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Biomedical subjects

K P Agrawal

Publications and source records attributed to K P Agrawal.

At least 19 recordsLinked to original sources

Body plethysmographic measurement of thoracic gas volume without panting against a shutter.

When a subject breathes through a pneumotachograph in a body box, the measured value of specific airway resistance (sRaw1) is equal to the product of thoracic gas volume (TGV) and the sum of the airway resistance (Raw) and the instrument resistance (Rins). If an additional resistance (Radd) is put in the breathing path, the measured specific, airway resistance (sRaw2) exceeds sRaw1 by the product of TGV and Radd and can be used for determining TGV. With the use of a device increasing Rins by a known amount (Radd) during normal breathing, sRaw1 and sRaw2 were measured in 3 normal subjects, 16 asthmatic patients, 2 patients with chronic obstructive pulmonary disease, and 1 patient with restrictive lung disease from the slopes of the x-y plots of airflow vs. box signals obtained before and after adding Radd. TGV was calculated by dividing (sRaw2-sRaw1) bu Radd. We also determined subjects' TGV by the panting method of A. B. DuBois, S. Y. Botelho, G. N. Bedell, and J. H. Comroe, Jr. (J. Clin. Invest. 35: 322-326, 1956) and functional residual capacity by the helium-dilution method. The results of the new method were quite reproducible (coefficient of variation = 5.6) and equivalent to those obtained by the other two methods.

Adolescent↗

Successful non-surgical embryo recovery from a goat.

Embryos from a Jamunapari goat were successfully recovered using non-surgical approach. The technique of Bessoudo et al [Theriogenology, 29 (1988) 221] was simplified and all the appliances used were fabricated indigenously. Fluid retrieval rate was 95%. Five excellent embryos (blastocysts) were recovered. The whole process of embryo collection was completed in 30 min.

Animals↗

Increased leukocyte phospholipase A2 activity and plasma lysophosphatidylcholine levels in asthma and rhinitis and their relationship to airway sensitivity to histamine.

This study was carried out to determine leukocyte phospholipase A2 (PLA2) activity and plasma lysophosphatidylcholine (LPC) levels in normal subjects and in patients with asthma and rhinitis and to examine their relationship to airway sensitivity to histamine. Leukocyte PLA2 activity and plasma LPC levels were highly correlated (rs = 0.90), and were found significantly raised in both the disease groups, more so in the asthmatics. Both PLA2 activity and LPC levels showed overall negative correlations with the log dose of histamine producing a 35% fall in specific airway conductance (PD35 histamine) when the patients and the normal subjects were examined together (rs = -0.77 and rs = -0.83, respectively). The patients with leukocyte PLA2 activity more than 1.8 U (nmol 14C-AA released/mg protein/10 min at 30 degrees C) or plasma LPC levels more than 8.7% of total phospholipids (mostly asthmatics) showed high airway sensitivity to histamine (PD35 histamine less than or equal to 1 mg/ml). On the other hand, the PD35 histamine values of patients with leukocyte PLA2 activity equal to or less than 1.96 U or plasma LPC levels equal to or less than 8.7% overlapped with those of normal subjects in the range of 1.2 to 10 mg/ml. Lowering of plasma LPC levels, which probably reflect tracheobronchial LPC content as shown in guinea pigs, seems to be an essential step in the return of airway reactivity toward normal.

Adult↗

Survival of mouse embryos after being frozen in glycerol-sucrose mixture.

Random bred female albino mice (6-8 weeks old) were used as a source of embryos. 8- to 16 cell embryos were dehydrated in glycerol-sucrose mixture in 0.25 ml straws at room temperature. Straws were cooled at the rate of 5 degrees C/min to -7 degrees C. Seeding was induced by touching the out side of the straw at -7 degrees C. Straws were further cooled at 0.5 degree C/min down to -35 degrees C and then plunged into liquid N2. Thawing of straws was done by direct transfer into water at 35 degrees C. Frozen-thawed embryos were cultured in a CO2 incubator maintained at 39 degrees C. Out 190 embryos (8-16 cell) initially frozen, 169 (88.94%) were recovered on thawing. 158 (93.5%) out of 169 were apparently normal and used for culture. 75 (47.46%) developed to morulae/early blastocysts and 72 (45.56%) to expanded blastocysts on 24 and 48 hr culture respectively. In conclusion, the incorporation of sucrose in the freezing medium at a concentration of 0.25 M has led us to propose a freezing, thawing and transfer method without dilution of glycerol. The technique being quite simple is worth trying in farm animals where importance of this technique in non-surgical transfer of frozen-thawed embryos will be a boon.

Animals↗

A protocol used for splitting mouse embryos into two halves.

