Hypoxia, temperature, and pH/CO2 effects on respiratory discharge from a turtle brain stem preparation

Johnson, Stephen M., Rebecca A. Johnson, and Gordon S. Mitchell. Hypoxia, temperature, andpH/CO₂ effects on respiratory discharge from a turtle brain stem preparation. J. Appl. Physiol. 84(2): 649-660, 1998.An in vitrobrain stem preparation from adult turtles (Chrysemyspicta) was used to examine the effects of anoxia andincreased temperature and pH/CO₂on respiration-related motor output. At pH ~7.45, hypoglossal (XII)nerve roots produced patterns of rhythmic bursts (peaks) of discharge(0.74 ± 0.07 peaks/min, 10.0 ± 0.6 s duration) that werequantitatively similar to literature reports of respiratory activity inconscious, vagotomized turtles. Respiratory discharge was stable for 6 h at 22°C; at 32°C, peak amplitude and frequency progressivelyand reversibly decreased with time. Two hours of hypoxia had no effecton respiratory discharge. Acutely increasing bath temperature from 22 to 32°C decreased episode and peak duration and increased peakfrequency. Changes in pH/CO₂increased peak frequency from zero at pH 8.00-8.10 to maxima of0.81 ± 0.01 and 1.44 ± 0.02 peaks/min at 22°C (pH 7.32) and32°C (pH 7.46), respectively;pH/CO₂ sensitivity was similar atboth temperatures. We conclude that1) insensitivity to hypoxiaindicates that rhythmic discharge does not reflect gasping behavior,2) increased temperature altersrespiratory discharge, and 3)central pH/CO₂ sensitivity isunaffected by temperature in this preparation (i.e.,Q₁₀ ~1.0).

     

Appropriate thermal manipulations eliminate tremors in rats recovering from halothane anesthesia

Grahn, D. A., M. C. Heller, J. E. Larkin, and H. C. Heller.Appropriate thermal manipulations eliminate tremors in ratsrecovering from halothane anesthesia. J. Appl.Physiol. 81(6): 2547-2554, 1996.Tremors arecommon in mammals emerging from anesthesia. To determine whetherappropriate thermal manipulations immediately before emergence fromanesthesia are sufficient to eliminate these tremors,electroencephalographic (EEG) and electromyographic (EMG) activities,hypothalamic temperature (T_hy),and O₂ consumption were monitoredin 12 rats recovering from halothane anesthesia under three thermalregimes. EEG and EMG activities were recorded throughout anesthesia andserved as feedback signals for controlling anesthetic depth. Duringanesthesia, T_hy was either1) allowed to fall to32-34°C, 2) maintained at37-39°C, or 3) allowed to fall to 32-34°C and then raised to 37-39°C. Whenhypothermic on emergence from anesthesia, all of the animals exhibitedpostanesthetic tremors that persisted untilT_hy values returned tonormothermia. None of the animals expressed postanesthetic tremors whennormothermic on emergence from anesthesia. In addition, the timebetween emergence from anesthesia (as determined by EEG/EMG parameters)and the initiation of coordinated motor activities was significantlydecreased in the normothermic animals.

     

Mechanisms of the acido- and thermo-phily of Cyanidium caldarium Geitler I. Effects of temperature, pH and light intensity on the photosynthetic oxygen evolution of intact and treated cells

Photosynthetic oxygen evolution in an acido- and thermo-philicunicellular alga, Cyanidium caldarium, was measured under variousconditions, using a Clark-type oxygen electrode. 1). Maximum Hill reaction activity with p-benzoquinone as theHill oxidant was obtained at 45°C in a wide pH range from1.0 to 7.0. 2) The pH activity curve showed two peaks at pH3.0 and 7.0. The Hill activity had an optimum at pH 3.0 in cellspreilluminated under strong light (300,000 lux, 30 min, 40°C).Sonication of algal cells abolished the pH 3.0 component ofthe Hill reaction producing an activity maximum at pH 7.0. 3)Endogenous O₂ evolution in the absence of the Hill oxidant,which lasted for several minutes after illumination, had a maximumat pH 7.0. 4) This endogenous O₂ evolution was abolished bysonication. 5) KCN inhibited endogenous O₂ evolution, but notthe Hill reaction in the presence of p-benzoquinone. (Received August 19, 1974; )     

Thermal and metabolic responses to cold-water immersion at knee, hip, and shoulder levels

