Independent evolution of monkeypox and variola viruses. Smallpox was eradicated more than 10 years ago, but infection with another Orthopoxvirus, monkeypox virus, can result in a clinical picture resembling smallpox. Human infection with monkeypox virus is extremely rare, not easily transmitted, and confined to the rain forest belt of Africa (Z. Jezek and F. Fenner, p. 81-102, in Human Monkeypox, 1988). Evidence that variola virus, the causative agent of smallpox, might be readily derived from monkeypox virus was presented [S. S. Marennikova and E. M. Shelukhina, Nature (London) 276:291-292, 1978; S. S. Marennikova, E. M. Shelukhina, N. N. Maltseva, and G. R. Matsevich Intervirology 11:333-340, 1979], but this was not confirmed [K. R. Dumbell and L. C. Archard, Nature (London) 286:29-32, 1980] and was subsequently discounted (J. J. Esposito, J. H. Nakano, and J. F. Obijeski, Bull. W.H.O. 63:695-703, 1985). Although enough difference between the genomes of monkeypox and variola viruses to rule out a simple interconversion has been demonstrated [K. R. Dumbell and L. C. Archard, Nature (London) 286:29-32, 1980; J. J. Esposito and J. C. Knight, Virology 143:230-251, 1985; J. J. Esposito, J. H. Nakano, and J. F. Obijeski, Bull. W.H.O. 63:695-703, 1985; M. Mackett and L. C. Archard, J. Gen. Virol. 45:683-701, 1979], the possibility that monkeypox virus was a more remote ancestor of variola virus remained. We have now identified a sequence in monkeypox virus DNA which is a homolog of a 1,065-bp open reading frame in the conserved region of the variola virus genome but which has multiple deletions. This is strong evidence that monkeypox virus is not ancestral to variola virus and strengthens confidence in the long-term success of smallpox eradication.


Analysis of the UL36 open reading frame encoding the large tegument protein (ICP1/2) of herpes simplex virus type 1. Using peptide antisera specific for regions within the N terminus and C terminus of the predicted UL36 gene product, immunoblotting experiments were performed to demonstrate definitively that ICP1/2 is encoded by the UL36 gene. These data also suggest that both the cell- and the virion-associated forms of ICP1/2 are colinear with the complete predicted amino acid sequence of the UL36 gene. Computer-assisted analyses of the predicted amino acid sequence of the UL36 gene revealed the presence of two putative leucine zipper-type motifs and a potential ATP-binding domain. The possible functions of these consensus domains will also be discussed.


Therapeutic use of ganciclovir for invasive cytomegalovirus infection in cadaveric renal allograft recipients. Between November 1987 and September 1989, 419 cadaveric renal transplants were performed at our university. Of the patients 36 (8.6%) had invasive cytomegalovirus infection documented by gastric or duodenal mucosal biopsy in 23 (64%), bronchoalveolar lavage in 12 (33%), allograft biopsy or nephrectomy specimen in 5 (14%) and/or liver biopsy in 1 (3%). Cytomegalovirus severity was defined as mild in 27 patients, moderate in 6 and severe in 3. Ganciclovir [9-(1,3-dihydroxy-2-propoxymethyl)-guanine] was begun once the diagnosis was confirmed by histology or culture at a median of 56 days from transplantation (range 28 to 133 days). Duration of ganciclovir therapy was a minimum of 7 days or until fever was absent for 5 consecutive days (mean 12.2 +/- 3.5 days, range 4 to 21). Ganciclovir was well tolerated and side effects were limited to de novo neutropenia (7 patients), thrombocytopenia (2) and rash (1). Initial clinical improvement was observed in all patients. Two patients had recurrent cytomegalovirus infections that responded to a second course of ganciclovir. The 1-year actuarial patient survival was 100%. At a mean followup of 12.7 +/- 6.2 months 19 patients retained allograft function with a mean serum creatinine of 2.5 mg./dl. (range 1.2 to 4.6). Ganciclovir appears to be a safe and effective drug for the treatment of tissue invasive cytomegalovirus infection in cadaver renal transplant recipients. Prompt institution of this drug at diagnosis of invasive cytomegalovirus may lower the mortality rate formerly associated with this disease.


Control of massive hematuria in idiopathic hemorrhagic cystitis after administration of conjugated estrogen. A renal transplant patient with severe hemorrhagic cystitis of idiopathic etiology was treated initially with intravenous administration of conjugated estrogen (1 mg./kg.), followed on day 2 and thereafter with 5 mg. per day orally for 3 weeks. Hematuria decreased in intensity within 10 hours and disappeared within 48 hours. Hematuria did not recur by 6 months after completion of oral doses of conjugated estrogen. Complications or other side effects were not observed. In our experience conjugated estrogen controls hematuria in patients with idiopathic hemorrhagic cystitis and this form of treatment must be considered in this condition.


Gonadotropin-releasing hormone specific binding sites in normal and malignant renal tissue. Renal cell carcinoma has been reported to contain estrogen and progesterone receptors. Thus, it has been suggested that these tumors are hormone dependent in a similar manner described for the breast and prostate cancers. It has been recently shown that mammary and prostate tumor cells contain gonadotropin-releasing hormone (GnRH) receptors and are growth inhibited directly by GnRH antagonists. In this study we examined for the presence of GnRH, estrogen and progesterone receptors in normal and malignant renal tissues. Estrogen receptors were found both in the normal and malignant kidney while progesterone receptors were present only in the normal tissue. Specific binding of [125I]buserelin, a GnRH agonist, was evident in renal carcinoma and in normal kidney and was displaced with equal efficiency by unlabeled buserelin and by D-Trp6-GnRH, but not by unrelated peptides such as thyrotropin releasing hormone and oxytocin. The non-linear scatchard curve obtained for buserelin binding, suggests the presence of at least two binding sites, one with high affinity in the nanomolar range and another in the micromolar range.


Dimercaptosuccinic acid renal scintigraphy for the evaluation of pyelonephritis and scarring: a review of experimental and clinical studies. Renal cortical scintigraphy has been reported to be useful in children for confirmation of the diagnosis of acute pyelonephritis. Subsequent experimental studies have demonstrated that dimercaptosuccinic acid (DMSA) scintigraphy, when compared directly with histopathology, is highly reliable for the detection and localization of parenchymal inflammatory changes associated with acute pyelonephritis. Recent clinical studies of acute pyelonephritis using DMSA scintigraphy reveal that the majority (50 to 91%) of children with febrile urinary tract infections have abnormal DMSA renal scan findings and that the majority of these children do not have demonstrable vesicoureteral reflux. However, when vesicoureteral reflux is present, renal cortical abnormalities are demonstrated by DMSA scintigraphy in 79 to 86% of the kidneys. In children with febrile urinary tract infections routine clinical and laboratory parameters are not reliable in the differentiation of acute pyelonephritis, documented by DMSA renal scan findings, from urinary tract infections without parenchymal involvement. Furthermore, the presence of P-fimbriated Escherichia coli associated with febrile urinary tract infections does not reliably predict those kidneys that have acute parenchymal inflammation demonstrated by DMSA renal scans. DMSA is also the isotope agent of choice for the detection of renal scarring. Clinical studies report greater sensitivity of DMSA renal scintigraphy for the detection of renal scarring when compared with the excretory urogram, particularly in infants and young children. In a recent prospective study of post-pyelonephritic renal scarring in children we found that acquired renal scarring only occurs in sites corresponding exactly to previous areas of acute pyelonephritis demonstrated by DMSA scintigraphy at the time of infection. Furthermore, once acute pyelonephritis occurs, ultimate renal scarring is independent of the presence or absence of vesicoureteral reflux. These findings provide convincing evidence that renal parenchymal infection, rather than vesicoureteral reflux, is the prerequisite for acquired (postnatal) renal scarring. Vesicoureteral reflux as a risk factor for acquired renal scarring is directly related to its role as a risk factor for acute pyelonephritis. We conclude that DMSA scintigraphy is a valid tool for confirming the diagnosis of acute pyelonephritis in children and for identifying kidneys at risk for subsequent renal scarring.


Evaluation of acute urinary tract infection in children by dimercaptosuccinic acid scintigraphy: a prospective study. A prospective study examining the incidence of dimercaptosuccinic acid (DMSA) abnormalities in children at the time of acute urinary tract infection, the progression of these abnormalities following treatment and their correlation with the presence of vesicoureteral reflux is reported. DMSA scans performed within 72 hours of presentation in 65 previously healthy children with acute urinary tract infection were abnormal in 34 (52%). The scan appearances of 30 of 36 (83%) initially abnormal kidneys improved or became normal on the repeat DMSA study performed at 3 to 6 months after the acute urinary tract infection. A cystogram demonstrated significant vesicoureteral reflux in 11 of 45 cases (24%). Of these 11 cases 10 had abnormal DMSA studies and 1 had dilated upper tracts on ultrasound. Several conclusions may be drawn from our study. The incidence of DMSA abnormalities at the time of acute urinary tract infection is high but these abnormalities tend to resolve with time. An abnormal DMSA study at the time of urinary tract infection identifies most children with significant vesicoureteral reflux, and in our series a combination of ultrasound and DMSA identified all cases. This study may have major implications for the clinical investigation of children with urinary tract infection.


Distribution of retroperitoneal metastases after chemotherapy in patients with nonseminomatous germ cell tumors. For patients with advanced nonseminomatous germ cell tumors a retroperitoneal lymph node dissection is routinely performed following chemotherapy if the serum tumor markers have returned to normal. Bilateral retroperitoneal lymph node dissection has been recommended because metastatic deposits may be widespread. The aim of this study was to describe the distribution of retroperitoneal metastases following chemotherapy in patients with nonseminomatous germ cell tumor and determine if the extent of the retroperitoneal lymph node dissection can be modified. We studied 113 patients who had initially bulky retroperitoneal disease and underwent retroperitoneal lymph node dissection following chemotherapy. For the purposes of this study teratoma and malignant germ cell tumor are referred to as tumor. The most common location of tumor was the para-aortic area (91%) in patients with a left primary tumor and the interaortocaval area (78%) in those with a right tumor. Tumor was located outside the boundaries of a modified retroperitoneal lymph node dissection in 14 of the 60 patients with residual disease but the tumor was present within a palpable mass in 6 of these 14 patients. If the residual mass was removed and a modified retroperitoneal lymph node dissection was performed only 9 of the 113 patients (8%) would have tumor left in the retroperitoneum. For a select group of patients with advanced nonseminomatous germ cell tumor treated with chemotherapy, resection of the residual mass combined with modified retroperitoneal lymph node dissection is appropriate.


