In vitro and in vivo evaluation of seven 50 mg and 100 mg nitrofurantoin tablets

Four 50 mg and three 100 mg marketed nitrofurantoin tablets were studied in 14 healthy male subjects. Urine was collected 1, 2, 3, 4, 6, 8, 12, and 23 h after each dose, and nitrofurantoin was assayed by HPLC. The in vitro dissolution of the tablets was determined using USP Apparatus 1 and 2, with 0.1 N hydrochloric acid and pH 7.2 buffer as the dissolution fluids. One of the 50 mg tablets was more rapidly and completely absorbed than the other six products. The incidence of side-effects for this product was as low or lower than the other products. It was determined that the use of the USP Apparatus 1, at 100 rev min-1, with sampling of the pH 7.2 fluid at 30 min, provided for the best overall relationship between the urinary excretion and in vitro dissolution.     

Non-invasive dynamic assessment of the elasticity of intracranial structures

The dynamics of the speed of transcranial propagation of ultrasonic impulses has been established in humans. The possibility of detecting the changes in mechanical state of intracranial structures by dynamic measurement of the time of transcranial propagation of ultrasonic impulses (TUPT) has been ascertained for 10 healthy persons and 11 patients with different neurological pathology. The cerebrospinal fluid pressure (CSFP) was simultaneously monitored for 4 of the patients. Spontaneous fluctuations in TUPT and the changes caused by jugular veins, inferior vena cava compression, hyperventilation and temporary retention of breathing were detected for all investigated subjects. Spontaneous TUPT changes in the range of 0.25-0.85 x 10(-4) of initial mean value, which are related with cardiac and respiratory cycles, have been found in healthy persons while in patients they ranged from 0.07 x 10(-4) to 4.5 x 10(-4) of mean time. In healthy persons bilateral jugular vein compression leads to a decrease in TUPT by 1.8-3.0 x 10(-4), while the compression of inferior vena cava gives an increase in TUPT by 1.6-2.6 x 10(-4). Changes in TUPT during these maneuvers in neurological patients were found to be of different amplitude and direction. The rise of CSFP by 8.4-20.2 mmHg (mean 12.4 mmHg) was observed in 4 investigated patients during the compression of jugular veins or during the inferior vena cava compression, while the changes in TUPT obtained during each of such maneuvers were of opposite character. The results suggest that dynamic evaluation of acoustic impedance of intracranial system reflects the changes in the mechanical state of brain parenchyma.     

Atypical hemolytic uremic syndrome in human immunodeficiency virus-1-infected children

We describe the clinical and pathological findings of the hemolytic uremic syndrome (HUS) in two children with human immunodeficiency virus (HIV) infection. Both patients presented with microangiopathic hemolytic anemia, thrombocytopenia, and subsequently developed renal failure. The diagnosis of HUS was confirmed by renal histopathology in both patients. None of these children presented with bloody diarrhea, evidence of circulating antibody response to Escherichia coli O157 lipopolysaccharide, or other known risk factors for HUS, except for the presence of HIV infection. Each patient was treated with intravenous plasma infusion and renal replacement therapy. Their clinical course was characterized by non-oliguria and lack of significant hypertension throughout the acute phase of the disease. Despite these favorable clinical parameters, both patients developed end-stage renal failure. The etiology of this atypical HUS characterized by poor renal survival remains unknown and the role of HIV infection in its pathogenesis, although possible, is unclear. Received March 5, 1996; received in revised form and accepted October 15, 1996     

A study of the interrupted REM episode

Brief interruptions of REM sleep are considered to be part of the REM episode. The maximum allowable duration of such an interruption, which is used to define the end of the REM episode, is currently a matter of debate. Making measurements on individual REM cycles, inter-REM interval analysis was carried out to determine whether the generally adopted 15 minute empirical rule for this maximum needs to be extended to 25 minutes as suggested by several including Kobayashi et al. Our results show that there is no reason to alter the 15 minute rule and that measurements which do not take into account the time-of-night effect may be misleading. The proportion of interrupted REM episodes observed in our population of healthy adults is high. We have therefore also examined in some detail the phenomenology of the temporal evolution of the structure and content of the interrupted REM episodes. Both showed a definite change over the night: the interruptions in the earlier episodes tend to return the system to slow wave sleep while those in the later episodes tend to return it to wake. It is hypothesized that these interruptions reflect a measure of REM sleep pressure and its interaction with both slow wave sleep and wake pressures.     

