Effects of the perfusion flow rate on skeletal muscle energy metabolism and a possible role of second messengers in this process

The metabolic turnover in the isolated in vitro perfused and superfused rat skeletal muscle (musculus gracilis cranialis) was enhanced by increasing the medium flow rate under relaxed conditions. In a recent study we have measured the tissue concentrations of second messengers: cyclic adenosine 3'5'-monophosphate (cAMP), cyclic guanosine 3'5'-monophosphate (cGMP), and D-myo-inositol 1,4,5-trisphosphate (IP3) under similar experimental conditions to analyze their potential role in the described stimulation of metabolic rate by changes of perfusion flow rate. The tissue levels of the two second messengers' cAMP and cGMP were not significantly changed after increasing the perfusion flow rate and they probably have no transduction role in the induced alteration of skeletal muscle metabolism. However, the IP3 content was extremely reduced after increasing flow rate. This decrease in the tissue concentration of IP3 induced by increasing the flow rate indicates the possible role of IP3 in this signal transduction, leading to changes in the cellular metabolic pathways.     

Design of a sterile organ culture system for the ex vivo study of aortic heart valves

The biological response of valves to mechanical forces is not well understood. The aim of this study was to design a pulsatile system to enable the ex vivo study of aortic valves when subjected to various hemodynamic conditions. A bioreactor was designed to subject porcine aortic valves to physiological and pathophysiological pressure and flow conditions, while maintaining viability and sterility. Pressure and flow rate could be independently controlled to produce clinically relevant mechanical conditions. The oxygen transfer rate was characterized and sterile operation was achieved over 96 hours. The oxygenation capabilities ensure sufficient oxygen transport to valves, allowing operation for extended periods.     

Evaluation of Filters for Removal of Bacteriophages from Air

Glass wool, nonabsorbent cotton, fiberglass filter medium, and a commercial absolute filter were tested for effectiveness in removing aerosolized bacterial viruses under low flow rate (1 ft(3)/min) and high flow rate (10 to 25 ft(3)/min) air-flow conditions. Special equipment was designed for measurement of filter efficiencies under the two air-flow conditions. Under low air-flow rate test conditions, glass wool was only 98.543 to 99.83% efficient, whereas cotton (five layers), fiberglass medium (three layers), and the commercial absolute filter were at least 99.900, 99.999, and 99.999 efficient, respectively. Glass wool and cotton were not used under higher air-flow conditions because they were difficult to assemble in leak-tight filters. The commercial absolute filter and fiberglass medium (three layers) were at least 99.990 and 99.999% efficient, respectively, under the higher air flow conditions. A stainless-steel filter of simple design and fitted with three layers of fiberglass medium was found to be greater than 99.999% efficient in removing high concentrations (20,000 to 70,000 plaque-forming units per cubic foot) of aerosolized bacteriophages from air moving at a low flow rate (1 ft(3)/min). Use of this filter on pressure-vacuum tanks in the fermentation industry is suggested. Several other uses of such a filter are proposed.     

营养及水力条件影响光合细菌生物膜生长特性实验

对平板式生物膜反应器内,流量及底物浓度范围分别为37.8~1080ml/h、0.05~10g/L的不同生长条件下光合产氢细菌生物膜生长特性进行了实验研究,讨论了不同水力及营养条件对沼泽红假单胞菌生物膜表面覆盖率、膜厚、干重和密度的影响。实验结果表明,不同水力及营养条件对生物膜生长速率及结构具有重要影响。在相同的时间间隔内,在高流速条件下光合细菌菌落生长较快,但过高的液体流速会导致部分生物膜脱落;高流速条件易使生物膜形成薄而致密的结构。光合细菌生物膜在循环液底物浓度较高时生长较快,密度也最高;而贫营养条件可以促成结构疏松生物膜在固液界面的形成,这种生物膜结构有利于微生物在低底物浓度条件下底物在生物膜内的传输。     

Influence of flow on pulmonary vascular surface area inferred from blue dextran efflux data.

