Evidence for motility and pinocytosis in ramified microglia in tissue culture

Ramified microglial cells were investigated in primary cultures of dissociated cerebral cortical tissue from rats. The identification of these cells was confirmed through immunohistochemical staining with 7 monoclonal antibodies selective for microglia. While there was significant variation in staining intensity with different antibodies, all stained the identified ramified cells; the antibodies OX-42 and ED1 yielded the most intense immunoreactivity. Based on distinctive morphological features, the microglia could be identified in living cultures where they were monitored using time-lapse video recording. This technique revealed extremely dynamic features of cellular plasticity and motility. Ramified microglia exhibited constant and rapid alterations in the size and shape of their cell body with an associated extension and retraction of processes; concomitantly, the cells moved about in a circumscribed area. These features of plasticity and motility were unique to this cell type, and correlated with OX-42 immunostaining. The microglia also possessed a differentially high level of pinocytotic activity; this too was correlated with OX-42 staining. From the nature of their morphological plasticity and motility, high pinocytosis, and cellular distribution, it is hypothesized that the ramified microglia specifically function as a system of fluid cleansing in normal brain tissue.     

应用人精子头-尾膜完整性试验对生育力组及不育组精液的评价

目的应用人精子头-尾膜完整性试验,比较生育力组与不育组男性精液4种精子膜完整性类型的差异.方法精液标本分为生育力组(n=32)和不育组(n=50),采用"低渗肿胀-伊红拒染"结合试验,检测人精子头-尾膜完整性.结果头膜尾膜均损伤的Ⅰ型精子和头膜损伤-尾膜完整的Ⅲ型精子,生育组和不育组存在明显差异(P<0.01);头膜完整-尾膜损伤的Ⅱ型精子,生育组和不育组无明显差异(P>0.05);头膜-尾膜均完整的Ⅳ型精子,生育组明显高于不育组(P<0.01);Ⅳ型精子率与生育组精子活动率(n=32, r=0.82,P<0.01)和不育组精子活动率(n=50,r=0.80,P<0.01)均呈显著相关性.结论生育组具有较多头膜-尾膜均完整的精子."低渗肿胀-伊红拒染"结合试验能够清晰显示包括过渡型膜损伤的4种膜完整性类型的精子,精子头-尾膜完整性可以作为评价精子生理学的一项检测指标.     

Oral administration of methylergometrine shows a late and unpredictable effect on the non-pregnant human menstruating uterus

Objective: To study the pharmacodynamic and pharmacokinetic properties of oral and intravenous methylergometrine upon uterine motility during menstruation. Study-design: Intra-uterine pressure was measured in six volunteers with a fluid-filled sponge-tipped catheter during menstruation. Methylergometrine was given orally (0.5 mg) or intravenously (0.2 mg) in a cross-over design. Results: After intravenous administration, a fast increase of the frequency of uterine contractions and basal tone occurred with a decrease of amplitude, lasting at least 30 min. Oral administration had a late and less marked effect on uterine motility. An intravenous dose administered 24 h after an oral dose had no effect on uterine motility. Pharmacokinetic data, such as the maximum plasma concentration (C_max), the time at which C_max is reached (t_max) and the half-life of absorption (t_1/2abs) also demonstrated large individual variations after oral administration. Conclusion: Oral administration of methylergometrine had an unpredictable and late effect on uterine motility on the menstruating uterus, probably due to an unpredictable bioavailability, in contrast with the fast and predictable effect after intravenous administration.     

Comparative study of two computerized semen motility analyzers

Semen analysis is one of the primary tests carried out to investigate the infertile male. Subjective evaluation of semen is often prone to observer bias and error. To eliminate this, a number of computerized semen analyzers have recently been introduced into the market and we have evaluated two of the more popular models, the Cell Soft Semen Analyzer and the Hamilton Thorn Motility Analyzer (HTM 2000). The Cell Soft identifies sperm on the basis of user defined values for cell size and luminosity whereas the Hamilton Thorn identifies sperm by motility, and then applies the computer-calculated average size and luminosity of all moving objects to non moving sperm cells. Semen samples from 25 normal donors and 25 subfertile patients were analyzed using these two models of computerized semen analyzers, and also by an experienced technician using both the Makler chamber and the hemocytometer. The results obtained from the two automated analyzers were compared with those obtained by subjective evaluation. Variation in sperm count and motility were analyzed according to the sperm density. Four groups, less than 30 million/ml with debris, less than 30 million/ml, 30-50 million/ml, and greater than 50 million/ml were studied. The majority of patients fit into the first two groups. We observed that the HTM 2000 is superior to the Cell Soft in evaluating sperm count within the patient population group. For our donor population with an average sperm count of greater than 85 million/ml both systems provide extremely accurate counts.(ABSTRACT TRUNCATED AT 250 WORDS)     

