the family of non-receptor tyrosine kinases. A neural-specific form of pp60c-src, pp60c-src+, is detected only in neurons of the central nervous system. pp60c-src+ contains a six amino acid insert (neural insert) in the SH3 region that is generated by alternative splicing. Previous reports indicate that the profiles of proteins phosphorylated on tyrosine in chick embryo fibroblast (CEF) cells by pp60c-src+ or pp60c-src are equivalent. In this report, the activities of pp60c-src+ and pp60c-src, as well as the activated variants, pp60(527F+) and pp60(527F), were compared in CEF cells by examining the steady-state levels of tyrosine phosphorylation of several known pp60src substrates. Most substrates examined were phosphorylated on tyrosine to equivalent levels in CEF cells expressing either the neural- or fibroblast-specific src gene products. However, the relative extent of tyrosine phosphorylation of a 120 kDa protein (p120) was increased in cells expressing the neuronal forms of either c-src or c-src527F. The increased tyrosine phosphorylation of p120 did not appear to be caused by the neural insert facilitating a specific interaction between pp60c-src+ and p120. These data indicate that preferential phosphorylation of p120 in neural cells may contribute to the specialized function of pp60c-src+ in neural cells.


Pharmacology of upper respiratory allergy. Patients with upper respiratory allergy may benefit from the use of many pharmacologic products. Using the simplest form of therapy that will relieve the problem is usually both clinically efficacious and cost efficient. Understanding the indications, benefits, and potential adverse effects of each group of drugs, particularly the classic and new antihistamines, decongestants, intranasal and systemic corticosteroids, and mast cell stabilizers (such as cromolyn) will provide direction for the best total patient care.


Atrial fixation leads--a visual aid confirming actual fixation. Various active fixation mechanisms are available for atrial lead implantation. Confirmation of actual fixation of the lead tip in the myocardium is sometimes difficult with standard techniques such as fluoroscopy. Our observation of organized clockwise/counterclockwise motions of the fixation stylet in synchrony with atrial systole confirms adequate positioning of the Accufix model #330-801 atrial lead in the right atrial appendage, which is helpful when the screw is difficult to visualize under fluoroscopy. This observation was confirmed in nine patients.


An unusual cause of carotid sinus syndrome: multiple symmetric lipomatosis. A 45-year-old man with multiple symmetric lipomatosis suffered recurrent syncope attributed to carotid sinus syndrome caused by extrinsic compression of the carotid body by the lipomatous masses. Surgical removal reduced but did not stop syncope, which was then controlled by implantation of a DDD pacemaker.


Evaluation of electrocardiographic leads for detection of atrial activity (P wave) in ambulatory ECG monitoring: a pilot study. The usual lead systems for ambulatory ECG monitoring (AECG) used in the evaluation of arrhythmias is a modified bipolar V-1 and V-5. A comparison of various lead systems to enhance the detection of atrial activity (p waves) has not been reported. We evaluated various surface lead systems in 12 subjects comparing p waves recorded at 20 mm/mV and 50 mm/sec. We compared p wave area, amplitude, and duration from modified bipolar V1 and V5 as well as seven nonstandard leads recorded on a AECG monitor. Of the seven nonstandard leads, a vertical sternal lead, with the negative pole just below the suprasternal notch and the positive pole at the xiphoid process, had the largest area (1.46 +/- 0.65 mm2), and also had a greater area than the standard V1 (0.88 +/- 0.45 mm) and V5 (1.06 +/- 0.49 mm2) lead system (P less than 0.01). We conclude that the bipolar vertical sternal lead system provides a larger p wave area than seven nonstandard bipolar lead systems and the two standard lead systems currently used in AECG monitoring. Replacement of the modified bipolar V1 lead with a vertical sternal lead should improve the recognition of atrial activity and, therefore, enhance the diagnosis of cardiac arrhythmias.


Atrial paralysis in a patient with Emery-Dreifuss muscular dystrophy. Emery-Dreifuss disease is a benign X-linked muscular dystrophy characterized by a distinct pattern of muscle weakness, which is of insidious onset and slow progression. It is associated with atrial paralysis that results in sudden death in early adulthood if left untreated. The authors report the documentation of electrical and mechanical silence confined to the atria in a patient with this disease. Electrocardiography and electrophysiological study document the absence of electrical atrial activity, and inability to pace the atria. Hemodynamic studies demonstrate the absence of A waves, and angiography revealed immobility of the atria. This patient has done well following the institution of permanent ventricular pacing. His brother, who also had muscular dystrophy, died a sudden cardiac death at the age of 29 after refusing medical intervention. Emery-Dreifuss muscular dystrophy is particularly worthy of recognition because of the preventable occurrence of sudden death in young patients with an otherwise excellent prognosis. Permanent ventricular pacing is indicated.


Survival in 1,431 pacemaker patients: prognostic factors and comparison with the general population. A total of 1,431 patients (mean age 63.4 +/- 14.1) with pacemakers (96.2% VVI) primoimplanted between 1967 and 1985 were followed for a mean duration of 78.2 +/- 40 pacing months, with 0.6% loss to follow-up. Cumulative survival for 1, 3, and 10 years was 0.9427, 0.9136, and 0.7536, respectively. There was no significant difference in survival between atrioventricular block (AVB) and sick sinus syndrome (SSS) patients. In addition to age and gender, factors existent prior to implantation that independently affected prognosis included manifest coronary heart disease (CHD), congenital/acquired heart lesions, heart failure, noncardiac internal disease, syncope, and generalized fatigue. After implantation, the most important factor was generalized fatigue, then age, stroke, myocardial infarct (MI), gender (male), heart failure, and syncope. Patients with no underlying disease showed an extremely high cumulative survival (0.9173 at 10 years). Compared to the general population of Yugoslavia, the pacemaker patients showed a similar yearly mortality rate until 1981. After that, elderly males (70+) had a significantly lower yearly mortality than the matched population. Thus, in this large series of pacemaker patients followed into the most recent period with an extremely low loss to follow-up, short- and long-term survival was very high. Pacemaker patients of any age who are otherwise in good health have an excellent prognosis.


Effects of oral propafenone therapy on chronic myocardial pacing threshold. The effects of oral propafenone therapy on pacing threshold were studied in 36 patients chronically paced for sick sinus syndrome or AV block. The pacemakers, all unipolar models and with noninvasive threshold measurement facilities, were: 9 VVI, 15 AAI, and 12 DDD. Each patient received an initial propafenone dose of 450 mg/day, that in 18 cases was increased to 900 mg/day. Threshold was tested at baseline and at each dosage after 7 days of therapy. With the lower propafenone dosage the threshold, measured at 2.5 V, rose from 0.14 +/- 0.10 to 0.21 +/- 0.16 msec (+55%) in the atrium (P less than 0.0001) and from 0.10 +/- 0.08 to 0.15 +/- 0.09 msec (+63%) in the ventricle (P less than 0.0001). In the 18 patients who received both dosages, the mean atrial and ventricular threshold increased from 0.12 +/- 0.10 to 0.17 +/- 0.14 msec with the lower dose and to 0.27 +/- 0.22 msec (+125%) with the higher dose (P less than 0.0001 for both increments). With the 900 mg/day dose, a threshold increment greater than or equal to 300% was observed in 15% of the stimulated chambers. A good linear correlation (r = 0.76) was found between the ventricular threshold increment and the drug induced QRS widening. In conclusion, treatment with oral propafenone increases atrial and ventricular stimulation threshold in pacemaker patients. Threshold increment is dose dependent and proportional to the drug induced QRS widening. In the majority of the cases the threshold increment is not clinically significant, but caution must be used in prescribing high doses of the drug to patients with high baseline threshold.


Is epicardial dual chamber pacing a realistic alternative to endocardial DDD pacing? Initial results of a prospective study. Seventeen patients, in whom an epicardial (n = 7) or a transvenous DDDM pacemaker system had been implanted between June 1988 and October 1990, were followed up for pacemaker and lead related complications, pacemaker longevity, and electrophysiological lead parameters. The mean follow-up interval was 18 +/- 12 months, maximum 34 months. There were no differences in chronic atrial and ventricular sensing thresholds between epicardial and endocardial stimulation, nor were there any differences concerning lead related complications between the two pacing modalities. However, atrial as well as ventricular chronic stimulation thresholds were significantly higher with epicardial stimulation resulting in a twofold increase in atrial energy consumption and a threefold increase in the ventricular energy consumption. Thus, in one patient with an epicardial DDD system, the pacemaker had to be replaced prematurely because of battery depletion. It is concluded that epicardial DDD stimulation can be reliably performed as far as atrial and ventricular sensing is concerned, but that the energy requirements of available myocardial leads are not satisfactory for making optimal use of modern pacemaker capability.


