Growth and differentiation of adult rat hepatocytes regulated by the interaction between parenchymal and non-parenchymal liver cells

We have devised a medium which supports the continuous growth of hepatocytes without losing their replicative potential and differentiation capacity for a longer period. The medium HCGM, contains four key substances in addition to foetal bovine serum. They are epidermal growth factor, nicotinamide, ascorbic acid 2-phosphate and dimethylsulphoxide. When a non-parenchymal cell fraction containing small hepatocytes and non-parenchymal cells was cultured in HCGM, small hepatocytes grew clonally and differentiated into cells expressing either mature hepatocyte marker proteins or biliary cell marker proteins. Thus, for the first time, we showed the presence of a small compartment of bipotent and highly replicative clonogenic hepatocytes in the rat adult liver. HCGM also supported the growth of stellate cells (Ito cells) which were in the original preparation, suggesting the important role of stellate cells for the successful cultivation of hepatocytes. Together, these results suggest that a microenvironment is produced as a result of cooperative interactions between hepatocytes and stellate cells: one which stimulates the growth and differentiation of clonogenic hepatocytes.     

Pharmacokinetic and Pharmacodynamic Properties of FK070 (KDI-792), A Novel Thromboxane Receptor Antagonist/Thromboxane Synthetase Inhibitor,After Single and Multiple Oral Administrations to Healthy Volunteers

FK070, a thromboxane A₂ (TXA₂) receptor antagonist/TXA₂ synthetase inhibitor, was given orally to healthy male volunteers in a single-and multiple-dose study. In the single-dose study (200, 300, 400 mg), the area under the plasma concentration—time curve (AUC) and the maximum plasma concentration (C_max) increased non-linearly with dose, while the mean elimination half-life (t¹_β) was essentially unchanged (3.9-7.3 h). Recovery of the unchanged drug in the urine was 12–25%. C_max and AUC as determined with 200 mg of drug after a meal decreased by about 60 and 30%, respectively. Ex-vivo platelet aggregation in the plasma by a TXA₂ analogue, U46619, was almost completely inhibited within 1 h, after all doses of drug, with a significant dose-dependent inhibition maintained for 8 h or more, which was much longer than was expected from drug plasma concentration. The aggregation by adenosine diphosphate (ADP) was inhibited to a lesser extent. FK070 also inhibited TXA₂ synthetase as evidenced by decreased production of TXB₂ and reciprocally increased production of 6-keto-prostaglandin F_1α in the serum during ex-vivo whole blood coagulation. These effects peaked 1 h after drug and lasted until 4 h with the higher doses. In the multiple-dose study (300 mg, twice a day, after meals for 6.5 days), drug concentrations in the plasma were well fitted to a three-compartment open model with first-order absorption. FK070 afforded extensive inhibition of platelet aggregation by U46619 throughout the administration period, with a significant inhibition lasting as long as 48 h after conclusion of administration. No clearly drug-related changes were found in routine laboratory tests, subjective and objective findings, or vital signs. FK070 was concluded to be well tolerated and to provide long-lasting blockade of TXA₂ receptors, and plasma concentration-dependent inhibition of TXA₂ synthetase in the platelets.     

A Case of In-Stent Neoatherosclerosis 10 Years after Carotid Artery Stent Implantation: Observation with Optical Coherence Tomography and Plaque Histological Findings

We report a patient''s case of slow progressive in-stent restenosis 10 years after bare-metal stent implantation to his carotid artery. We treated the patient with an additional stent placement under a distal filter protection device. Optical coherence tomographic assessment and plaque histology during the carotid artery stenting (CAS) revealed atheromatous change at in-stent neointima, which contained lipid-rich plaque and calcification deposits. These findings suggest that in-stent neoatherosclerosis may play an important role in the pathogenesis of very late stent restenosis after CAS.     