Neuronal adrenergic and muscular cholinergic contractile hypersensitivity in canine jejunum after extrinsic denervation

Extrinsic denervation may be responsible for motor dysfunction after small bowel transplantation. The aim of this study was to examine the role of extrinsic innervation of canine jejunum on contractile activity. An in vitro dose response of cholinergic and adrenergic agonists was evaluated in canine jejunal strips of circular muscle at 0, 2, and 8 weeks in a control group and after jejunoileal extrinsic denervation (EX DEN). Neurons in circular muscle were quantitated by means of immunohistochemical techniques. Adrenergic and cholinergic responses did not differ at any time in the control group. However, at 2 and 8 weeks, extrinsic denervation caused an increased sensitivity to the procontractile effects of the cholinergic agonist bethanechol at the level of the smooth muscle cells, and increased sensitivity to the inhibitory effects of the adrenergic agent norepinephrine mediated at the level of the enteric nervous system. Immunohistochemical analysis showed a reduction in all neurons and a complete lack of adrenergic fibers in the EX DEN group after 2 and 8 weeks. Extrinsic denervation induces enteric neuronal cholinergic and adrenergic smooth muscle hypersensitivity in canine jejunal circular muscle. Presented in part at the annual meeting of the American Gastroenterological Association, Orlando, Florida, May 18, 1999 (poster presentation), and published as an abstract in Gastroenterology 116:A1075, 1999. Supported by United States Public Health Service grant DK39337 from the National Institutes of Health (M.G.S.); the Swiss National Science Foundation; the Swiss Society of Gastroenterology and Hepatology; the Swiss Foundation for Medical and Biological Science; the Novartis Foundation; Astra Zeneca Pharmaceuticals, Switzerland; and the Department of Visceral and Transplantation Surgery, University of Bern, Switzerland.     

人T淋巴母细胞（Jurkat）移动性的研究

利用人Ｔ淋巴母细胞（Ｊｕｒｋａｔ）细胞系作为研究对象。企图探探索细胞外基质成份及肌醇磷脂细胞信息传递系统与细胞移动的关系。研究结果表明：当细胞被伏波醇酯（ＰＭＡ）处理过后，其粘连性与移动件对纤维连结蛋白（ＦＮ）底物的反应在已是高水平的基础上有更进一步提高的作用。与ＦＮ刊相反的是，未经ＰＭＡ处理处的Ｊｕｒｋａｔ细胞对层粘连蛋白（Ｌｍ）底物却无明显的粘连性及移动性增加的反应，尽管在ＰＭＡ处理过４小时内亦无任何的增强。但是，当ＰＭＡ作用于细胞２４一４８小时后则明显池增加其Ｌｍ底物的粘连性及移动性反应，表明了其明显的协同作用。分别用抗ＦＮ、抗Ｌｍ及蛋白激酶Ｃ抑制剂Ｓｔａｕｒｏｓｐｒｉｎｅ（ＳＰ）时均起到特异性的抑制作用。可见，协同作用的产生是通过蛋白激酶Ｃ激活后的－段时间内，使细胞表面Ｌｍ受体密度增加而对底物Ｌｍ反应增强所致。     

Entrainment of Intestinal Slow Waves with Electrical Stimulation Using Intraluminal Electrodes