Effects of verapamil on ventricular tachycardia induced by ouabain in guinea pigs. Calcium ions appear to play a central role in the development of ventricular arrhythmias associated with digitalis intoxication. Therefore, the effects of the calcium channel antagonist, verapamil, on ouabain-induced ventricular tachycardia were investigated. Ventricular tachycardia (three or more consecutive wide QRS complexes) was induced in guinea pigs by intravenous infusion of ouabain (1 microgram/min) and bursts of rapid ventricular stimulation. Of seven guinea pigs given the infusion of ouabain, all developed ventricular tachycardia at a dose of 82 +/- 17 micrograms/kg (mean +/- SD) and none developed heart block or asystole. Eight guinea pigs were treated with verapamil (2 micrograms/min for 30 minutes) prior to exposure to ouabain. Of those eight animals, only two developed ventricular tachycardia but six developed heart block or asystole at a significantly higher dose of ouabain (154 +/- 47 micrograms/kg). None of three control guinea pigs given an infusion of normal saline for 90 minutes developed ventricular tachycardia. These results show that pretreatment with verapamil inhibits ouabain-induced ventricular tachycardia in guinea pigs. Combined treatment with verapamil and ouabain is associated with a high incidence of heart block and asystole, which may limit the usefulness of verapamil.


A new pacemaker algorithm for continuous capture verification and automatic threshold determination: elimination of pacemaker afterpotential utilizing a triphasic charge balancing system. A new pacemaker algorithm designed to automatically verify pacemaker capture and determine pacing threshold by detection of a stimulus evoked potential was studied in 20 patients undergoing permanent pacemaker implantation. To eliminate pacing stimulus afterpotential and detect an evoked response, a hardware feedback circuit and a software template matching algorithm were used to produce a triphasic charge-balanced pacing pulse. After charge balancing the pacing lead, a residual artifact is measured. A capture window is defined as the area integral of the first 24 msec of the evoked depolarization, and a capture threshold as one third the amplitude of the capture window. The maximum allowable residual artifact is one eighth the amplitude of the capture window. Once the stimulus afterpotential is eliminated and the evoked response detected, capture threshold is automatically and continuously determined and the algorithm adds a 0.8-V safety margin to the pacemaker output. This algorithm was run automatically and after simulated loss of capture, produced by manually decreasing pacer output below threshold, in the bipolar (13 patients) and unipolar (20 patients) pacing modes. In each patient loss of capture was immediately detected. The data were consistent (P = NS) between algorithm runs. During unipolar pacing the area integral of the first 24 msec of the evoked response was 412 +/- 137 versus 413 +/- 144 and the residual artifact 5.8 +/- 4.8 versus 8.1 +/- 7.5. The resulting ratio (signal/noise) of the two parameters was 150 +/- 141 versus 145 +/- 181. Automatically determined threshold was 0.69 +/- 0.43 V versus 0.69 +/- 0.42.(ABSTRACT TRUNCATED AT 250 WORDS)


Long-term therapy of antitachycardia pacing for supraventricular tachycardia. Long-term antitachycardia pacing therapy with the InterTach 262-12 and 262-16 was evaluated in 32 consecutive patients (mean age 50 +/- 13 years) with recurrent, drug refractory supraventricular tachycardia. AV nodal reentrant tachycardia was present in 20 patients, Wolff-Parkinson-White syndrome in ten patients, and a reentrant tachycardia due to Mahaim fibers in one patient. During follow-up of 39 +/- 17 months, 250 persistent tachycardia episodes occurred in 22 patients. By adjusting detection and termination mode, recurrent supraventricular tachycardia could be controlled in 19 of 32 patients (60%) by antitachycardia pacing alone. Concomitant antiarrhythmic drug therapy was required in ten of 32 patients (30%). During follow-up antitachycardia pacing became ineffective in three patients (10%). Thus, chronic antitachycardia pacing proved to be safe and effective in selected patients with drug refractory supraventricular tachycardia and could significantly improve quality of life by rapid termination of recurrent supraventricular tachycardia episodes.


Activity-based pacing: comparison of a device using an accelerometer versus a piezoelectric crystal. The EXCEL VR, an accelerometer-based pacemaker (AC), and the Legend, a pacemaker utilizing a piezoelectric crystal (PZ), were compared under ergometric conditions and during stair climbing to assess the appropriateness of their rate responses. The pacemakers, programmed to the manufacturers' nominal settings in order to compare different technologically based sensors under identical conditions, were strapped over subjects' left mid-pectoral region. Placement of the devices was randomized to control for positional effects. Ten healthy subjects (55-72 years) completed a graded exercise treadmill test to 80% of maximum predicted heart rate (HR). An additional group of ten subjects (50-66 years) completed exercise protocols involving bicycle ergometry and stair climbing. Throughout all tests, pacemaker pulse rates and subjects' intrinsic HR were monitored continuously. For the treadmill exercise, the average correlations between the AC and PZ pacemakers' pulse rate and HR for the group as a whole were r = 0.92 and r = 0.82, respectively. Individual subject comparisons were also made between each pacemaker rate and intrinsic HR. The mean difference from intrinsic rate was 11 ppm for the AC pacemaker and 24 ppm for the PZ pacemaker. In addition, the PZ pacemaker's maximal pulse rate was significantly lower (105 +/- 9.6 ppm) than the other two rates (AC 137 +/- 6 ppm; intrinsic HR 129 +/- 2 beats/min). Throughout the bicycle ergometry testing, the intrinsic HR was higher than the AC and PZ pacing rates. However, the AC's rate was significantly higher than the PZ's rate. When subjects ascended stairs, the intrinsic HR and AC rate were closely correlated, but the PZ rate was significantly lower.(ABSTRACT TRUNCATED AT 250 WORDS)


Propagation versus delayed activation during field stimulation of cardiac muscle. This modeling study seeks to explain the experimentally detected delay between the application of an electric field and the recorded response of the transmembrane potential. In this experiment, conditions were deliberately set so that the field should excite all cells at once and so that no delay should be caused by a propagating wave front. The explanation of the observed delay may lie in the intrinsic properties of the membrane. To test this hypothesis, the strength latency curves were determined for three cases: (1) for a membrane patch model, in which the membrane is uniformly polarized and its intrinsic properties can be studied; (2) for the cardiac strand directly excited by the electric field; and (3) for the cardiac strand excited by a propagating wave front. The models of the membrane patch and the directly excited strand yield excitation delays that are comparable to those observed experimentally in magnitude and in the overall shape of the strength latency curves. The delays resulting from propagation are, in general, dependent on the position along the strand, although for some positions the strength latency curves for propagation are similar to those obtained from the directly activated strand and from the patch model. Therefore, the delay in excitation does not necessarily imply the presence of propagating wave fronts and can be attributed to intrinsic membrane kinetics.


First derivative of right ventricular pressure, dP/dt, as a sensor for a rate adaptive VVI pacemaker: initial experience. Ten patients underwent implantation of a rate adaptive ventricular pacing system with a new pulse generator and lead. The unipolar lead has a steroid eluting tip and a pressure sensor. The first derivative of the signal from this sensor, dP/dt, is determined and the pacemaker rate is varied in response to changes in the right ventricular dP/dtMAX. During implantation, dP/dt values were in the range of 180-720 mm Hg/sec. The autothreshold for pacing at 2.5 V remained unchanged 1 month after implantation (0.065 +/- 0.045 msec, range 0.05-2.00 msec) and only slightly increased after 3 months (0.075 +/- 0.045 msec, range 0.05-2.00 msec). A significant correlation existed between the dP/dt measured during implantation and the right ventricular pressure measured by telemetry at follow-up visits (r = 0.93, P = 0.0001). Initial pacemaker programming was performed on the second day after implantation following a short walk and was adjusted subsequent to follow-up visits according to the patient's subjective assessment and in accordance with the results of exercise tests and Holter monitoring. Exercise and Holter tests did not significantly change initial programming. There was a significant correlation between right ventricular systolic pressure and the rate response setting (r = -0.66, P less than 0.05). During dP/dt pacing, all patients felt well, and eight of these reported an improvement compared to nonrate adaptive pacing. The heart rate response to effort and recovery was appropriate. It was concluded that: (1) right ventricular dP/dt is a suitable parameter for controlling the pacing rate; (2) appropriate programming of the dP/dt pacemaker results in a suitable heart rate response to exercise and recovery.