Lee, Dae T., Michael M. Toner, William D. McArdle, IoannisS. Vrabas, and Kent B. Pandolf. Thermal and metabolic responses tocold-water immersion at knee, hip, and shoulder levels.J. Appl. Physiol. 82(5):1523-1530, 1997.To examine the effect of cold-water immersion atdifferent depths on thermal and metabolic responses, eight men (25 yrold, 16% body fat) attempted 12 tests: immersed to the knee (K), hip(H), and shoulder (Sh) in 15 and 25°C water during both rest (R) orleg cycling [35% peak oxygen uptake; (E)] for up to 135 min. At 15°C, rectal (T_re)and esophageal temperatures(T_es) between R and E were notdifferent in Sh and H groups (P > 0.05), whereas both in K group were higher during E than R(P < 0.05). At 25°C,T_re was higher(P < 0.05) during E than R at alldepths, whereas T_es during E washigher than during R in H and K groups.T_re remained at control levels inK-E at 15°C, K-E at 25°C, and in H-E groups at 25°C,whereas T_es remained unchanged inK-E at 15°C, in K-R at 15°C, and in all 25°C conditions (P > 0.05). During R and E, themagnitude of T_re change wasgreater (P < 0.05) than themagnitude of T_es change in Sh andH groups, whereas it was not different in the K group(P > 0.05). Total heat flow wasprogressive with water depth. During R at 15 and 25°C, heatproduction was not increased in K and H groups from control level(P > 0.05) but it did increase in Shgroup (P < 0.05). The increase inheat production during E compared with R was smaller(P < 0.05) in Sh (121 ± 7 W/m² at 15°C and 97 ± 6 W/m² at 25°C) than in H (156 ± 6 and 126 ± 5 W/m²,respectively) and K groups (155 ± 4 and 165 ± 6 W/m², respectively). These datasuggest that T_re andT_es respond differently duringpartial cold-water immersion. In addition, water levels above knee in15°C and above hip in 25°C cause depression of internal temperatures mainly due to insufficient heat production offsetting heatloss even during light exercise.

     

Skeletal muscle chemoreflex and pHi in exercise ventilatory control

Oelberg, David A., Allison B. Evans, Mirko I. Hrovat, PaulP. Pappagianopoulos, Samuel Patz, and David M. Systrom. Skeletal muscle chemoreflex and pH_i inexercise ventilatory control. J. Appl.Physiol. 84(2): 676-682, 1998.To determinewhether skeletal muscle hydrogen ion mediates ventilatory drive inhumans during exercise, 12 healthy subjects performed three bouts ofisotonic submaximal quadriceps exercise on each of 2 days in a 1.5-Tmagnet for ³¹P-magnetic resonancespectroscopy(³¹P-MRS). Bilaterallower extremity positive pressure cuffs were inflated to 45 Torr duringexercise (BLPP_ex) or recovery(BLPP_rec) in a randomized orderto accentuate a muscle chemoreflex. Simultaneous measurements were madeof breath-by-breath expired gases and minute ventilation, arterializedvenous blood, and by ³¹P-MRS ofthe vastus medialis, acquired from the average of 12 radio-frequencypulses at a repetition time of 2.5 s. WithBLPP_ex, end-exercise minuteventilation was higher (53.3 ± 3.8 vs. 37.3 ± 2.2 l/min;P < 0.0001), arterializedPCO₂ lower (33 ± 1 vs. 36 ± 1 Torr; P = 0.0009), and quadricepsintracellular pH (pH_i) more acid (6.44 ± 0.07 vs. 6.62 ± 0.07; P = 0.004), compared withBLPP_rec. Bloodlactate was modestly increased withBLPP_ex but without a change inarterialized pH. For each subject, pH_i was linearly relatedto minute ventilation during exercise but not to arterialized pH. Thesedata suggest that skeletal muscle hydrogen ion contributes to theexercise ventilatory response.