Fluorescence behaviour of human arterial tissue

Total fluorescence from arterial tissue is influenced by three factors: the absorption coefficient of tissue at a specific excitation wavelength, the laser excitation power and the fluorescence coefficient which is related to chemical species in tissue. These various influences were demonstrated by the following experimental results in vitro: (1) the effect of increasing power on fluorescence intensity, (2) the total fluorescence intensity in normal aorta and plaque and (3) the effect of a chromophore such as β-carotene on total fluorescence intensity. The fluorescence intensity of normal artery is an incremental function of laser excitation power, and the fluorescence emission from normal artery compared to fluorescence emission from plaque is significantly different at the same excitation power. The total fluorescence of normal artery was measured to be twice as great as that of atheromatous plaque (relative mean ratio of 2.58±0.46 compared to unity,p<0.0002 at 488 nm; relative mean ratio of 2.57±0.51 compared to unity,p<0.0009 at 514 nm). The total fluorescence emission decreases with the increase of β-carotene content in arterial tissue (R=0.97). These emission differences, when intensified by an exogenous chromophore of β-carotene, may provide an improved guidance signal for diagnosis of plaque from normal artery during laser angioplasty procedures.     

Identification of human chorionic gonadotropin (HCG) secreting cells and other cell types using antibody to HCG and a new monoclonal antibody (mABlu-5) in cultures of human placental villi

Summary We have identified cells which secrete human chorionic gonadotropin (HCG) of cultures if first trimester placental villi. As a first step, we identified epithelial cells using a new monoclonal antibody. We then added HCG antibodies to the cultured cells. We found that syncytiotrophoblast (and not cytotrophoblast), Hofbauer cells and some mesenchymal cells stained with HCG antibodies.     

Preparation and evaluation of temperature sensitive liposomes containing adriamycin and cytarabine

Temperature sensitive liposomes (TSL) containing adriamycin (ADM) and cytarabine (Ara-C) were prepared. ADM and Ara-C were selected as model compounds of amphiphilic and hydrophilic drug, respectively. Encapsulation efficiency of ADM entrapped into TSL was about twice greater than that of Ara-C. It might be due to different polarity of the drugs. Lipid compositions of TSL had no effect on the encapsulation efficiency of drugs. Thermal behavior of TSL using a differential scanning calorimetry (DSC) was also investigated. Phase transition temperature (T_c) of TSL was dependent on the lipid compositions of TSL.ADM broadened thermogram of TSL but Ara-C did not. However, T_c of TSL was not changed by any drug. Release rate of drugs was highly dependent on temperature. The release profile of ADM was similar to that of Ara-C. The maximum release rate of drugs from TSL was occurred at the near T_c and observed at 39–41°C for DPPC (Dipalmitoylphosphatidylcholine) only, 52–54°C for DSPC (Distearoylphosphatidylcholine) only, 41–43°C for DPPC and DSPC (3∶1), and 43–45°C for DPPC and DSPC (1∶1), respectively. Effect of human serum albumin (HSA) on the release rate of ADM was investigated. HSA had no significant effect on the release of ADM below T_c. However, ADM release from TSL was increased at the near and above T_c. The HSA-induced leakage of drug may result from the interaction of liposomal constituents with HSA structure at the near T_c. From the fact that the release profiles of ADM from freshly prepared TSL and stored TSL for 1 week at 4°C was not changed, the TSL was considered to be stable for at least 1 week at 4°C. Based on these findings, TSL may be useful to deliver drugs to preheated target sites due to its thermal behaviors.     