AQP3、7、9在人下颌下腺的免疫组化定位

采用免疫组化ABC法检测源自10例颈淋巴清扫患者的正常下颌下腺组织AQP3、7、9蛋白的表达。AQP3在浆液性腺泡和黏液性腺泡细胞均有表达,分布于腺泡细胞的基底膜和侧膜,在浆液性腺泡细胞中的表达明显高于黏液性腺泡细胞,导管系统未见AQP3表达;AQP7在极少数肌上皮细胞有表达;AQP9在部分浆液性腺泡腔、混合性腺泡腔以及部分闰管管腔和浆液性半月体上有表达。     

Comparative study of the stability of the folding intermediates of the calcium-binding lysozymes

Unfolding profiles of two calcium-binding lysozymes, equine milk lysozyme and pigeon egg-white lysozyme, were obtained by circular dichroism and proton NMR measurements. Equine lysozyme unfolds through a stable molten globule intermediate. The molten globule of equine lysozyme was characterized as more ordered than that of bovine α-lactalbumin. On the other hand, pigeon lysozyme unfolds by a two-state mechanism and the intermediate could not be observed in guanidine or thermal unfolding, the same as with conventional non-calcium-binding lysozymes. Thus, from the point of view of the unfolding profile, equine lysozyme belongs to the group of α-lactalbumin, but pigeon lysozyme belongs to the conventional lysozyme group.     

QUALITATIVE DIFFERENCES BETWEEN PROTEIN IN BLOOD SERUM AND IN CEREBROSPINAL FLUID OF PSYCHOTICS —THE POLAROGRAPHIC AND ELECTROPHORETIC STUDIES—

SUMMARY By the use of the polarographic method and paper electrophoresis, the proteins in the serum and in the CSF of schizophrenic, epileptic, and general paretic groups were examined. The examinations for the serum were carried out on 157 cases and for the CSF on 139 cases with the use of PLG; for the serum on 157 cases and for CSF on 105 cases with the use of PEP and a normal group were used as the controls. A comparison of the results from the serum and the CSF according to each symptom type group, the qualitative difference of the proteins is discussed.

• 1) In the normal group the difference between the quantity of the proteins in the serum and in the CSF is great with marked qualitative difference. The PLG findings show that the SH group activity of the protein surface is higher in the CSF than in the serum, while those of all three types are higher in the serum. The PEP findings show that the v-fraction is contained only in the CSF and that there is a difference between the fraction patterns in terms of the percentage of the protein and glycoprotein.
• 2) As to the PLG findings, the schizophrenic group as a whole showed protein surface SH group activity in the CSF and mucoprotein SH group activity in the serum higher than the normal group. Consequently, there was a disparity between the CSF/serum ratio of each and that of the normal group. The PEP findings showed an increase of the u-fraction of the CSF above the normal and in most cases a decreases in Al and α₂-Gl in the glycoproteins of both the fluids. As to the CSF/serum ratio, that of the γ- and β-Gl is lower and of α₁-Gl higher than the normal. As to the glycoprotein, Al and α₂-Gl are smaller and α₁-, β-, and γ-Gl greater than the normal. The differences according to the case development and symptom types were discussed.
• 3) In the intermittent period between epileptic seizures, the protein surface SH group activity is higher in the CSF and lower in the serum than in the normal, while the mucoprotein SH group activity is lower in the CSF and higher in the serum than in the normal; with a resulting variation in the CSF/serum ratio. As regards the PEP findings, many cases showed an v-fraction and α₂-Gl increase in the CSF, and an α₁-Gl increase and Al, and α₂-Gl decreases of the glycoproteins. In comparison with the normal group CSF/serum ratio, Al, α₁ and α₂-Gl showed a higher ratio and γ-Gl, a lower ratio, whereas in glycoprotein α₁-, and γ-Gl were higher and α₂- and β₂-Gl lower.
• 4) The general paresis groups examined were chronic cases. In them, both the serum and the CSF showed an increase in the proteins. In spite of the PLG finding that the protein SH group activity in each is higher than the normal, the CSF/serum ratio is nearly the same as the normal. The PEP findings showed many cases of v-fraction, γ-, and α₂-Gl increases in the CSF, and α₁-Gl increase in the serum. As to the glycoprotein, many cases showed α₁-, α₂- and γ-Gl increases and Al, α₂-, and α₂-Gl decreases in the CSF, and a β-Gl increase in the serum. Al and γ-Gl were high in the CSF/serum ratio, and α₁, and β-Gl were low; α₁- and γ-Gl of the glycoprotein, were high, and Al, α₁- and β-Gl low.
• 5) In general, the CSF showed greater changes than the serum in spite of the fact that the former is smaller in quantity than the latter. A discussion was attempted concerning the qualitative difference between the serum and the CSF.