     

Alterations in the surface properties of lung surfactant in the torpid marsupial Sminthopsis crassicaudata

Lopatko, Olga V., Sandra Orgeig, Christopher B. Daniels, andDavid Palmer. Alterations in the surface propertiesof lung surfactant in the torpid marsupial Sminthopsiscrassicaudata. J. Appl.Physiol. 84(1): 146-156, 1998.Torpor changes thecomposition of pulmonary surfactant (PS) in the dunnartSminthopsis crassicaudata [C.Langman, S. Orgeig, and C. B. Daniels. Am. J. Physiol. 271 (Regulatory IntegrativeComp. Physiol. 40): R437-R445, 1996]. Herewe investigated the surface activity of PS in vitro. Five micrograms ofphospholipid per centimeter squared surface area of whole lavage (frommice or from warm-active, 4-, or 8-h torpid dunnarts) were applieddropwise onto the subphase of a Wilhelmy-Langmuir balance at 20°Cand stabilized for 20 min. After 4 h of torpor, the adsorption rateincreased, and equilibrium surface tension (ST_eq), minimal surface tension(ST_min), and the %areacompression required to achieveST_min decreased, compared with thewarm-active group. After 8 h of torpor,ST_min decreased [from 5.2 ± 0.3 to 4.1 ± 0.3 (SE) mN/m]; %area compressionrequired to achieve ST_min decreased (from 43.4 ± 1.0 to 27.4 ± 0.8); the rate ofadsorption decreased; and ST_eqincreased (from 26.3 ± 0.5 to 38.6 ± 1.3 mN/m). ST-areaisotherms of warm-active dunnarts and mice at 20°C had a shoulderon compression and a plateau on expansion. These disappeared on theisotherms of torpid dunnarts. Samples of whole lavage (from warm-activeand 8-h torpor groups) containing 100 µg phospholipid/ml were studiedby using a captive-bubble surfactometer at 37°C. After 8 h oftorpor, ST_min increased (from 6.4 ± 0.3 to 9.1 ± 0.3 mN/m) and %area compressiondecreased in the 2nd (from 88.6 ± 1.7 to 82.1 ± 2.0) and 3rd(from 89.1 ± 0.8 to 84.9 ± 1.8) compression-expansion cycles, compared with warm-active dunnarts. ST-area isotherms ofwarm-active dunnarts at 37°C did not have a shoulder oncompression. This shoulder appeared on the isotherms of torpiddunnarts. In conclusion, there is a strong correlation between in vitrochanges in surface activity and in vivo changes in lipid composition of PS during torpor, although static lung compliance remained unchanged (see Langman et al. cited above). Surfactant from torpid animals ismore active at 20°C and less active at 37°C than that ofwarm-active animals, which may represent a respiratory adaptation tolow body temperatures of torpid dunnarts.

     

Intravascular oxygen distribution in subcutaneous 9L tumors and radiation sensitivity

Cerniglia, George J., David F. Wilson, Marek Pawlowski,Sergei Vinogradov, and John Biaglow. Intravascular oxygendistribution in subcutaneous 9L tumors and radiation sensitivity.J. Appl. Physiol. 82(6):1939-1945, 1997.Phosphorescence quenching was evaluated as atechnique for measuring PO₂ in tumors and for determining the effect of increasedPO₂ on sensitivity of the tumors toradiation. Suspensions of cultured 9L cells or small pieces of solidtumors from 9L cells were injected subcutaneously on the hindquarter ofrats, and tumors were grown to between 0.2 and 1.0 cm in diameter.Oxygen-dependent quenching of the phosphorescence of intravenouslyinjected Pd-meso-tetra-(4-carboxyphenyl) porphine was used to image thein vivo distribution of PO₂ in thevasculature of small tumors and surrounding tissue. Maps (512 × 480 pixels) of tissue oxygen distribution showed that thePO₂ within 9L tumors was low(2-12 Torr) relative to the surrounding muscle tissue (20-40Torr). When the rats were given 100% oxygen or carbogen (95%O₂-5%CO₂) to breathe, thePO₂ in the tumors increasedsignificantly. This increase was variable among tumors and was greaterwith carbogen compared with 100% oxygen. Based on irradiation andregrowth studies, carbogen breathing increased the sensitivity of thetumors to radiation. This is consistent with the measured increase inPO₂ in the tumor vasculature. It isconcluded that phosphorescence quenching can be used for noninvasivedetermination of the oxygenation of tumors. This method for oxygenmeasurements has great potential for clinical application in tumoridentification and therapy.

