Haematological improvement by long-term administration of recombinant human granulocyte-colony stimulating factor and recombinant human erythropoietin in a patient with severe aplastic anaemia

A 6-year-old girl with post-hepatitic severe aplastic anaemia was referred to our hospital. Haematological examination showed a haemoglobin level of 5.2 g/dl, platelet count of 8,000/l, and white blood cell count of 130/l with 17% neutrophils. She was treated with recombinant human granulocytecolony stimulating factor (15 g/kg/day i.v.) and cyclosporin A (6 mg/kg/day p.o.). The absolute neutrophil count gradually increased, but Hb and platelets were not improved. The intravenous administration of recombinant human erythropoietin (100 U/kg three times a week) was started, and the reticulocyte count reached 20000/l on day 12. The platelets increased to 81000/l after 16 months of combined administration of recombinant human granulocyte-colony stimulating factor, recombinant human erythropoietin and cyclosporin A. After 20 months of combined administration, the haematological results were: Hb, 13.1 g/dl; platelets 80000/l: WBC, 9500/l with 40% neutrophils. After recombinant human granulocyte-colony stimulating factor treatment, the myeloid elements of the bone marrow and the number of granulocyte-macrophage colony forming units increased. Bone marrow erythropoiesis and erythroid colonies also increased after recombinant human erythropoietin administration. The clinical course suggested a beneficial effect of haemopoietic growth factors and cyclosporin A in post-hepatitic aplastic anaemia.     

Evaluation of the pressure transfer system in the intracranial cavity by coherency

Coherency provides a method to evaluate model linearity. The characteristics of pressure wave transmission in the intracranial cavity were studied by coherency in 16 cats with hydrostatic pressure loading to assess the linearity of the system, which is an assumption for use of the transfer function. Linearity was observed in only the fundamental waves of the respiration-induced component and the cardiac-induced component of intracranial pressure oscillation, and in the second harmonic wave of the latter. Linearity at the other frequencies was close to zero. The pressure transfer system in the intracranial cavity was basically a non-linear system. As intracranial pressure rose, the increase in the pressure transfer efficiency was largest in the low-frequency domain and smallest in the high-frequency domain, indicating that the cerebral blood vessels are characterized by inferior transmission of high frequency due to increased intracranial pressure. In addition, the correlation between the coherencies of the cardiac-induced fundamental wave component and intracranial pressure, and between those of the cardiac-induced second harmonic wave component and intracranial pressure, showed that the slope of the straight line was greater between 45 and 70 mmHg than between 10 and 45 mmHg. This suggests that there is a break point, located between 45 and 70 mmHg, where the increase in the coherency values is accelerated, caused by an increase in the intracranial elastance, as well as an increase in the cerebrovascular compliance due to the reduced vascular transmural pressure.     

Aprindine blocks the sodium current in guinea-pig ventricular myocytes

Summary Aprindine is a class Ib antiarrhythmic agent. We studied effects of aprindine (3 µmol/l) on the Na⁺ current using whole cell voltage clamp (tip resistance = 0.5 , [Na]_i and_o = 10 mmol/l at 18°C). Aprindine revealed tonic block (Kd_rest = 37.7 µmol/l, Kd_i = 0.74 µmol/l; n = 4). Aprindine, shifted inactivation curve to hyperpolarizing direction by 11.4 ± 3.5 mV (n = 4) without changes in slope factor. In the presence of 3 µmol/l aprindine, aprindine showed phasic block, i.e., duration-dependent block at 2 Hz (64% ±3070 at 1.5 ms, 82%±6% at 20 ms, 93%±7% at 200 ms; n = 4). Short single prepulse also produced aprindine-induced phasic block (12% at 1.5 ms, 22% at 100 ms; n = 2). After removal of fast inactivation of Na⁺ current by 3 mmol/l tosylchloramide sodium, aprindine revealed phasic block, independent of holding potential. The recovery time constant from aprindine-induced phasic block was 4.8 s at holding potential = –100 mV and 5.0 s at holding potential = –140 mV. This use-dependent block of aprindine had pH dependency. Under acidic condition (pH 6.0), 3 µmol/l aprindine showed smaller use-dependent block (14% ± 7% at 2 Hz; n = 4) comparing with either at pH 7,4 (68% ± 13%; n = 4) or at pH 8.0 (90% ±12%; n = 4).The results suggest that aprindine could bind to the receptor via activation process through channel pore, resulting in decrease of Na⁺ current, and egress from the receptor through the lipid bilayer. These effects might be attenuated under acidic condition due to changes in intracellular ratio of charged to neutralized form of drug molecule. Send offprint requests to: R. Sato at the above address     

