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Biomedical subjects

D L Reid

Publications and source records attributed to D L Reid.

At least 19 recordsLinked to original sources

Cell attachment, collagen binding, and receptor analysis on bovine articular chondrocytes.

The purpose of this study was to investigate collagen receptors on primary bovine articular chondrocytes from full-thickness and different layers of bovine articular cartilage. Cytometric studies with antibodies showed that approximately 56% of the chondrocytes from the superficial layer and 29% of the chondrocytes from the deep layer bound anti-annexin V. A similar tendency was found for alpha5 and beta1 integrin antibodies. Flow cytometric analysis initially detected annexin V on chondrocytes following isolation; the level of detection subsequently decreased by 24 hours, whereas that of alpha5 and beta1 integrins increased. Treatment of chondrocytes with collagenase at 24 hours restored the initially high epitope recognition of annexin V, indicating masking of annexin V by newly formed collagen matrix. There was little effect on detection levels for beta1 integrin. Contrary to the specific matrix receptor expression, chondrocytes from superficial and deep layers differed little in attachment to immobilized types I and II collagens. However, the attachment was more effectively inhibited with anti-annexin V than with integrin antibodies. Competition studies with preparations of soluble collagens revealed a preferential binding of bovine type-II collagen compared with bovine type-I collagen. Anti-annexin V antibodies inhibited binding of type-II collagen more effectively than anti-alpha5 or anti-beta1 integrin antibodies. Evidently, under the in vitro conditions of this study, annexin V is the quantitatively predominant type-II collagen receptor on bovine articular chondrocytes. This opens a discussion of the possibly dualistic metabolic/mechanical annexin V-integrin receptor elements.

Animals↗

Absence of cell-surface annexin V is accompanied by defective collagen matrix binding in the Swarm rat chondrosarcoma.

Annexin V has been characterized as a major collagen type II binding cell-surface component of normal chondrocytes and is also called anchorin CII in chondrogenic populations. Herein we present evidence that in vitro cultured Swarm rat chondrosarcoma cells are not capable of binding collagen type II in significant quantities to their surfaces, as compared to normal rat chondrocytes. This finding coincides with a deficiency of annexin V on the surface of these cells. A small quantity of an intracellular polypeptide could be detected which is immunologically cross-reactive with annexin V but displayed a mobility in SDS-PAGE of less than 34 kD compared to the M(r) 36 kD of intact rat annexin V. By immunohistochemistry the protein could be localized in the cytoplasm of in vitro and in vivo grown tumor cells. By reverse transcription-polymerase chain reaction and Northern blot analysis, a regular-sized mRNA for annexin V could be detected in the chondrosarcoma cells that is expressed in only slightly lower quantities than in normal chondrocytes. Taken together, the data suggest a modified processing or turnover for annexin V in the chondrosarcoma excluding it from being a functionally active collagen type II binding protein. The findings support the hypothesis of cell-surface annexin V as a key component for the formation of the pericellular matrix of chondrocytes.

Amino Acid Sequence↗

Effects of ductus arteriosus occlusion on pulmonary artery pressure during in utero ventilation in fetal sheep.

Seven fetal sheep were prepared to study the short-term effects of in utero ventilation and ductus arteriosus occlusion on pulmonary artery pressure and on fetal right ventricular function assessed using the right atrial pressure-right ventricular stroke volume relationship. Nine days post-surgery (140 days gestation), blood gas and haemodynamic values were obtained before and during in utero ventilation with 100% O2, and during ventilation with the ductus arteriosus occluded. Oxygen content increased significantly from 7.2 to 14.5 ml dl-1 with ventilation and remained elevated at 14.4 ml dl-1 with ventilation with the ductus arteriosus occluded. In utero ventilation produced a left to right atrial pressure gradient and depression of the right atrial pressure-right ventricular stroke volume relationship. Ductus arteriosus occlusion during in utero ventilation reduced the left to right atrial pressure gradient, and along with a decrease in pulmonary artery pressure, resulted in an upward shift of the right atrial pressure-right ventricular stroke volume relationship, but only to the preventilation level. This study indicates that the fetal right atrial pressure-right ventricular stroke volume relationship is significantly altered, both by changes in the left to right atrial pressure gradient and by changes in pulmonary artery pressure seen with in utero ventilation and subsequent ductus arteriosus occlusion.

