基于渐变波导COHb与O2Hb混合浓度传感技术研究

研制了折射率渐变光波导传感器,在灵敏度与波导参数理论模型基础上,进行了波导传感厚度及模式的优化设计.在此基础上,搭建了混合物浓度测试回路测试装置,实现了COHb和O2Hb的混合物浓度的测试,并对渐变波导包层吸收模谱特性进行了理论分析和实验测试,测试了包层吸收损耗.结果表明溶液包层吸收特征的折射率虚部与溶液浓度之间具有十分显著的线性相关性.利用该装置测试了11种不同浓度混合物的溶液样品,浓度测试值与实际配制浓度值的均方差小于0.054%.当COHb＞1%时,测定结果较为理想,适用于诊断性定量测定,具有重要的临床应用价值.     

Another look at: fuel + O2 --> CO2 + H2O. Developing a water-oriented perspective.

Water plays many vital roles which must be maintained despite constant threat of water stress from the environment. Mechanisms to maintain these roles over the long-term involve intermediates of many metabolic pathways, such as glycerol, sucrose, free amino acids and their derivatives. In addition to viewing metabolic products as intermediates along energy-related pathways, nutrition researchers and epidemiologists should consider them as determinants of intracellular fluid maintenance. Satiety processes, oxidative-fuel selection, hormonal control, and intracellular-signaling processes can all be interpreted in terms of water-oriented metabolism. Water intake and hydration status can vary considerably, and may be inadequate for a sizeable proportion of Western adults. Due to the metabolic adaptations required to compensate for perpetually inadequate water, chronic sub-optimal water intakes may be highly relevant to chronic disease etiology. Preliminary evidence links water-oriented metabolism to obesity, diabetes, cardiovascular disease, hypertension and cancer. Development of a water-oriented perspective may reveal an important new area of research in human nutrition and epidemiology.     

Transcutaneous measurement of PO2 and PCO2 in the dermis at the site of the tuberculin reaction in healthy human subjects

The respiration of the skin at the site of a delayed hypersensitivity reaction in tuberculin skin tests was studied by transcutaneous measurement of dermal oxygen and carbon dioxide tensions in normal individuals who had been immunized with BCG: six reactions were strong positives, four were weak positives, and four without induration were regarded as negative. The tcPO2 fell over the first 2 days of the reaction and remained low for the next 2 days: the severity of the changes was greater in the 'strong' reactions than in the 'weak' reactions. The tcPCO2 showed a reciprocal rise over the first 2 days and, although still high, tended to recover over the fourth day. These results indicate that local hypoxia and hypercapnia are prominent features of the positive tuberculin test, probably as a consequence of the respiration of the infiltrating lymphocytes and monocytes. It is likely that similar respiratory changes occur in those chronic inflammatory diseases where delayed hypersensitivity reactions make a contribution to the pathogenesis of the lesion.     

Interstitial PCO2 and pH in rat hippocampal slices measured by means of a novel fast CO2/H+-sensitive microelectrode based on a PVC-gelled membrane

We describe here the construction and properties of a double-barrelled microelectrode (tip diameter 4–10 m) which permits simultaneous measurements of PCO₂ and pH, and which has a 90% response time of only one or a few seconds for a step change in PCO₂. The fast response of the CO₂-sensitive barrel is due to (i) the use of a PVC-gelled (tridodecylamine-containing) membrane solution which enables the construction of extremely short ( 4 m), yet mechanically stable, membrane columns, and (ii) the presence of carbonic anhydrase in the filling solution. Recordings made in the pyramidal layer of area CA1 in rat hippocampal slices showed that the deviation in the acid direction of the basal interstitial pH (pH₀) from that of the perfusion solution was attributable to a higher PCO₂ level within the tissue. Most of the late acid shift evoked by stimulation of the Schaffer collaterals (5- to 20-s trains at 10 Hz) could also be explained on the basis of an accumulation of interstitial CO₂ at a constant HCO ₃ ^– concentration. This conclusion was supported by the finding that inhibition of extracellular carbonic anhydrase activity by 10 M benzolamide completely abolished the activity-induced fall in pH₀, but not the increase in PCO₂. The initial stimulus-induced alkalosis was accompanied by a slight decrease in PCO₂ only, implying a parallel increase in the interstitial HCO ₃ ^– concentration. Benzolamide produced a dramatic enhancement of the early alkaline shift as well as of the simultaneous fall in PCO₂. The latter effect of the drug unmasks a cellular CO₂ sink that is induced by neuronal activity.     