Proliferative activity in primary ovarian carcinoid tumors.

The proliferative potential of six primary ovarian carcinoids with different clinical outcome and histogenetic origin was examined immunohistochemically. The results showed that two cases with extremely high level of proliferative activity were associated with metastatic spread. In the remaining tumors, the examined factor was found to be at low level comparable with excellent prognosis of typical carcinoids in other locations. The preliminary results showed a possibility of a prognosis prediction according to typing of the ovarian carcinoids into two categories, i.e., tumors of low and intermediate malignancy. Topoisomerase II-alpha and Ki-67 are suitable markers giving valuable information about this phenomenon.     

Adsorption of microorganism components by lixelle beads

We reported previously that Lixelle, which was used for beta-2 microglobulin (BMG) adsorption columns, could adsorb not only BMG but also inflammatory cytokines. We then were interested in the application of Lixelle to patients with systemic inflammatory response syndrome (SIRS) and tried to find out its ability to adsorb microorganism components in vitro using lipopolysaccharide (LPS) (E. coli: B8), endotoxin (ET) containing water, and peptidoglycan (PG: Micrococcus luteus). The initial concentrations of each solution were LPS (ET: 29,135 EU/L), contaminated water (ET: 3,523 EU/L), and PG (67.1 ng/ml) and 2.5 ml of each of the stock solutions and adjusted diluted solutions contained 0.5 ml of Lixelle beads. After shaking at 37 degrees C for 2 h, ET in the solutions was determined by the ET specific-limulus amebocyte lysate (ES-LAL) method and PG by the silkworm larbae plasma (SLP) method. The results revealed that even when ET concentrations in LPS and contaminated water were high, the samples containing Lixelle beads showed significant decreases. There was some adsorption of PG but no significant differences. Thus, Lixelle beads can adsorb not only BMG but also microorganism components such as ET and PG. These findings, together with the ability to adsorb inflammatory cytokines by Lixelle, show the possibility of application for the treatment of infectious SIRS.     

Overexpression of the aldo-keto reductase family protein AKR1B10 is highly correlated with smokers' non-small cell lung carcinomas.

PURPOSE: Squamous cell carcinoma (SCC) and adenocarcinoma of the lung are currently subject to similar treatment regimens despite distinct differences in histology and epidemiology. The aim of this study is to identify a molecular target with diagnostic and therapeutic values for SCC. EXPERIMENTAL DESIGN: Genes specifically up-regulated in SCC were explored through microarray analysis of 5 SCCs, 5 adenocarcinomas, 10 small cell lung carcinomas, 27 normal tissues, and 40 cancer cell lines. Clinical usefulness of these genes was subsequently examined mainly by immunohistochemical analysis. RESULTS: Seven genes, including aldo-keto reductase family 1, member B10 (AKR1B10), were identified as SCC-specific genes. AKR1B10 was further examined by immunohistochemical analysis of 101 non-small cell lung carcinomas (NSCLC) and its overexpression was observed in 27 of 32 (84.4%) SCCs and 19 of 65 (29.2%) adenocarcinomas. Multiple regression analysis showed that smoking was an independent variable responsible for AKR1B10 overexpression in NSCLCs (P < 0.01) and adenocarcinomas (P < 0.01). AKR1B10 staining was occasionally observed even in squamous metaplasia, a precancerous lesion of SCC. CONCLUSION: AKR1B10 was overexpressed in most cases with SCC, which is closely associated with smoking, and many adenocarcinoma cases of smokers. These results suggest that AKR1B10 is a potential diagnostic marker specific to smokers' NSCLCs and might be involved in tobacco-related carcinogenesis.     