     

Rate of uptake of CO by hemoglobin in pig erythrocytes as a function of PO2

Te Nijenhuis, Francis C. A. M., Lydia Lin, Gerko H. Moens,Adrian Versprille, and Robert E. Forster. Rate of uptake of CO byhemoglobin in pig erythrocytes as a function ofPO₂. J. Appl.Physiol. 81(4): 1544-1549, 1996.This study wasinitiated to obtain data on the rate of carbon monoxide (CO) uptake(_CO) by hemoglobin in pigerythrocytes to derive, in a later study, the pulmonary capillary bloodvolume (Qc) in pigs from the Roughton-Forster relationship. Blood fromfive different female pigs was used. The_CO, the milliliters of CO takenup by 1 ml of whole blood per minute per Torr CO tension, wasdetermined on each blood sample with a continuous-flow rapid-mixingapparatus and double-beam spectrophotometry at 37°C and pH 7.4 atfour or five different PO₂ values.Because the individual regression lines of _CO vs.PO₂ were not significantly different,a common regression equation was calculated:1/_CO = 0.0084 PO₂ + 0.63. The slope of thisregression line is significantly steeper than the reported slopes ofthe regression lines for human and dog erythrocytes measured under thesame conditions. Our results revealed that calculation ofQc in pigs by using _CO valuesfor human or dog erythrocytes would result in an underestimation of 51 and 50%, respectively.

     

Skeletal muscle ECF pH error signal for exercise ventilatory control

Evans, Allison B., Larry W. Tsai, David A. Oelberg, HomayounKazemi, and David M. Systrom. Skeletal muscle ECF pH error signalfor exercise ventilatory control. J. Appl.Physiol. 84(1): 90-96, 1998.An autonomic reflexlinking exercising skeletal muscle metabolism to central ventilatorycontrol is thought to be mediated by neural afferents having freeendings that terminate in the interstitial fluid of muscle. Todetermine whether changes in muscle extracellular fluid pH(pH_e) can provide an errorsignal for exercise ventilatory control,pH_e was measured duringelectrically induced contraction by³¹P-magnetic resonancespectroscopy and the chemical shift of a phosphorylated, pH-sensitivemarker that distributes to the extracellular fluid (phenylphosphonicacid). Seven lightly anesthetized rats underwentunilateral continuous 5-Hz sciatic nerve stimulation in an 8.45-Tnuclear magnetic resonance magnet, which resulted in a mixed lacticacidosis and respiratory alkalosis, with no net change in arterial pH.Skeletal muscle intracellular pH fell from 7.30 ± 0.03 units atrest to 6.72 ± 0.05 units at 2.4 min of stimulation and then roseto 7.05 ± 0.01 units (P < 0.05), despite ongoing stimulation and muscle contraction.Despite arterial hypocapnia, pH_eshowed an immediate drop from its resting baseline of 7.40 ± 0.01 to 7.16 ± 0.04 units (P < 0.05)and remained acidic throughout the stimulation protocol. During the on-and off-transients for 5-Hz stimulation, changes in the pH gradientbetween intracellular and extracellular compartments suggestedtime-dependent recruitment of sarcolemmal ion-transport mechanisms.pH_e of exercising skeletal musclemeets temporal and qualitative criteria necessary for a ventilatorymetaboreflex mediator in a setting where arterial pH doesnot.

     

Thermal Dependence of Air Convection Requirement and Blood Gases in the Snake Coluber constrictor

In this report I discuss ventilatory and circulatory adjustmentsthat prov for increased O₂transport associated with increasedbody temperature in the snake Coluber constrictor. Also includedis the effect of temperature upon acid-base status. Minute ventilationincreases with rising body temperature but does not keep pacewith the increment in resting O₂ consumption. The decrease inair convection requirement (i.e., ventilation ÷ oxygenconsumption) causes lung pO₂ and arterial oxygen contentto falland lung pCO₂ to rise. With the rise in lung pCO₂, systemicarterial pCO₂ and H⁺; concentration increase while plasma bicarbonateconcentration does not change. The effect of temperature uponair convection requirement, arterial pCO₂, and pH are most pronouncedat body temperatures above about 27°C whereColuber behavesapproximately as an alphastat pH regulator. Despite the inverserelationship between temperature and lung pO₂, systemic arterialpO₂ is about 80 torr lower at 15°C than at 35°C. Thisdecline in arterial pO₂ as temperature falls is explained byleft shifting the oxygen dissociation curve in the presenceof aconstant right-to-left intracardiac shunt.     