Sequential androgen blockade: a biological study in the inhibition of prostatic growth. We attempted to determine the effects of the combination of a 5-alpha reductase inhibitor and an antiandrogen on rat ventral prostate and seminal vesicle weight. We also attempted to determine whether the prostatic cell death gene TRPM-2 would be expressed using this combination of drugs. Adult male Sprague-Dawley rats were randomly assigned to 7 groups of 15 animals. Four groups served as controls: an intact group sacrificed at the initiation of the trial (group 1), a castrate control group (group 2), an intact control group (group 3), and a group treated with the combination of an LHRH agonist plus antiandrogen (group 7). Three other groups were treated with daily subcutaneous injections of 5 alpha reductase inhibitor (group 5), a nonsteroidal pure antiandrogen (group 4) or both (group 6). After 5 days of treatment 5 animals in each group were sacrificed and prostatic tissue was assayed for the androgen repressed prostatic cell death gene TRPM-2. At 30 days (35 days for group 7) the remaining animals were sacrificed and their ventral prostates, seminal vesicles, and testes (except group 3) were weighed. The combination group (group 6) had a significantly lower prostate weight than either of the monotherapy groups (4, 5), or intact control groups, was equivalent to group 7 but was significantly heavier than the castrate group 2. The seminal vesicle weights of the combination group 6 were significantly lower than the monotherapy groups (4, 5), intact control group, castrate group (3) and was equivalent to group 7. Only castration was able to induce expression of the cell death gene TRPM-2. In this model, the combination of 5 alpha reductase inhibitor and an antiandrogen is as effective a mode of androgen ablation as combination therapy of LHRH agonist plus antiandrogen. Clinically, this combination may translate into adequate androgen blockade without impotence or other side effects of testosterone deprivation. Clinical trials appear warranted to assess this hypothesis.


Local motor responses to bradykinin and bacterial chemotactic peptide formyl-methionyl-leucyl-phenylalanine (FMLP) in the guinea-pig isolated renal pelvis and ureter. The local motor response to bradykinin and the bacterial chemotactic peptide, formyl-methionyl-leucyl-phenylalanine (FMLP) was investigated in the guinea-pig isolated renal pelvis and ureter in relation to possible activation of capsaicin-sensitive primary afferent nerves and release of sensory neuropeptides. Both bradykinin (1 nM-10 microM) and FMLP (10 nM-10 microM) produced a concentration-dependent positive inotropic effect in the isolated renal pelvis which was unaffected by in vitro capsaicin desensitization. The response to bradykinin was antagonized by HOE 140, a bradykinin receptor antagonist, while it was unaffected by MEN 10,376, a tachykinin receptor antagonist, hCGRP(8-37) a calcitonin gene-related peptide (CGRP) receptor antagonist and N-t-BOC-Phe-DLeu-Phe-DLeu-Phe (BPLPLP), an FMLP antagonist. The response to FMLP was blocked by BPLPLP while it was unaffected by HOE 140, MEN 10,376 or hCGRP(8-37). Indomethacin (10 microM) enhanced the response to both bradykinin and FMLP. Bradykinin transiently activated rhythmic contractions in the isolated ureter. The response to bradykinin was blocked by HOE 140 and was unaffected by in vitro capsaicin desensitization, indomethacin, MEN 10,376 or BPLPLP. FMLP had no motor effect on the resting ureter but when rhythmic background contractions were evoked by the addition of 100 nM endothelin 1, it produced a transient suppression of ureteral motility. This inhibitory effect was unchanged by in vitro capsaicin desensitization or HOE 140 while it was abolished by indomethacin or BPLPLP pretreatment. Both bradykinin and FMLP evoked the release of CGRP-like immunoreactivity in the renal pelvis. The effect of bradykinin but not that of FMLP was abolished by indomethacin. By contrast neither bradykinin nor FMLP did evoke a significant CGRP-LI release in the ureter. It is concluded that bradykinin and FMLP affect pyeloureteral motility through specific and independent pathways. The local motor responses produced by these chemical stimulants are independent from the release of sensory neuropeptides from capsaicin-sensitive primary afferent neurons. Direct neurochemical evidence was obtained for activation of capsaicin-sensitive primary afferents in the renal pelvis: such a mechanism could be involved in the genesis of ureteral pain whenever bradykinin or FMLP come into contact with sensory nerves in the pyeloureteral wall.


The effect of experimental urethral obstruction and its reversal on changes in passive electrical properties of detrusor muscle. The passive electrical properties of guinea pig detrusor muscle were studied in order to determine how bladder outflow obstruction and reversal might modify the electrical activity of the bladder and, thus, contractility. Experimental bladder outflow obstruction was produced in guinea pigs and resulted in an increase in bladder weight with a decrease in spontaneous electrical activity, membrane time constant and space constant. In addition, the membrane Na-K pump activity increased with obstruction. Following reversal of obstruction, bladder weight gain associated with obstruction was only partially reversible. The decrease in the membrane time constant induced by obstruction was almost fully reversible following release of obstruction. In contrast, the membrane space constant which reflects spread of current, in addition to spontaneous electrical activity, were only partially reversible. The membrane Na-K pump activity of the detrusor muscle decreased to control levels following reversal of bladder outflow obstruction. There was no significant change in the resting membrane potential of detrusor smooth muscle with either obstruction or following reversal of obstruction. These results suggest, that, the changes in the bladder smooth muscle membrane electrical properties induced by experimental bladder outflow obstruction are only partially reversible following release of obstruction. Furthermore, the results suggest that, the dysfunctional cystometric patterns associated with bladder outflow obstruction might not only be due to changes in detrusor innervation but, fundamental reorganization of the detrusor's electrical syncytium with irreversible suppression of cell-to-cell transfer of electrical activity.


Pedigree analysis and involvement of peroxidase in sickle cell disease. Routine hematological tests were performed in a family which was at risk for sickle cell disease. Cellulose acetate electrophoresis and Triton PAGE were employed to differentiate between various variants of hemoglobin. Based on the data a pedigree was constructed which indicated that few members of the respectively had received the S gene, some of them were sickle cell disease while few were sickle cell trait. Elevated levels of peroxidase enzyme in affected individuals reflect its involvement in RBCs destruction. Statistical analysis strengthen this statement.


Cellular and molecular bases for the immunopathology of the myocardial cell damage involved in acute viral myocarditis with special reference to dilated cardiomyopathy. Cell-mediated autoimmunity has been strongly implicated in the pathogenesis of the myocardial cell damage involved in viral myocarditis. To investigate the cellular and molecular bases of both target cells and effector cells for cell-mediated cytotoxicity involved in viral myocarditis and dilated cardiomyopathy, we first examined the expression of major histocompatibility complex (MHC) antigens and a cell adhesion molecule, intercellular adhesion molecule-1 (ICAM-1) in myocardial cells of a murine model of viral myocarditis and in patients with acute myocarditis and dilated cardiomyopathy. Secondly, we analyzed the characteristics of the infiltrating cells in the heart, especially the expression of a cytolytic factor, perforin. We found that Coxsackievirus B3 (CVB3)-induced murine acute myocarditis resulted in enhanced expression of MHC (class I) antigens and ICAM-1 on myocardial cells, and that perforin was abundantly expressed in NK (natural killer)-like large granular lymphocytes (LGL), which represent the main infiltrating cell type in the early stage. Immunoelectron microscopic study showed killer cells directly damaging cardiac myocytes by the release of perforin. Perforin was also expressed in the infiltrating cells in the heart of a patient with acute myocarditis. Both MHC antigens and ICAM-1 were clearly expressed in the hearts of patients with acute myocarditis and dilated cardiomyopathy. These data provided direct evidence that cell-mediated cytotoxicity plays a critical role in the myocardial cell damage involved in viral myocarditis.


Genomic detection of enteroviruses in the myocardium--studies on animal hearts with coxsackievirus B3 myocarditis and endomyocardial biopsies from patients with myocarditis and dilated cardiomyopathy. We examined myocardial tissues for the presence of enteroviral RNA in animal models with experimental coxsackievirus B3 myocarditis and in endomyocardial biopsy samples obtained from patients clinically diagnosed as having dilated cardiomyopathy or myocarditis using polymerase chain reaction (PCR) gene amplification with enterovirus-generic primers and/or coxsackievirus B3-specific primers. In animal models, coxsackievirus B3 was detected in myocardial tissues up to 28 days, 56 days and 180 days after inoculation, in C3H/He mice, A/J mice and Syrian golden hamsters, respectively. The viral genomes were identified by in situ hybridization in myocardial cells and some interstitial cells in and around the myocarditic lesions in animals. In human endomyocardial biopsy samples, enteroviral RNA sequences were detected in 8 (32%) out of 25 patients with clinical dilated cardiomyopathy and in 3 (33%) out of 9 patients with clinical myocarditis. The patients showing histologic findings of myocarditis and clinical features resembling dilated cardiomyopathy had a high incidence (83%) of positive PCR result for enteroviral RNA sequences. Additionally, 25% of patients with dilated cardiomyopathy showing no histologic findings of myocarditis had positive PCR result. This study supports a link between viral myocarditis and dilated cardiomyopathy.


[Cardiovascular effects of, and catecholamine response to, high dose fentanyl or NLA in patients for valve replacement]. We measured the cardiovascular effect of, and catecholamine and other hormonal responses to, anesthetic doses of fentanyl and original NLA in 25 patients for open heart surgery. The patients were randomly divided into three groups (group N, F30, F75). During induction, in group N; droperidol 0.25 mg.kg-1 and fentanyl 5 micrograms.kg-1, in group F30; fentanyl 30 micrograms.kg-1, and in group F75; fentanyl 75 micrograms.kg-1 were administered intravenously. Additional fentanyl was administered at a rate of 100 to 200 micrograms.h-1. Droperidol 0.25 mg.kg-1 was administered in group N when cardiopulmonary bypass (CPB) was disconnected. Plasma samples were assayed for norepinephrine, epinephrine, ACTH and cortisol before and after induction, during sternotomy, 60 minutes after institution of CPB, after weaning from CPB, and before as well as after extubation. Heart rate (HR), systolic blood pressure (SBP), diastolic blood pressure (DBP) and rate pressure product (RPP) were calculated simultaneously at the blood samplings. In all groups, no remarkable change in cardiovascular dynamics was observed. CPB was associated with marked increases in catecholamines, but high dose fentanyl in dose of 75 micrograms.kg-1 was able to suppress epinephrine level more than in group N. In high dose fentanyl group (F30, F75) ACTH was within normal ranges, even during CPB. The results suggest that high dose fentanyl is a complete anesthetic in patients for cardiac surgery. But a large dose of fentanyl causes small decreases in heart rate and arterial blood pressure. Our data indicate that group F30 is an attractive anesthetic technique for patients with valvular disease.


[ACE Index in patients with respiratory failure]. To investigate whether ACE Index (ACE. Cardiac output-1) which excludes the influence of pulmonary perfusion volume is more useful than angiotensin converting enzyme (ACE) as an early marker of pulmonary endothelial injury and repair, we measured ACE, ACE index, and PaO2.FIO2-1 ratio several times during the stay in the ICU in two patients with respiratory failure. ACE index had a closer relationship to clinical course of respiratory condition and PaO2.FIO2-1 (r = 0.806, 0.889, respectively), in comparison with ACE. We conclude that ACE index could be more sensitive than ACE as an early marker of pulmonary endothelial injury and repair.