Human lymphoblastoid cells produce extracellular matrix-degrading enzymes and induce endothelial cell proliferation, migration, morphogenesis, and angiogenesis

Human lymphoproliferative diseases can be hypothesized to invade locally and to metastatize via mechanisms similar to those developed by a variety of solid tumors, i.e., the secretion of extracellular matrix-degrading enzymes and stimulation of angiogenesis. To assess this hypothesis, Namalwa, Raji, and Daudi cell lines (Burkitt’s lymphoma), LIK and SB cell lines (B-cell lymphoblastic leukemia), CEM and Jurkat cell lines (T-cell lymphoblastic leukemia), and U266 cell line (multiple myeloma) were evaluated for their capacity to produce matrix metalloproteinase-2 and -9, and urokinase-type plasminogen activator. These cell lines were also assessed for their ability: (1) to produce the angiogenic basic fibroblast growth factor and vascular endothelial growth factor; (2) to induce an angiogenic phenotype in cultured endothelial cells, represented by cell proliferation, chemotaxis, and morphogensis; (3) to stimulate angiogenesis in different in vivo experimental models. All cell lines expressed the mRNA for one or both metalloproteinases. Namalwa, Raji, LIK, SB, and U266 cells secreted the active form of both metalloproteinases, while Daudi, CEM, and Jurkat cells produced metalloproteinase-2 but not -9. In contrast, urokinase-type plasminogen activator was secreted only by SB cells. While Raji, LIK, SB, CEM, and Jurkat cells secreted both basic fibroblast growth factor and vascular endothelial growth factor, Daudi and U266 cells produced only the former, and Namalwa cells only the latter. Accordingly, the conditioned medium of all cell lines stimulated cell proliferation and/or chemotaxis in cultured endothelial cells, with the exception of that of Namalwa cells which was ineffective. The conditioned medium of CEM and Jurkat cells induced morphogenesis in cultured endothelial cells grown on a reconstituted basement membrane (Matrigel). Lastly, Namalwa, Raji, LIK, SB, U266, CEM, and Jurkat cells induced angiogenesis and mononuclear cell recruitment in the murine Matrigel sponge model and in a chick embryo chorioallantoic membrane assay. The extent of angiogenesis in both models was strictly correlated with the density of the mononuclear cell infiltrate. The results indicate that human lymphoproliferative disease cells possess both local and remote invasive ability via the secretion of matrix-degrading enzymes and the induction of angiogenesis which is fostered by host inflammatory cells and by an intervening ensemble of angiogenic factors.     

人乳头瘤病毒多肽与人免疫球蛋白G融合表达

目的 将人乳头瘤病毒16型(Human papillomavirus type 16，HPV-16)的晚期表达蛋白E7上的抗原24肽(从第38位氨基酸到第61位氨基6病毒感染防治酸)与人免疫球蛋白G的重链恒定区融合表达，并以此融合蛋白作为抗原，可能为HPV-1提供免疫治疗方法。方法 利用PCR方法分别扩增HPV-16 E7(38-61)24肽的DNA片段和人免疫球蛋白G的重链恒定区DNA片段，并构建到pEV21a表达载体上，转化入E．coli中表达，利用SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质免疫印迹(Western-blotting)的方法对表达结果进行鉴定。结果 构建的表达载体HPV16E7e／hIgGHCCR-pET21a经酶切鉴定和测序显示序列正确；通过SDS-PAGE和Western-blotting的鉴定，重组融合蛋白Mr约40000，表达量可占菌体蛋白的20％左右。结论 成功构建HPV16-E7的抗原多肽片段和人免疫球蛋白G的重链恒定区的融合蛋白，并可在E．coli中高效表达。