Evaluation of stress on the autonomic nervous system of patients on mechanical circulatory assist

PURPOSE: The power density values of high (HF; 0.15 to 0.4 Hz) and low frequency (LF; 0.04 to 0.15 Hz) components of arterial pressure were adopted for evaluation of stress on the autonomic nervous system of patients supported by mechanical circulatory assist. METHODS: Power spectral analysis (PSA) of arterial pressure signals was carried out, and trends in changes in the spectral components were observed in 12 patients on mechanical circulatory support (IABP and/or PCPS) and without the support (n=10). RESULTS: The normalized LF was depressed initially and increased gradually in both groups of patients except for four patients on mechanical circulatory assist. Three patients among them consequently died. CONCLUSIONS: Depressed LF represented marked stress on ANS and prolongation of the depression was related to poor outcome of patients. PSA of systemic blood pressure offers a reasonable tool for evaluation of stress on the ANS and for prediction of the prognosis of patients with circulatory assist devices.     

Suppression of arthritis by forced expression of cyclin-dependent kinase inhibitor p21(Cip1) gene into the joints

Rheumatoid synovial fibroblasts (RSF) express cyclin-dependent kinase (CDK) inhibitors p16(INK4a) and p21(Cip1) when they are growth-inhibited in vitro. The induction of p16(INK4a) is characteristic of RSF and intra-articular p16(INK4a) gene therapy has been shown to suppress adjuvant arthritis (AA) of rats. The other inducible CDK inhibitor, p21(Cip1), has multiple functions depending on the cell type. They include inhibition of CDK as well as promotion of active CDK complex formation and induction of apoptosis. This study is to discern the biological effects of p21(Cip1) gene transfer into RSF and its therapeutic effects on AA. A recombinant adenovirus containing a human p21(Cip1) gene and control adenoviruses were prepared. RSF infected with these viruses were examined for their cell growth. Apoptotic cell death was evaluated by nuclear staining and DNA fragmentation analysis. In vivo gene therapy of rat AA was carried out by intra-articular injection of the viruses. Severity of the arthritis was clinically scored. The treated joints were examined histologically and proliferating cell nuclear antigens (PCNA) were detected immunohistochemically. The adenoviral p21(Cip1) gene transfer inhibited growth of RSF without inducing apoptosis. p21(Cip1) gene therapy suppressed AA clinically and histologically. The effects were comparable to p16(INK4a) gene therapy. PCNA expression was reduced in the p21(Cip1)-treated joints. The adenoviral gene transfer of p21(Cip1) ameliorated rat AA. The effect was attributable to inhibition of proliferation. Because p21(Cip1) is induced more easily by many chemicals than p16(INK4a), it also appears to be a feasible target in developing anti-rheumatic drugs.     

Changes in the plasma histamine concentration after the administration of vecuronium bromide

Clinical symptoms of anaphylactoid reaction to muscle relaxants vary from localized flush to cardiovascular collapse. Vecuronium bromide is reported to have very little histamine releasing property. However, there are some reports of anaphylaxis or anaphylactoid reaction to vecuronium. We studied plasma histamine concentration after the intravenous injection of vecuronium to confirm the histamine release. Twenty patients were randomly allocated to one of two groups, each group comprising of 10 patients: one group was to receive vecuronium 0.1mg·kg^–1 and the other 0.2mg·kg^–1 using the priming principle. Blood samples were taken prior to and 1, 3, 5, 8 and 13min after the administration of vecuronium. The plasma histamine concentration was measured by radioimmunoassay with monoclonal antibody.There were no significant changes in plasma histamine concentration over 13min after the administration of vecuronium compared with the baseline value. There were also no significant differences between these two groups. We concluded that vecuronium up to 0.2mg·kg^–1 did not change the plasma histamine concentration in the patients having no previous history of allergy or atopic tendencies.(Mitsuhata H, Matsumoto S, Enzan K, et al.: Changes in the plasma histamine concentration after the administration of vecronium bromide. J Anesth 5: 24–29, 1991)     

Oncostatin M inhibits proliferation of rat oval cells, OC15-5, inducing differentiation into hepatocytes