Colonic fermentation of ispaghula, wheat bran, glucose, and albumin to short-chain fatty acids and ammonia evaluated in vitro in 50 subjects. The production of short-chain fatty acids and ammonia was measured in 16.6% fecal homogenates from 50 subjects incubated at 37 degrees C for 6 and 24 hours. All 50 homogenates produced ammonia and short-chain fatty acids of any chain length (C2-C5). Incubation for 24 hours with dietary fiber (ispaghula husk or wheat bran), albumin, or glucose (10 mg/mL) increased the short-chain fatty acid production (43.6 +/- 2.8, 45.4 +/- 2.0, 60.3 +/- 3.2, and 65.8 +/- 3.1 mmol/L, respectively) compared with controls (21.4 +/- 1.3 mmol/L). The degradation of different substrates was associated with the production of different amounts of ammonia and short-chain fatty acids. Ispaghula, wheat bran, albumin, and glucose were fermented to acetate (> 2 mmol/L; 24-hour incubations) in 86%, 96%, 98%, and 98% of the homogenates, to propionate in 80%, 76%, 100%, and 68%, and to butyrate in 32%, 94%, 88%, and 54% of the homogenates, respectively. Isobutyrate, valerate, and isovalerate were produced from albumin in all (100%) of the homogenates, but only in 2 to 4% of the homogenates incubated with ispaghula or glucose. Ammonia was always (100%) produced after the addition of albumin and always (98%) consumed (assimilated) when glucose was fermented. Surgery (sigmoid or right- or left-sided colonic resection) did not change the pattern of ammonia and short-chain fatty acid production from these substrates. This study suggests that the different colonic flora from a large number of subjects share general biochemical characteristics, which metabolize different substrates to specific patterns of ammonia and short-chain fatty acids.


[Acute undifferentiated leukemia from the viewpoints of diagnosis and therapy]. Myeloperoxidase (MPO)- and Sudan Black B-not more than 3%-positive, esterase staining-negative, lymphoid, megakaryocyte lineage and erythroid surface marker-negative and electron microscopic platelet peroxidase-negative acute leukemia (AL) was diagnosed as acute undifferentiated leukemia (AUL), and myeloid marker (CD13, CD33), electron microscopic MPO (EMMPO), and DNA analysis of immunoglobulin heavy chain and T cell receptor as well as chemotherapy and its reactivity were examined. Of 239 cases of AL, 10 (4.2%) were AUL, and of these 10 cases, 9 were CD13 or CD33-positive AML-MO (MO) cases. Of 9 cases examined for EMMPO, 4 (44%) were positive, and of 3 cases of MO subjected to DNA analysis, 1 and 1 showed rearrangements of immunoglobulin heavy chain and T cell receptor beta chain, respectively. Of 6 cases of MO on myeloid induction therapy, 1 and 1 showed complete remission (CR) and partial remission (PR), respectively, each having lymphoid genotype, and 4 showed no remission (NR), being 3 of them EMMPO-positive. Of 2 cases on lymphoid induction therapy, 1 and 1 showed CR and NR, respectively, the former being EMMPO-positive MO. BHAC-EM therapy with behenoyl cytosine arabinoside, VP-16 and mitoxantrone performed on 2 cases refractory to any one of both these myeloid and lymphoid induction therapies led to CR in all these 2 cases.


[Morphological and immunohistochemical analysis of Ki-1 lymphoma]. We investigated 9 cases of Ki-1 lymphomas morphologically and immunohistochemically. The lymphoma cells infiltrated in the lymph nodes in 7 cases, in the skin in 6 cases, in both in 4 cases and in the bone marrow in 2 cases. In lymph nodes, the giant tumor cells infiltrated paracortical areas, showing a sheet-like appearance or free cell growth pattern and in the marginal and medullary sinuses as well. In the skin, they formed subcutaneous tumor and infiltrated diffusely also in dermis. All cases were positive for Ber-H2, Ki-1 and HLA-DR and 8 cases revealed some of the T cell phenotypes. But Leu-4 and beta F1 for which all 11 peripheral T cell lymphomas were positive, were only positive in 2 cases, respectively. In the present study, the tumor cells were positive for a few antibodies for T lymphocytes and there were few cases showed positivity for Leu-4 and beta F1. The results suggest that it was questionable whether the tumor cells were derived from T lymphocytes. There may be few cases of Ki-1 lymphomas of distinct T cell lineage.


[Anti-Pr2 cold agglutinin disease with polyneuropathy evolving to malignant lymphoma]. A 53-year-old male was diagnosed as having ataxic polyneuropathy associated with IgM-kappa monoclonal gammopathy in January 1988. Plasmapheresis and chemotherapy with chlorambucil and Melphalan-Prednisolone were effective for his neuropathy, but hemolytic anemia appeared in February 1989. The diagnosis of low-titer cold agglutinin disease (IgM-kappa) with anti-Pr2 specificity was made. Hemolytic anemia became refractory to high-dose corticosteroids, and fever, hepatosplenomegaly and severe pancytopenia appeared in January 1990. Bone marrow involvement of malignant lymphoma (mu, kappa) was found, and he died of pneumonia and gastrointestinal bleeding after the start of chemotherapy. Postmortem examination revealed a widespread infiltration of malignant lymphoma, diffuse, large cell (B-cell) type. Erythrophagocytic histiocytes also increased in bone marrow, liver, spleen and lymph nodes, as if there were hemophagocytic syndrome associated with lymphoma present. In addition to the high thermal amplitude of cold agglutinin in this case, the systemic activation of histiocytes induced by the development of malignant lymphoma may be responsible for progressive hemolysis and severe pancytopenia.


[Function, molecular structure and gene expression regulation of hepatocyte growth factor and its receptor]. Hepatocyte growth factor (HGF), a potent mitogen of mature hepatocytes in primary culture, is composed of 69 kDa alpha-chain with four kringle domains and 34 kDa beta-chain. HGF is currently thought to be a pleiotropic factor, that stimulates the growth and motility of various epithelial cells, and that inhibits the growth of some carcinoma cells. These cells have a single class of high affinity receptor with a dissociation constant (Kd) of 20-30 pM. The maximum number of binding sites (Bmax) was determined to be 100-1500 sites/cell. After 70% partial hepatectomy and unilateral nephrectomy, HGF activity in serum and their tissues increased markedly and HGF receptors of the injured organ rapidly decreased, but not in other normal organs. These results suggest that HGF acts only in a regenerating organ after injury through the control mechanisms governing HGF receptor.


[Function, molecular structure and gene expression regulation of tumor necrosis factor and lymphotoxin]. Tumor necrosis factor (TNF, also called TNF-alpha) and lymphotoxin (LT, TNF-beta) have 32% homology in their primary structures, and it has recently been clarified that they also have closely related tertially structures. These two related cytokines share various biological functions, such as cytotoxic effect, immunomodulating effect, etc. although they produce different effects on several lymphoid, endothelial, and other cellular targets. In this paper, the structures of the two cytokines and their receptor binding sites are described. Furthermore, the structures of the two types of TNF receptors, as well as their role in TNF mediated signal transduction, and possible second messengers responsible for TNF action are also discussed.


[Function, molecular structure and gene expression regulation of erythroid differentiation factor (EDF/activin A)]. Erythroid differentiation factor (EDF), initially found as a differentiation inducer of murine erythroleukemia cells, also acts on normal erythroid progenitors in vitro and in vivo. Furthermore, it is produced endogenously and supporting in vivo erythropoiesis. EDF is structurally identical to activin A, a gonadal protein with follicle stimulating hormone releasing activity, and belongs to TGF beta superfamily. Its activity could be regulated by follistatin, a binding protein with neutralizing activity against EDF/activin A. Molecular cloning of EDF/activin A receptor cDNA has revealed its domain structure characteristic to serine/threonine kinase.


[H(+)-ATPase and H(+), K(+)-ATPase].

[H(+)-ATPase and H(+), K(+)-ATPase]. Two different types of proton transporting ATPases, v-and p-type H(+)-ATPases engage in epithelial ion transport. Properties, function, molecular structure and distribution of these H(+)-ATPase (v-type) and H+, K(+)-ATPase (p-type) are summarized here. Intraorganellar spaces, such as lysosome, synapse, multivesicular body, are acidified by the vacuole-type H(+)-ATPases. Secretion of proton by some types of intercalated cells of kidney collecting tubules is due to H(+)-ATPase. Gastric proton secretion is due to H+, K(+)-ATPase. Proton secretion and absorption of potassium by distal colon is due to p-type ATPase.


[Na/H antiporter].

[Na/H antiporter]. Na/H antiporter located in the plasma membrane is mandatory to maintain cell pH. Two isoforms with different sensitivities to amiloride have been identified. They seem to be products from different mRNAs. The amiloride-sensitive form exists in the non-epithelial cells and the basal membrane of the epithelial cells. The amiloride-resistant form is confined in the apical membrane of the epithelial cells and is important for the vectorial transport of HCO3-. The amiloride-sensitive form has been cloned and sequenced. It is a glycoprotein composed of 815 amino acids. Functional analysis of Na/H antiporter by expressing mutant Na/H antiporter cDNA is in progress.


[Mechanism of acid production and secretion by osteoclasts]. Osteoclasts are primary cells responsible for bone resorption. The most characteristic feature of osteoclasts is the presence of ruffled borders and clear zones. The resorbing area under the ruffled border of osteoclasts is acidic, which favors dissolution of bone mineral. In bone-resorbing osteoclasts, hydrogen ions are provided by carbonic anhydrase II, which catalyzes the hydration of CO2 to H2CO3. Recently, it has been shown that the proton pump of the vacuolar H(+)-ATPase type exists in the ruffled border membranes of osteoclasts. Secretion of hydrogen ions by osteoclasts generates an equal amount of cytoplasmic base equivalents, principally as HCO3-. Osteoclasts have a chloride/bicarbonate exchanger, which normalizes the intracellular pH when osteoclasts actively resorb bone. In this paper, we review the mechanism of the acid secretion by osteoclasts.


Tumor-infiltrating lymphocytes and prognosis of hepatocellular carcinoma. Tumor-infiltrating lymphocytes (TIL) were isolated from 17 human hepatocellular carcinomas (HCC). The proliferation of TIL cultured with recombinant interleukin-2 (rIL-2) was evaluated. We also examined prognosis in relation to TIL. Successful expansion of TIL was achieved in 16 of the 17 lesions. In 10 of the 16, TIL increased more than 100-fold. Cytotoxicity to the allogeneic HCC cell line, HC-4, was demonstrated in all 13 TIL cultures tested. Maximum cytotoxicity was noted two to four weeks after culture. No statistical difference was observed either with respect to prognosis based upon growth rate or the cytotoxicity demonstrated in vitro. The initial number of TIL per unit weight of tumor was, however, significantly greater in the group for which the prognosis was good (19.0 +/- 5.1 x 10(6) vs. 5.6 +/- 1.6 x 10(6), P < 0.05). It would appear that greater lymphocytic tumor infiltration is a prognostic marker.