Development of a rate adaptive pacemaker based on the maximum rate-of-rise of right ventricular pressure (RV dP/dtmax). The maximum rate of rise of right ventricular pressure (RV dP/dtmax) may change in response to physiological stress and thereby provide an appropriate parameter upon which to base rate adaptive pacing. Initial feasibility testing was carried out in six patients using externally closed loop rate adaptive pacing with a pressure sensing lead (Model 6220) and an investigational VVI pulse generator (Medtronic, Model 2451). During exercise, maximum positive RV dP/dtmax increased from 223 +/- 55 to 405 +/- 181 mmHg.sec.1 (P less than 0.05). Based on these results, rate adaptive pulse generators using maximum positive RV dP/dt were implanted in 12 patients (Medtronic, Model 2503). Exercise treadmill testing in the VVI mode resulted in heart rates ranging from 69 +/- 6 beats/min at rest to 79 +/- 14 beats/min (n = 12; P greater than 0.05). In contrast, VVIR mode pacing rates ranged from 71 +/- 11 beats/min to 115 +/- 24 beats/min (n = 17; P less than 0.05). Holter recording showed heart rates ranging from 51 +/- 6 to 110 +/- 22 beats/min during activities of normal daily living (n = 9; P less than 0.05). Passive postural tilt resulted in rates of 69 +/- 8 beats/min in the supine position increasing to 74 +/- 14 beats/min with 60 degrees upright tilt (n = 16; P greater than 0.05). With up to 5-year follow-up data, there have been no late failures of pacing but one lead showed insulation failure with over- and undersensing after 4.5 years. A number of deficiencies were identified in the prototypes leading to modifications of a subsequent generation of rate responsive pacemaker based on RV dP/dtmax. These initial data demonstrate that rate adaptive pacing based on RV dP/dtmax responds in a physiological manner. This rate responsive system is of particular interest as it is based on a beat-to-beat parameter of cardiac mechanical function.


Effect of vagal nerve electrostimulation on the power spectrum of heart rate variability in man. The power spectrum of heart rate variability contains low frequency (LF = 0.08-0.12 Hz) and high frequency (HF = 0.18-0.30 Hz) components said to represent neurocardiac rhythms. To verify whether such a relationship exists we report a unique study where the heart rate autospectrum was determined in a 28-year-old epileptic male patient with an implanted vagal electrical stimulator. The stimulator was activated at 20 Hz, 300 microseconds pulse, and 1.25 V. Continuous ECG and respiratory waveform records were obtained over 45 minutes every 8 hours (7-8 AM; 3-4 PM; 11-12 PM) with the stimulator ON, then 24 hours OFF and then 24 hours ON again. The overall LF:HF peak ratio increased from 0.64 to 1.99 (P less than 0.001) after the stimulator was turned OFF. There was a dramatic increase in the LF peak power (greater than 60%) and a corresponding decrease in the HF peak power (greater than 65%) when the stimulator was turned OFF. These values were reversed when the stimulator was turned ON again. In the early morning and late evening hours, there was a significant rightward shift in the LF peak power frequency (average 0.057 to 0.075 Hz) whenever the stimulator was ON. Otherwise, there were no significant circadian variations in any of the autospectral components. The results demonstrate an unequivocal relationship between selective vagal nerve electrostimulation and alterations in the heart rate autospectrum.


Evaluation of very rapid emit Qst methods for measuring serum procainamide and N-acetylprocainamide concentrations. This study assessed the Emit Qst procainamide (PA) and N-acetylprocainamide (NAPA) assays. Accuracy and intraday precision were evaluated by repeatedly measuring PA and NAPA concentrations in spiked serum samples using Qst and high-performance liquid chromatography methods. Interday precision was evaluated by measuring concentrations in spiked samples over 4 weeks. Correlation between methods was assessed in patient samples, and proportional, constant, and random errors were estimated. Intraday coefficients of variation (CVs) were below 6.4% for PA and NAPA for both methods; interday CVs were below 7.8%. The proportional, constant, and random errors of the PA Qst assay in patient samples were 5.7%, -0.224 mg/L, and +/- 0.574 mg/L, respectively. The same errors in the NAPA Qst assay were 17.2%, 0.229 mg/L, and +/- 1.79 mg/L, respectively. The Qst assays are rapid, accurate, and precise methods for routine clinical measurement of PA and NAPA, although the proportional error in the NAPA assay should be recognized.


Prostate specific antigen and prostatitis. I. Effect of prostatitis on serum PSA in the human and nonhuman primate. Prostate specific antigen (PSA) has become a mainstay in the diagnosis and management of patients with prostate cancer. We have found, as have others, that it may be elevated in patients with prostatic inflammation. Ten patients had clinical evidence of prostatitis and elevated PSA levels. Six of these had persistently elevated levels after antibiotic treatment. After transrectal ultrasonography and biopsy, two had findings of adenocarcinoma, and the rest had a pathologic diagnosis of acute or chronic prostatitis. We studied this process in an experimental model of prostatitis using a nonhuman primate. We infected six cynomolgus monkeys and followed their PSA levels until resolution of the infection. The PSA peaked between 5 and 7 days after inoculation and gradually returned to baseline in 8 weeks. The dramatically elevated serum PSA levels in bacterial prostatitis can cause confusion in the diagnosis of prostatic carcinoma.


Prostate specific antigen and prostatitis. II. PSA production and release kinetics in vitro. Elevations in serum prostate specific antigen (PSA) levels in patients with prostatitis are well known, but the pathophysiologic mechanisms involved with this phenomenon are poorly understood. We have recently evaluated the effect of prostatitis on PSA levels in primates. This data demonstrated a rapid rise in PSA, which subsequently fell along the biological decay curve. This suggested a release of sequestered PSA. We evaluated the effect of infection upon PSA production for the prostatic adenocarcinoma cell line LNCaP. The cell line was determined to produce 9.6 +/- 2.7 fg/cell/hr PSA with essentially linear kinetics. No effect was seen with dead bacteria, bacterial supernatant or complement upon the PSA production. Live E. coli had no effect for 4-8 hours at which time the cells sloughed and PSA production ceased. No release of stored PSA was seen. These data do not elucidate the reasons for increased serum PSA levels found with acute bacterial prostatitis, but indicate that it is not a storage phenomenon.


Evaluation of a new tumor marker for localized prostate cancer. Adenocarcinoma associated antigen (ACAA) is a large molecular weight protein that is normally found in low serum levels. Recent data have revealed elevations in patients with adenocarcinomas, including prostate cancer. To evaluate the relationship of ACAA levels with prostate cancer, we measured the cytosol content in malignant and nonmalignant prostate tissue and compared these results to those of the standard markers, prostatic acid phosphatase (PAP) and prostatic specific antigen (PSA). Enzyme solid phase immunoassay was used to quantitate PSA and ACAA levels, and the enzymatic method was used to measure PAP. Wedge resection from the right and left posterior lobes of 50 fresh radical retropubic prostatectomy specimens were used for cytosol analysis. All foci of within each prostate gland were carefully mapped by a single pathologist. When all malignant wedges (N = 74) were compared to all the benign wedges (N = 21), only the PSA levels showed significant elevation (p less than 0.02). However, when benign and malignant tissue from the same prostate were available for comparison, both PSA (N = 17) and ACAA (N = 16) showed significant elevations in the cytosol of the malignant tissue (p less than 0.002 and p less than 0.03, respectively). Although not statistically significant, the cytosol PAP did show a consistent trend to be greater in malignant tissue. It appears that there is an association of increased cytosol ACAA and PSA with prostate cancer.


Studies on the proliferation, secretory activities, and epidermal growth factor receptor expression in benign prostatic hyperplasia explant cultures. Short term explant cultures of benign prostatic hyperplasia (BPH) tissues were studied immunohistochemically to characterise both the morphological changes within the explant tissue and the cellular origin of the epithelial cell outgrowth. Altered patterns of expression of cytokeratins, prostate specific antigen (PSA) prostatic acid phosphatase (PAP), and epidermal growth factor (EGF) receptor were observed. After sloughing of the secretory epithelium in the majority of the acini repopulation and outgrowth of a monolayer was accomplished by cells which were strongly positive for stratifying keratin and EGF receptor and negative for PAP and PSA, indicative of a basal cell phenotype. The peak of proliferation in the acini, as assessed by Ki-67 immunohistochemistry, occurred after 2-4 days in culture. Preliminary studies on BPH tissue xenografts in nude mice indicated that better preservation of normal morphology, secretory activity, and antigen expression could be achieved.