8-16 cell embryos and early blastocysts were obtained from the oviducts and anterior portion of uterine horns of albino mice at 70 and 90 hr after LH injection respectively. Splitting of embryos was done by using two microtools attached to a micromanipulator unit (Research Instruments Ltd, UK). After bisection, each pair of the half embryos is transferred to a dish containing 2 ml of T-6 medium and cultured in CO2 incubator (at 39 degrees C, 95% RH and 5% CO2 in air mixture). Splitting of blastocysts as compared to 8-16 cell embryos was found difficult (35.48% vs 52.44%, respectively). 38.88% of bisected 8-16 cell embryos and 11.36% of bisected blastocysts developed on 48 hr culture. Information on splitting mouse embryos and their subsequent development in culture are significant in view of using the technique for commercial application and for research in developmental biology of animal embryos.

Animals↗

Cryopreservation of mouse embryos at -196 degrees C by vitrification.

Embryos (8-16 cell) were obtained from random bred albino mice (6-8 weeks old) that were induced to superovulate by injections of 5 I.U. PMSG and 5 I.U. hCG given 48 hr apart. Embryos were exposed to intracellular cryoprotecting medium (glycerol 10%, 1-2 propanediol 20% in PBS) for 10 min and then transferred to extracellular vitrification medium (25% glycerol, 25% 1-2 propanediol in PBS). Vitrification medium containing embryos, and diluent (1 M sucrose) were loaded in a straw and immediately plunged into liquid N2. After thawing at 20 degrees C, the contents of the straw were mixed by shaking (1 step dilution) and emptied in a petri dish. After 3 washings in culture medium the embryos were kept in CO2 incubator for further development. In 3-step dilution procedure the dilution of cryoprotectants was done in 0.5 and 0.25 M sucrose before culture. Embryos in 3-step dilution of cryoprotectants exhibited high survival as compared to 1-step dilution (20.23% vs 6.55%).

Animals↗

Metabolic basis of asthma. A united hypothesis.

Asthma is a heterogenous disease triggered by a large number of different stimuli. This article presents a theory of the metabolic mechanisms of asthma. The theory is based on the growing understanding of the activity of lysophosphatidylcholine (LPC). Since the effect of LPC on cell membranes, membrane bound enzymes and the various types of cells involved in the pathogenesis of asthma, this may represent a unifying link between the various types of asthma.

Animals↗

Airway responses to inhaled ouabain in subjects with and without asthma.

Challenges with ouabain and histamine were performed a week apart in 10 patients with asthma and 5 normal subjects. Concentrations were increased cumulatively until specific airway conductance decreased by 30% or the maximal concentration of 1.0% was reached. At low concentrations, ouabain induced bronchodilatation in six patients who had asthma. Bronchodilatation gradually decreased with increasing concentrations and was followed by bronchoconstriction in two patients with asthma who had high airway sensitivity to histamine. Ouabain caused only bronchoconstriction in three patients with severe asthma. The normal subjects showed mild bronchodilatation or no response to ouabain. Several possible biochemical mechanisms may be responsible for the bronchodilatory response to low doses of ouabain, such as stimulation of adenylate cyclase or (Na+,K+)-adenosine triphosphatase. The absence of a bronchodilatory response to ouabain in patients with severe asthma suggests an impairment in the activity of these enzymes.

Adult↗

Airway responses to inhaled ouabain and histamine in conscious guinea pigs.

Tracheal Na+-K+-ATPase activity is positively correlated with in vivo airway responsiveness to histamine. We wondered whether this were a chance association or whether it was directly related to the mechanism of hyperreactivity. Therefore, we obtained dose-response curves to aerosols of histamine and ouabain in guinea pigs to determine whether an in vivo relationship existed between the excitatory effects of histamine and the enzyme-inhibiting effect of ouabain. Airway responsiveness to ouabain was measured as the ouabain concentration producing a 30% decrease in specific airway conductance (ED30) or that producing a half-maximal response (ED50). Responsiveness to histamine was measured either as ED30 or as ED50. Significant positive correlations were noted between the log ED50 of ouabain and log histamine ED30 or ED50 (r = 0.81 and 0.83, respectively; P less than 0.001), and between log ouabain ED30 and log histamine ED30 and ED50 (r = 0.76 and 0.77, respectively; P less than 0.002). Pretreatment with ouabain increased airway responsiveness to histamine (P less than 0.05). We suggest that in hyperreactive airways Na+-K+-ATPase serves a homeostatic function of preventing Na+ and Ca2+ loading of the cell and that it is not directly responsible for the hyperreactivity.

Aerosols↗

Quiet-breathing vs. panting methods for determination of specific airway conductance.