Dehydration markedly impairs cardiovascular function in hyperthermic endurance athletes during exercise

González-Alonso, José, RicardoMora-Rodríguez, Paul R. Below, and Edward F. Coyle.Dehydration markedly impairs cardiovascular function inhyperthermic endurance athletes during exercise. J. Appl. Physiol. 82(4): 1229-1236, 1997.Weidentified the cardiovascular stress encountered by superimposingdehydration on hyperthermia during exercise in the heat and themechanisms contributing to the dehydration-mediated stroke volume (SV)reduction. Fifteen endurance-trained cyclists [maximalO₂ consumption(O_{2 max}) = 4.5 l/min] exercised in the heat for 100-120 min and either became dehydrated by 4% body weight or remained euhydrated by drinkingfluids. Measurements were made after they continued exercise at 71%O_{2 max} for 30 minwhile 1) euhydrated with anesophageal temperature (T_es) of38.1-38.3°C (control); 2)euhydrated and hyperthermic (39.3°C);3) dehydrated and hyperthermic withskin temperature (T_sk) of34°C; 4) dehydrated withT_es of 38.1°C and T_sk of 21°C; and5) condition4 followed by restored blood volume. Compared withcontrol, hyperthermia (1°C T_esincrease) and dehydration (4% body weight loss) each separatelylowered SV 7-8% (11 ± 3 ml/beat;P < 0.05) and increased heart ratesufficiently to prevent significant declines in cardiac output.However, when dehydration was superimposed on hyperthermia, thereductions in SV were significantly (P < 0.05) greater (26 ± 3 ml/beat), and cardiac output declined 13% (2.8 ± 0.3 l/min). Furthermore, mean arterialpressure declined 5 ± 2%, and systemic vascular resistanceincreased 10 ± 3% (both P < 0.05). When hyperthermia wasprevented, all of the decline in SV with dehydration was due to reducedblood volume (~200 ml). These results demonstrate that thesuperimposition of dehydration on hyperthermia during exercise in theheat causes an inability to maintain cardiac output and blood pressurethat makes the dehydrated athlete less able to cope with hyperthermia.

     

Modification of active cutaneous vasodilation by oral contraceptive hormones

Charkoudian, Nisha, and John M. Johnson. Modificationof active cutaneous vasodilation by oral contraceptive hormones. J. Appl. Physiol. 83(6):2012-2018, 1997.It is not clear whether the alteredthermoregulatory reflex control of the cutaneous circulation seen amongphases of the menstrual cycle also occurs with the synthetic estrogenand progesterone in oral contraceptive pills and whether any suchmodifications include altered control of the cutaneous activevasodilator system. To address these questions, we conducted controlledwhole body heating experiments in seven women at the end of the thirdweek of hormone pills (HH) and at the end of the week of placebo/nopills (LH). A water-perfused suit was used to control body temperature.Laser Doppler flowmetry was used to monitor cutaneous blood flow at acontrol site and at a site at which noradrenergic vasoconstrictorcontrol had been eliminated by iontophoresis of bretylium (BT),isolating the active cutaneous vasodilator system. The oral temperature(T_or) thresholds for cutaneousvasodilation were higher in HH at both control [37.09 ± 0.12 vs. 36.83 ± 0.07°C (LH), P < 0.01] and BT-treated [37.19 ± 0.05 vs. 36.88 ± 0.12°C (LH), P < 0.01]sites. The T_or threshold forsweating was similarly shifted (HH: 37.15 ± 0.11°C vs. LH: 36.94 ± 0.11°C, P < 0.01). Arightward shift in the relationship of heart rate toT_or was seen in HH. Thesensitivities (slopes of the responses vs.T_or) did not differstatistically between phases. The similar threshold shifts at controland BT-treated sites suggest that the hormones shift the function ofthe active vasodilator system to higher internal temperatures. Thesimilarity of the shifts among thermoregulatory effectors suggests acentrally mediated action of these hormones.

     

Posthemorrhagic antipyresis: what stage of fever genesis is affected?