Inadvertent tumor violation during musculoskeletal sarcoma surgery and risk of recurrence. During the limb preserving procedure for musculoskeletal sarcoma, the course of action that should be taken when a tumor or tumor-cell-contaminated adjacent tissue is violated remains controversial. From January, 1973, to July, 1989, 120 patients with musculoskeletal sarcoma were treated by limb salvage surgery and, in 40 of them, such violations inadvertently occurred during surgery. Follow-up studies on the patients have been conducted for at least one year after the violation. Soft tissue sarcoma was noted in 24 cases (low grade, 4; high grade, 20) and bone sarcoma in 16 (low grade, 6; high grade, 10). To treat the violation, re-excision with a clean margin and copious lavage was generally conducted after careful closure of the violated site. Sixteen cases in the soft tissue sarcoma group and eight in the bone sarcoma group were treated in this way. Local recurrences were found in 13 of the 40 cases (33%), but the percentage was reduced to 13 in cases treated by re-excision with a clean margin. In the present study, there was no significant effect of adjuvant therapy after violation. The survival rate in cases of local recurrence was very low, and not at all improved by adjuvant therapy.


Adenocarcinoma in Barrett's esophagus following total resection of the gastric remnant: a case report. We report a case of adenocarcinoma in Barrett's esophagus following a total resection of the gastric remnant. A 52-year-old man had undergone a distal gastrectomy for gastric cancer at 33 years of age and a total resection of the gastric remnant for local recurrence of the gastric cancer at 35 years of age. Repeated endoscopic examinations revealed the sequence of reflux esophagitis and Barrett's esophagus. Furthermore, adenocarcinoma in Barrett's esophagus was detected in December, 1989. A subtotal esophagectomy was performed in January, 1990. The elevated lesion in the lower esophagus showed coarse lobulation and measured 7.4 x 3.2 cm. The histologic type was that of well-differentiated adenocarcinoma, with the invasion limited to the muscularis mucosae without lymph node involvement. Severe dysplasia was seen adjacent to the definite carcinoma. The case supports the acquired theory of pathogenesis for Barrett's esophagus and suggests that reflux esophagitis after total gastrectomy may result in a dysplasia-carcinoma sequence.


[Fundamental studies of hepatitis C virus: a review]. In 1974, Prince et al. reported the existence of posttransfusion hepatitis with a long incubation period which was not related to hepatitis B virus (HBV). These cases were named "non-A, non-B" (NANB) hepatitis. The genome of NANB hepatitis virus was discovered recently using a recombinant complementary DNA (cDNA) approach. It was termed the hepatitis C virus (HCV), and a specific diagnostic tool for the circulating HCV antibody (anti-HCV) was developed using a purified viral polypeptide derived from recombinant yeast expressing a small part of the HCV genome. HCV is believed to be the main cause of blood-borne non-A, non-B hepatitis worldwide, which frequently evolves to chronic hepatitis and cirrhosis, and which may also be involved in the development of hepatocellular carcinoma. HCV is classified as part of the flaviviridae family and contains a positive-stranded RNA molecule by approximately 10 kb nucleotides. The HCV genome encodes a large polyprotein precursor, which is processed in structural nucleocapsid and envelope proteins and in non-structural proteins (NS1-NS5). Nucleotide sequence comparisons of distinct HCV isolates have shown a significant genetic variability between the different HCV strains. At present the diagnosis of HCV infection depends on various anti-HCV tests including second generation HCV Ab. Antigenic markers for HCV are being developed but the concentrations of HCV antigens in serum are at the lower limit of detectability by existing immunoassay technology. A polymerase chain reaction has been used to detect HCV RNA in the serum and liver. Serum HCV RNA disappears from serum after effective IFN treatment.(ABSTRACT TRUNCATED AT 250 WORDS)


[Epidemiological study of hepatitis B and C virus in Okinawa and Kyushu, Japan]. Epidemiological studies of hepatitis B virus (HBV) and hepatitis C virus (HCV) infection in Okinawa and Kyushu have been conducted since 1970. Early in the study period, the prevalence of HBsAg on Iriomote and Hateruma islands in Okinawa was 9.0% and 6.1%, respectively, which were significantly higher than the 2.4% in Kyushu. Prevalence of anti-HBc on Iriomote island was 59.8% and that on Hateruma island 70.9%, also which were significantly higher than the 30.9% in Kyushu. Prevalence of HBV infection has been observed in both Okinawa and Kyushu with prevalences of HBsAg in Okinawa decreasing from 11.6% in 1970 to 3.8% in 1988 and in Kyushu from 2.1% in 1980 to 0.7% in 1991. Prevalence of anti-HCV in Okinawa was only 0.7%, while the prevalence was 3.4% in Kyushu when measured by C100-3 and anti-GOR. In Kyushu, the prevalence of anti-HCV advanced with age and reached a peak in the population 60 years of age. HCV transmission occurs mainly by blood transfusion. We investigated the systems of blood supply in Okinawa and Kyushu. In Japan, including Kyushu, paid blood was mainly used from 1955 to 1969, but in Okinawa it was donated by relatives and friends. The difference in blood supply between these areas may be a cause of the differing of HCV infection.


[Complex formation of ulinastatin with alpha-thrombin]. The inhibition mode of ulinastatin was indicated noncompetitive by the Lineweaver-Burk's double reciprocal plotting method using the production rate of fibrinopeptide A. Consequently Km of alpha-thrombin could be calculated 2.8 mM and its Vmax was reduced from 24 U/l to 15 U/l by the addition of ulinastatin and Ki of ulinastatin was 1.05 x 10(-2) M. The complex of ulinastatin and alpha-thrombin were found. Furthermore, the mixing of alpha-thrombin, AT-III and ulinastatin produced a larger complex on SDS-PAGE at pH 7.0, and it was evident by the use of Western blotting method, that the complex was consisted of alpha-thrombin, AT-III and ulinastatin. Although ulinastatin and thrombomodulin showed multiple similarities in inhibitory functions on alpha-thrombin, still there are some differences functioning on alpha-thrombin, because of the different binding sites of ulinastatin and thrombomodulin on alpha-thrombin, indicating no crossreaction for antithrombomodulin monoclonal antibody relating to alpha-thrombin binding site of thrombomodulin and it could be contributed to form noncompetitive or uncompetitive inhibitors.


Monoclonal antibody to human plasma gelsolin. Human plasma gelsolin was purified by column chromatography. The method yielded a protein of high purity and activity. Using this protein, we produced monoclonal antibody (Mab H6B11) against human plasma gelsolin by somatic cell fusion. This monoclonal antibody reacted in a dose-dependent manner with gelsolin derived from human plasma and platelets and neutralized depolymerizing activity to F-actin. It differed from the commercially available substance (Mab G4896; Sigma) in that the time required for the reaction between the antigen and antibody in the enzyme-linked immunosorbent assay could be shortened by one-third. The antibody was judged to be useful in assays for elucidating the physiological role of plasma gelsolin.


[Effects of ouabain on 201Tl and 99mTc MIBI kinetics in rat myocardium--in vivo study using miniature CsI detector]. The aim of this study is to investigate the effect of ouabain (inhibitor of Na-K ATPase) in the myocardial uptake and clearance of 201Tl and 99mTc MIBI using rats. Time activity curves of three groups were measured by CsI miniature detector for 15 min after the administration of each radiopharmaceutical. Groups were divided as follows; 1) control group (CON group), 2) early phase as 15 min after the administration of ouabain (OU15 group), and 3) late phase as 120 min after the administration of ouabain (OU120 group). Main organs of rats including myocardium were resected at 1 min and 15 min after the administration of 201Tl or 99mTc MIBI under the anesthesia of pento-barbital sodium. The uptake (%ID/g) of 201Tl or 99mTc MIBI was measured by well type scintillation counter. The uptake of 201Tl of OU15 group was significantly higher than CON group (OU15: 10.81 +/- 1.90, CON: 1.86 +/- 0.42) and cleared more rapidly. On the contrary, OU120 group showed lower uptake (1.70 +/- 0.21) and slower clearance than OU15 group. Time activity curve of OU15 group indicated the accelerated washout compared with CON group. However, OU120 group showed significantly the reduced washout (CON group: 19%, OU15 group: 22%, OU120 group: 9%). On the other hand, the uptake and time activity curves of 99mTc MIBI were not influenced by ouabain administration. In conclusion, myocardial uptake and clearance of 99mTc MIBI were not related to Na-K ATPase activity, while those of 201Tl were markedly influenced by Na-K ATPase.


Block of sodium channels by tyramine and its analogue (N-feruloyl tyramine) in frog ventricular myocytes. Pharmacological effects of tyramine and its analogue, N-feruloyl tyramine (NFT), on sodium and calcium currents in frog ventricular myocytes were examined using the whole-cell voltage-clamp technique. To improve the temporal and spatial control of the membrane potential, sodium currents (INa) were recorded in 45.5 mM [Na+]o at 10 degrees C. Both tyramine and NFT (1-100 microM) induced a concentration-dependent decrease in INa evoked from a holding potential of -80 mV without affecting a change in either the time to peak or the time constant for the falling phase of INa. Similarly the reversal potential for INa remained unchanged at a value close to that predicted from the Nernst equation. The finding that both tyramine and NFT decreased INa when activated maximally, from a holding potential of -120 mV, indicates that the amplitude of INa can be reduced independently of a change in the kinetics of the current. In addition, tyramine (100 microM) shifted the membrane potential for half maximal inactivation (Vh) of the steady-state inactivation (h infinity)-curve from -74 to -84 mV without affecting its slope. In contrast, NFT failed to affect the h infinity-curve. The calcium current (ICa) recorded in the presence of 0.3 microM TTX was not affected by either 100 microM tyramine or NFT. We concluded that tyramine directly blocks Na channel by shifting h infinity-curve and by suppressing maximum Na channel conductance, while NFT suppresses only maximum Na channel conductance.


Activation of Ca(2+)-dependent K+ channel and Cl- conductance in canine pancreatic acinar cells through a cyclic AMP pathway. Effects of adenosine 3',5'-cyclic monophosphate (cAMP) on Ca(2+)-dependent K+ channel and Cl- conductance in the plasma membrane of isolated canine pancreatic acinar cells were studied by patch-clamp methods. In whole-cell current recordings on isolated cells dialyzed with K(+)-rich solution containing 0.5 mM EGTA, addition of 0.5 mM dibutyryl cAMP (dbcAMP), or 50 microM forskolin to the bath increased outward K+ and inward Cl- currents associated with depolarizing and hyperpolarizing voltage jumps, respectively. In intact cells (cell-attached configurations), addition of 0.5 mM dbcAMP or 50 microM forskolin to the bath increased the opening of single K+ channel. In Ca(2+)-free external solution (bath and pipette) 50 microM forskolin or 0.5 mM dbcAMP application evoked an increase in the opening of single K+ channel in intact cells. Addition of 0.5 mM dbcAMP to the bath solution containing 10 mM EGTA without Ca2+ increased the currents of whole-cell dialyzed with K(+)-rich solution containing 10 mM EGTA. When cell was dialyzed with 20 mM EGTA, dbcAMP, or forskolin application did not increase the whole-cell currents. In excised inside-out patches, addition of the catalytic subunit of cAMP-dependent protein kinase (16 U/ml) in the presence of 0.3 mM ATP to the cytoplasmic face of membrane activated the K+ channel, but 0.1 mM cAMP did not. These results suggest that cAMP-dependent phosphorylation can activate Ca(2+)-dependent K+ channels without increase in intracellular free Ca2+ and cAMP-dependent mechanism can activate Ca(2+)-dependent Cl- conductances without the increase in Ca2+ in canine pancreatic acinar cells.