The kinetics of the reaction of carbon monoxide with fully oxygenated haemoglobin in solution and erythrocytes

1. The effect of electrical stimulation of the motor nerve supplying the whiskers on the activity of single cells in the vibrissal region of the ventrobasal complex of the thalamus has been studied in rats under urethane anaesthesia.2. The stimulation caused protraction of the ipsilateral whiskers. 60% of the cells which fired to mechanical movements of the whiskers were found to respond to this electrical stimulus with 1-2 impulses at short latency (average 7.7 msec), provided the stimulus was sufficient to move the whiskers.3. When the moving whiskers hit a barrier, 92% of the cells responded to the stimulus. The most effective position of the barrier was in front of the whiskers, although other positions often produced a response as well. Static displacement of the whiskers, particularly in the forward direction, could abolish the response or increase its latency.4. The following-frequencies for these cells were 5-10 stimuli/sec. Combinations of electrical stimuli with mechanical ramp movements of the whiskers showed that similar recovery times followed both types of stimuli.5. These results are compared with those reported from studies in the afferent nerve fibres after electrical stimulation of the motor nerve and also with responses in the thalamus following mechanical movements of the whiskers. The possible importance of the latency of these sensory responses is considered.     

Numerical solution of partial differential equations describing the simultaneous O2 and CO2 diffusions in the red blood cell

To describe the overall gas exchange rates in red blood cells (RBC), a computer program for solving the diffusion equations for O2, CO2, and HCO3-that accompany the chemical reactions of Bohr- and Haldane-effects was developed. Three diffusion equations were solved alternatively and repeatedly in an increment time of 2 ms. After solving the diffusion equations the Po2, O2 saturation (So2), Pco2, pH, and HCO3-content were corrected by using the Henderson-Hasselbalch equation, where the buffer value was newly derived from the CO2 dissociation curve. In computing the Haldane effect, the buffer value was taken to be 44mmol X l(RBC)-1 X pHc-1, so that the change in intracellular dissolved CO2 caused by the So2 change was fully compensated by the subsequent CO2 diffusion. The oxygenation and deoxygenation rate factors of hemoglobin were assumed to be 2.09 X (1-S)2.02 and 0.3s-1 X Torr-1, respectively. The Po2 change due to the Bohr-shift was computed from Hill's equation, in which the K value was given by a function of the intracellular pH. When the parameter values thus far measured were used, the computed Bohr- and Haldane-effects coincided well with the experimental data, supporting the validity of the equations. The overall gas exchange profiles calculated in the pulmonary capillary model showed that the CO2 equilibration time was significantly longer than the oxygenation time.     

The impact of PCO2 and H+ on the release of acetylcholine from the cat carotid body

The carotid body (CB) is a sensor of oxygen, carbon dioxide, hydrogen ion, and glucose in the arterial blood. Many studies of the CB's responses to low oxygen (hypoxia) have been reported. Recently attention has been increasingly focused on its responses to elevated CO2 (hypercapnia). An increase in ventilation or carotid body neural output (CBNO) can result from stimulating the CB with blood or perfusion fluids having an elevated CO2 or H+. The increase in ventilation seen with a hypoxic stimulus is accompanied with an increase in CBNO and an increased release of both acetylcholine (ACh) and ATP from the CB. The present in vitro study using both CBs harvested from six cats was undertaken to determine if hypercapnia also provoked an increased release of ACh from the incubated CBs. The anesthetizing, handling, and euthanizing of the animals were according to the guidelines of the Johns Hopkins Animal Care and Use Committee which are totally consonant with those of the NIH. CBs, once harvested and prepared for the experimental protocol, were subjected to the following steps each lasting 10 min: (1) control; (2) stress; (3) recovery. The stresses were respiratory acidosis (RAC; acidic hypercapnia), compensated respiratory acidosis (CRAC; isohydric hypercapnia), and metabolic acidosis (MtAC). The first and last forms of acidosis generated small but significant increases in the release of ACh from the CBs; the second generated a very small and insignificant increase in ACh release. Since it is generally accepted that ACh is a key excitatory neurotransmitter in the CB along with ATP, these data are consistent with other studies measuring the increase in ventilation in response to a small increase in CO2 and those studies recording CBNO in response to hypercapnia. In five of the six animals the responses to RAC and MtAC were compared to the responses to hypoxia. The latter were statistically indistinguishable from the former two.     