The aim of this study was to investigate whether the intestinal stimulation would be feasible using a less invasive method: intraluminal electrodes. The study was performed in nine healthy hound dogs (15–26 kg). Four pairs of electrodes were implanted on the serosa of the jejunum at an interval of 5 cm with the most proximal pair 35 cm beyond the pylorus. An intestinal fistula was made 20 cm beyond the pylorus. Simultaneous recordings of intestinal myoelectrical activity were made for 2 h in the fasting state from both intraluminal and serosal electrodes. Various pacing parameters were tested. The frequency of the intestinal slow wave recorded from the intraluminal electrodes was identical to that from the serosal electrodes , p < 0.001), and so was the percentage of normal 17–22 cycles/min waves (95.8±33.9% vs 98.16±1.33%, r=0.96, p<0.01).p < 0.01). A complete entrainment of the intestinal slow wave was achieved in every dog with electrical stimulation using intraluminal ring electrodes. The effective pacing parameters were pulse width of 70 ms, amplitude of 4 mA and frequency of 1.1 IF (intrinsic frequency). The time required for the entrainment of the intestinal slow wave with intraluminal pacing was 25.0±2.1s. The maximum driven frequency was found to be 1.43±0.01 IF. The results reveal that intraluminal pacing is an effective and efficient method for the entrainment of intestinal slow waves. It may become a potential approach for the treatment of intestinal motor disorders associated with myoelectrical abnormalities. © 2000 Biomedical Engineering Society. PAC00: 8754Dt, 8719Ff, 8717Nn     

Detection and analysis of gastrointestinal sounds in normal and small bowel obstructed rats

This study is aimed at detecting gastrointestinal sounds (GIS) and correlating their characteristics with gastrointestinal (GI) conditions. The central hypotheses are that GIS generation depends on the motility patterns and the mechanical properties of the gut, and that changes in those result in measurable differences in GIS. An animal model which included both healthy rats and those with small bowel obstruction (SBO) was developed. The acoustic bursts, of GIS were detected by amplitude thresholding the signal envelope. Three methods of envelope estimation were proposed and evaluated. Envelope estimation using a Hilbert transform was found to produce the best results in the current application. The duration and dominant frequency of each detected GIS event was estimated and clear differences between healthy and diseased rats were discovered. In the control state, GIS events were found to consistently be of relatively short duration (3–65ms). Although the majority of events in the SBO state had similar short duration, infrequent longer events were also detected and appeared to be pathognomonic. Long duration events (>100 ms) occurred in each of seven obstructed, but in none of 14 non-obstructed, cases (p<0.001). It is concluded that GIS analysis may prove useful in the non-invasive, rapid, and accurate diagnosis of SBO.     

生长抑素对奥迪括约肌运动的作用机制

目的了解徨长抑素影响败类迪括约肌运动的神经通路成分。方法建立奥迪括约股测压慢性实验动物模型,研究Ｍ受体阻断剂、α受体阻断剂、β受体阻断剂、５－ＨＴ４受体阻断剂、一氧化氮合酶抑制剂和河豚毒素对生长抑素调节奥迪括约肌运动的影响。结果阿托品、二乙胺盐酸甲酯和河豚毒素阻断生长抑素对括约肌的兴奋作用;Ｌ－精氨酸甲酯阻断生长抑素对括约肌的抑制作用。结论生长抑素通过胆碱能和５－羟色胺神经元组成的兴奋性神经通路,     

Extracellular acidification decreases the basal motility of cultured mouse microglia via the rearrangement of the actin cytoskeleton

The present study was undertaken to examine the effect of extracellular pH (pH(0)) on the locomotor function of murine microglial cells in vitro. We have found that basal motility of microglia, as measured by a computer-assisted video assay, decreased in an acidic, but not in an alkaline environment. Extracellular acidification affected the architecture of F-actin cytoskeleton, inducing bundling of actin and the formation of stress fibers. The change in intracellular pH (pH(i)) resulting from the change in pH(0) seems to be a prerequisite for the motility decrease since other means to decrease pH(i), namely Na(+)-free solution (in the absence of HCO(-)(3)) and nigericin-containing solution, mimicked the extracellular acidification. In contrast to its pronounced effect on basal motility of microglial cells, the motility increase, as induced by the chemoattractant complement 5a (C5a), was not affected by the acidic environment. The relationship of pH(0) to the locomotor function was also studied in a long-term microchemotaxis assay where microglia migrated within a pH gradient. Intracellular acidification induced by lowering pH(0) to 6.0 or removal of Na(+) from the assay medium decreased basal microglial cell migration. The C5a-induced chemotactic migration was moderately decreased by the acidic environment. In conclusion, our results suggest that acidification of the microglial extracellular milieu leads to a decrease in pH(i) and thereby reduces the basal microglial motility and C5a-induced chemotaxis via a rearrangement of the cytoskeleton. We would therefore like to speculate that changes in pH(i) constitute an important control mechanism in regulating the locomotor function of microglia in culture and probably also in the intact tissue.