Romanovsky, Andrej A., and Yelena K. Karman.Posthemorrhagic antipyresis: what stage of fever genesis isaffected? J. Appl. Physiol. 83(2):359-365, 1997.It has been shown that hemorrhage leads to adecreased thermal responsiveness to lipopolysaccharide (LPS). The aimof this study was to clarify what stage of fever genesis[production of endogenous pyrogens such as interleukin-1 (IL-1),increase of the prostaglandin E₂(PGE₂) concentration in braintissue, activation of cold-defense effectors] is deficient inposthemorrhagic antipyresis. In adult rabbits, we evaluated the effectof acute hemorrhage (15 ml/kg) on the rectal temperature (T_re) responses to LPS fromSalmonella typhi (200 ng/kg iv),ethanol-purified preparation of homologous IL-1 (1 ml from 3.5 × 10⁷ cells, 1.5 ml/kg iv), andPGE₂ (1 µg,intracisternal injection). The effect of hemorrhage onT_re was also studied in afebrilerabbits, both at thermoneutrality (23°C) and during ramp cooling(to 7°C). The hemorrhage strongly attenuated the biphasicLPS-induced fever (a T_re rise of0.4 ± 0.1 instead of 1.2 ± 0.2°C at the time of the secondpeak), the monophasic T_re responseto IL-1 (by ~0.5°C for over 1-5 h postinjection), and thePGE₂-induced hyperthermia (0.4 ± 0.1 vs. 0.9 ± 0.1°C, maxima). In afebrileanimals, the hemorrhage neither affectedT_re at thermoneutrality norchanged the T_re response to coldexposure. The data suggest that neither insufficiency of cold-defenseeffectors nor lack of endogenous mediators of fever (IL-1,PGE₂) can be the only or eventhe major cause of posthemorrhagic antipyresis. Wespeculate that fever genesis is altered at a stage occurring after theintrabrain PGE₂ level is increasedbut before thermoeffectors are activated.

     

Hypothyroidism alters diaphragm muscle development

Sieck, Gary C., Louise E. Wilson, Bruce D. Johnson, andWen-Zhi Zhan. Hypothyroidism alters diaphragm muscle development. J. Appl. Physiol. 81(5):1965-1972, 1996.The impact of hypothyroidism (Hyp) onmyosin heavy chain (MHC) isoform expression, maximum specific force(P_o), fatigability, and maximumunloaded shortening velocity(V_o) wasdetermined in the rat diaphragm muscle (Dia) at 0, 7, 14, 21, and 28 days of age. Hyp was induced by treating pregnant rats with6-n-propyl-2-thiouracil (0.05% indrinking water) beginning at gestational day10 and was confirmed by reduced plasma levels of3,5,3-triiodothyronine and thyroxine. MHC isoforms wereseparated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels and analyzed by densitometry. IsometricP_o and fatigue resistance of theDia were measured in vitro at 26°C, andV_o was determined at 15°C with the slack test. Compared with control muscles,expression of MHC-slow was higher and expression of adult fast MHCisoforms was lower in Hyp Dia at all ages. The neonatal isoform of MHC continued to be expressed in the Hyp Dia until day28. At each age,P_o and fatigability were reducedand V_o was slowerin the Hyp Dia. We conclude that Hyp-induced alterations in MHC isoform expression do not fully predict the changes in Dia contractile properties.

     

Lactate efflux from exercising human skeletal muscle: role of intracellular PO2

It remainscontroversial whether lactate formation during progressive dynamicexercise from submaximal to maximal effort is due to muscle hypoxia. Tostudy this question, we used direct measures of arterial and femoralvenous lactate concentration, a thermodilution blood flow technique,phosphorus magnetic resonance spectroscopy (MRS), and myoglobin (Mb)saturation measured by ¹H nuclearMRS in six trained subjects performing single-leg quadriceps exercise.We calculated net lactate efflux from the muscle and intracellularPO₂ with subjects breathing room airand 12% O₂. Data were obtained at50, 75, 90, and 100% of quadriceps maximalO₂ consumption at each fraction ofinspired O₂. Mb saturation wassignificantly lower in hypoxia than in normoxia [40 ± 3 vs. 49 ± 3% (SE)] throughout incremental exercise to maximalwork rate. With the assumption of aPO₂ at which 50% of Mb-binding sitesare bound with O₂ of 3.2 Torr,Mb-associated PO₂ averaged 3.1 ± 0.3 and 2.3 ± 0.2 Torr in normoxia and hypoxia, respectively. Netblood lactate efflux was unrelated to intracellular PO₂ across the range of incrementalexercise to maximum (r = 0.03 and 0.07 in normoxia and hypoxia, respectively) but linearly related toO₂ consumption(r = 0.97 and 0.99 in normoxia andhypoxia, respectively) with a greater slope in 12%O₂. Net lactate efflux was alsolinearly related to intracellular pH(r = 0.94 and 0.98 in normoxia andhypoxia, respectively). These data suggest that with increasing workrate, at a given fraction of inspiredO₂, lactate efflux is unrelated tomuscle cytoplasmic PO₂, yet theefflux is higher in hypoxia. Catecholamine values from comparablestudies are included and indicate that lactate efflux in hypoxia may bedue to systemic rather than intracellular hypoxia.