Phototransduction in cones as examined in excised membrane patch. Phototransduction mechanisms have been so far investigated mostly in rods, whereas those in cones are much less known. In the present experiment, we investigated phototransduction mechanisms in inside-out patches excised from cone outer segments of the carp. Cyclic GMP-activated channels on the patch became light-sensitive when both GTP and Mg2+ were supplied by perfusion. When the channels were activated by a hydrolysis-resistant analogue of cGMP, activities were not suppressed by light even though both GTP and Mg2+ were present. Thus activation of transducin and phosphodiesterase (PDE) were involved in the transduction processes, indicating that phototransduction mechanisms in cones are qualitatively similar to those in rods. In cone patches, however, light responses fully terminated even though ATP was absent, opposing to the report that ATP was indispensable for light response termination in rods. The response termination in the cone patch might result from activation of guanylate cyclase and/or inactivation of PDE. Under the perfusion of GTP together with Mg2+ and 3-isobutyl-1-methyl xanthine, no channel activities were observed, indicating that no guanylate cyclase activity was present in cone patch preparations. Therefore, termination of the light response in the patch might be caused by inactivation of PDE which resulted from inactivation of photopigment and transducin. Based on these observations, differences in light response kinetics between the rod and cone are discussed.


Extracellular ATP induces rapid cell rounding in cultured human Chang liver cells. Extracellular ATP (7 microM-14 mM) induced monolayer Chang liver cells (ATCC CCL 13) to retract and round up in dose-dependent and time-dependent manners. ATP-induced rounding was concomitant with raised intracellular pH (7.3 to 7.8) and suppressed (a) in sodium-free medium (250 mM sucrose), (b) with the addition of 1 mM quinidine, an inhibitor of Na+/H+ exchanges, and (c) with the addition of 10 nM staurosporine, a potent inhibitor of protein kinase C that is known to activate Na+/H+ exchanges. These findings suggest that ATP-induced cell rounding is due to activation of Na+/H+ antiporters.


Effects of ouabain, DBcAMP, caffeine, and high [Ca2+]o on twitch tension, intracellular Na+ activity, and action potential of guinea pig papillary muscles. To explore ionic mechanisms of the positive inotropic process in guinea pig ventricular papillary muscles, we simultaneously measured twitch tension, intracellular Na+ activity (aiNa) and action potential at a stimulus rate of 60/min. The aiNa was 5.0 +/- 0.1 mM in the steady state. When the Na-K pump was inhibited by ouabain (1 microM), the twitch tension was increased in proportion to the aiNa, and the action potential was decreased at diastolic state and at duration for 90% of repolarization (APD90). While exposed to caffeine (3 mM), the aiNa kept increasing even when the twitch tension achieved a peak level. Concomitantly, the action potential slightly depolarized at diastolic state with a prolonged APD90. An application of DBcAMP (1 mM) or an increase in [Ca2+]o (from 1.8 to 3.6 mM) enhanced the twitch tension with a fall of aiNa and a shortening of APD90. These results suggest that in guinea pig papillary muscles Na-Ca exchange plays an important role in the regulation of contractile force and membrane potential, and that the Na influx should be balanced by the activation of the sarcolemmal Na-K pump, a negative feedback mechanism, to prevent calcium overload and abnormal pacemaker activity in electrical excitation.


Resistance of the snake neuromuscular junction to the blocking effect of beta-bungarotoxin. beta-Bungarotoxin at a concentration of 0.05 microM was enough to block successively the nerve-evoked intracellular action potential of muscle fiber, the end-plate potential, and miniature end-plate potentials within 60, 90, and about 180 min, respectively, in the isolated nerve-muscle preparation of the frog. On the other hand, at the snake neuromuscular junction beta-bungarotoxin caused an initial transient reduction and a following sustained facilitation of the miniature end-plate potential discharge, but did not block the nerve-evoked intracellular action potential of muscle fiber at a concentration of as high as 50 microM over 4 h. The results show that beta-bungarotoxin binds to snake motor nerve ending without leading to the transmission block, probably due to lacking the target of action.


Dibutyryl cyclic AMP and forskolin inhibit phosphatidylinositol hydrolysis, Ca2+ influx and contraction in vascular smooth muscle. Dibutyryl cyclic AMP and forskolin inhibited the contraction induced by norepinephrine (NE) more strongly than the high K(+)-induced contraction in isolated rat aorta. These inhibitors inhibited the 45Ca2+ influx stimulated by NE but not that by high K+, and they inhibited NE-induced inositol monophosphate accumulation. These results suggest that cAMP inhibits NE-induced contraction, at least partly, by inhibiting the alpha-adrenoceptor-mediated signal transduction and high K(+)-induced contraction by decreasing Ca2+ sensitivity but not Ca2+ influx.


Alpha 1B-adrenoceptor mechanisms in rabbit iris dilator. Rabbit isolated iris dilator strips were contracted by norepinephrine, an alpha 1A- and alpha 1B-nonselective agonist, but not by methoxamine, an alpha 1A-selective agonist. The concentration-response curve for norepinephrine was considerably inhibited by chloroethylclonidine. The pA2 values for WB4101 and 5-methylurapidil were 8.16 +/- 0.09 and 7.84 +/- 0.08 (means +/- S.E. of 8-12 experiments), respectively, and significantly smaller than the values reported in the rat renal artery and thoracic aorta, and rabbit bronchus, where the alpha 1A-subtype is predominant. These results suggest that the rabbit iris dilator contains primarily the alpha 1B-subtype. Clonidine and tizanidine did not contract the rabbit iris dilator but shifted the curve for norepinephrine in a parallel manner, suggesting that they interact with the alpha 1B-subtype and act as competitive antagonists in this muscle. Methoxamine (up to 10(-3) M) had no effect on the contractile response to norepinephrine, suggesting that methoxamine does not interact with the alpha 1B-subtype.


Central alpha 2-adrenoceptor-mediated pressor response to clonidine in conscious, spontaneously hypertensive rats. Pressor responses to intracerebroventricular (i.c.v.) injection of clonidine were investigated in conscious spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY). Clonidine (1-10 micrograms, i.c.v.) caused a dose-dependent pressor response and decrease in heart rate in both SHR and WKY. In SHR, low doses (1, 2.5 micrograms) but not high doses (5, 10 micrograms) of i.c.v.-clonidine induced a depressor response following the pressor response. Both pressor and depressor responses to i.c.v.-clonidine were significantly greater in SHR than in WKY. In both SHR and WKY, pressor responses to i.c.v.-clonidine were abolished by pentobarbital anesthesia, pretreatment with i.v.-furosemide (5 mg/kg), 24-hr water deprivation and pretreatment with i.c.v.-yohimbine (100 micrograms), but not by pretreatment with i.v.-yohimbine (100 micrograms) and i.c.v.-prazosin (10 micrograms). On the 1st day after surgery for arterial catheter implantation, SHR reduced their water intake, and i.c.v.-clonidine (5 micrograms) caused a slight pressor response, whereas the same dose of clonidine on the 7th day after surgery resulted in a marked pressor response. These results suggest that clonidine caused a central alpha 2-adrenoceptor-mediated pressor response, which is greater in SHR than in WKY and is sensitive to body fluid volume changes and anesthesia.


Experimental models of acute and chronic sialoadenitis in the guinea pig. Acute and chronic sialoadenitis were induced in ovalbumin-immunized guinea pigs by a single or repeated (once a day for 5 days) instillation of antigen into the parotid gland via the parotid duct. The acute sialoadenitis was characterized by infiltration of inflammatory polymorphonuclear leukocytes and the chronic one, by extensive tissue destruction together with infiltration of mononuclear leukocytes. In acute sialoadenitis, myeloperoxidase activity in the parotid gland, which was a marker of accumulation of neutrophils, was elevated, but in the chronic stage, it returned nearly to the control level. This observation is in accord with the histological findings that infiltrating cells in acute and chronic sialoadenitis were mainly polymorphonuclear and mononuclear leukocytes, respectively. Although cyclophosphamide suppressed the inflammation, both in acute and chronic sialoadenitis, indomethacin exerted its anti-inflammatory effect only in the acute stage. Our experimental models of acute and chronic sialoadenitis were easy to prepare, and had a high incidence. As the typical features of inflammatory development from acute to chronic phases were observed in these models, these models may be useful for studying the mechanism of the chronic course in immunologically induced inflammation and the effects of drugs on each phase and the chronic course of inflammation.


Effects of nifedipine and nicardipine on glucagon-stimulated gluconeogenesis in primary cultures of rat hepatocytes. The effects of nifedipine and nicardipine on glucagon-stimulated gluconeogenesis from lactate were examined in primary cultures of rat hepatocytes. Nifedipine and nicardipine (10(-7)-10(-5) M) significantly potentiated the glucagon-stimulated gluconeogenesis from lactate by increasing intracellular cAMP levels. In contrast, diltiazem and verapamil did not potentiate the glucagon-stimulated gluconeogenesis. 1-Methyl-3-isobutylxanthine and papaverine also potentiated the glucagon-stimulated gluconeogenesis from lactate. On the basis of these results, possible mechanisms by which nifedipine and nicardipine potentiate the glucagon-stimulated gluconeogenesis will be discussed.


Antagonistic activity of Y-25130 on 5-HT3 receptors. This paper describes the 5-hydroxytryptamine3 (5-HT3) receptor antagonism of Y-25130 ((+-)-N-(1-azabicyclo[2.2.2]oct-3-yl)-6-chloro-4-methyl-3-oxo-3,4-dih yd ro- 2H-1,4-benzoxazine-8-carboxamide monohydrochloride) in the rat cerebral cortex, isolated rabbit heart and isolated guinea pig ileum. In an in vitro binding assay, Y-25130 inhibited the specific binding of [3H]quipazine to 5-HT3 receptors at the synaptic membranes of the rat cerebral cortex with a Ki value of 2.9 nM, the same as that of ondansetron. Metoclopramide, 5-HT and 2-methyl-5-HT also showed an inhibitory effect, but their affinities for 5-HT3 receptors were lower than that of Y-25130. Y-25130 showed low affinity for histamine H1 receptors (IC50 = 4.4 microM) but it could not reveal any affinities for the other receptors (5-HT1A, 5-HT2, dopamine D1, dopamine D2, alpha 1-adrenoceptor, alpha 2-adrenoceptor, muscarine and benzodiazepine) even at a 10 microM concentration. In the isolated rabbit heart, Y-25130 antagonized the indirect sympathomimetic responses to 5-HT (pA2 value = 10.06) and this effect was more potent than that of metoclopramide. In the isolated longitudinal smooth muscle of the guinea pig ileum, concentration-contraction effect curves for 5-HT were biphasic in the presence of ketanserin. Y-25130 shifted to the right only in the second phase of concentration-effect curves for 5-HT (pA2 value = 7.04) and its activity was more potent than that of metoclopramide. These results indicate that Y-25130 is a potent and selective 5-HT3 receptor antagonist.


