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Biomedical subjects

J Mitra

Publications and source records attributed to J Mitra.

At least 37 records · Page 2Linked to original sources

Purification and characterization of rat uterine glycerylphosphorylcholine diesterase and its tissue-specific induction by 17 beta-estradiol.

The rat uterine enzyme glycerylphosphorylcholine (GPC) diesterase found in the proestrous secretions was purified and characterized biochemically with respect to its subunit mol wt, native size, pI, and amino acid and carbohydrate composition. The 30-kDa protein was assessed to have a native mol wt of 105 kDa, as determined by analytical gel filtration. It had a basic pI with an unusually high carbohydrate/protein ratio (0.83:1). The estrogen inducibility of the protein was identified by its de novo synthesis in vitro after in vivo 17 beta-estradiol administration. For this experiment, uteri from estradiol-treated immature rats were incubated in the presence of [14C]glucosamine, and the 14C-labeled total glycoconjugates were then subjected to the enzyme purification procedure. The peak enzyme fraction was observed to correspond to one of the 14C-labeled protein peaks in the elution profile of the glycoconjugates. Assay of GPC diesterase activity showed significant enhancement only in the uterus, not in other organ systems, after in vivo estradiol treatment, and this hormone-stimulated increase was inhibited by nafoxidine, a potent estrogen antagonist. The enzyme activity profile of the three main separated uterine cellular types showed that the enzyme was secreted by epithelial cells. No antigenic homology of the enzyme was observed with GPC diesterase from nonmammalian sources. The results suggested that GPC diesterase was one of the estrogen-regulated proteins of the preovulatory uterine fluid of rats, and the availability of a purified preparation of the enzyme could serve as a useful tool to study the mechanism of estrogen action.

Amino Acids↗

Type I protein S deficiency and skin necrosis.

A kindred with Type I protein S deficiency is described in which the index case developed skin necrosis during induction of oral anticoagulant therapy for deep venous thrombosis. Two other family members with protein S deficiency have been detected, and demonstrate the clinical variability of this condition.

Acenocoumarol↗

Immunological status of women with prolonged oral contraceptives and occurrence of giardiasis.

Immunological status of 14 women taking oral contraceptive for prolonged period (more than 6 months, low dose pill) was studied. Phytohaemagglutinin induced lymphocytes stimulation (PILT) was found to be significantly depressed in these subjects though T-lymphocyte subpopulation was found to be normal. Serum IgA and IgG levels in these cases were found to be normal but IgM level was increased. Though incidence of G lamblia infection was reported to be high in immunocompromised subjects, it was found only one out of 14 subjects taking oral contraceptive harboured the protozoa.

Adolescent↗

Glycerylphosphorylcholine diesterase activity of uterine fluid in conditions inducing secondary sex ratio change in the rat.

Food restriction or decreasing the ratio of [Na+, K+] to [Ca++, Mg++] in the diet of female rats before conception favoured the production of female offspring. Seven days of food deprivation decreased uterine fluid GPC diesterase activity in female rats, whereas long-term food restriction (21 days), rather than decreasing the enzyme activity, apparently stimulated it. Dietary Ca and Mg supplementation, likewise, produced significant decrease in GPC diesterase activity. A significantly positive correlation was observed between the levels of uterine GPC diesterase and secondary sex ratio change, which indicates that these dietary techniques of sex-ratio manipulation may modulate the uterine fluid GPC diesterase activity and influence sex determination in utero.

Animals↗

Effects of DDT on the growth of crop plants.

The effects of DDT on the germination and growth of plants were studied using many crop species. Of the species tested, oil-rich seeds of plants, such as peanut (Arachis hypogaea) and mustard (Brassica juncea), were more prone to DDT induced inhibition of germination and subsequent plant growth than cereals, pulses and fibre crops, like rice (Oryza sativa), barley (Hordeum vulgare), mung bean Vigna radiata), pigeon pea (Cajanus cajan) and cotton (Gossypium hirsutum). Studies with (14)C labelled DDT showed that insecticide uptake by seeds was directly proportional to seed size. However, there was no direct relationship between DDT uptake by the seeds and its subsequent translocation to the growing regions or the degree of growth inhibition. Data suggest that oil content of the seeds per se has a bearing on the susceptibility or tolerance of a plant to DDT. It is suggested that lipids of the plant cell solubilize and disperse DDT in the cytoplasm, which, in turn, affects normal metabolism within the cell.

Journal Article↗

Integration of cardiorespiratory responses in the ventrolateral medulla.

Interventions confined to the region adjacent to the VMS can produce both respiratory and circulatory effects. Although it has been suggested that both breathing and vasomotor changes arise from the same neural elements near the VMS, our own investigations indicate that the neurons involved are closely linked but not identical. This belief is supported by recent studies which show that AII and angiotensin antagonists microinjected into the rostral portion of the VMS can significantly modify blood pressure and respiration but can produce effects of different sign. These observations, coupled with previous studies of the VMS, indicate the possibility that regions near the VMS may contribute to integration of circulatory and respiratory responses.

Animals↗

Sustained hypertransfusion and induction of a transplantable myeloid leukemia in RLV-A-infected BALB/c mice.

Infection of BALB/c mice with the RLV-A virus typically results in an erythropoietic dysplasia characterized by hepatosplenomegaly, erythroblastosis, erythroblastemia and severe anemia without reticulocytosis. Mice hypertransfused weekly with 75%-packed red cells for 42 days prior to RLV-A infection and viral potency controls manifested this typical RLV-A response. Mice that were hypertransfused prior to and following RLV-A infection never developed the "typical" RLV-A pathogenesis. Instead, a transplantable myeloid leukemia was established. Although the reason for altered pathogenesis remains uncertain, it seems plausible that continued hypertransfusion, presumably after establishment of an altered granulopoietic microenvironment, resulted in a completely different viral expression and development of the transplantable myeloid leukemia.