Blue dextran (BD), which binds to proteins on the pulmonary endothelial surface and to plasma albumin, was used in isolated perfused dog lung lobe experiments to address the question: do changes in perfusate flow rate cause changes in perfused vascular surface area? When BD was added to a protein-free perfusate under zone 3 conditions at a high flow rate (15.8 +/- 0.7 ml/s), it was adsorbed by the endothelial surface. Then by changing the perfusate entering the lobe to an albumin-containing perfusate, the BD was eluted from the perfused surface by competitive binding to the perfusate albumin. The amount of BD eluted was measured in three experiments. In experiment 1, elution of the BD by the perfusate albumin was initiated after a balloon had been inflated within the lobar arterial tree to occlude a portion of the lobar vascular bed containing BD. Then the balloon was deflated, permitting albumin perfusate to perfuse the previously occluded part of the lobe. In experiment 2, BD elution began at a flow rate of 3 +/- 0.1 ml/s under zone 3 conditions and continued after the high-flow zone 3 conditions were reestablished. In experiment 3, the BD elution began at a flow rate of 4.2 +/- 0.7 ml/s under zone 2 conditions and continued after the high-flow zone 3 conditions were reestablished. Balloon inflation reduced the amount of BD recovered by 43%, demonstrating that a decrease in perfused vascular surface area could decrease BD recovery.(ABSTRACT TRUNCATED AT 250 WORDS)     

Experimental optimization of a step aeration waste treatment process

Evolutionary operation (EVOP) was used to experimentally investigate the optimum steady state operating conditions for a step aeration activated sludge waste treatment process. A laboratory scale two tank step aeration activated sludge unit with fixed total volume, total influent flow rate, recycle flow rate, and sludge wasting rate was employed. The volume ratio and flow rate ratio which minimized effluent chemical oxygen demand were determined. The results indicate that EVOP is a useful technique for improving the performance of biological processes.     

Production of 6-APA using a recirculated packed bed batch reactor

Summary A recirculated packed bed batch reactor has been designed for the production of 6-aminopenicillanic acid. It was observed that the flow rate of penicillin G solution is a rate limiting step for its hydrolysis. Under the conditions used, the maximum rate of hydrolysis of penicillin G was observed at a flow rate of 3.0 L/min.     

一种制备新生隐球菌单细胞的方法

【目的】建立流式细胞仪分选新生隐球菌单细胞的方法,确定新生隐球菌单细胞恢复生长的条件和能力。【方法】利用Moflo XDP流式细胞分析分选仪,体外测定不同条件下新生隐球菌恢复生长的比率。【结果】建立了流式细胞仪新生隐球菌单细胞分选流程。确定流式细胞仪分选得到的新生隐球菌单细胞具有恢复生长的能力,恢复生长的能力受营养条件和菌株差异的影响。在营养丰富的条件下,新生隐球菌JEC21和H99单细胞恢复生长比率分别为74%和89%。在寡营养条件下,JEC21和H99单细胞恢复生长比率分别为37%和80%。JEC21生长比率随细胞数的增加而升高,细胞数为100个时,生长比率为55%;细胞数为1000个时,生长比率为97%。【结论】流式细胞仪分选得到新生隐球菌单细胞具有恢复生长的能力,生长能力受营养条件、菌株差异的影响。     

Detection of Bacillus anthracis spores and a model protein using PEMC sensors in a flow cell at 1 mL/min

Piezoelectric-excited millimeter-sized cantilever (PEMC) sensors of 4mm(2) sensing area were immobilized with antibody specific to Bacillus anthracis (anti-BA) spores or bovine serum albumin (anti-BSA). Detection of pathogen (Bacillus anthracis (BA) at 300 spores/mL) and BSA (1 mg/mL) were investigated under both stagnant and flow conditions. Two flow cell designs were evaluated by characterizing flow-induced resonant frequency shifts. One of the flow cells labeled SFC-2 (hold-up volume of 0.3 mL), showed small fluctuations (+/-20 Hz) around a common resonant frequency response of 217 Hz in the flow rate range of 1-17 mL/min. The total resonant frequency change obtained for the binding of 300 spores/mL in 1h was 90+/-5 Hz (n=2), and 162+/-10 Hz (n=2) under stagnant and flow conditions, respectively. Binding of antibodies, anti-BA and anti-BSA, were more rapid under flow than under stagnant conditions. The sensor was repeatedly exposed to BSA with an intermediate release step. The first and second responses to BSA were nearly identical. The total resonant frequency response to BSA was 388+/-10 (n=2) Hz under flow conditions. Kinetic analysis is carried out to quantify the effect of flow rate on antibody immobilization and the two types of detection experiments.     