Tizanidine may discriminate between imidazoline-receptors and alpha 2-adrenoceptors. The selectivity of tizanidine to the imidazoline-receptor and alpha 2-adrenoceptor recognized by 3H-p-aminoclonidine was examined in rat kidney membranes and was compared with those of other imidazoline compounds. Tizanidine bound to the imidazoline-receptors with approximately 20 times higher affinity than the alpha 2-adrenoceptors. The order of relative selectivity to imidazoline receptor was tizanidine greater than oxymetazoline greater than clonidine greater than naphazoline, where clonidine showed an equal affinity to both receptors. Tizanidine may act more potently on the imidazoline-receptors than the alpha 2-adrenoceptors.


[Clinical significance of oncogene product expression in human lung cancer]. The clinical importance of ras oncogene product p21 was evaluated in surgically treated non-small cell lung cancer patients. Paraffin sections of tumors were analysed immunohistochemically using anti-ras p21 monoclonal antibody rp35. The ras p21 expression was correlated with clinicopathological parameters and survival. Survival analysis demonstrated significantly longer survival times in patients with p21-negative tumors than those with p21-positive tumors. In Cox's multivariate analysis, ras p21 expression was a major and independent prognostic determinant of survival. On the other hand, in small cell lung cancer, L-myc gene is known to be frequently amplified and overexpressed. Immunoprecipitation analysis of two small cell lung cancer cell lines (classic type) revealed three major L-myc proteins (p60, p66 and p68), all of which were derived from extensive phosphorylation of a p59 protein. Expression and phosphorylation of L-myc protein, as well as the autocrine growth mechanism of gastrin-releasing peptide (GRP), is thought to be involved in the malignant behavior of small cell lung cancer.


[Effects of angiotensin peptides on airway epithelial ion transport and its modulation by angiotensin converting enzyme]. To study the effects of angiotensin (ANG) peptides on airway epithelial ion transport function, we evaluated the bioelectric properties of canine cultured tracheal epithelium under short-circuit conditions in vitro. Addition of ANG I, II and II dose-dependently increased short-circuit current (Isc) and transepithelial potential difference, an effect that was more pronounced with addition to the submucosal solution than to the mucosal solution, with rank order of potency of ANG II greater than or equal to ANG II much greater than ANG I. The ANG-induced increase in Isc was not altered by the Na channel blocker amiloride, but was greatly reduced by the CI channel blocker diphenylamine-2-carboxylate and Cl-free medium. The response of Isc to ANG I was reduced by MK422, an angiotensin converting enzyme inhibitor, in a dose-dependent fashion. These results suggest that ANG II and III selectively stimulate Cl secretion across airway epithelium and that ANG I may exert its effect after its conversion to ANG II by angiotensin converting enzyme.


[Syphilitic coronary ostial obstruction and aortic regurgitation associated with advanced gastric cancer: successful 2-stage surgery]. We report a case of syphilitic coronary ostial obstruction and aortic regurgitation associated with advanced gastric cancer for which subtotal curative gastrectomy and AVR with CABG was successfully performed in a two-stage operation. A 65-year-old male was admitted complaining of tarry stool and angina. A serological test for syphilis was strongly positive and angiography demonstrated a left coronary ostial obstruction accompanied by moderate aortic regurgitation. An endoscopic examination of the stomach revealed a Borrmann type II advanced gastric cancer with active bleeding. In the first stage, a subtotal gastrectomy with wide margin was performed, under hemodynamic monitoring because of the active tumoral bleeding. After the second-stage AVR, the postoperative course was uneventful.


[Endoscopic closure of the postoperative bronchopleural fistula]. A case of successful endoscopic closure of a bronchopleural fistula with fibrin glue was reported. Fifty-eight-year-old man underwent right lower lobectomy for lung carcinoma. On the 21st postoperative day, he suffered from high fever and diagnosed as right side pyothorax. Then tube thoracotomy was done under UCG-guide. As a fistula in the right lower bronchial stump was found tiny by BFS, we tried to close the fistula bronchoscopically with fibrin glue. The fibrin glue was injected into the fistula via the ERCP tube, which was passed through the bronchoscope. This procedure was performed twice for two weeks interval. The fistula was completely closed about a month after the trial. This method is useful to close postoperative small bronchopleural fistula without surgical intervention.


[Granulocyte functions of chronic hemodialysis patients]. With increasing number of chronic hemodialysis (HD) patients, urologists share much more chance to encounter HD patients who need urological operations. In these patients healing of surgical wound was sometimes hampered by intractable infection. In this paper we investigated granulocyte functions of HD patients. Thirty two male HD patients (20-50 years of age) and 18 healthy volunteers, who had no infectious foci and had no anti-inflammatory drugs were adopted. Heparinized blood samples were drawn between 8:00 an 9:00 am from the volunteers, and just before HD from the HD patients. The oxygen dependent microbicidal response of granulocytes was evaluated by luminol dependent chemiluminescence which measured the amount of superoxide produced by opsonized zymosan stimulated granulocytes. Phagocytic function of granulocytes was evaluated by counting the number of granulocytes which phagocytized fluorescent monodisperse carboxylated microspheres by flow cytometry. Superoxide production per 1000 granulocytes of HD patients was significantly higher than that of normal control. During HD superoxide production was temporally decreased for the first one hour, became maximal at the end of HD and returned to the baseline level on the second post HD day. Phagocytosis was significantly depressed in HD patients. This functional deficit could lead to prolonged healing of infection in HD patients.


[A preliminary study to determine the activity of topoisomerase II in human kidney cancer cells with DNA unknotting method]. For the purpose of examining the Topoisomerase II (Topo II) activity in human kidney cancer cells, we performed experiments with using DNA unknotting method. This method check relative Topo II activity with its conversion of knotted form P4 phage DNA to unknotted form. Our preliminary results demonstrate remarkable activity of Topo II with specific conversion of knotted form P4 DNA to unknotted form in human kidney cancer cells, YCR and ACHN. Moreover, addition of etoposide to the same experiment suppressed Topo II activity in a dose dependent manner. Our results suggest that kidney cancer has a certain amount of Topo II, a target for etoposide. We believe this method is useful to measure Topo II activity in cancer cells and to estimate chemotherapeutic potential of Topo II inhibitors including etoposide in human kidney cancers.


[Antimycobacterial activity of a newly synthesized fluoroquinolone, Y-26611]. Y-26611, a newly developed fluoroquinolone having a morpholine moiety at the 7 position was examined for in vitro antimycobacterial activity by the agar dilution method using 7H11 medium. The MIC90 values of Y-26611 were as follows: Mycobacterium tuberculosis (25 strains), 0.4 micrograms/ml; M. kansasii (19 strains), 6.25 micrograms/ml; M. marinum (10 strains), 25 micrograms/ml; M. scrofulaceum (19 strains), 50 micrograms/ml; M. avium (18 strains), 50 micrograms/ml; M. intracellulare (31 strains), greater than 100 micrograms/ml; M. fortuitum (20 strains), 0.4 micrograms/ml; M. chelonae subsp. abscessus (15 strains), greater than 100 micrograms/ml; and M. chelonae subsp. chelonae (20 strains), 100 micrograms/ml. The MICs against M. tuberculosis and M. fortuitum were lower than those of ofloxacin (OFLX), although it had somewhat higher MICs against M. avium complex than OFLX. Antimicrobial activity of Y-26611 against M. tuberculosis phagocytosed in cultured murine peritoneal macrophages were somewhat lower, as compared to that of OFLX. When M. fortuitum-infected (iv) A/J mice were treated with Y-26611 by gavage at doses of 0.5-2 mg/mouse, once daily, six times per week, from day 1 for up to 4 weeks after infection, mice were protected from death and the number of CFU recovered from their visceral organs, such as the lungs, spleen and kidneys were reduced. The therapeutic efficacy of Y-26611 was similar as that of OFLX.


Transmission of sialodacryoadenitis virus (SDAV) from infected rats to rats and mice through handling, close contact, and soiled bedding. Thirty mice and six rats were exposed through handling, soiled bedding, or close contact to rats previously inoculated with sialodacryoadenitis virus (SDAV). All exposed rats developed coronaviral antibody without clinical signs or lesions of SDAV infection. Exposed mice had no lesions or clinical signs of coronavirus infection. Mice exposed by handling or by soiled bedding did not develop coronavirus antibody. Two of 10 mice exposed to SDAV-inoculated rats by close contact were coronavirus seropositive when tested 3 weeks postexposure. SDAV-inoculated rats and mice developed coronavirus lesions and antibody. These results suggest that rat-to-rat transmission of SDAV is likely via fomites or handling; however, rat-to-mouse transmission is unlikely when animals are housed and husbanded using modern techniques. Results also suggest that coronavirus antibody in mice is due to exposure to mouse coronavirus and not to rat coronaviruses.


Duration and strain-specificity of immunity to enterotropic mouse hepatitis virus. We examined the duration and strain-specificity of immunity to enterotropic mouse hepatitis virus (MHV). Two strains of enterotropic MHV (MHV-Y and MHV-RI) were determined to be distinct virus strains by serum neutralization and by enzyme immunoassay. BALB/cByJ mice immunized by oral infection with either MHV-Y or MHV-RI developed serum MHV IgG titers that remained stable for more than 6 months. The animals were protected from reinfection with the homologous virus strain at 1 and 6 months after an initial immunizing infection, based on intestinal histology and polymerase chain reaction for a 375-base-pair segment of the membrane glycoprotein gene. Immunity was also fully protective against challenge with the heterologous strain 1 month after initial immunization and partly protective after 6 months. Maternally-derived passive immunity prevented MHV infection in 1-week-old pups challenged with the homologous strain of MHV, and pups challenged with the heterologous virus strain were partially protected.


Investigation of enzyme defects in children with lactic acidosis. Screening for enzyme deficiencies was carried out in cultured skin fibroblasts and leukocytes of 19 patients with lactic acidosis and neurological problems. Pyruvate carboxylase deficiency was demonstrated in three cases. Reduced pyruvate oxidation was found in seven cultures; six showed no significant stimulation of the oxidation rate by methylene blue and in three a decreased pyruvate dehydrogenase complex activity was confirmed. Methylene blue restored a near normal oxidation rate in the seventh culture which had decreased cytochrome c oxidase activity.