The relative efficacy of terazosin versus terazosin and flutamide for the treatment of symptomatic BPH. Pharmacotherapy for the treatment of BPH is currently being targeted to relax prostate smooth muscle (alpha blockade) and decrease prostate volume (androgen suppression). The objective of the present study was to determine the relative efficacy of terazosin vs. terazosin and flutamide (combination therapy) for the treatment of symptomatic BPH. Twenty-nine males with symptomatic BPH were enrolled into this 6-month open label study. The entry criteria included peak urinary flow rate between 4-15 ml/sec, a total Boyarsky symptom score greater than 7, and postvoid residual less than 300 ml. The daily dosage of terazosin was titrated to 5 mg over a 2-week interval. A 750 mg daily dosage of flutamide was added following 1 month of terazosin monotherapy. The dosages were lowered if significant adverse events developed. Efficacy assessments were performed at 1 month (terazosin alone), 6 months (combination therapy), or at the time of early withdrawal from the study. The efficacy of combination therapy was evaluable in 24 patients receiving combination therapy. At one month, the mean peak urinary flow rate and mean total Boyarsky symptom score improved 38% and 56% relative to baseline, respectively. The present study confirmed the previously observed efficacy and safety of terazosin for BPH. The addition of flutamide did not result in statistically significant improvements in the peak urinary flow rate or total Boyarsky symptom score. The adverse events related to flutamide resulted in 16 dose reductions and 14 premature withdrawals. The role of combination therapy will require a randomized placebo controlled study sufficiently powerful to identify clinically and statistically significant improvements in objective outcome parameters relative to the monotherapies.


Temperature mapping in the canine prostate during transurethrally-applied local microwave hyperthermia. Although the therapeutic effects of heat on tumors have been known for more than a century, only recently has hyperthermia been applied in the treatment of various solid tumors in a scientific fashion. In preliminary clinical trials, heat applied by transurethrally or transrectally placed microwave antennas has been used in the treatment of benign prostatic hyperplasia (BPH) with some success. However, basic information about prostatic temperature distribution and the cellular effects of hyperthermia is lacking. In an attempt to establish the safety of local hyperthermia of the prostate and the precise temperature distribution within the gland, we performed temperature mapping studies in canine prostates. Eight mongrel dogs were anesthetized, and a 16 Fr catheter with three helical coil microwave antennas was placed in the bladder through a perineal urethrostomy. Laparotomy was performed and the bladder opened. The antennas were placed under direct control in the prostatic urethra. Linear array and single point thermometers (Clini-Therm TS1200 thermometry system) were placed (1) within the catheter alongside the antennas, (2) alongside the Foley catheter in the urethra, and (3) longitudinally and radially in the prostate for mapping of tissue temperature. Heating was performed with a 915-MHz ISM frequency Z-80 microprocessor controlled microwave power generator (Prostek 3000, Clini-Therm Corporation, Dallas, TX) using 2-9 watts per channel (6-27 watts total) for 30 min to 1 hr. Baseline body temperatures varied between the individual dogs from 37 degrees to 38 degrees C, but temperature distribution within the prostate was even prior to heating. With relatively low power (6 watts total), temperatures of greater than or equal to 45 degrees C were reached within the catheter. Therapeutic temperatures of approximately 43 degrees C were achieved in the periurethral prostate. Intraurethral temperatures were in general 1-2 degrees C lower than within the catheter. The radial temperature dropoff in the prostate was sharp and about 1 degrees C per 3-mm distance from the catheter. This would limit the area of effectiveness to a periurethral zone of 1 cm in diameter in this model. No unsafe temperature peaks were noted either intraurethrally or between the prostate and the rectum during steady-state conditions. Histologic studies demonstrated an intact rectal wall and varying degrees of prostatic inflammation and/or necrosis despite uniform treatment regimens administered. These studies demonstrate the short-term safety of microwave hyperthermia in the canine prostate. Further studies will be necessary to determine the clinical efficacy and toxicity in men with BPH.


[Exogenous allergic alveolitis: assessment of antigen-specific IgG antibodies using a new chemiluminescence method]. The demonstration of elevated levels of antigen-specific IgG antibodies is of importance for the diagnostic workup of patients with a suspected hypersensitivity pneumonitis. We established a solid phase immunoassay for the determination of these antibodies based on chemiluminescence (CLIA). The CLIA demonstrated a low level of unspecific binding. The reproducibility of the CLIA was evaluated by calculating the intra- and inter-variation; the variation coefficient was 4% (intra-variation) resp. 13% (inter-variation). Using the CLIA we quantified IgG antibodies against Aspergillus fumigatus (A.f.), Micropolyspora faeni (M.f.) and Thermoactinomyces vulgaris (T. v.) in the sera of patients with farmer's lung (FL, n = 10), healthy farmers (n = 13) and normal volunteers without prior hay contact (n = 12). For all 3 antigens the antibody levels of patients with FL were significantly higher than the titers of the other groups. As a comparison all sera were tested with an established solid phase radioimmunoassay (RIA). Using the RIA a statistically significant difference could only be demonstrated between the antibody levels of patients with FL and of normal volunteers without prior hay contact and was limited to the antigens A.f. and M.f. Comparing the results achieved with both assays on a point by point basis there was a good correlation for the antigens A.f. (r = 0.69; p less than 0.0001) and M.f. (r = 0.74; p less than 0.0001), whereas the correlation for T.v. was not satisfying (r = 0.42, p less than 0.02). In summary the CLIA is a promising new method for the determination of antigen specific IgG antibodies with the advantages of a radioimmunoassay without its potential hazards.


Induction of nitric oxide synthase activity by toxic shock syndrome toxin 1 in a macrophage-monocyte cell line. Toxic shock syndrome toxin 1 (TSST-1) is a Mr 22,000 protein produced by Staphylococcus aureus. It is thought to be the cause of toxic shock syndrome. We investigated the hypothesis that TSST-1 induces nitric oxide (NO) synthase and that the NO formed may be involved in the pathogenesis of toxic shock syndrome. We used the murine monocyte-macrophage cell line J744.2 that responds to TSST-1 and also expresses NO synthase activity upon immunological stimulation. J774.2 macrophages stimulated with TSST-1 (10-100 nM) generated nitrite, a breakdown product of NO, and induced concentration-dependent elevations of cGMP in the pig kidney epithelial cell line (LLC-PK1). This latter effect was due to the generation of L-arginine-derived NO for it was (i) abolished by oxyhemoglobin (10 microM), a scavenger of NO, or by methylene blue (10 microM), an inhibitor of NO-activated guanylate cyclase; (ii) potentiated by superoxide dismutase (100 units/ml), which prolongs the life of NO; (iii) inhibited by NG-monomethyl-L-arginine (0.3 mM), an inhibitor of NO synthase; (iv) significantly decreased when L-arginine (0.4 mM) in the medium was replaced by D-arginine (0.4 mM). Moreover, TSST-1 (100 nM) enhanced the activity of cytosolic NO synthase in J774.2 cells. Hydrocortisone (1 microM) but not indomethacin (5 micrograms/ml) or salicylic acid (5 micrograms/ml) prevented the generation of NO2- and the increases in cGMP levels in LLC-PK1 cells induced by J774.2 cells stimulated with TSST-1. The effects of hydrocortisone were partially reversed by coincubation with RU 486 (1 microM), an antagonist of glucocorticoid receptors. Thus, TSST-1 and perhaps other exotoxins produced by Gram-positive bacteria induce NO synthase and the increased NO formation may contribute to toxic shock syndrome and possibly to changes in the immune responses that accompany infection.