Inhibitory action of Ca2+ on spontaneous transmitter release at motor nerve terminals in a high K+ solution

The inhibitory effect of a high external Ca²⁺ ([Ca²⁺]_o) on spontaneous transmitter release in a high K⁺ solution (Gage and Quastel 1966; Birks et al. 1968) was studied at the frog neuromuscular junction, based on the hypothesis that an increased intracellular free Ca²⁺ ([Ca²⁺]_i) in the nerve terminal plays a key role in the depression. Three procedures were employed to increase [Ca²⁺]_i; increasing [Ca²⁺]_o, application of caffeine and tetanic nerve stimulation. All of these procedures increased m.e.p.p. frequency in normal Ringer. However, as the basic m.e.p.p. frequency was increased by raising the external K⁺ concentration (7–15 mM), their facilitatory effects on m.e.p.p. frequency decreased, disappeared and eventually reversed to depressant actions. Since a rise in the external K⁺ concentration would increase the steady state level of [Ca²⁺]_i, it is suggested that when the [Ca²⁺]_i is preset at a high level, manipulations so as to further increase [Ca²⁺]_i depress spontaneous release of transmitter. Possible mechanisms for this inhibition was discussed in relation to a question whether or not the rate of spontaneous transmitter release is a monotonic function of [Ca²⁺]_i.     

A method for making solution changes in the sub-millisecond range at the tip of a patch pipette

A method is described here for making multiple fast external solution changes at the tip of a patch pipette. The time for the change, 0.2 ms, has been established by measuring changes in liquid junction potential at the tip of an open patch pipette. This technique of producing an abrupt change in solution allows agonist/receptor reactions to be studied under non-equilibrium conditions. We have applied this technique to the nicotinic receptors in outside-out patches from skeletal muscle cell line C2 (Jaffe and Saxel 1979) and from bovine adrenal chromaffin cells. The application of step changes in acetylcholine concentration produces current traces with a characteristic shape, which may be compared with the predictions of established models for the activation and desensitisation of the nicotinic receptor. The results of making single steps and also short pulses in acetylcholine concentration are demonstrated. The direct comparison of two different cholinergic agonists is demonstrated.     

Arterial PCO2 and pH in man during 3 days' exposure to 2.8 kPa CO2 in the inspired gas

It has not been firmly established how respiration adapts to long-term CO2 exposure in man. We have therefore exposed five healthy human subjects to 2.8 kPa CO2 in the inspired gas for about 70 h in a chamber with controlled atmospheric conditions at ambient pressure PCO2 and pH were determined in arterial or arterialized venous blood drawn before, during and after the exposure. One subject was studied twice. We found that PaCO2 increased acutely and then increased further within the 5- to 24-h period of exposure to 2.8 kPa CO2. No consistent change was observed during the following 2 days. At the end of exposure the PaCO2 was 0.5 kPa above the pre-exposure level. When the breathing gas was switched back to room air, PaCO2 promptly returned to pre-exposure values. The secondary rise in PaCO2 within the first day would correspond to a decrease in alveolar ventilation of about 10% assuming constant production and elimination of CO2. Arterial pH remained slightly below the pre-exposure level during the entire exposure period. A slight renal compensation resulting in an increase in base excess of about 1 mmol l-1 may have occurred in the middle part of the exposure period. We conclude that a significant, but moderate, respiratory adaptation takes place during the first day of exposure to an increased inspired load of CO2.     

Activation of the contractile system of insect fibrillar muscle at very low concentrations of Mg2+ and Ca2+