Mevalonate kinase assay using DEAE-cellulose column chromatography for first-trimester prenatal diagnosis and complementation analysis in mevalonic aciduria. Mevalonic aciduria due to mevalonate kinase deficiency, an inherited defect of cholesterol biosynthesis, has presented with clinical variability in 10 patients from 7 families. We sought to define a genetic basis for this heterogeneity by determining mevalonate kinase activity in fibroblast heterokaryons obtained by polyethylene glycol fusion. To this end we developed a DEAE-cellulose (Cl-) column chromatography procedure for assessing mevalonate kinase in cell extracts that would allow multiple rapid analyses. Fusion of control fibroblasts with those from affected patients from six families with mevalonate kinase deficiency yielded 37% of the mean control activity. None of the fusions between the six cell lines of patients resulted in measurable mevalonate kinase activity. Using the chromatographic procedure, we developed an optimized assay for mevalonate kinase in biopsied chorionic villi. Km values for chorionic villi were similar to those obtained in fibroblasts. Mevalonate kinase activity in biopsied chorionic villi showed a linear increase (0.75-4.3 nmol/min per mg protein) with gestational age from 7 to 14 weeks. Using the optimized assay in biopsied chorionic villi we performed a first-trimester prenatal diagnosis in a pregnancy at risk for mevalonate kinase deficiency and correctly diagnosed an unaffected fetus. The availability of an optimized assay for mevalonate kinase in biopsied chorionic villi should allow reliable first-trimester prenatal diagnosis for families at risk.


Evolution of clonality and invasive behavior of Epstein-Barr virus immortalized lymphoblastoid cell lines in SCID mice brains. Recently established Epstein-Barr virus immortalized lymphoblastoid cell lines express polyclonal immunoglobulins, are diploid, and grow into invasive tumors when injected intracerebrally into mice with severe combined immunodeficiency (SCID). It is unclear whether clonal selection of neurotropic cell lines occurs during long-term growth in the brain and the effect of this selection on brain invasiveness. Epstein-Barr immortalized lymphoblastoid cell lines from a normal Epstein-Barr negative donor were serially passaged seven times intracerebrally within groups of SCID/SCID CB 17 mice. Each cell line was injected into five or more animals during each passage. Clonality of the rescued cell lines, genotype, and brain invasiveness were examined. All mice developed extensive intracerebral lymphoproliferative disease within 10-18 days after injection. Intracerebral, subarachnoid, intraventricular, and perivascular lymphoid lesions were noted. Infiltrates were similar in all animals studied regardless of the passage number. Clonal B cell populations were detectable in lesions after the first passage by Southern blot hybridization using JH probe. Immunohistochemically, polyclonal tumors were seen initially, but after the fourth passage, monoclonal cytoplasmic immunoglobulin was predominantly expressed by all tumors. Minor bands seen in the early passages disappeared subsequently. Random chromosomal abnormalities appeared in the rescued cell lines after the third passage; however, after the sixth passage, the abnormalities became more consistent. Clonability in agarose was very low initially in both cell lines and increased significantly after the sixth passage. These experiments demonstrate that within the immunoprivileged conditions of the SCID mouse brain, the evolution of Epstein-Barr immortalized lymphocytes from polyclonal to oligo- and monoclonal cell lines with chromosomal abnormalities occurs very early. This evolution is not paralleled by increased invasiveness in vivo.


Comparison of in situ hybridization using different nonisotopic probes for detection of Epstein-Barr virus in nasopharyngeal carcinoma and immunohistochemical correlation with anti-latent membrane protein antibody. The detection of Epstein-Barr virus (EBV) in nasopharyngeal carcinoma (NPC) may be of diagnostic importance, particularly in cases from nonendemic areas. For cellular localization of viral genomes, cold in situ hybridization methods for the demonstration of EBV-associated NPC remain difficult and relatively insensitive for routinely processed tissues. The aim of the present study was to assess the importance of tissue processing and the hybridization targets to improve the sensitivity of the cold in situ hybridization method. In situ hybridization was performed in six cases of NPC using three biotinylated EBV cDNA probes (BamHI W/IR1, BamHI Y/EBNA2, XhoI/latent membrane protein) and two cocktails of EBER and BHLF1 oligonucleotides labeled with fluorescein isothiocyanate on routinely fixed and paraffin embedded sections. In two cases, in situ hybridization was also performed on specially processed (ModAMeX) sections. Immunohistochemistry was used to detect EBV-induced antigens using monoclonal antibodies against latent membrane protein, EBNA2 and ZEBRA (BZLF1). All cases showed EBV nucleic acids regardless of the tissue preparation with the three cDNA probes and on routinely processed sections with EBER oligonucleotides. By using cDNA probes, the best EBV DNA signal was obtained with BamHI W without heating of slides in tissue sections processed by ModAMeX, which probably gives rise to large amounts of single stranded DNAs. All cases positive with cDNA probes were found to be positive with EBER oligonucleotides and negative with BHLF1. However, on routinely processed paraffin sections, the signals with EBER oligonucleotides were stronger than with BamHI W cDNA probe. Dual labeling with in situ hybridization and immunohistochemistry showed that the hybridization signals were restricted to malignant epithelial cells. Latent membrane protein expression was detectable in four of six EBV nucleic acid-positive cases on both ModAMeX and routinely processed sections. The anti-EBNA2 and anti-ZEBRA antibodies were found to be negative on the two cases processed by ModAMeX. Cold in situ hybridization, in particular with EBER oligonucleotides, appears to be more reliable than immunohistochemistry with anti-latent membrane protein antibody to detect EBV in NPC in routine pathology. These findings confirm a distinctive phenotype (latent membrane protein +/-, EBNA2-, ZEBRA-) of EBV-positive NPC. The negative staining for BHLF1 oligonucleotides further supports the viral latency.


Establishment and characterization of a new human synovial sarcoma cell line, HS-SY-II. The line of differentiation in synovial sarcoma still remains controversial. Thus far, only a few human synovial sarcoma cell lines have been described. However, their morphologic characteristics have not been fully established. We established a new synovial sarcoma cell line (HS-SY-II) from pleural effusion with lung metastasis in a typical example of the monophasic spindle cell type. The HS-SY-II cells, in vitro and in vivo, were examined by light microscopy, immunohistochemistry, electron microscopy, and cytogenetics. The HS-SY-II cells showed a hypertriploid karyotype with complex chromosome abnormalities including pathognomonic t(X;18)(p11;q11), and have been stably maintained for more than 40 months in vitro, showing rather small spindle or polygonal shape without conspicuous pleomorphism. Histologic features of initially and serially transplanted tumors in nude mice were essentially the same as those of the original sarcoma, corresponding to the monophasic spindle cell variant with a prominent palisading pattern and calcified foci in parts. The HS-SY-II cells in vitro and in vivo similarly expressed vimentin and cytokeratin by immunohistochemistry, and also exhibit the same ultrastructural features such as irregularly shaped nuclei with prominent nucleoli, many paranuclearly running intermediate filaments, and filopodia-like processes. This HS-SY-II cell line retaining the distinct morphological characteristics as the monophasic spindle cell type of synovial sarcoma therefore will be extremely useful for various pathomorphologic investigations on synovial sarcoma.


Ultrastructural localization of human papilloma virus by nonradioactive in situ hybridization on tissue of human cervical intraepithelial neoplasia. A nonradioactive in situ hybridization was developed to localize human papilloma virus (HPV) at the ultrastructural level. Cervical biopsies from human uterine cervices clinically suspicious of condyloma were embedded in Lowicryl K4M at low temperature. Postembedding in situ hybridization was performed with DNA probes specific for HPV types 6/11, 16, and 18. The hybrids were detected by anti-horseradish peroxidase antibodies conjugated with 10 nm colloidal gold particles. Localization for HPV 16 and 18 both was to intranuclear and cytoplasmic sites. Cytoplasmic detected HPV signals were between masses of intermediate filaments and in vacuoles; other organelles were devoid of positive signal. Within the nucleus the precise localization of the viral nucleic acid was episomal, vacuolar, and chromosomal. In situ hybridization with plasmid control DNA confirmed the specificity of the HPV positive signals. This study helps define the subcellular compartmentalization of HPV DNA in infected human cells.


Possible involvement of Na(+)-H+ exchange in the early phase of reperfusion in myocardial stunning. Calcium overload during reperfusion after prolonged ischemia has been associated with the Na(+)-Ca2+ exchange system. It has been proposed that the promotion of Na(+)-Ca2+ exchange at reperfusion may be mediated by Na(+)-H+ exchange. To evaluate whether this hypothesis is applicable for stunned myocardium, we examined the influence of temporary suppression of Na(+)-H+ and/or Na(+)-Ca2+ exchange during early reperfusion in isolated rat hearts. Myocardial stunning was produced by global ischemia for 15 min at 37 degrees C. The initial reperfusate was given during the subsequent 10 min after ischemia, and followed by reperfusion with normal Krebs-Henseleit buffer solution for 40 min. Hemodynamic indices, creatine kinase in coronary effluent, and myocardial water content were measured during reperfusion. The functional recovery of stunned myocardium was improved with higher extracellular Na+ concentration and/or lower Ca2+ concentration of the initial reperfusate. Aortic flow recovery of group II (135 mM Na(+)-0.5 mM Ca2+) was 77.0 +/- 3.4%, which was substantially greater (P < 0.05) than that of other groups: group I (control, 135 mM Na(+)-1.5 mM Ca2+), 68.2 +/- 2.4%; group III (25 mM Na(+)-0.5 mM Ca2+), 48.7 +/- 2.9%; group IV (25 mM Na(+)-1.5 mM Ca2+), 21.6 +/- 1.5%. Administration of amiloride, an inhibitor of Na(+)-H+ exchange, in the initial reperfusate ameliorates cardiac damage and improved aortic flow recovery in a dose-dependent manner (10(-6) M, 70.1 +/- 3.7%; 10(-5) M, 77.3 +/- 1.7%; 10(-4) M, 82.0 +/- 2.1% vs control 68.2 +/- 2.4%).(ABSTRACT TRUNCATED AT 250 WORDS)


When do alcoholics first discuss drinking problems? Data from a sample of 338 alcoholics who were identified from medical record reviews and diagnosed as alcohol dependent according to DSM-III criteria by means of a structured psychiatric interview were analyzed to determine when in the course of alcohol disorder development first discussion with a health professional about their drinking problems had occurred. The data support an image of alcoholics as experiencing many drinking problems before seeking professional help and, particularly, before attending AA. These data point to early identification as a possible fruitful strategy to alleviate some of the social costs of drinking problems.


Self-control concerns and drinking loss of control in general and clinical populations. In popular conceptions, loss of control over drinking involves a double loss of control, over one's life as well as over one's drinking. A measure of "control worries," concerning the experience of difficulties in controlling one's life, was developed. The relationships between measures of control worries, loss of control over drinking and heavy-drinking behavior were examined in samples both of alcohol treatment clients and of the general population in a California county. Alcohol treatment clients reported greater control worries and loss of control over drinking than members of the general population. Although control worries were only slightly correlated with drinking loss of control, this relationship tended to be conditional in the general population. The findings suggest that concerns about personal control in general may be a useful construct to consider in conjunction with the drinking loss of control concept.


A follow-up study of problem drinkers offered a goal choice option. Patients (N = 100) who had been admitted to behaviorally oriented residential treatment for their drinking problems were followed up for 1 year. A treatment goal option of controlled drinking was explicitly catered for. Overall outcome, in which 27% of those available for follow-up were categorized as "successful," 35% as "equivocal" and 38% as "failure," does not appear to be markedly dissimilar to that reported from other agencies. The distribution of approximately equal abstinent and nonabstinent successful outcomes is similar to that found following treatment programs that promote a single goal. Sociodemographic variables were less influential in predicting outcome than were treatment variables--with frequency of aftercare attendance being particularly significant. Those who had received previous hospital treatment for their problem, those who habitually drank in company and those who had abnormal blood test results prior to entering treatment had poorer outcome.