Continued growth of Escherichia coli after stopping medium addition to a K+-limited chemostat culture

The steady-state bacterial dry wt of Escherichia coli, growing under K+-limited conditions in the chemostat, was inversely dependent on the growth rate. This phenomenon was more carefully investigated in medium-flow stop experiments. Growth did not stop immediately but continued for a time, initially at the same rate as before. The dry wt increased to a value corresponding to a steady-state growth rate near zero, independent of the initial specific growth rate. This was observed in both the wild-type strain and a mutant that lacked the high-affinity K+ uptake system. The wild-type strain maintained a low extracellular K+ concentration both in the chemostat under steady-state conditions and after stopping the medium flow. The mutant, on the other hand, maintained a much higher extracellular K+ concentration in the steady state, which decreased to much lower values after stopping the medium flow. From the increase in bacterial dry wt and the low external K+ concentration after stopping the medium flow it is concluded that the intracellular K+ is redistributed among the cells, including new cells. The growth yield on K+ was highest in the stationary growth phase of a batch culture and all steady-state cultures converged ultimately to this yield value after the medium flow had been stopped. It is proposed that the growth rate of E. coli under K+-limited conditions is determined by the intracellular K+ concentration.     

静水与流水下气泡幕对异齿裂腹鱼的阻拦效应

为了探究不同水流条件下不同形式的气泡幕对异齿裂腹鱼(Schizothorax oconnori)趋避行为的影响。在黑暗环境下测试了两种流速条件(静水与流水)、三种气量(15、30、45 L/min)及两种摆放角度(与水流方向呈90°和45°)的气泡幕对异齿裂腹鱼的阻拦效果,在静水和流水条件下各设置一组空白对照(气量为0 L/min)。结果显示: (1)在静水条件下, 工况2阻拦率最高(50%); 在流水条件下, 工况7阻拦率最高(50%)。(2)气量15 L/min时, 流水及90°摆放阻拦时间显著大于其他工况; 30 L/min时, 静水及90°摆放阻拦时间远大于其他工况。(3)在静水和流水中, 当异齿裂腹鱼尝试次数达到6次左右时对气泡幕表现出适应性, 并在48min前通过气泡幕; (4)在流水中各工况下气泡幕的影响距离显著大于静水(P<0.05), 即异齿裂腹鱼产生逃逸行为时距离气管的距离显著大于静水中。研究可得出结论: 推荐阻拦效果最佳的组合方式为流水条件下15 L/min 90°摆放(工况7)、静水条件下30 L/min 90°摆放(工况2); 气泡幕连续阻拦时间不宜超过48min。实验结果可为实际工程中气泡幕的布置提供参考。     

Inversion of sucrose in a continuous process with β-d-fructofuranosidase (invertase) immobilized on bead DEAHP-cellulose

Inversion of sucrose with β-d-fructofuranosidase (EC 3.2.1.26) immobilized by the ionic bond on bead DEAHP-cellulose has been studied under flow conditions. Under these conditions, the inversion of sucrose is affected by the concentration and flow rate of the substrate and by the reaction temperature. The effect of substrate concentration on the reaction was investigated in the range 19.5–64.2 wt %; the effect of flow rate was examined in the range 0.25–5.57 g solution per min, and the temperature range used was 25–50°C. It was found that the activities of immobilized β-d-fructofuranosidase in stirred and flow reactors were the same. The lower activities of β-d-fructofuranosidase in the case of concentrated solutions, and of immobilized β-d-fructofuranosidase compared with the native enzyme are attributed to more difficult diffusion through the beads of the ion exchanger, especially of the strongly viscous substrate. A long-term investigation of the enzyme activity over a period of three months demonstrated the stability of the β-d-fructofuranosidase immobilized by the ionic bond on bead DEAHP-cellulose; the half-life of the enzyme was 215 days. It was also found that the immobilization of the enzyme on a carrier was more effective under flow conditions, i.e. through an ion exchanger in the column, than under the equilibrium conditions of a stirred reactor.     