     

Phosphorescence quenching method for measurement of intracellular PO2 in isolated skeletal muscle fibers

Values of skeletal muscle intracellularPO₂ during conditions ranging fromrest to maximal metabolic rates have been difficult to quantify. Amethod for measurement of intracellular PO₂ in isolated single skeletalmuscle fibers by using O₂-dependent quenching of aphosphorescent-O₂probe is described. Intact single skeletal muscle fibersfrom Xenopus laevis were dissectedfrom the lumbrical muscle and mounted in a glass chamber containingRinger solution at 20°C. The chamber was placed on the stage of aninverted microscope configured for epi-illumination. A solutioncontaining palladium-meso-tetra(4-carboxyphenyl) porphine bound to bovine serum albumin was injectedinto single fibers by micropipette pressure injection.Phosphorescence-decay curves (average of 10 rapid flashes) wererecorded every 7 s from single cells(n = 24) in which respiration had beeneliminated with NaCN, while the PO₂of the Ringer solution surrounding the cell was varied from 0 to 159 Torr. For each measurement, the phosphorescence lifetime was calculatedat the varied extracellular PO₂ byobtaining a best-fit estimate by using a monoexponential function. Thephosphorescence lifetime varied from 40 to 70 µs at an extracellularPO₂ of 159 Torr to 650-700 µsat 0 Torr. The phosphorescent lifetimes for the variedPO₂ were used to calculate, by usingthe Stern-Volmer relationship, the phosphorescence-quenching constant(100 Torr¹ · s¹),and the phosphorescence lifetime in azero-O₂ environment (690 µs) forthe phosphor within the intracellular environment. This techniquedemonstrates a novel method for determining intracellular PO₂ in isolated single skeletalmuscle fibers.     

Raising P50 increases tissue PO2 in canine skeletal muscle but does not affect critical O2 extraction ratio

Curtis, Scott E., Thomas A. Walker, W. E. Bradley, andStephen M. Cain. Raising P₅₀increases tissue PO₂ in canineskeletal muscle but does not affect criticalO₂ extraction ratio.J. Appl. Physiol. 83(5):1681-1689, 1997.Affinity of hemoglobin (Hb) forO₂ determines in part the rate ofO₂ diffusion from capillaries tomyocytes by altering capillary PO₂.We hypothesized that a decrease in HbO₂ affinity (increasedP₅₀) would increase capillary and tissue PO₂(Pti_O2) andimprove O₂ consumption duringischemia. To test this hypothesis, blood flow to the pump-perfused lefthindlimb of 18 anesthetized and paralyzed dogs was progressively decreased over 90 min while hindlimb O₂ consumption andO₂ delivery (O₂)and Pti_O2 weremeasured at the muscle surface. Arterial PO₂ was maintained at 150 ± 10 Torr in all dogs. We increased P₅₀by 12.3 ± 0.9 (SE) Torr in nine dogs with RSR-13, an allosteric modifier of Hb. This decreased arterialO₂ saturation to 90-92% butincreased meanPti_O2 from 35.5 ± 11.6 to 44.1 ± 15.2 (SD) Torr(P < 0.05) with no change incontrols (n = 9).O₂ extraction ratio at criticalO₂was 74 ± 2% in controls and 79 ± 1% in RSR-13-treated dogs(P = not significant).Pti_O2 was30-40% higher in the RSR-13-treated group at anyO₂above critical but did not differ between groups below criticalO₂.Perfusion heterogeneity and convergence of the dissociation curvesnear criticalO₂ may have mitigated any effect of increasedP₅₀ onO₂ diffusion. Still, increasingP₅₀ by 12 Torr with RSR-13significantly increased Pti_O2 atO₂values above critical.

     