Elderly versus younger problem drinker profiles: do they indicate a need for special programs for the elderly? To address the question, "Do elderly problem drinkers differ from younger ones and therefore might they need special treatment programs?", the descriptive profiles of a representative sample of older and younger persons arrested for drinking and driving in Iowa were compared. Subjects were interviewed by telephone or mail using a structured, clinical interview schedule that was designed to obtain a comprehensive self-report picture of the role of alcohol in their lives. Younger persons (18-54 years old) were compared with two overlapping elderly age groups (55 and over and 65 and over). The elderly subjects were also dichotomized as "early onset" (at least one problem-drinking indicator occurred prior to age 55) and "late onset" (all problem drinking indicators occurred at age 55 or later). Although there were several statistically significant (p < or = .01) differences between the elderly and younger problem drinkers, there was a much, or more, heterogeneity within the elderly groups as there were differences between the elderly and their younger counterparts. Also, the descriptive profile of these at-large elderly problem drinkers differed, depending on whether their alcohol abuse was early- or late-onset.


Alcohol use and high-risk behavior by intravenous drug users in an AIDS education paradigm. The relationship between alcohol consumption and high-risk behavior for HIV infection was examined among 313 in-treatment intravenous drug users (IVDUs) by random assignment of these subjects to one of three interventions: (1) structured interview regarding risk behavior; (2) interview plus one group AIDS education session; (3) interview, AIDS education plus optional HIV testing. Alcohol users (n = 148) had more needle sharing and sexual partners than did nondrinkers. Follow-up interviews revealed no significant behavioral changes as a function of intervention condition or alcohol use. Better interventions, including more vigorous treatment of alcohol use, are needed to reduce risk behaviors among IVDUs.


Impaired neutrophil bactericidal activity correlates with the infection occurring after surgery for esophageal cancer. We examined whether or not preoperative impaired bactericidal activities of polymorphonuclear neutrophils (PMN) are associated with infections following surgery for esophageal cancer. Intracellular killing (KI), superoxide anion-producing capacity (SOP), and myeloperoxidase (MPO) activity were measured in 22 patients with esophageal cancer, 27 with gastric cancer, and 13 age-matched controls. The average level of KI was significantly depressed in patients with esophageal cancer or with gastric cancer, to a similar extent, compared to findings in controls, but SOP was not. In esophageal cancer patients, the SOP level was significantly higher in those with postoperative septic complications than in those without such problems, whereas the KI level was depressed to a similar extent in both. Therefore, a depression of KI with elevation of SOP of PMN may serve to predict complications of infection following surgery in patients with esophageal cancer.


Minimal model for Ca(2+)-dependent oscillations in excitable cells. A minimal model for calcium controlled oscillations is presented. The model considers only an exchange of potassium and calcium ions over the plasma membrane. Calcium ions leak into the cell through a potential dependent channel and is extruded by a pump. Potassium leaks out through a calcium dependent, but voltage independent, channel. The cytosolic calcium concentration is buffered, so a fixed fraction is free. Inactivation, membrane capacity, and time delays for the conductance changes are not included, so the time dependence is solely introduced through the temporal changes of the intracellular Ca(2+)-concentration. With continuous parameter changes the model can switch between five states: (1) a non-excitable, stable state; (2) single-spike excitability; (3) slow, spontaneous oscillations; (4) reverse-spike excitability; and (5) another non-excitable, stable state. One of the key parameters for this switching behavior is the rate constant for the calcium pump.


Ras oncogene point mutation: an infrequent event in bronchioloalveolar cancer. Ras oncogene point mutation, primarily activating the K-ras gene, has been reported in approximately one third of lung adenocarcinomas. This identifies a subset of early stage tumors clinically associated with smoking and an aggressive clinical course. Because of these findings, this study was undertaken to determine the occurrence of ras point mutations in bronchioloalveolar carcinoma. This uncommon form of lung adenocarcinoma is usually indolent but can sometimes present as a rapidly growing, multifocal tumor. Twenty tumor samples obtained at thoracotomy were examined for H-ras, K-ras, and N-ras oncogene mutational activation involving codons 12, 13, or 61. This was performed by an oligonucleotide hybridization technique following polymerase chain reaction amplification of these specific sequences. K-ras point mutation involving codon 12 was observed in two tumors, but not in the adjacent histologically benign lung tissue. These mutations were confirmed by direct sequencing of these polymerase chain reaction products. Both patients were smokers, had stage I tumors, and remain disease-free at 27 and 40 months postoperatively. No H-ras or N-ras point mutations were found. These findings suggest that ras activation is an infrequent event in bronchioloalveolar carcinoma. We speculate that ras activation is not a common transformational event in this form of lung adenocarcinoma.


Prognostic value of deoxyribonucleic acid aneuploidy in primary non-small-cell lung carcinomas and their metastases. The ploidy status of the deoxyribonucleic acid of a malignant lung tumor provides additional information besides histologic grading and tumor staging according to lymph node infiltration and tumor metastasis. Ninety-nine surgical specimens from patients with non-small-cell lung carcinoma were investigated by flow cytometry. Deoxyribonucleic acid aneuploidy was found in 48% of the primary tumors. Patients with deoxyribonucleic acid-euploid tumors showed better survival (p < 0.01) than those with deoxyribonucleic acid-aneuploid carcinomas independent of tumor stage. Deoxyribonucleic acid ploidy status of the primary tumor was compared with that of N2 lymph node metastases in 29 cases. Seven samples showed a change from deoxyribonucleic acid aneuploidy in the primary tumor to deoxyribonucleic acid euploidy in the lymph node metastases. Survival was significantly better for patients with euploid primary tumors and lymph node metastases, followed by patients with deoxyribonucleic acid-aneuploid primary tumors and euploid lymph node metastases. Survival was poorest in patients with deoxyribonucleic acid-aneuploid primary tumors and lymph node metastases. It was observed that only the simultaneous determination of deoxyribonucleic acid ploidy of primary tumors and lymph node metastases permits accurate prognostic evaluation in case of lymph node infiltration.


Quantal analysis using maximum entropy noise deconvolution. When applying quantal analysis to synaptic transmission it is often unclear how much of the measured postsynaptic signal fluctuation arises from random sampling and noise rather than from the probabilistic transmitter release process. Unconstrained noise deconvolution methods do not overcome this because they tend to overfit the data, often giving a misleading picture of the underlying process. Instead, maximum entropy deconvolution provides a solution which is the smoothest, or most featureless, distribution that is still compatible with the data, taking noise and sample size into account. A simple way of achieving this is described, together with results of Monte Carlo simulations which show that the features present in the maximum entropy solution usually reflect the process underlying the data and not random sampling or noise.


Systemic administration of platelet-activating factor in rat reduces specific pulmonary uptake of circulating monoclonal antibody to angiotensin-converting enzyme. The biodistribution of radiolabeled mouse monoclonal antibody (MoAb) to angiotensin-converting enzyme (ACE) and control, nonimmune mouse IgG in platelet activating factor (PAF)-treated rats was studied. The blood level of both preparations was slightly decreased (90% of the control) in PAF-treated rats. Specific pulmonary accumulation of anti-ACE MoAb was reduced to 50% of control in contrast to a doubling in nonspecific pulmonary uptake of non-immune IgG. The changes in anti-ACE MoAb biodistribution were lung-specific and were accompanied by decrease in the pulmonary ACE activity (to 60% of control) and increase in serum ACE activity (to 170% of control). Thus anti-ACE MoAb reveals PAF-induced changes in the status of the pulmonary ACE and therefore can be used for the studies of pathology of the pulmonary endothelium.


Myeloid leukaemogenicity and transmission of the HPRS-103 strain of avian leukosis virus. The HPRS-103 strain of avian leukosis virus (ALV) was isolated recently from meat-type chickens and represents a new envelope subgroup. Its oncogenicity has been studied in three meat-type and five Leghorn strains of chickens. In the meat-type strains, the virus, following embryonal inoculation, induced an overall incidence of 27% myelocytic myeloid leukosis (myelocytomatosis) and 12% renal adenomas, with long median latent periods. Amongst the Leghorn lines, these tumors occurred with similar incidence in line 0, but with lower or zero incidences in the other lines. A variety of other tumours occurred with low incidence. Embryonal infection resulted in a permanently tolerant viraemic state with shedding of ALV group specific (gs)-antigen to egg albumen; contact infection resulted mainly in the development of non-shedder birds with serum virus-neutralising antibodies. Contact infection in a meat-type line was associated with the development of transient or permanent viraemia in some birds, and a low tumour incidence. A viraemic phase was not detected following contact infection in a Leghorn line and no tumours developed. The long latent period between embryo infection and tumour mortality, apparently differing from the consequences of infection with acutely transforming ALVs, and the inability of HPRS-103 ALV to transform cultured bone marrow cells, suggests that this virus may lack a viral oncogene and exert its oncogenic properties by some other mechanism such as promoter insertion activation of a cellular oncogene.


Antibodies to human herpes virus-6 in patients with acute lymphocytic leukemia. Human herpesvirus-6 (HHV-6), a ubiquitous virus that causes exanthem subitum and occasional cases of infectious mononucleosis, hepatitis and other viral syndromes, has also been associated with acute lymphocytic leukemia (ALL) in children. To further investigate this association, we obtained sera from 50 patients with ALL and 50 age-sex matched controls. Antibodies to HHV-6 were determined using ELISA and indirect immunofluorescent antibody (IFA) tests. No significant difference between antibody titers in the cases and controls was observed. Since seroepidemiologic studies have demonstrated higher HHV-6 antibody titers in young children than in adults, this serologic study suggests that the previous association reported for HHV-6 and ALL was a result of the age of the population rather than a relationship between the virus and the disease.


Biochemical evaluation of adrenal dysfunction: the laboratory perspective. In this study, we reviewed the diagnostic efficiency of laboratory tests that are performed for assessment of patients with Cushing's syndrome or adrenal insufficiency. Baseline laboratory data from patients subsequently diagnosed with adrenal dysfunction were analyzed for tests performed between 1987 and 1989 at our institution. Results were analyzed for 36 patients diagnosed with pituitary-dependent Cushing's syndrome, 15 with ectopic Cushing's syndrome, 12 with adrenal-dependent Cushing's syndrome, 20 with primary adrenal insufficiency, and 7 with secondary adrenal insufficiency. Tests reviewed were plasma cortisol, plasma corticotropin, urinary free cortisol, urinary 17-ketosteroids, urinary ketogenic steroids, low-dose and high-dose dexamethasone suppression, and metyrapone stimulation. Our findings suggest that a substantial proportion of diagnoses could be based on the results of three tests--plasma corticotropin, plasma cortisol, and urinary free cortisol. We present a nomogram that combines the results of plasma corticotropin and plasma cortisol testing to enhance the diagnostic efficiency of these tests.