Flow patterns at the major T-junctions of the dog descending aorta

Using a novel technique developed in our own laboratory, an isolated transparent arterial segment containing the whole descending aorta and its four major branches was prepared from a dog. The flow patterns at each aortic T-junction were studied in detail under the conditions of steady flow by means of flow visualization and cinemicrographic techniques. It was found that a standing recirculation zone consisting of a pair of thin-layered spiral secondary flows located symmetrically about the common median plane of the aorta and side branches was formed at each T-junction over a wide range of flow conditions including the time-averaged estimated mean values of physiological flow rates and flow rate ratios. The results support the recent in vivo findings by other investigators that flow reversal occurs at some junctions of the dog abdominal aorta during each cardiac cycle. The flow patterns at the aortic T-junctions were very much similar to those previously observed in various glass model T-junctions. However, due to the particular anatomical structure of the vessel wall at each branching site (the curvature of the wall was very sharp at the flow divider, but gently rounded at the bend opposite to it) no recirculation zone was formed in the side branches. At a given flow rate ratio, the measured critical Reynolds numbers for the formation of spiral secondary flows and fully developed disturbed flows were much higher in aortic T-junctions than those in glass model T-junctions having equivalent branching angles and diameter ratios. These results indicate that, in the circulation, conditions at arterial T-junctions appear to be optimal for minimizing the formation of disturbed flows.     

Effect of oxygen on denitrification in continuous chemostat culture withComamonas sp SGLY2

Continuous cultures ofComamonas sp SGLY2 were grown anaerobically prior to establishing steady states at different oxygen flow rates. At a low oxygen transfer rate, no dissolved oxygen accumulated in the medium and all nitrate was reduced to dinitrogen. Concurrently with the increase of dissolved oxygen concentration in the liquid phase, the rate of denitrification decreased. However, at a dissolved oxygen concentration near saturation (33 mg L^–1), a part of the electron flow always diverted to nitrate with production of dinitrogen: the aerobic denitrification rate was equivalent to 35% of that calculated under anaerobic conditions. These experiments reflected the co-utilization of oxygen and N-oxides and the production of dinitrogen, up to saturated conditions, which implied synthesis and activity of the four denitrifying enzymes under various aeration conditions.     

Specific heat flow rate: an on-line monitor and potential control variable of specific metabolic rate in animal cell culture that combines microcalorimetry with dielectric spectroscopy

One of the requirements for enhanced productivity by the animal culture systems used in biotechnology is the direct assessment of the metabolic rate by on-line biosensors. Based on the fact that cell growth is associated with an enthalpy change, it is shown that the specific heat flow rate is stoichiometrically related to the net specific rates of substrates, products, and indeed to specific growth rate, and therefore a direct reflection of metabolic rate. Heat flow rate measured by conduction calorimetry has a technical advantage over estimates for many material flows which require assays at a minimum of two discrete times to give the rate. In order to make heat flow rate specific to the amount of the living cellular system, it would be advantageous to divide it by viable biomass. This requirement has been fulfilled by combining a continuous flow microcalorimeter ex situ with a dielectric spectroscope in situ, the latter measuring the viable cell mass volume fraction. The quality of the resulting biosensor for specific heat flow rate was illustrated using batch cultures of Chinese hamster ovary cells (CHO 320) producing recombinant human interferon-gamma (IFN-gamma) during growth in a stirred tank bioreactor under fully aerobic conditions. The measuring scatter of the probe was decreased significantly by applying the moving average technique to the two participant signals. It was demonstrated that the total metabolic rate of the cells, as indicated by the specific heat flow rate sensor, decreased with increasing time in batch culture, coincident with the decline in the two major substrates, glucose and glutamine, and the accumulation of the by-products, ammonia and lactate. Furthermore, the specific heat flow rate was an earlier indicator of substrate depletion than the flow rate alone. The calorimetric-respirometric ratio showed the intensive participation of anaerobic processes during growth and the related IFN-gamma production. Specific heat flow rate was monotonically related to specific cell growth rate and associated with specific IFN-gamma production. Specific heat flow rate is potentially a valid control variable for the growth of genetically engineered cell lines producing target proteins.     