Oval cells of the liver participate in liver regeneration when hepatocytes are prevented from proliferating in response to liver damage. To clarify the role of oncostatin M (OSM) in the liver regeneration involving oval cells, we examined the expression of OSM and OSM-specific receptor (OSM-R) in the liver undergoing regeneration in the 2-acetylaminofluorene/partial hepatectomy model. Expression levels of OSM-R changed in correlation to the number of oval cells, and its expression was exclusively observed in oval cells. On the other hand, OSM was expressed in both oval cells and Kupffer cells. To examine the effect of OSM on the growth and differentiation of oval cells, rat oval cells (OC15-5) were incubated in conditioned medium of 293T cells expressing rat OSM cDNA. This resulted in suppression of growth, changes in morphology (microvilli and large cytoplasm with developed organelles), and expression of hepatocyte markers (albumin, tyrosine amino transferase, and tryptophan oxygenase). The effects of the conditioned medium with rat OSM were abrogated by introducing a small interfering RNA specifically targeting rat OSM-R into OC15-5 cells. These results indicate that OSM is a key mediator for inducing differentiation of OC15-5 cells into hepatocytes and suggest that the OSM/OSM-R system is pivotal in the differentiation of oval cells into hepatocytes, thereby promoting liver regeneration.     

S-100-positive nerve fibers in hepatocellular carcinoma and intrahepatic cholangiocarcinoma: an immunohistochemical study

The aim of the present study was to determine whether or not liver carcinomas are innervated, since there have been no previous reports on the distribution of nerve fibers in human hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (ICC). We investigated nerve fibers by immunohistochemical and morphometric methods in 63 cases of HCC and 28 cases of ICC. An antibody to S-100 protein was used to visualize nerve fibers. In HCC, S-100-positive nerve fibers were absent in the tumoral region, including the sinusoids, and tumoral fibrous septa, while in the capsule of HCC some S100-positive nerve fibers (density: 0.08 +/- 0.03/mm2) were present in contact with vasculatures. In ICC, a few nerve fibers were noted in the tumoral stroma (density: 0.02 +/- 0.01/mm2). No nerve fibers were seen in the neovasculized vessels (tumor vessels) in both HCC and ICC. In invasive regions of HCC and ICC, there were S-100-positive nerve fibers in pre-existing residual portal tracts (density: HCC, 0.11 +/- 0.03/mm2; ICC, 0.13 +/- 0.04/mm2). In non-tumor regions of HCC and ICC, there were many S100-positive nerve fibers (density: 0.41 +/- 0.13/mm2) in portal tracts and, to a much lesser degree, in the sinusoids. These results suggest that tumor cells and vasculatures in HCC and ICC are rarely influenced by nerve fibers.     

Maintenance by androgen and thyroid hormone of androgen-induced convoluted tubular cells in mouse submandibular glands

Injection of 5α-dihydrotestosterone (DHT) into castrated adult female mice stimulated the proliferation of a small proportion of the convoluted tubular cells in the submandibular glands. We investigated the effects of DHT and thyroxine (T₄) on the maintenance of these proliferated convoluted tubular cells. For this, castrated adult female mice that had been treated daily with DHT for 3 days and then once with [³H]thymidine, received a first series of daily injections of DHT for various periods or T₄ for 10 days, and then a second series of injections of treatment with DHT or T₄, or no further treatment. The second series of treatments with DHT or T₄ maintained the percentages of ³H-labeled convoluted tubular cells at similar or slightly lower levels than those at the end of the first series of treatments. In mice that did not receive the second series of treatments, the percentages of ³H-labeled convoluted tubular cells decreased markedly, becoming significantly lower than those at the end of the second series of treatment with DHT or T₄. We also examined the effect of DHT on the proliferation of convoluted tubular cells of castrated adult female mice that had received 10 daily injections of DHT and then no treatment for 28 days. In these mice, the cells did not proliferate markedly on stimulation with DHT. These results suggest that androgen and thyroid hormone maintain convoluted tubular cells that have proliferated in response to androgen, and that the convoluted tubular cells may become unresponsive to androgen in terms of proliferation after their exposure to androgen.     

The role of chemical mediators and mucosal hyperreactivity in nasal hypersecretion in nasal allergy

This study was designed to elucidate, first, the degree of participation of direct effects of histamine on the nasal glands and the nasal vasculature in clinical manifestation of hyperrhinorrhea in nasal allergy and, second, the existence of hyperreactivity of the nasal glands to acetylcholine in nasal allergy. The study demonstrates that histamine released by degranulation of basophilic cells in the nasal mucosa causes nasal hypersecretion mostly by way of the reflexive pathway. Approximately 4% of the amount of nasal secretion induced by an antigen challenge in subjects with house-dust nasal allergy was due to a leakage of plasma. There were almost no direct effects of histamine on the nasal glands. Nasal secretion induced by nasal challenge with acetylcholine after vidian neurectomy comes from the nasal glands by its direct effects on the nasal glands, the amount of which indicates degree of reactivity of the nasal glands to acetylcholine independent of hypersensitivity of the mucosal sensory system. The nasal glands of vidian neurectomized subjects having nasal allergy reacted more excessively to extrinsic acetylcholine than nasal glands of subjects of the control group. This verifies the existence of hyperreactivity in the nasal glands to acetylcholine in nasal allergy. The nasal glands of nasal allergy patients may respond more excessively to a given amount of acetylcholine released from parasympathetic terminals.     