Animals↗

Spectral analysis of antepartum fetal heart rate variability from fetal magnetocardiogram recordings.

Fetal heart rate variability was derived from fetal magnetocardiogram recordings in ten subjects at gestation ages 32-38 weeks. Maternal interference was negligible and R-wave detection was highly reliable. Oscillations suggestive of respiratory sinus arrhythmia (RSA) were prominent in many of the heart rate tracings. Spectral analysis was used to quantify heart rate variability and to examine the influence of the RSA-like oscillations on heart rate variability. The oscillations were associated with increased power in the frequency range 0.4-1.0 Hz (P < or = 0.05). Magnetic recording appears to offer significant advantages for investigation of beat-to-beat fetal heart rate throughout the latter stages of pregnancy.

Adult↗

Differential effects of intravenous hydralazine on myoendometrial and placental blood flow in hypertensive pregnant ewes.

OBJECTIVE: The differential vasoactive effects of hydralazine on the uteroplacental vascular bed were studied. STUDY DESIGN: After control measurements were taken, near-term chronically prepared pregnant sheep were continuously infused with angiotensin II. Maternal arterial pressure was increased by 32 mm Hg. Hydralazine was then administered; the effects on regional resistance and blood flow were evaluated with a radionuclide-labeled microsphere technique. Analysis of variance for repeated measures was used to compare observations. RESULTS: When compared with the hypertensive state, hydralazine caused the following changes by 40 minutes (mean +/- SEM): Although maternal blood pressure fell 31% +/- 5% (p = 0.0005), placental blood flow was unchanged, total uteroplacental blood flow increased 24% +/- 8% (p = 0.03), total uteroplacental resistance decreased 43% +/- 4% (p = 0.0002), placental resistance decreased 19% +/- 9% (p = 0.01), myoendometrial blood flow increased 390% +/- 82% (p = 0.0005), and myoendometrial resistance decreased 82% +/- 4% (p = 0.0005). CONCLUSIONS: In angiotensin II-induced hypertensive ewes, hydralazine is an effective dilator of the uteroplacental vascular bed and can maintain placental blood flow while blood pressure.

Adrenal Glands↗

Influence of terbutaline on ovine uterine response to prostaglandin E2 challenge.

OBJECTIVE: Our purpose was to test the effects of terbutaline on uterine electric and contractile responses to prostaglandin E2. STUDY DESIGN: In five late-gestation ewes, prostaglandin E2 (22.9 +/- 2.3 micrograms/min for 3 minutes) was given twice at 30-minute intervals during control. Terbutaline sulfate (2 micrograms/min) was then infused for 30 minutes. Prostaglandin E2 challenge was repeated 10 minutes after the onset of terbutaline infusion and thereafter at 30-minute intervals. Two-way analysis of variance for repeated measures revealed a significant main effect for time (p less than or equal to 0.0001) and between time and response (p less than or equal to 0.05). RESULTS: Both electric (p less than or equal to 0.0001) and intrauterine pressure (p less than or equal to 0.0001) responses were suppressed during terbutaline. The influence on intrauterine pressure persisted 10 minutes after terbutaline (p less than or equal to 0.01) while the electric response was not different from control. CONCLUSIONS: Terbutaline initially diminishes both uterine contractile and electric activity, but electric recovery precedes contractile recovery.

Animals↗

Fetal heart rate variability and behavioral state: analysis by power spectrum.