大孔树脂纯化岩高兰多酚的工艺研究

为获得大孔树脂纯化岩高兰多酚的最佳工艺,以岩高兰的地上部分为原料,通过考察6种不同类型树脂(HPD-100、X-5、AB-8、D101、HPD-600、NKA-II)的含水率、吸附率和解吸率的大小,筛选出一种最适合纯化岩高兰多酚的树脂。在此基础上,选择对纯化工艺影响较大的4种因素(上样浓度、乙醇浓度、洗脱流速、洗脱体积),进行响应面法分析得到最佳工艺。结果表明:HPD-600型大孔树脂对岩高兰多酚的纯化效果最佳,其最优工艺参数为:上样浓度0.84 mg·mL^-1;乙醇浓度62.15%;洗脱流速0.67 mL·min^-1;洗脱体积2.71 BV。该条件下,岩高兰多酚的提取率为229.18 mg·g^-1,岩高兰多酚的纯度由8.11%提高到22.56%,回收率为67.78%。本研究为岩高兰多酚的纯化工艺提供了新的技术路线,也可为岩高兰提取物的研究和应用提供参考。     

Production of proteolytic enzyme by Rhizopus oryzae in a bubble column bioreactor

Production of proteolytic enzyme in a bubble column bioreactor was carried out using the microorganism Rhizopus oryzae. Optimization of the production conditions were done adopting the EVOP (evolutionary operation technique). Studies on two variable and three variable parameters were carried out. The variable parameters were air flow rate, aeration time and incubation period. 8 hours aeration time per day, 1.66 vvm air flow rate and 2 days incubation was found to be optimum.     

非硬化土路径流侵蚀产沙动力参数分析

采用野外径流冲刷试验的方法,模拟研究非硬化路面土壤剥蚀率与各水动力学参数之间的关系,并建立各自的定量关系式.结果表明： 不同流量和坡度下,平均土壤剥蚀率可以用放水流量和坡度的幂函系数关系进行描述,并随放水流量和坡度的增大而增大,流量对土壤剥蚀率的影响大于坡度;土壤剥蚀率与水流流速呈幂函数关系;土壤剥蚀率与径流动能呈幂函数关系,径流动能对土壤剥蚀率有重要作用;土壤剥蚀率与单宽径流能耗呈线性函数关系,土壤可蚀性参数和临界单宽径流能耗的均值分别为0.120 g·m^-1·J^-1和2.875 g·m^-1·J^-1.放水流量和坡度、单宽能耗可准确地描述道路土壤侵蚀过程并对土壤侵蚀量进行测算.     

Water and Sulphate Uptake at Root Reduction in Ricinus communis

The aim of the investigation was to study the influence of the rate of water uptake on the uptake of sulphate at supernormal rates of water flow. This was achieved by reducing the size of the root system of 42 days old Ricinus plants. The rate of water flow through the root increased 3 times by reducing the root system to 20 percent. This did not change the retention of sulphate in the roots. The uptake of sulphate was proportional to the size of the root system and thus independent of the rate of water flow while the water uptake (transpiration) was a function of the size of the shoot and the resistance of the root. This was contrary to the conditions at a moderate rate of water flow, when water and sulphate uptake followed each other. The results are discussed in terms of the salt uptake as a series of active and passive processes.     

The relationship among flow rate, chamber volume and calculated rate of metabolism in vertebrate respirometry

A relationship exists among the calculated rate of metabolism of an animal enclosed in a chamber, chamber volume and air flow rate. A "critical" flow rate, defined as the minimal flow rate that produces a complete mixture of chamber gases, characterizes each chamber/animal combination. At flow rates below the critical flow rate, calculated rates of metabolism decrease with flow rate and approach zero as flow rate approaches zero. These calculated rates are unreliable estimates of rate of metabolism. At flow rates greater than the critical flow rate, calculated rates of metabolism are independent of flow rate because a reciprocal relationship exists between flow rate and the differential in oxygen content found between the air entering and exiting the chamber; they represent an accurate estimate of the animal's rate of metabolism under the conditions to which the animal is exposed. The critical flow rate increases with chamber volume, the animal's rate of metabolism and with any factor that increases rate of metabolism, including body mass, activity and in endotherms ambient temperatures below thermoneutrality, although chamber volume is its single most important determinant. Some evasions of the critical flow rate are discussed.