The mechanical properties of single fibres and fibre bundles of glycerinated dorsal longitudinal muscle from lethocerus maximus were investigated in ATP-salt solutions containing only trace concentrations of free Ca²⁺ (pCa>9). A reduction in the magnesium concentration (pMg 7) resulted in an increase in the instantaneous stiffness of glycerinated insect flight muscle fibres, though very little accompanying tension was developed. Stiffness was measured either using small amplitude sinusoidal length changes of high frequency (1 kHz) or rapid rectangular form length changes. The ratio of stiffness to tension in solutions free of added magnesium and calcium was equal to or greater than that obtained from the tissue in the rigor state, and much larger than that obtained in the presence of both magnesium and calcium. Extrapolation of the linear part of the change-tension relationship (obtained during rapid length changes completed within 0.3 ms) back to zero tension indicated that the elastic elements of attached crossbridges were less extended under conditions of Mg²⁺-deprivation than during Ca-activation in Mg²⁺-rich solution. Following a quick stretch a delayed tension development similar to that obtained in the presence of magnesium and calcium ions was observed. The rise in tension was delayed with respect to the accompanying rise in stiffness and reached a peak value after about 2 s. Similar tension transients followed a subsequent release. The possibility that an unusually slow corss-bridge cycle might be responsible for these slow transients was suggested by the finding that the fibres showed a very low ATPase activity under these conditions which could be slightly activated by stretches. On increasing the free Ca²⁺ concentration during magnesium deprivation, the time course of the stretch induced tension transients became faster, while stiffness and the steady state tension rose to reach a high tension state at aboutpCa 6.5.This work was supported by a grant from the Deutsche Forschungs-gemeinschaft (RU 154/12). One of us (P.J.G.) was supported by the Royal Society European Science Exchange Programme     

Reciprocal modulation of O2 and CO2 cardiorespiratory chemoreflexes in the tambaqui

This study examined the effect of acute hypoxic and hypercapnic cardiorespiratory stimuli, superimposed on existing cardiorespiratory disturbances in tambaqui. In their natural habitat, these fish often encounter periods of hypoxic hypercapnia that can be acutely exacerbated by water turnover. Tambaqui were exposed to periods of normoxia, hypoxia, hyperoxia and hypercapnia during which, externally oriented O2 and CO2 chemoreceptors were further stimulated, by administration into the inspired water of sodium cyanide and CO2-equilibrated water, respectively. Hyperoxic water increased the sensitivity of the NaCN-evoked increase in breathing frequency (f(R)) and decrease in heart rate. Hypoxia and hypercapnia attenuated the increase in f(R) but, aside from blood pressure, did not influence the magnitude of NaCN-evoked cardiovascular changes. Water PO2 influenced the magnitude of the CO2-evoked cardiorespiratory changes and the sensitivity of CO2-evoked changes in heart rate and blood flow. The results indicate that existing respiratory disturbances modulate cardiorespiratory responses to further respiratory challenges reflecting both changes in chemosensitivity and the capacity for further change.     

Relationship between O2 consumption,high energy phosphates and the kinetics of the O2 debt in exercise

Summary The oxygen consumption together with lactic acid production and concentration of ATP, ADP, and creatinephosphate was measured during exercise and recovery on an isolated dog gastrocnemius.Oxygen debt contraction and payment follow an exponential path with a half reaction time of about 20 sec. The concentration of ATP and ADP at steady state seem to be unaffected by the intensity of the exercise when this is submaximal and no appreciable production of lactic acid takes place. The concentration of creatinephosphate in muscle at steady state decreases with the intensity of the exercise. The ratio of the oxygen consumption at steady state to the alactic oxygen debt is identified with the speed constant of the resynthesis of phosphagen in muscle; the half reaction time of this process is 17–20 sec. The total alactic oxygen debt amounts to about 50 ml/kg of muscle. These figures are in good agreement with earlier data found in man.We are greatly indebted to Dr.J. Piiper for his useful criticism and help during the preparation of this paper.     

Arterial PCO2 and lung ventilation in man exposed to 1–5% CO2 in the inspired gas

Conflicting results have been published on the shape of the curve relating the change in lung ventilation to the change in alveolar or arterial PCO₂ induced by increased inspired CO₂ (the CO₂ sensitivity). In this study eight human subjects with in-dwelling arterial cannulae were each exposed to five different levels of increased inspired CO₂ (1–5%). Arterial PCO₂ and ventilation were measured in the 7th minute of each period of CO₂ exposure. Each CO₂ exposure period was flanked by control periods in which similar measurements were carried out during air breathing. We found non-linear increases in both ventilation and arterial PCO₂ with increasing levels of inspired CO₂. When 5 % CO₂ in air was inspired the arterial PCO₂ increased by about 15% of the inspired CO₂ load. There was no significant non-linearity in the relation between change in alveolar ventilation (normalized to body surface) and change in arterial P_CO2. The inter-individual variation in CO₂ sensitivity was less when alveolar ventilation was normalized to the CO₂ output rather than to body surface area. We conclude that the sensitivity to CO₂ is close to constant within the range 0–5 % CO₂ in the inspired gas.