Specific airway conductance (sGaw) was measured during quiet breathing and during panting in 21 normal subjects and 10 patients with obstructive lung disease. The direct method used does not require measuring thoracic gas volume (TGV). Coefficients of variation were 5.5% for panting and 5.1% for quiet breathing. Interobserver variability was 4.7% in the quiet-breathing method and 6.3% in the panting method. The two methods gave equivalent results for sGaw. A slightly greater sGaw was found by the panting method in normal subjects with the highest sGaw values, probably due to widening of the oropharynx-glottis during panting. In six normal subjects studied for intrasubject variability over time, no significant diurnal or day-to-day variability was seen by either method. We conclude that the quiet-breathing method is a simple valid means of determining sGaw and utilizes a physiological respiratory maneuver. Obviation of the need to measure TGV is advantageous. Results are equivalent to those of the panting method and variability is similar.

Adult↗

Biochemical correlates of airway hyperreactivity in guinea pigs: role of lysophosphatidyl choline.

To elucidate the biochemical basis of airway hyperreactivity, we studied the relationships between in vivo airway sensitivity of guinea pigs to histamine and their tracheal beta-adrenergic binding sites, Ca++- and (Na+-K+)-ATPase activities, and composition of phospholipids. The relationships between tracheal and plasma phospholipids were also examined. beta-Adrenergic receptor binding with 3H-dihydroalprenolol in tracheal tissue showed an inverse relationship with in vivo airway sensitivity to histamine. Among the phospholipids, tracheal phosphatidyl ethanolamine content varied inversely with in vivo airway sensitivity, whereas tracheal and plasma lysophosphatidyl choline contents showed a direct correlation with airway sensitivity. A significant direct correlation was also observed between tracheal and plasma lysophosphatidyl choline levels. Both Ca++-ATPase and (Na+-K+)-ATPase activities increased with increasing airway sensitivity. These enzymes showed inverse correlations with phosphatidyl ethanolamine content and direct correlations with lysophosphatidyl choline content. Our data suggest that increased lysophosphatidyl choline may cause various biochemical changes associated with airway hyperreactivity.

Animals↗

Specific airways conductance in guinea pigs: normal values and histamine induced fall.

A differential body plethysmograph was developed for direct determination of specific airway conductance (SGaw) in intact conscious guinea pigs, breathing spontaneously at room temperature. Box pressure changes were measured as differences between chest volume (delta V1) and air volume respired at atmospheric pressure (delta V2) and fed to X-axis of an X-Y recorder. Airflow signal was fed to its Y-axis. A loop was formed. The slope of the rising limb of this loop provided the ratio of airflow change during transition from expiration to inspiration (delta V2) and corresponding delta (V1-V2), which is practically free from "temperature-humidity artifact" because of minimal lung volume change over this period. This ratio delta V2/delta (V1-V2), when divided by barometric pressure after subtracting water vapour pressure at body temperature (PB-PH2O), gave the value of SGaw at normal end expiration, i.e., at functional residual capacity (FRC). In 42 male guinea pigs, used in this study, SGaw was found to be 0.48 +/- 0.06 sec-1 (cm H2O)-1, comparable to the value obtained by using available data regarding lung resistance and FRC. It was independent of body weight (250-750 g). Histamine induced fall in SGaw was determined by comparing the slopes of the rising limbs of the X-Y loops obtained after and before exposure to histamine aerosol. Log-normal plots of histamine base (W/V%) and SGaw, as % of control, were used to assess airway reactivity.

Airway Resistance↗

Fall in specific airway conductance at residual volume in small airway obstruction.

A technique of body plethysmography has been developed for measuring specific airway conductance (SGaw) at residual volume (RV). Box pressure variations were recorded as difference between chest volume change and air volume respired at atmospheric pressure, delta(V1--V2). They could easily be related to alveolar pressure change per liter of lung volume and SGaw could directly be calculated without breathing against a closed shutter. To eliminate 'temperature-humidity artifact', calculations were made near end-tidal points where air movement in and out of lungs is minimal. For that purpose airflow changes were related to box pressure changes over a period of 0.2 sec from the beginning of inspiratory effort. SGaw was measured at FRC as well as at RV. Excessive air trapping at RV in some cases resulted in some chest expansion unaccompanied by any airflow at mouth. This has been termed pre-flow work (PFW). Any abnormal fall in SGaw at RV or the presence of PFW could be used as an indicator of small airway obstruction. Using this method obstruction in small airways was found in all smokers with smoking history of 5 to 30 packs years (mean 13.1 pack years) and in 4 patients who had a history of cough and/or dyspnoea or exertion. Simultaneous measurements of SGaw at FRC and SGaw 0.2 sec and PFW at RV could be of great help in determining the degree and site of airflow obstruction.

Adult↗