OBJECTIVE: We attempted to determine the relationship between the fetal heart rate power spectrum and fetal state. STUDY DESIGN: Interbeat intervals, electrocortical activity, and fetal breathing movements were recorded from five near-term fetal lambs. Interbeat intervals were taken from epochs of low-voltage electrocortical activity with breathing, low-voltage electrocortical activity without breathing, and high-voltage electrocortical activity without breathing. Power spectral techniques were applied to determine the underlying frequencies contributing to fetal heart rate variability. Spectral analysis was also performed on fetal breathing data from three animals. RESULTS: Significant differences were found between low-voltage electrocortical activity with breathing and high-voltage electrocortical activity without breathing at 0.62 Hz and from 1.09 to 1.56 Hz. There was no clear relationship between the breathing and heart rate spectra. CONCLUSIONS: Fetal heart rate is mediated by both state and respiratory variables. The respiratory component is not strictly related to respiratory rate.

Animals↗

Effects of severe reduction in maternal placental blood flow on blood flow distribution in the sheep fetus.

To test the hypothesis that fetal lambs are able to maintain oxygen delivery to myocardial, brain and adrenal tissues during reduction in uterine blood flow to 25% of control, we performed experiments on five ewes and their fetuses. A snare occluder was placed around the maternal common hypogastric artery and catheters were placed for measurement of blood pressures, flows, blood gas tensions, pH and oxygen content. After a five day recovery period, control measurements were made. The snare occluder was then closed until the artery was fully occluded. The arterial occlusion caused uteroplacental blood flow to fall to 32 +/- 4% and maternal placental blood flow to fall to 25 +/- 3% of control values. This level of asphyxia was maintained for 19 +/- 3 minutes, when maternal and fetal blood flows were measured again. In response to occlusion, fetal ascending aortic PO2 fell from 21 +/- 2 (SEM) to 13 +/- 2 mmHg (P less than or equal to 0.01), oxygen content from 4.3 +/- 0.3 to 1.4 +/- 0.2 mM (P less than or equal to 0.01) and pH from 7.37 +/- 0.01 to 7.21 +/- 0.05 (P less than or equal to 0.01). PCO2 rose from 48 +/- 1 to 62 +/- 3 mmHg (P less than or equal to 0.01). Fetal arterial blood pressure increased from 51 +/- 3 to 61 +/- 3 mmHg (P less than or equal to 0.001) and heart rate decreased from 172 +/- 10 to 104 +/- 4 beats.min-1 (P less than or equal to 0.01). The heart, brain and adrenals showed vasodilation in response to the asphyxic stimulus.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenal Glands↗

The pathophysiology of the anophthalmic socket. Part I. Analysis of orbital blood flow.

A wide variety of complications of the anophthalmic socket develop in patients after enucleation, including enophthalmos, superior sulcus deformities, eyelid malpositions, implant migration and extrusion, poor prosthetic motility, and socket contraction. Changes in the orbital blood flow and metabolic activity of the socket tissues and atrophy of the orbital fat occurring after enucleation have been suggested as two theoretical mechanisms that result in the development of these clinical conditions. Lack of scientific evidence and a limited understanding of the pathophysiologic basis of the features of anophthalmos led us to evaluate the validity of these proposed mechanisms in an animal model. Selected parameters of the normal orbits were compared with the contralateral anophthalmic orbits at different time intervals after surgery. Orbital blood flow was studied with selective ophthalmic artery angiography and radioactive microsphere techniques. Ophthalmic arteriography demonstrated symmetric caliber and filling characteristics of the major orbital vessels of the control and experimental orbits, although their topographic course was slightly more tortuous in the anophthalmic socket. Results of radioactive microsphere analysis of capillary blood flow per weight of the different orbital tissue compartments of the animals in the long-term group showed no significant difference between the normal and anophthalmic sockets. These findings provide evidence that the circulation dynamics and blood flow to orbital tissues do not change after enucleation surgery.

Angiography↗

The pathophysiology of the anophthalmic socket. Part II. Analysis of orbital fat.

