Regulation of phosphatidylinositol kinases by arachidonic acid in rat submandibular gland cells

Phosphoinositide kinases were characterized in membrane extracts of rat submandibular gland cells. Both phosphatidylinositol (PI) 4-kinase and phosphatidylinositol-4-phosphate (PI(4)P) 5-kinase phosphorylated endogenous substrates in reactions that were linear for up to 5 min, were activated by Mg²⁺ and showed maximal activity around neutral pH. PI 4-kinase was stimulated by Triton X-100 at an optimal concentration of 0.22%, but the detergent had an inhibitory effect on PI(4)P 5-kinase. Arachidonic acid (AA), at concentrations greater than 100 M, inhibited the activity of both enzymes in a dose-dependent manner. The inhibitory effect was replicated by other unsaturated fatty acids, but not by a saturated fatty acid of the sn-20 series. The nature of AA inhibition of the kinases was examined in enzyme kinetic studies with exogenous phosphoinositide and adenosine 5-triphosphate (ATP) substrates. Lineweaver-Burk plots of PI 4-kinase activity showed that AA had no effect on the apparent K _m for either PI or ATP, but that the fatty acid significantly reduced V _max (PI) from 331 to 177 pmol.mg^–1.min^–1 and V _max (ATP) from 173 to 59 pmol.mg^–1.min^–1. This inhibitory action was consistent for PI(4)P 5-kinase kinetics, where again, AA did not alter apparent K _m values, but lowered V _max for both PI(4)P and ATP by around 50%. Since the combination of a reduced V _max and an unchanged K _m value indicates noncompetitive enzyme inhibition, it is proposed that AA regulates phosphoinositide cycle activity in submandibular gland cells by acting as a noncompetitive inhibitor of PI 4-kinase and PI(4)P 5-kinase.     

Neuromuscular function and mechanical efficiency of human leg extensor muscles during jumping exercises

The influence of prestretch amplitude on the mechanical efficiency was examined with 5 subjects, who performed 5 different series of vertical jumps, each of which differed with respect to the mechanics of the knee joint action during the prestretch (eccentric) phase of the contact on the floor. Electromyographic activity was recorded from the major extensor muscles during the entire work period of 1 min per series. In addition, expired air was collected during the test and recovery for determination of energy expenditure. Mechanical work was calculated from the vertical displacement of the body during the jumps. The results indicated that high net efficiency of 38.7% was observed in condition where amplitude of knee bending in eccentric phase was small. In large range motion the corresponding net efficiency was 30.1%. In jumps where no prestretching of extensor muscles ocurred the net efficiency was 19.7%. The high efficiency of small amplitude jumps was characterized by low myoelectrical activity of the leg extensor muscles during the positive (concentric) work phase. In addition, the small amplitude jumps had shorter transition time in the stretch-shortening cycle, high average eccentric force and high stretching speed. Therefore the results suggest that the restitution of elastic energy, which was also related to the length change and stiffness of the muscles during stretch, plays an important role in regulating the mechanical efficiency of work.     

PENA方法的建立及与ELISA IgM检测CMV的比较

目的　介绍一种敏感、稳定、快速、简便的实验室检测CMV的方法 ,同时探讨该种新方法与ELISA检测CMV方法的优、缺点。方法　对 5 5 2例病人应用间接荧光免疫法测定细胞核中的特异病毒早期抗体 (PENA)和ELISA法测定IgM抗体。结果　PENA方法 :强阳性 88例 ,阳性率 15 4 9% ,弱阳性 2 73例 ,阳性率 5 9 4 6 % ;ELISA -IgM方法 :阳性 34例 ,阳性率6 16 %。结论　PENA方法操作简便 ,与ELISA方法相比较 ,可对CMV感染进行早期测定及诊断 ,并可区分既往感染和即时感染 ,具有敏感性和稳定性 ,是测定小儿CMV感染的一种较好的方法     

2450MHz介质复介电常数的测量

目的测量2450MHz频率下蒸馏水和不同浓度NaCl溶液的复介电常数,用以判定系统的可靠性和稳定性.方法根据微扰法得到复介电常数的测量公式,然后通过实验对已知参数的蒸馏水和NaCl溶液进行测量,并计算其复介电常数和电导率,验证计算结果的可靠性和稳定性.结果对计算结果进行统计分析和误差分析,其均值、标准差和变异系数表明,最终结果可与M.I.T标准较好地吻合.结论这种系统可较好地测量2450MHz频率下高损耗介质的复介电常数,测量系统的主要误差是由介质体积的误差引起的,因此精确测量介质体积是测量生物组织等高损耗介质复介电常数的关键.     