Immunocytochemical study of membrane skeletons in abnormally shaped erythrocytes as revealed by a quick-freezing and deep-etching method

Summary Ultrastructures of membrane skeletons in spherocytic and elliptocytic erythrocytes were investigated immunocytochemically. Erythrocytes obtained from patients with hereditary spherocytosis (HS) and hereditary elliptocytosis (HE) were split open mechanically to obtain exposed cytoplasmic sides of erythrocyte membranes and were immunostained with anti-spectrin antibody. Replica membranes were prepared by a quick-freezing and deep-etching method and were checked by electron microscopy. The in situ membrane skeletons of normal erythrocytes consisted mainly of reticular patterns of spectrin filaments, which formed networks on the cytoplasmic sides of the cell membrane. In contrast, the membrane skeletons of abnormally shaped erythrocytes (HS and HE) were much less filamentous and more granular than those of normal erythrocytes. This abnormal organization in erythrocyte membrane skeletons may be one of the factors that induce abnormally shaped erythrocytes in HS and HE patients.     

Inhibitory activity in saliva of cell-to-cell transmission of human T-cell lymphotropic virus type 1 in vitro: evaluation of saliva as an alternative source of transmission.

Human T-cell lymphotropic virus type 1 (HTLV-1) is known to be transmitted vertically through breastfeeding and horizontally by blood transfusion and sexual contact. Our intervention study has suggested the presence of additional alternative maternal transmission pathways. To explore the possibility of transmission through saliva, we used PCR to quantify the HTLV-1 provirus in saliva samples from 18 carrier mothers and 10 patients with HTLV-1-associated myelopathy/tropical spastic paraparesis. The provirus was detected in 60 and 90%, respectively, of the samples, with estimated copy numbers in the range of 10 to 10(4)/ml. However, the saliva, regardless of the presence or absence of antibodies to the virus, showed a strong tendency to inhibit the cell-to-cell transmission of HTLV-1 in vitro, as examined by a syncytium inhibition assay. The natural inhibitory activity in saliva of seronegative volunteers was heat sensitive, and most of the activity was recovered by ultrafiltration in the fraction of macromolecules with a molecular weight of more than 100,000. In addition to this natural activity, saliva of HTLV-1-infected individuals contained immunoglobulin G molecules capable of neutralizing syncytium formation. These results strongly suggested that HTLV-1-infected cells in the carriers' saliva, which contains neutralizing antibodies in addition to the natural activity inhibiting cell-to-cell viral infection, barely transmit the virus. Transmission of HTLV-1 through the saliva would thus seem to be rare, if it occurs at all.     

An 8‐cM interstitial deletion on 4q21‐q22 in DNA from an infant with hepatoblastoma overlaps with a commonly deleted region in adult liver cancers

We performed molecular analysis of a germline interstitial deletion of chromosome 4 [del(4)(q21.22q23)], which had been observed in a male infant manifesting early‐onset hepatoblastoma (HBL). The chromosomal anomaly in this child was associated with a unique congenital syndrome including HBL, atrial septal defect, ventricular septal defect, patent ductus arteriosus, mental retardation, and seizures. However, the patient did not exhibit a megalencephaly typical of 4q21‐22 deletions. His HBL was associated with an increasing serum α‐fetoprotein level and rapid growth. To define the chromosomal deletion at the molecular level in this child, we analyzed his lymphoblasts with fluorescence in situ hybridization, using as probes a panel of BAC/PAC genomic clones containing STS markers covering the 4q12‐27 region. The analysis revealed that the affected chromosome had an 8‐cM deletion within 4q21‐q22, flanked by markers D4S2964 and D4S2966. This microdeletion overlaps with the commonly deleted region at 4q21‐q22 that was recently defined in adult hepatocellular carcinomas. © 2001 Wiley‐Liss, Inc.