The pathophysiologic mechanisms responsible for the clinical features of the anophthalmic socket are poorly understood. Atrophy of orbital fat has been thought to be a major contributing cause of enophthalmos and the superior sulcus deformities that develop after enucleation, but it has never been demonstrated histopathologically or confirmed by scientific analysis. This study was undertaken to investigate the changes that occur in the orbital fat compartment of the anophthalmic socket in an animal model by measuring orbital soft tissue mass and evaluating adipocyte cell size. Instead of reduction in the tissue mass, a statistically significant greater weight of the fat and connective tissue compartment was found in the anophthalmic orbit by nearly 13% compared to the control orbit in the animals in the long-term group. No significant change in the mean maximal diameter of adipocytes developed 7 months after enucleation. These analyses do not support the concept that orbital fat atrophy or a reduction of metabolic activity occurs in the anophthalmic socket in this animal model. From these results and our previous findings that the circulation dynamics and blood flow to orbital tissues do not change after enucleation, we propose that the pathophysiologic basis of the problems associated with anophthalmos is a disturbance in the spatial architecture and interrelationships of the multiple tissue components of the orbit, not a change in the orbital blood flow or development of fat atrophy.

Adipose Tissue↗

Pulmonary pressure-flow relationships in the fetal lamb during in utero ventilation.

Pressure-flow relationships in the ventilated lung have not been previously determined in undelivered fetal sheep. Therefore we studied 11 late-gestation chronically prepared fetal sheep during positive-pressure ventilation with different gas mixtures to determine the roles of mechanical distension and blood gas tensions on pressure-flow relationships in the lung. Ventilation with 3% O2-7% CO2 produced a substantial fall in pulmonary vascular resistance even though arterial blood gases were not changed. Increases in pulmonary arterial PO2 during ventilation were associated with falls in pulmonary vascular resistance beyond that measured during mechanical distension. Decreases in pulmonary arterial PCO2 and associated increases in pH were also associated with falls in pulmonary vascular resistance. Pulmonary blood flow ceased at a pulmonary arterial pressure that exceeded left atrial pressure, indicating that left atrial pressure does not represent the true downstream component of driving pressure through the pulmonary vascular bed. The slope of the driving pressure-flow relationship in the normal mature fetal lamb was therefore different from the ratio of pulmonary arterial pressure to pulmonary arterial flow. We conclude that mechanical ventilation, increased PO2 and decreased PCO2, and/or increased pH has an important influence on the fall in pulmonary vascular resistance elicited by positive pressure in utero ventilation of the fetal lamb and that the downstream driving pressure for pulmonary blood flow exceeds left atrial pressure.

Animals↗

High frequency differences between high and low voltage electrocorticograms in the ovine fetus.

Dawes (1986) has stated that, "The difference between high and low voltage activity depends solely on the presence in the latter of higher amplitude oscillations with relatively low frequency superimposed on the low voltage components as shown by spectral analysis". We have tested the constancy of the high frequency section of the power spectrum of the electrocorticogram in 7 near-term sheep fetuses. Under sterile conditions we implanted biparietal electrodes in the dura and a ground lead subcutaneously on the scalp. Five days after the surgery, with the animal standing quietly in the laboratory, we acquired the fetal electrocorticograms. Data were acquired during several high and low voltage electrocorticographic cycles in each animal. Two hundred power spectra were obtained during high and low voltage fetal electrocortical activity and statistically analyzed by paired t-test to discern differences in power between the high and low voltage pattern at each frequency (n = 7). We found that at all frequencies between 4 and 12 cycles/s the high voltage electrocorticogram had significantly more power than the low voltage electrocorticogram (P less than 0.05). This is in accordance with the established literature. We also observed that from 17 through 24 cycles/s the low voltage electrocorticogram is significantly higher than the high voltage electrocorticogram (P less than 0.05). In this frequency range the power of the high voltage expressed as a percentage of the power of the low voltage were respectively, 80, 74, 71, 66, 64, 64, 67, 64. These differences are of considerable magnitude and may be physiologically important.

Animals↗

Adenosine causes a biphasic response in the ovine fetal placental vasculature.