QUANTIFICATION OF THE REM SLEEP CYCLE AS A RHYTHM

The goal of this study was to develop an objective quantitative method for representing the temporal organization of sleep in terms of the period and rhythmicity of REM sleep occurrences. Data on normative human sleep, already scored for stage REM and not stage REM, were subjected to a “binary autocorrelation.” The mean period over 92 nights of sleep for 10 Ss was 101.5 min and quite stable. Data is also presented on variability of the rhythm in terms of an “index of rhythmicity.” Measures of temporal organization may prove to be as significant for sleep research as amount of the various sleep stages.     

Determination of glucokinase and hexokinase activity in liver extracts previously purified on Molselect G-50 dextran gel

Hexokinase and glucokinase activity in the supernatant of a rabbit liver homogenate obtained at 18,000g was determined by a spectrophotometric method. Preliminary purification to remove low-molecular-weight components by gel filtration on Molselect G-50 dextran was shown to prevent reduction of NADP unconnected with the hexokinase reaction.Presented by Academician of the Academy of Medical Sciences of the USSR V. S. Il'in.Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 7, pp. 891–892, July, 1976.     

A method for assessing muscle fatigue during sprint exercise in humans using a friction-loaded cycle ergometer

This study investigated the mechanical changes induced by muscle fatigue caused by repeated sprints and determined whether a friction-loaded cycle ergometer has any advantages for assessing muscle fatigue. Nine subjects performed 15 sprints, each of 5 s with a 25-s rest, on a friction-loaded cycle ergometer. The averaged force, power and velocity of each push-off were calculated. Maximal power decreased by 17.9%, with a concomittent slowing of muscle contraction, but without any change in the maximal force. These results demonstrated that repeated sprints slow down muscle contraction, leading to a fall in maximal power without any loss of force. This would suggest that fast twitch fibres are selectively fatigued by repeated sprints. However, the ergometer used in the present study made it difficult to evaluate the relative influences of contraction velocity and sprinting time. This was certainly the most important limitation. On the other hand, it showed the advantage of measuring instantaneous power and total work dissipated in the environment simultaneously. It also permitted a force-velocity relationship to be obtained from a single sprint and this relationship is known to be closely related to the muscle fibre composition. Accepted: 5 March 1998     

Production of genetically modified cells expressing specific transgenes by retroviral vectors for gene therapy

Summary Techniques and protocols are described for the generation of genetically modified cells that can be used for gene therapy. Primary fibroblast cultures are established from skin biopsies, maintained in culture, frozen for long-term storage, and retrieved when necessary. Retroviral packaging cell lines are generated by transfection of DNA into retroviral packaging cells by calcium-phosphate precipitation method or by lipofection method. To generate cell lines expressing high titer virus, individual colonies of cells are cloned and the virus titer is determined. Virus collected from packaging cells expressing high titer virus is then used to infect primary fibroblasts. To obtain fibroblast cell lines expressing high amounts of transgenes, individual cells can be cloned to generate clonal cell lines. Although the methods described here are for fibroblasts, the same methods or modification of the methods can be used for other cell types.     

Superior cycle control with a contraceptive vaginal ring compared with an oral contraceptive containing 30 microg ethinylestradiol and 150 microg levonorgestrel: a randomized trial

BACKGROUND: This trial was conducted to compare cycle control with vaginal ring a combined contraceptive vaginal ring, and a combined oral contraceptive (COC) delivering 30 mug ethinylestradiol (EE) and 150 mug levonorgestrel. METHODS: This open-label, randomized, multi-centre, Phase III study involved adult women from 11 countries. Subjects were treated with either vaginal ring or a COC for 13 cycles (12 months). RESULTS: A total of 1030 subjects (vaginal ring, n=512; COC, n=518) comprised the intention-to-treat (ITT) population. The percentage of women in the ITT population who completed the trial was 70.9% for vaginal ring and 71.2% for the COC group. The incidence of breakthrough bleeding and spotting over cycles 2-13, the primary efficacy parameter, was lower with vaginal ring (range 2.0-6.4%) than the COC (range 3.5-12.6%), and for cycles 2 and 9 the lower incidence with vaginal ring was confirmed as statistically significant (P=0.003 and P=0.002 respectively). The incidence of intended bleeding was significantly higher over all cycles with vaginal ring (58.8-72.8%) than with the COC (43.4-57.9%). CONCLUSIONS: Cycle control with vaginal ring was excellent and superior to that of a COC containing 30 mug EE.