Phenolic glycosides from berries of Pimenta dioica

Four new phenolic glycosides, (2-hydroxy-3-methoxy-5-allyl)phenyl beta- d-(6-O-E-sinapoyl)glucopyranoside (1), (1' R,5' R)-5-(5-carboxymethyl-2-oxocyclopentyl)-3 Z-pentenyl beta-D-(6-O-galloyl)glucopyranoside (2), (S)-alpha-terpinyl [alpha-L-(2-O-galloyl)arabinofuranosyl]-(1-->6)-beta-D-glucopyranoside (3), and (R)-alpha-terpinyl [alpha-L-(2-O-galloyl)arabinofuranosyl]-(1-->6)-beta-D-glucopyranoside (4), were isolated from the berries of Pimenta dioica together with eight known flavonoids. The structures of 1-4 were elucidated on the basis of MS and NMR data and enzymatic hydrolysis. All four glycosides showed radical-scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals.     

Ureteral catheter placement for prevention of ureteral injury during laparoscopic hysterectomy

AIM: Ureteral injury is among the most devastating complications of gynecologic surgery. Estimated incidence of ureteral injury during laparoscopic hysterectomy is 2.6-35 times (0.2-6.0%) that in abdominal hysterectomy. We investigated preoperative ureteral catheter (UC) placement as a way to prevent ureteral injury in laparoscopic hysterectomy. METHODS: Clinical records of 94 women who underwent laparoscopic hysterectomy between February 2006 and January 2007 in Yazaki Hospital, Kanagawa, Japan, were reviewed retrospectively. Thirty-four patients between February and June 2006 underwent the surgery without ureteral catheterization and 60 patients between July 2006 and January 2007 underwent surgery with ureteral catheterization. Clinical outcomes were statistically compared between the two groups. RESULTS: The average time required for catheter insertion was 9.35 min. The ureter in which the catheter was placed was visualized clearly. In one patient, whose left ureter was deviated by a massive myoma, catheter insertion was not possible. No complications arose from catheter placement except for minor complaints including low back pain, urinary discomfort, and transient hematuria. While one injury occurred in a patient without ureteral catheterization (1/34), no ureteral injury occurred in any patient with ureteral catheterization (0/60). Operative time, total blood loss, and hospital stay were not significantly different between the two groups. CONCLUSIONS: UC placement is simple, helping to prevent ureteral injury during laparoscopic hysterectomy and enhancing safety of this procedure.     

Developmentally regulated release of intraretinal neurotrophic factors in vitro

The effects of conditioned media either from aggregates or from explants of embryonic chick retinae and of recombinant neurotrophins were tested upon the survival in vitro of ganglion cells in dissociated cell cultures from the retina of newborn rats. Ganglion cells were identified by the detection of retrogradely transported horseradish peroxidase injected bilaterally into the superior colliculus. Conditioned media increased significantly the survival of ganglion cells after 2 days in culture, at a wide range of plating densities, and had no effect upon adhesion of rat retinal cells. Media conditioned by cell ensembles from chick retinae from embryonic day 8 (E8) to E16 had neurotrophic effects. Release of neurotrophic activity peaked at E10-E12, irrespective of the numbers of cells or total concentration of protein in the conditioned media. The active molecules were non-dialyzable and were released either in the presence or in the absence of fetal calf serum. The neurotrophic activity was abolished by trypsinization, and recovered by salting-out with 25–75% ammonium sulfate. NT-4, BDNF and, to a lesser extent, NT-3, increased the survival of ganglion cells in our assay, while NGF had no effect. The data show that chick retinal cells release soluble trophic proteins according to a developmentally regulated pattern. These neurotrophic factors may be involved in local competitive interactions that help control naturally occurring neuron death among ganglion cells of the vertebrate retina.