Activation of the cytotactin promoter by the homeobox-containing gene Evx-1. Cytotactin is a morphoregulatory molecule of the extracellular matrix affecting cell shape, division, and migration that appears in a characteristic and complex site-restricted pattern during embryogenesis. The promoter region of the gene that encodes chicken cytotactin contains a variety of potential regulatory sequences. These include putative binding sites for homeodomain proteins and a phorbol 12-O-tetradecanoate 13-acetate response element (TRE)/AP-1 element, a potential target for transcription factors thought to be involved in growth-factor signal transduction. To determine the effects of homeobox-containing genes on cytotactin promoter activity, we conducted a series of cotransfection experiments on NIH 3T3 cells using cytotactin promoter-chloramphenicol acetyltransferase (CAT) reporter gene constructs and plasmids driving the expression of mouse homeobox genes Evx-1 and Hox-1.3. cotransfection with Evx-1 stimulated cytotactin promoter activity whereas cotransfection in control experiments with Hox-1.3 had no effect. To localize the sequences required for Evx-1 activation, we tested a series of deletions in the cytotactin promoter. An 89-base-pair region containing a consensus TRE/AP-1 element was found to be required for activation. An oligonucleotide segment containing this TRE/AP-1 site was found to confer Evx-1 inducibility on a simian virus 40 minimal promoter; mutation of the TRE/AP-1 site abolished this activity. To explore the potential role of growth factors in cytotactin promoter activation, chicken embryo fibroblasts, which are known to synthesize cytotactin, were first transfected with cytotactin promoter constructs and cultured under minimal conditions in 1% fetal bovine serum. Although the cells exhibited only low levels of CAT activity under these conditions, cells exposed for 12 h to 10% (vol/vol) fetal bovine serum showed a marked increase in CAT activity. Cotransfection with Evx-1 and cytotactin promoter constructs of cells cultured in 1% fetal bovine serum was sufficient, however, to produce high levels of CAT activity. These findings are consistent with the hypothesis that Evx-1, a homeobox-containing gene, may activate the cytotactin promoter by a mechanism involving a growth-factor signal transduction pathway. More generally, the results support the hypothesis that the place-dependent expression of morphoregulatory molecules may depend upon local cues provided by homeobox genes and their encoded proteins.


cDNA cloning of a liver isoform of the phosphorylase kinase alpha subunit and mapping of the gene to Xp22.2-p22.1, the region of human X-linked liver glycogenosis. We have cloned cDNA molecules encoding another isoform of the alpha subunit of phosphorylase kinase (ATP:phosphorylase-b phosphotransferase, EC 2.7.1.38). Sequence comparison with the previously characterized muscle isoform reveals a pattern of highly conserved and variable domains and demonstrates that the isoforms are the products of distinct genes. In contrast to the muscle isoform gene, PHKA1, the gene of this additional isoform, PHKA2, is predominantly expressed in liver and other nonmuscle tissues. It was mapped to the distal short arm of the human X chromosome (Xp22.2-p22.1), the same region to which human X-linked liver glycogenosis due to phosphorylase kinase deficiency has been mapped. Thus, X-linked liver glycogenosis is probably caused by mutations affecting PHKA2.


Paradoxical transcriptional activation of rat liver cytochrome P-450 3A1 by dexamethasone and the antiglucocorticoid pregnenolone 16 alpha-carbonitrile: analysis by transient transfection into primary monolayer cultures of adult rat hepatocytes. The family 3A cytochromes P-450, among the most abundant members of this supergene family of microsomal hemoproteins expressed in animal and human liver, are inducible by glucocorticoids but also by such antiglucocorticoids as pregnenolone 16 alpha-carbonitrile (PCN). To investigate the mechanism for this nonclassical glucocorticoid effect, we analyzed the ability of 1.5 kilobases of DNA or of its successive subsegments isolated from the 5' flanking region of the rat CYP3A1 structural gene to modulate transcription of a reporter gene consisting of a viral promoter coupled to the chloramphenicol acetyltransferase (CAT) structural gene (expression vector pBLCAT2) and transiently expressed in a homologous cell system consisting of primary monolayer cultures of adult rat hepatocytes in which CYP3A1 mRNA and protein are inducible. The CAT activity measured after chimeric gene constructions were transferred into the cultured rat hepatocytes by lipofection increased as much as 7.2-fold if the cells were treated with dexamethasone (DEX). One CYP3A1 fragment (positions -220 to -56; 164 base pairs), which does not contain a traditional glucocorticoid responsive element, conferred dose-dependent DEX responsiveness independent of its orientation but not its position in pBLCAT2. This construction was activated by addition of PCN to the cultures and was synergistically induced by PCN plus DEX. In contrast, induction of CAT activity in cultures containing MMTVCAT, a plasmid containing the CAT gene controlled by the mouse mammary tumor virus long terminal repeat, was unaffected by PCN treatment, required lower concentrations of DEX for a maximal response, and was inhibited by treatment with DEX plus PCN. We conclude that a primary mechanism for induction of CYP3A1 is stimulated transcription through a pathway activated by steroid hormones.


Molecular cloning and expression of chicken C-terminal Src kinase: lack of stable association with c-Src protein. Cloning and sequencing of chicken C-terminal Src kinase (CSK), a tyrosine kinase that phosphorylates the regulatory C-terminal tyrosine residue present on cytoplasmic tyrosine kinases of the Src family, demonstrated a high degree of interspecies conservation as well as src homology 2 and 3 domains N-terminal to the kinase domain. The lack of autophosphorylation sites distinguishes CSK from other tyrosine kinases. CSK is unique and does not belong to a gene family, suggesting that it may phosphorylate other members of the Src family of tyrosine kinases in addition to c-Src. Since complex formation between c-Src and CSK seemed a likely regulatory step in the control of c-Src kinase activity, such an association was investigated by immunoprecipitation and Western blotting as well as intracellular localization studies. Although some portions of CSK were found in a membrane fraction, no complex formation between CSK and c-Src was observed, suggesting that the src homology 2 domain of CSK does not play a role in the direct interaction of c-Src.


Chromosomal position of rearranging gene segments influences allelic exclusion in transgenic mice. Formation of a complete immunoglobulin heavy-chain transcription unit involves the ordered rearrangement of variable (V), diversity (D), and joining (J) region gene segments. In antibody-producing cells, this process is regulated such that only one of two antibody genes is expressed. Experiments with transgenic mice suggest that this mechanism, known as allelic exclusion, is mediated through the membrane-bound form of the immunoglobulin heavy chain. However, in all transgenic lines produced to date exclusion of the endogenous genes by the transgene is incomplete. To characterize the molecular basis for this escape from regulation, we have examined the rearrangements of endogenous immunoglobulin heavy-chain genes. We find that a transgene that encodes the membrane-bound form of human IgM efficiently inhibits rearrangements of endogenous gene segments located at the 5' end of the heavy-chain locus. However, recombining elements found at the 3' end of the locus escape and continue to undergo recombination. A transgene that encodes the secreted form of the same immunoglobulin protein has no effect on recombination, regardless of position of the recombining segment in the chromosome. These results have important implications for our understanding of the control of allelic exclusion.


Granule membrane protein 140 (GMP140) binds to carcinomas and carcinoma-derived cell lines. The glycoproteins granule membrane protein 140 (GMP140), endothelial-leukocyte adhesion molecule 1 (ELAM-1), and Leu-8 are members of a family of glycoprotein receptors (selectins or LEC-CAMs) that play an important role in adhesive interactions between circulating leukocytes and vascular endothelium. Recently it has been reported that ELAM-1 is able to mediate the binding of the colon carcinoma cell line HT-29 to cytokine-activated vascular endothelium, suggesting that tumor cell adhesion to vascular endothelium, a prerequisite for tumor extravasation and metastasis, is in part the result of adhesive interactions between blood-borne tumor cells and cell surface proteins expressed by vascular endothelium. Here, using an approach in which soluble immunoglobulin chimeras of the GMP140 and ELAM-1 receptors were prepared and used to carry out immunohistological studies, we establish that GMP140 binds to tumor cells in a variety of human carcinoma tissue sections (colon, lung, and breast), whereas ELAM-1 binds exclusively to tumor cells in colon carcinoma tissue sections. In addition, GMP140 was found to bind to the cell surface of a number of cell lines derived from various carcinomas but not from melanomas, whereas ELAM-1 bound only colon carcinoma cell lines. We further investigated the nature of the ligands of GMP140 and ELAM-1 on the surface of the carcinoma cells and found that the GMP140 ligand on the surface of tumor cells appears to be distinct from that expressed on the myeloid cell line HL-60. Neuraminidase treatment of a breast carcinoma cell line does not affect, or in some instances increases, GMP140 binding, whereas it completely abolishes GMP140 binding to HL-60 cells. On the other hand, the ligand of ELAM-1 on both the colon carcinoma and HL-60 cells is neuraminidase sensitive in accord with its identification as sialyl-CD15. Parallel results were obtained with neuraminidase-treated frozen carcinoma tissue sections. The present findings form the basis for investigating the role of GMP140 in tumor invasiveness and metastasis.