     

A new antimicrobial quinolone (AM-1155) analysed in hair as an index of drug exposure and as a time-marker

Abstract— Scalp hair samples were obtained at one-month intervals for up to four months after the administration from each of twelve healthy male volunteers participating in a phase I study of a new antimicrobial quinolone, AM-1155, (±)-1-cyclopropyl-6-fluoro-1, 4-dihydro-8-methoxy-7-(cis-3,5-dimethyl-1-piperazinyl)-4-oxo-3-quinoline carboxylic acid. After hair was sectioned into 1 cm lengths from the scalp end, corresponding portions from five pieces of hair were dissolved in 1 m NaOH and assessed for AM-1155 by HPLC. In all subjects who had taken a single dose (600 mg, n = 6) or repeated doses (300 mg twice daily for 6·5 days, n = 6), the drug was detected in hair. The hair portions containing the drug were shown in most subjects to move outwards month by month at the rate of about 1 cm month^?1. A single hair, which was obtained from each subject of the repeated-dose study 3 months after the completion of administration, was cut into 2·5-mm lengths from the scalp side and analysed for AM-1155. The drug was shown to be contained in 4 to 6 consecutive 2·5-mm lengths, showing that there was no large axial diffusion of the drug along the hair shaft even after 3 months. These findings indicate the utility of measuring this quinolone derivative in hair as an index of drug exposure and, furthermore, as a time marker for analysing other drugs in hair.     

Predicting optimal continuous positive airway pressure in Japanese patients with obstructive sleep apnoea syndrome

Background and objective: Several algorithms that predict the optimal CPAP have been developed for Caucasian patients with OSA syndrome, but these algorithms do not allow for racial differences in craniofacial anatomy. We investigated whether an equation that included data on craniofacial structure, physique and severity of OSA could more accurately predict the optimal CPAP for Japanese patients with OSA syndrome. Methods: In 170 Japanese patients with OSA syndrome, the optimal CPAP was determined by manual titration during polysomnography. An equation predicting the optimal pressure was derived from anthropometric, polysomnographic and cephalometric data. This equation was validated in another 110 Japanese patients with OSA syndrome. Results: Stepwise multiple regression analysis identified AHI, BMI, mean SaO₂ and a cephalometric parameter: the angle between a line from point B to the menton (Me) and a line from Me to the hyoid bone (H) (BMeH), as independent predictors of optimal CPAP. The following equation was constructed to predict the optimal CPAP: 27.78 + (0.041 × BMeH) + (0.141 × BMI) + (0.040 × AHI) ? (0.312 × mean SaO₂). This equation accounted for 47% of the variance in optimal pressure (R² = 0.47, P < 0.0001). The measured optimal pressure and the pressure calculated using this equation were very similar in the other 110 patients with OSA syndrome (9.5 ± 3.0 and 9.2 ± 2.1 cmH₂O, respectively). Conclusion: Optimal CPAP was more accurately predicted by combining a cephalometric parameter with BMI and polysomnographic data in Japanese patients with OSA, suggesting that craniofacial structure may be important in the pathogenesis of OSA syndrome among Asians.     

Immunohistochemical analysis of development of desmin-positive hepatic stellate cells in mouse liver

Development of desmin-positive hepatic stellate cells was studied in mice using double immunofluorescent techniques and in vitro cultures with special attention given to their cell lineages. Several studies recently reported on the presence of cells that are immunologically reactive with both antidesmin and anticytokeratin antibodies in young fetal rat livers, and suggested the possibility that these cells give rise to hepatocytes and hepatic stellate cells. At early stages of mouse liver development, stellate cells with desmin-positive filaments were scattered in the liver parenchyma. However, the stellate cells definitely differed from hepatoblasts and hepatocytes in terms of their morphology and expression of desmin and hepatoblast and hepatocyte-specific E-cadherin in the liver. Fetal hepatoblasts and hepatocytes did not react with antidesmin antibodies, nor did desmin-positive stellate cells express E-cadherin in vivo and in vitro. Thus it is likely that desmin-positive stellate cells and hepatoblasts belong to different cell lineages. In the fetal liver, the desmin-positive stellate cells surrounded blood vessels, and extended their processes to haematopoietic cells and megakaryocytes. Many, but not all, hepatoblasts and hepatocytes were observed to be associated with the stellate cells. At fetal stages, cellular processes positive for desmin in the stellate cells were also thick compared with those in the adult liver, in which desmin-positive stellate cells lay in Disse's space and were closely associated with all hepatocytes. These developmental changes in the geography of desmin-positive cells in the liver parenchyma and their morphology may be associated with their maturation and interactions with other cell types.