We have reported in a previous study that adenosine infusion causes fetal placental vascular resistance to increase after 2 min. To determine whether this action is followed by a more prolonged vasodilation, we studied 7 mature fetal lambs. At surgery, catheters were inserted into the fetal hindlimb arteries and veins. After a five day recovery period, control blood flow measurements were made by radiolabeled microsphere technique immediately after an infusion of 0.9% NaCl, (vehicle, 1.03 ml.min-1) into a fetal vein for 2 min. Within 5 min of the control blood flow measurement, adenosine (10 mg/min) was infused for 2 min. Blood flow measurements were repeated 5, 10, 15, 20 and 30 min after the end of the infusion period. Fetal arterial blood pressure dropped from 50 +/- 1 to 34 +/- 5 mmHg immediately after the adenosine infusion and returned to the control value within 5 min after the infusion. No further blood pressure response was detected. However, placental vascular resistance fell from 0.334 +/- 0.040 to 0.269 +/- 0.027 (P less than 0.05) at the 15 min measurement, remained low through the 20 min measurement (P less than 0.001) and was not different from control levels 30 min after the adenosine infusion. We conclude that the fetal placental vasculature responds to systemic adenosine infusion in a biphasic manner. The immediate reaction to adenosine is a transient vasoconstriction in the fetal placental vasculature followed by vasodilation 15 to 20 min after the initial exposure to adenosine.

Adenosine↗

Relationship between plasma catecholamine levels and electrocortical state in the mature fetal lamb.

In late pregnancy the electrocortical activity (ECoG) in the ovine fetus starts to cycle between high and low voltage states. During the high voltage states of this activity fetal regional blood flows are decreased, and heart rate and fetal arterial blood pressure are both increased. Jensen et al. (1986) have postulated that these changes may be mediated by changes in autonomic tone. To test this hypothesis we placed catheters in 6 near-term sheep fetuses (gestational age = 128 days) and implanted electrodes to measure electrocortical activity. Five days after the surgery, fetal arterial blood was withdrawn during the first 3-5 min of each high and low voltage ECoG for 5 full cycles in each fetus. Plasma samples were analyzed for epinephrine, norepinephrine, and dopamine. Dopamine levels were not different in high and low voltage electrocortical state. In the high voltage ECoG state, epinephrine levels were 75 +/- 7 pg/ml and fell to 34 +/- 4 pg/ml during low voltage ECoG (P less than 0.01). During the high voltage state plasma norepinephrine was 623 +/- 85 pg/ml and fell during the low voltage period to 462 +/- 99 pg/ml (P less than 0.01). These data demonstrate that in the near-term sheep fetus plasma catecholamine levels fluctuate with ECoG state.

Animals↗

Uncoupling of excitation from contraction in uterine smooth muscle in near-term ewes.

We infused forskolin in 30% ethanol or 30% ethanol systemically in seven chronically catheterized near-term sheep to determine the response of ovine uterine musculature to cyclic adenosine monophosphate stimulation. Maternal and fetal arterial pressure, fetal venous pressure, intrauterine pressure, and uterine electromyogram activity were monitored continuously. Prostaglandin E2 was infused at a delivery rate that caused a minimal 5 mm Hg increase in intrauterine pressure with definite contraction-like pressure spikes and associated uterine electromyogram activity. Forskolin (10 mg in 30% ethanol) or ethanol (vehicle) was then infused for 20 minutes. The prostaglandin E2 challenge was repeated 3 minutes later and again every 30 minutes over a 2 1/2-hour period. Data were analyzed by repeated measures analysis of variance. Infusion of ethanol (n = 2) had no apparent effect on either the intrauterine pressure or the uterine electromyogram response to prostaglandin E2 challenge. Forskolin infusion (n = 5) caused an attenuation of the intrauterine pressure response to prostaglandin E2 for 93 minutes. This effect was greatest 33 minutes after the infusion ended when the integrated pressure signal was 37% of the initial prostaglandin E2 response. The forskolin infusion had no effect on uterine electromyogram response to prostaglandin E2. We conclude that forskolin causes an uncoupling of excitation from contraction in the intact near-term ovine uterus.

Animals↗