Isolation and characterization of human cDNA clones encoding a high mobility group box protein that recognizes structural distortions to DNA caused by binding of the anticancer agent cisplatin. Human cDNA clones encoding a structure-specific recognition protein, SSRP1, that binds specifically to DNA modified with cisplatin have been isolated and characterized. The SSRP1 gene maps to human chromosome 11q12. The cDNA clones, obtained by using partial-length cDNAs described previously, predict an 81-kDa protein containing several highly charged domains and a stretch of 75 amino acids 47% identical to a portion of the high mobility group (HMG) protein HMG1. This HMG box most likely constitutes the structure recognition element for cisplatin-modified DNA, with the probable recognition motif being the local duplex unwinding and bending toward the major groove that occurs upon formation of intrastrand cis-[Pt(NH3)2]2+ d(GpG) and d(ApG) cross-links. Although the DNA recognition properties of members of the HMG-box family of proteins have been characterized with respect to their sequence specificity, the present work demonstrates that proteins with this domain can recognize particular DNA structures as well. The Pt-DNA SSRP described here is the human homolog of a recently identified mouse protein that binds to recombination signal sequences [Shirakata, M., Hüppi, K., Usuda, S., Okazaki, K., Yoshida, K. & Sakano, H. (1991) Mol. Cell. Biol. 11, 4528-4536]. These sequences have been postulated to form stem-loop structures, further implicating local bends and unwinding in DNA as a recognition target for HMG-box proteins. Expression analysis in a variety of tissues and cisplatin-resistant cell lines and the inability of cisplatin to induce the message in HeLa cells argue against a direct link between SSRP1 mRNA levels and the response of cells to the drug.


A conditional yeast mutant deficient in mRNA transport from nucleus to cytoplasm. Transport of mRNA from nucleus to cytoplasm is critical for eukaryotic gene expression; however, the mechanism of export is unknown. Selection and screening procedures have therefore been used to obtain a family of temperature-sensitive conditional mutants of Saccharomyces cerevisiae that accumulate poly(A)+ RNA in the nucleus when incubated at 37 degrees C, as judged by in situ hybridization. In one such mRNA transport mutant, mtr1-1, RNA synthesis continues, the export of poly(A)+ RNA is inhibited, intranuclear poly(A)+ is remarkably stable, and protein synthesis gradually stops. Thus, there is no tight coupling between RNA synthesis and export. The export lesion is reversible. Although mRNA export is clearly not a default option, neither inhibition of protein synthesis, inhibition of mRNA splicing, nor inhibition of poly(A)-binding protein function blocks export of the average poly(A)+, as judged by in situ hybridization. Further analysis of the family of mtr mutants should help map the path of RNA transport.


Characterization of the cystic fibrosis transmembrane conductance regulator in a colonocyte cell line. An anti-peptide antibody raised to the C-terminal sequence of the cystic fibrosis transmembrane conductance regulator (CFTR) was used to examine CFTR immunoreactivity in the T84 colonocyte cell line. Immunoblots of T84 cell lysates detected CFTR as a 170-kDa protein that appeared as a broad band or doublet in SDS/PAGE. This protein comigrated with the predominant immunoblot signal detected in human pancreas and colon. An equivalent protein was detected as a prominent substrate for protein kinase A and for protein kinase C in T84 cell immunoprecipitates with this antibody. The immunoprecipitated protein resembled the protein detected by immunoblot in that both proteins showed the same change in electrophoretic mobility after digestion by N-Glycanase. The precipitated protein was indentified as CFTR by two criteria. First, the same protein was immunoprecipitated with an antibody to a different CFTR peptide, [Lys102]CFTR-(102-116). Second, two-dimensional phosphopeptide mapping was used to compare the immunoprecipitated protein with a bacterially expressed protein known to contain most of the predicted protein kinase A phosphorylation sites in CFTR. Because the six most prominent peptides in each map were equivalent, these maps confirm that the precipitated protein is CFTR. By using these antibodies for immunofluorescence and immunoperoxidase staining, CFTR was localized to the apical region of T84 cells grown in tumors and in monolayers. Thus, T84 cells express CFTR at sufficient levels to permit identification and immunochemical studies of this protein in its endogenously occurring form.


Preproenkephalin promoter "cassette" confers brain expression and synaptic regulation in transgenic mice. The preproenkephalin A gene is a neurotransmitter gene whose expression can be modulated "trans-synaptically" by changes in neuronal activity. DNA sequences lying within 200 base pairs of this gene's transcription start site resemble consensus binding sites for several transcription factor families. In nonneuronal cell cultures, this promoter region is sufficient to mediate gene responses to depolarization, phorbol esters, adenylate cyclase, and calcium fluxes. To assess the role that these cis-acting elements could play in preproenkephalin expression and regulation in vivo, the expression of a construct containing this 200-base-pair region fused to the chloramphenicol acetyltransferase gene was examined in transgenic mice. This promoter confers modest expression in brain, adrenal, and small intestine, with substantially higher levels in testis. These elements confer trans-synaptic regulation in two well-studied models of trans-synaptic preproenkephalin upregulation but not in a third system, underscoring the specificity of the regulatory sequence elements implicated in the synaptic regulation of neuronal genes.


The Na+/H+ antiporter cytoplasmic domain mediates growth factor signals and controls "H(+)-sensing". The amiloride-sensitive Na+/H+ exchanger (NHE1 human isoform) is activated in response to diverse mitogenic and oncogenic signals presumably through phosphorylation. To get insight into the activating mechanism, a set of deletion mutants within the C-terminal cytoplasmic domain of NHE1 has been generated. These mutant forms expressed in antiporter-deficient fibroblasts revealed that deletion of the complete cytoplasmic domain (i) preserves amiloride-sensitive Na+/H+ exchange and activation by intracellular H+, (ii) reduces the affinity of the internal "H(+)-modifier site" in a manner mimicked by cellular ATP depletion, and (iii) abolishes growth factor-induced cytoplasmic alkalinization. We conclude that NHE1 can be separated into two distinct functional domains. One is an N-terminal transporter domain (T) that has all the features required to catalyze amiloride-sensitive Na+/H+ exchange with a built-in H(+)-modifier site. The other is a C-terminal cytoplasmic regulatory domain (R) that (i) determines the set point value of the exchanger and (ii) mediates growth factor signals by interacting with the "H(+)-sensor" in a phosphorylation-dependent manner.


Cloning of a D-type cyclin from murine erythroleukemia cells. We report the complete coding sequence of a cDNA, designated CYL2, derived from a murine erythroleukemia cell library. CYL2 is considered to encode a D-type cyclin because (i) there is cross hybridization with CYL1 (a murine homolog of human cyclin D1) and the encoded protein has 64% amino acid sequence identity with CYL1 and (ii) murine erythroleukemia cell-derived CYL2 contains an amino acid sequence identical to that previously reported for the C-terminal portion of a partially sequenced CYL2. Transcripts of murine erythroleukemia cell CYL2 undergo alternative polyadenylylation like that of human cyclin D1. A major 6.5-kilobase CYL2 transcript changes its expression during the cell cycle with a broad peak through G1 and S phases and a decrease in G2/M phases. The present findings suggest that CYL2 plays a role in the G1 to S phase progression.


Down-regulation of a member of the S100 gene family in mammary carcinoma cells and reexpression by azadeoxycytidine treatment. A cDNA clone, designated CaN19 (originally called clone 19), isolated by subtractive hybridization, contains sequences that are preferentially expressed in normal mammary epithelial cells but not in breast tumor cells. Comparison of its deduced amino acid sequence with sequences in the GenBank data base revealed similarity with the S100 protein family, a group of small Ca(2+)-binding modulator proteins involved in cell cycle progression and cell differentiation. CaN19 expression is down-regulated in normal cells by A23187, a calcium ionophore, suggesting that its regulation is calcium-dependent. We have assigned CaN19 to human chromosome 1q21-q24, a region containing four other S100-related genes. In contrast to CaN19 mRNA expression, most members of the S100 protein family are activated or overexpressed in tumor cells. Synchronization experiments by growth-factor deprivation demonstrated a biphasic induction of CaN19 expression in normal cells, approximately 2-fold in early G1 phase and another 2- to 3-fold at the G1/S boundary. Exposure of mammary tumor cells to 5-aza-2'-deoxycytidine, an inhibitor of DNA methylation, reactivated the expression of CaN19 mRNA.


Induction of broadly cross-reactive cytotoxic T cells recognizing an HIV-1 envelope determinant. An immunodominant determinant for cytotoxic T lymphocytes (CTLs) exists in the hypervariable portion of human immunodeficiency virus-1 (HIV-1) gp160. Three mouse CTL lines (specific for isolates MN, RF, and IIIB) were examined for recognition of homologous determinants from distinct isolates. Only MN-elicited CTLs showed extensive interisolate cross-reactivity. Residue 325 played a critical role in specificity, with MN-elicited CTLs responding to peptides with an aromatic or cyclic residue and IIIB-induced cells recognizing peptides with an aliphatic residue at this position. CTL populations with broad specificities were generated by restimulation of IIIB-gp160 primed cells with MN-type peptides that have an aliphatic substitution at 325. This represents an approach to synthetic vaccines that can generate broadly cross-reactive CTLs capable of effector function against a wide range of HIV isolates.