Effects of hyperchloremia on blood oxygen binding in healthy calves

Three different levels of hyperchloremia wereinduced in healthy Friesian calves to study the effects of chloride onblood oxygen transport. By infusion, the calves received either 5 ml/kg of 0.9% NaCl (low-level hyperchloremia; groupA), 5 ml/kg of 7.5% NaCl (moderate hyperchloremia;group B), or 7.5 ml/kg of 7.5% NaCl(high-level hyperchloremia; groupC). Blood was sampled from the jugular vein and thebrachial artery. Chloride concentration, hemoglobin content, arterialand venous pH, PCO₂, and PO₂ were determined. At each timepoint (0, 15, 30, 60, and 120 min), the whole blood oxygen equilibriumcurve (OEC) was measured under standard conditions. Ingroups B andC, hyperchloremia was accompanied by asustained rightward shift of the OEC, as indicated by the significantincrease in the standard PO₂ at 50%hemoglobin saturation. Infusion of hypertonic saline also inducedrelative acidosis. The arterial and venous OEC were calculated, withbody temperature, pH, and PCO₂ valuesin arterial and venous blood taken into account. The degree of blooddesaturation between the arterial and the venous compartments[O₂ exchange fraction(OEF%)] and the amount of oxygen released at tissue level by 100 ml of bovine blood (OEF vol%) were calculated from the arterial andvenous OEC combined with the PO₂ andhemoglobin concentration. The chloride-induced rightward shift of theOEC was reinforced by the relative acidosis, but the alteredPO₂ values combined with the lowerhemoglobin concentration explained the absence of any significantdifference in OEF (% and vol%). We conclude that infusion ofhypertonic saline induces hyperchloremia and acidemia, which canexplain the OEC rightward shift observed in arterial and peripheralvenous blood.

     

Chronic hormone replacement therapy alters thermoregulatory and vasomotor function in postmenopausal women

Brooks, E. M., A. L. Morgan, J. M. Pierzga, S. L. Wladkowski, J. T. O'Gorman, J. A. Derr, and W. L. Kenney. Chronic hormone replacement therapy alters thermoregulatory and vasomotor function in postmenopausal women. J. Appl.Physiol. 83(2): 477-484, 1997.This investigationexamined effects of chronic (2 yr) hormone replacement therapy (HRT),both estrogen replacement therapy (ERT) and estrogen plus progesteronetherapy (E+P), on core temperature and skin blood flow responses ofpostmenopausal women. Twenty-five postmenopausal women [9 not onHRT (NO), 8 on ERT, 8 on E+P] exercised on a cycle ergometer for1 h at an ambient temperature of 36°C. Cutaneous vascularconductance (CVC) was monitored by laser-Doppler flowmetry, and forearmvascular conductance (FVC) was measured by using venous occlusionplethysmography. Iontophoresis of bretylium tosylate was performedbefore exercise to block local vasoconstrictor (VC) activity at oneskin site on the forearm. Rectal temperature (T_re) was ~0.5°C lower forthe ERT group (P < 0.01) comparedwith E+P and NO groups at rest and throughout exercise. FVC: mean body temperature (T_b) and CVC:T_b curves were shifted~0.5°C leftward for the ERT group(P < 0.0001). Baseline CVC wassignificantly higher in the ERT group(P < 0.05), but there was nointeraction between bretylium treatment and groups once exercise wasinitiated. These results suggest that1) chronic ERT likely acts centrally to decrease T_re,2) ERT lowers theT_re at which heat-loss effector mechanisms are initiated, primarily by actions on active cutaneous vasodilation, and 3) addition ofexogenous progestins in HRT effectively blocks these effects.

     

Redox behavior of cytochrome oxidase in the rat brain measured by near-infrared spectroscopy

Hoshi, Yoko, Osamu Hazeki, Yasuyuki Kakihana, and MamoruTamura. Redox behavior of cytochrome oxidase in the rat brain measured by near-infrared spectroscopy. J. Appl.Physiol. 83(6): 1842-1848, 1997.Usingnear-infrared spectroscopy, we developed a new approach for measuringthe redox state of cytochrome oxidase in the brain under normalblood-circulation conditions. Our algorithm does not require theabsorption coefficient of cytochrome oxidase, which differs from studyto study. We employed this method for evaluation of effects of changesin oxygen delivery on cerebral oxygenation in rats. When fractionalinspired oxygen was decreased in a stepwise manner from100 to <10%, at which point the concentration of oxygenatedhemoglobin([HbO₂])decreased by ~60%, cytochrome oxidase started to be reduced.Increases in arterial PO₂ underhyperoxic conditions caused an increase in[HbO₂], whereas further oxidation of cytochrome oxidase was not observed. The dissociation of the responses of hemogloblin and cytochrome oxidase wasalso clearly observed after the injection of epinephrine under severelyhypoxic conditions; that is, cytochrome oxidase was reoxidized withincreasing blood pressure, whereas hemoglobin oxygenation was notchanged. These data indicated that oxygen-dependent redox changes incytochrome oxidase occur only when oxygen delivery is extremelyimpaired. This is consistent with the in vitro data of our previousstudy.