Surgical treatment of gastric cancer. Proximal, mid, and distal stomach. At present, only radical surgical resection of gastric cancer offers a chance for cure. The objective of an operation for patients with disease that is confined locally is to maximize the potential for cure. The objective for patients with advanced incurable disease, obstruction, hemorrhage, and intractable pain is to provide the best palliation. The evaluation, staging, and surgical management of gastric cancer are described.


Sputum sol phase proteins and elastase activity in patients with clinically stable bronchiectasis. Inflammatory and proteolytic activity occurs in sputum from patients with stable purulent bronchiectasis and has been proposed as the main pathogenetic mechanism of the disease. This study was designed to define further the role of inflammation and proteolysis in bronchiectasis. Neutrophil elastase activity, sputum concentrations of the serum derived inhibitors alpha 1 antiproteinase and alpha 2 macroglobulin, and the sputum to serum ratios of albumin and C reactive protein concentration were measured in 26 patients with bronchiectasis. Free elastase activity was found in 15 sputum samples. A trend to higher proteolytic and inflammatory activity was found between mucoid and purulent sputum samples, suggesting that inflammatory and proteolytic activities are related to the macroscopic degree of purulence. Purulent sputum had a high sputum to serum ratio of C reactive protein, suggesting local production or active transport of this protein into bronchial secretions. C reactive protein was more sensitive than albumin in detecting a higher degree of inflammation in elastase positive samples. The finding of greater concentrations of alpha 2 macroglobulin in purulent and elastase positive samples than in mucopurulent, mucoid and elastase negative sputum samples suggests that this inhibitor may have a role in the proteolysis-antiproteolysis balance in bronchial secretions.


A comparison of endocrine and exocrine function after pancreatic fragment autotransplantation into splenic pulp, portal vein, and hepatic parenchyma. Three sites were evaluated for potential pancreatic fragment autotransplantation. Both endocrine and exocrine functions were evaluated following autotransplantation into splenic pulp, portal vein, or hepatic parenchyma in 44 pancreatectomized dogs. Cholecystic bile amylase concentrations in the hepatic parenchyma group surviving more than 2 months were elevated significantly, and choledochal bile amylase concentrations increased markedly following pancreozymin-secretin injection. In contrast, bile amylase concentrations in dogs with intrasplenic or intraportal implants were low and did not respond to PS injection. Histologically pancreatic autografts in hepatic parenchyma revealed marked proliferation of exocrine tissue with abundant zymogen granules and reconstruction of the acinar lobules with a few islets. These acinar cells in the hepatic parenchyma were ultrastructurally normal. Transplant endocrine function, estimated by K values, was significantly better after splenic pulp and portal vein than after a hepatic parenchyma implantation, but no group improved during 1-year follow-up. Glucose-stimulated initial insulin responses were abnormally low in all recipients. Islet B cells lacked mature insulin granules, such as seen in normal resting B cells. This ultrastructural finding implies a persistent demand on the B cells and may explain the spontaneous recurrence of hyperglycemia and the diminished initial insulin response to a glucose load. This study indicates that euglycemic recipients of pancreatic fragment autotransplantation remain unstable and prediabetic.


Evidence that nitric oxide production by in vivo allosensitized cells inhibits the development of allospecific CTL. The oxidative metabolism of L-arginine to its bioactive product, nitric oxide (.N = O) has been shown to inhibit rat splenocyte mixed lymphocyte reactions. To determine if alloantigen-induced .N = O production might be operative in vivo, cells that had infiltrated a rat sponge matrix allograft were tested for de novo .N = O production as well as .N = O production upon restimulation with the sensitizing alloantigen. When graft-infiltrating cells were placed in culture, a peak in de novo .N = O production was observed by day 6 graft-infiltrating cells, the time when donor-specific CTL activity by the graft-infiltrating cells was first observed. Upon restimulation with alloantigen, allograft-infiltrating cells produced greatly increased levels of .N = O, and this production was associated with inhibition of lymphocyte cytolytic function. The addition of NG-monomethyl-L-arginine (NMA), the competitive inhibitor of oxidative L-arginine metabolism, inhibited .N = O production and promoted allospecific CTL development. Both observed effects of NMA were reversed by addition of excess L-arginine. Cytokine(s) able to induce proliferation of the IL-2-dependent T cell line CTLL-2 could be detected in alloantigen-stimulated cultures in both the presence and absence of NMA. However, proliferation of the graft-infiltrating cells in response to these cytokines was observed only in the presence of NMA. The immunosuppressive macrolide FK506 was a potent inhibitor of .N = O production in these cultures, presumably acting by inhibiting the production of those cytokines that induce the oxidative L-arginine pathway.


Acute experimental pancreatitis in rat induced by sodium taurocholic acid: objective quantification of pancreatic necrosis. Retrograde injection of 5% sodium taurocholic acid (TA) in Wistar rat pancreatic duct is followed by acute pancreatitis, resulting in 100% mortality within 36 h. Biochemical determinations show raised levels of amylase in ascites and blood. Necrosis has been measured using seven morphometric characteristics of pathological changes that add precise information on the type and extension of the pancreatic lesion. The percentage of necrotic tissue (by area) seems to be the most objective parameter. Necrosis appears 6 h after TA infusion, being 5.77% in extent after 12 h, 14.9% after 24 h and animals die with an area of 29.5% necrosis. This experimental model seems to one in which physiopathological and therapeutic trials on acute pancreatitis may be tried out.


Microcytic variant of thymoma: histological and immunohistochemical findings in two cases. Two cases of microcytic variant of thymoma are presented. Both tumours were well encapsulated with a yellow-whitish colour and soft consistency. Microscopically, they consisted of round cells, having ample vacuolated cytoplasm. Fat droplets were not detected in one case where fat staining was performed. Immunohistochemically, the tumour cells were strongly positive for AE1/AE3, MB1, MB2, and LN1 and faintly positive for epithelial membrane antigen. They lacked any other leucocyte antigens. Leu 7 showed a positive immunoreaction in a ring-like or homogeneous pattern, compatible with the cytoplasmic vacuoles or cytoplasm. Ultrastructurally, the vacuoles resembled cystically dilated rough-surfaced endoplasmic reticulum. Desmosome-like structures (case 1) and intermediate junctions (case 2) were identified between adjacent cells. These findings indicate that the present tumours belong to a category of microcystic thymoma. The vacuoles were attributed to excess accumulation of Leu-7-positive material, probably in the cystically dilated endoplasmic reticulum.


Primary thymic carcinoid with Cushing's syndrome. In a 52-year-old Caucasian man osteopoikilosis had been misdiagnosed roentgenologically 2 years before his death. Gradually he developed Cushing's syndrome and ultimately superior vena caval obstruction. At autopsy a primary thymic carcinoid with extensive osteoblastic bone metastasis was found. Immunohistochemically the tumor was shown to be positive for adrenocorticotropic hormone (ACTH), cytokeratin (KL1), neuron-specific enolase, synaptophysin, chromogranin and glucagon. Remarkably the tumour was negative for serotonin despite high urinary hydroxyindolacetic acid levels. Bilateral hyperplasia of the adrenal cortex was found. The adenohypophysis showed a considerable reduction of ACTH-producing cells and numerous Crooke's cells with a characteristic immunohistochemical pattern.


Synergistic action of postextrasystolic potentiation and diperdipine on left ventricular wall motion abnormalities in coronary heart disease. The effects on left ventricular function of postextrasystolic potentiation and the dihydropyridinic calcium antagonist diperdipine, alone or in combination, were studied in 14 patients with coronary heart disease by means of two consecutive left ventricular angiographies. To ensure the reproducibility of coupling intervals of the extrasystolic and postextrasystolic beats, the heart was paced during both angiographies. Results showed that postextrasystolic potentiation and diperdipine improved left ventricular ejection fraction to the same degree, but the mechanisms of such an improvement were different and consisted, respectively, of a positive inotropic effect associated with an increase in preload and a slight increase in preload coupled with a marked decrease in afterload. Diperdipine did not abolish the inotropic component of postextrasystolic potentiation, and a combination of the two interventions had additive effects on the improvements in left ventricular ejection fraction and regional wall motion analyzed by the centerline method. Ventricular segments that were normokinetic or hypokinetic during the control basal cycle responded equally to postextrasystolic potentiation and to diperdipine, whereas the former intervention alone had no significant effect on akinetic segments. Diperdipine restored the responsiveness of akinetic segments to postextrasystolic potentiation, a finding that, although it remains to be confirmed by independent techniques, may be interpreted as a possible consequence of improved calcium metabolism or coronary flow in ischemic but still viable myocardium. However, it is concluded that the calcium antagonist revealed a contractile reserve in most of the severely asynergic areas, which would have otherwise been judged to be irreversibly damaged on the basis of the unresponsiveness to postextrasystolic potentiation alone.


[Significance of pathological alpha-1-fetoprotein levels in the framework of prenatal diagnosis]. Merkatz (1984) showed that in 68% of live birth with Down Syndrome maternal age was 34 years or less. It would be desirable to define a high risk collective of women younger then 35 years with the help of a screening test, which could further on lead to a prenatal diagnostics. Several studies showed that low maternal serum alpha-fetoprotein levels (SAFP) in pregnancy is associated with fetal chromosomal abnormalities. The results of a retrospective study at the Department of Obstetrics and Gynecology at University of Mainz showed that maternal SAFP-level does not accomplish the demand of a prenatal screening test, because only 17% of the Down Syndrome could be diagnosed by this parameter. In case of trisomy 13 and 18 SAFP-concentrations were normal or even increased, maybe on account of the combination with other malformations like neural-tube or abdominal-wall defects.


Weekly dose-intensive chemotherapy in patients with small-cell lung cancer: a pilot study. The study was aimed to evaluate the feasibility of dose-intensive chemotherapy given on a weekly basis for 12 weeks. Seventeen [7 with limited disease (LD) and 10 with extensive disease (ED)] previously untreated patients with small-cell lung cancer (SCLC) were treated with the cisplatin, vincristine, doxorubicin, and etoposide (CODE) regimen. Recombinant human granulocyte colony-stimulating factor (rhG-CSF) was given to eight patients for the purpose of increasing the dose intensity. Overall response rate was 88%, with a 29% complete response. The median survival times were greater than 20.5 months for LD patients and 8.1 months for ED patients. Overall actual dose intensity was 88% of planned protocol. The major toxicity was myelosuppression. Fifteen patients (88%) had grade 3 or 4 leukopenia. Other problems were weight loss and worsening of performance status during the treatment. RhG-CSF significantly reduced leukopenic nadirs and shortened the neutropenic period. Our preliminary results indicate that 12 cycles of the CODE regimen on a weekly schedule is effective for SCLC, but is also associated with significant toxicity.


Treatment and prognosis of orbital non-Hodgkin's lymphomas. Twenty patients with orbital lymphomas were treated and followed over a 14-year period. Ten of these patients had well-differentiated lymphocytic lymphoma (WDL), and all of them were clinical stage IE. Five patients had nodular poorly differentiated lymphocytic lymphomas (NPDL), three patients had diffuse histiocytic lymphomas (DHL), one had nodular mixed-cell lymphoma, and one had diffused mixed-cell lymphoma. The patients with WDL received local radiation therapy, and all of them entered completed remissions. The projected survival at 10 years was 100% for these patients. The patients with low-grade lymphomas with advanced disease were treated with chlorambucil and prednisone or cytoxan and vincristine. The patients with high-grade lymphomas received treatment with methotrexate, cyclophosphamide, doxorubicin, vincristine, bleomycin, dexamethasone (mBACOD) or cyclophosphamide, vincristine, methotrexate, cytosine arabinoside, leucovorin (COMLA). The overall survival estimate for all patients was 63% at 10 years. The disease-free survival was 49% at 10 years. The patients with high-grade lymphomas had a poor prognosis, with no survivors after 4 years. This study suggests that patients with WDL usually present with localized disease to the involved orbit, and have an excellent prognosis with radiation therapy alone. Patients with high-grade orbital non-Hodgkin's lymphomas have a poor outcome despite use of aggressive combination chemotherapy regimens.


Erythropoietin affects the maturation pattern of fetal G-CSF-responsive progenitors. A transient hyporegenerative neutropenia has been reported in neonates, but not in older children or adults, undergoing treatment with recombinant erythropoietin (epo). Monocytopenia has not been reported. We postulated that epo might selectively reduce the responsiveness of neonatal progenitors to Granulocyte Colony-Stimulating Factor (G-CSF), while not similarly affecting their responsiveness to Macrophage Colony-Stimulating Factor (M-CSF). To test this hypothesis two types of experiments were performed. First, progenitors of adult or fetal origin were pre-incubated with epo (or control), then washed, and their responsiveness to G-CSF and M-CSF evaluated in clonogenic culture assays. Second, clonogenic maturation was initiated using either G-CSF or M-CSF, after which the effect of a late addition of epo to the developing clones was evaluated. Indeed, pre-incubation with epo resulted in production of fewer neutrophils from fetal progenitors grown in G-CSF (P less than 0.001), but it did not reduce the number of macrophages generated from progenitors grown in M-CSF. Adding epo to the already-developing G-CSF-responsive and M-CSF-responsive adult and fetal clones did not alter colony development. Thus, epo appears to have an action on G-CSF-responsive, but not-M-CSF-responsive fetal progenitors, resulting in reduced production of neutrophils. This effect is no longer apparent, however, when progenitors have matured to the 8-cell clone stage.


Fluctuations in serum cytokine levels in the patient with cyclic neutropenia. The fluctuations of the levels of serum cytokines in a patient with cyclic neutropenia were studied. The greatest fluctuation was found in granulocyte colony-stimulating factor (G-CSF). Tumor necrosis factor-alpha level fluctuated inversely with fluctuation of G-CSF, and oscillation of interleukin (IL)-6 level preceded that in G-CSF level. It is likely that the variations of the levels of cytokines play a significant role in regulating hematopoiesis in cyclic neutropenia. Our results suggested that the stage of the granulocyte-committed stem cell is the major step of the defect, and the defect at the stage of the multipotent stem cell or bone marrow microenvironment was also suggested.


[Distal post-traumatic edema--symptom of a sympathetic reflex dystrophy (Sudeck's disease)?]. The present paper describes various mechanisms, possibly being involved in the development of the posttraumatic, distally generalized edema. New ideas point to a special importance of the sympathetic vasoconstrictor system for this clinical phenomenon, since this system could induce an enhanced venoconstriction at the exit of the capillary bed, which would result in an edema producing diminished venous return. Since the distally generalized edema is an initially and very commonly occurring symptom of reflex sympathetic dystrophy (M. Sudeck), the observation of such an edema should lead one to look for further symptoms of this disorder, especially for the typical triad of autonomic (sympathetic), motor, and sensory disturbances.


Circulating CD34+ cells: an adverse prognostic factor in the myelodysplastic syndromes. As part of an epidemiological survey of myelodysplastic syndromes (MDS) in southern Tasmania, 62 MDS patients identified over a 2 year period were tested for the presence of CD34, the human progenitor cell antigen (HPCA), in their peripheral blood. The results were correlated with transformation to acute myeloid leukemia (AML) and patient survival, and CD34+ status was compared as a prognostic indicator with Bournemouth score, cytogenetics, and CFU-GM colony growth which were also assessed. Circulating CD34+ cells were found in 23 of the 62 MDS patients; 9 of the 23 patients with circulating CD34+ cells transformed to AML, as compared with none of the 39 CD34 negative patients (P less than 0.0001); and 11 of the 23 patients with circulating CD34+ cells were dead at the end of the 2 year period, as opposed to 6 of the 39 with no CD34+ cells (P less than 0.03). The Bournemouth score was also significantly associated with transformation to AML (P less than 0.0001) and poor survival (P less than 0.04). These were the only significant associations of the possible prognostic factors studied with either transformation or survival. In summary, the presence of circulating CD34+ cells was significantly associated with both progression to AML and poor survival and was found to be a better prognostic indicator than cytogenetics or CFU-GM colony growth.


Xeroderma pigmentosum and Cockayne syndrome: overlapping clinical and biochemical phenotypes. Two siblings are described whose clinical presentation of cutaneous photosensitivity and central nervous system dysfunction is strongly reminiscent of the DeSanctis-Cacchione syndrome (DCS) variant of xeroderma pigmentosum. An extensive clinical evaluation supported a diagnosis of DCS and documented previously unreported findings. In vitro fibroblast studies showed UV sensitivity that was two to three times that of normal controls. However, neither a post-UV-irradiation DNA excision-repair defect indicative of XP nor a semiconservative DNA replication defect indicative of XP variant was found. Rather, a failure of RNA synthesis to recover to normal levels after UV exposure was observed, a biochemical abnormality seen in Cockayne syndrome (CS), one of the premature-aging syndromes with clinical UV sensitivity. These patients, therefore, clinically have XP, but their biochemical characteristics suggest CS. The reason(s) for the severe neurologic disease, in light of the relatively mild cutaneous abnormalities, is unclear. Other cases with unusual fibroblast responses to irradiation have been noted in the literature and, along with the data from our patients, reinforce the notion of the complexity of DNA maintenance and repair.