Animals↗

Involvement of ventral medullary surface in respiratory responses induced by 2,4-dinitrophenol.

We examined the contribution of the neural elements near the ventral medullary surface (VMS) to the respiratory response caused by 2,4-dinitrophenol (DNP). Two series of experiments were performed on 12 vagotomized and sinoaortic denervated cats. The first series examined the effect of focal cooling of the VMS on the respiratory response to DNP in four spontaneously breathing, anesthetized cats. When the VMS temperature was 37 degrees C, systemic administration of DNP increased minute ventilation under nearly isocapnic conditions, and focal cooling of the intermediate area of VMS to 20 degrees C attenuated the ventilatory augmentation caused by DNP. To eliminate the influence of anesthetics, a second group of experiments was performed on eight decerebrate, artificially ventilated cats while phrenic nerve activity was monitored as an index of respiration. AgNO3 (10%) was topically applied to the VMS until the respiratory response to inhaled CO2 was abolished. Apnea occurred in seven of eight cats after AgNO3, whereas in the remaining one animal, tidal phrenic activity decreased substantially. Systemic administration of DNP produced no respiratory excitation in any of the animals. On the other hand, rhythmic respiratory activity could be provoked by electrical stimulation of the mesencephalic locomotor area and carotid sinus nerve and by excitation of somatic afferents. Histological examination of the brain stem showed that the AgNO3 had penetrated no more than 350 microns from the ventral medullary surface. These results indicate superficial structures of the VMS are of potential importance in mediating the respiratory responses to hypermetabolism.

2,4-Dinitrophenol↗

Respiratory effects of N-methyl-D-aspartate on the ventrolateral medullary surface.

We studied the central effects of N-methyl-D-aspartate (NMDA) on respiration in 18 artificially ventilated cats anesthetized with alpha-chloralose. Unilateral topical application of NMDA (1 x 10(-8) mol) to the intermediate region of the ventrolateral medulla exaggerates the phrenic response to CO2 at end-tidal PCO2 levels of less than 50.0 Torr. At higher end-tidal PCO2 levels, however, such differences disappear. Unilateral NMDA application increases the activity of the right and left phrenic nerves equally. Furthermore, the magnitude of the phrenic response after unilateral application of NMDA was not different from that after bilateral application. NMDA also had a vasopressor action when applied to the ventrolateral medullary surface. In contrast to respiratory responses, bilateral application of NMDA caused a significant increase in blood pressure compared with unilateral application of NMDA. Application of the NMDA antagonist 2-amino-5-phosphonovaleric acid abolished both the blood pressure and respiratory effects of NMDA. These results suggest that CO2 and NMDA may act on a common respiratory premotoneuron to produce stimulation of breathing. Because blood pressure responses, unlike respiratory responses, were greater after bilateral application than after unilateral application of NMDA, it is suggested that the neural substrates for the two effects of NMDA seem to be different.

2-Amino-5-phosphonovalerate↗

Cytogenetic analyses of somatic chromosomes in a transplantable monomyelocytic leukemia in BALB/c mice.

In vivo cytogenetic analyses have been performed using G-, C-, and nuclear organizing region (NOR)-banding techniques, and sister chromatid exchanges (SCE) on a transplantable monomyelocytic leukemia (MML) initially induced in female BALB/c mice by the Rauscher leukemia virus (RLV). Centromeric associations have been found to be greatly increased in MML transplanted mice. Transplantability of the disease has been demonstrated at the cytogenetic level by the presence of female cells in males transplanted with MML cells. G-banding analysis has shown the existence of a marker deleted chromosome 18 in all tissues examined (bone marrow, spleen, and peripheral blood) restricted to female transplanted cells. The NOR-banding analysis has shown a slight increase in the number of Ag-NOR sites per metaphase in MML transplanted mice compared with controls and the existence of an extra chromosome having NOR in MML transplanted mice. No differences were found in C banding between controls and MML transplanted mice. In MML transplanted males, female transformed cells showed a significant reduction in SCE frequency compared with host male cells or controls.

Animals↗

Correlation of polarized absorption spectroscopic and X-ray diffraction studies of crystalline cytosolic aspartate aminotransferase of pig hearts.

Absorption spectra of large, well-formed crystals of cytosolic aspartate aminotransferase have been recorded using plane polarized light. Making use of measurements of crystal thickness we have calculated extinction coefficients with the electric vector of the light parallel to both the a and c axes of the crystals of the enzyme in space group P2(1)2(1)2(1). The spectra have been resolved into components with lognormal distribution curves and the resulting integrated intensities have been used to calculate the c/a polarization ratios for the absorption bands of the bound co-enzyme pyridoxal 5'-phosphate. From the polarization ratio and the co-ordinates of the co-enzyme ring atoms, provided by X-ray crystallography, we have assigned principal molecular directions of the transition dipole moment within the plane of the co-enzyme ring. Of two possible orientations, only one predicts the correct crystal extinction coefficients for the 436 nm band. In this orientation, when viewed from the B face of the ring (i.e. looking into the active site of the enzyme), the transition moment is related to the N-1-C-4 axis of the ring by counterclockwise rotation by 27 degrees. A tentative assignment of the principal molecular directions of the transition moment has also been made for the 368 nm band of the high pH form of the enzyme. In each case, the plane of the co-enzyme ring was located from the atomic co-ordinates of the ring atoms and of those atoms attached directly to the ring. The projection of the N-1 to C-4 axis on to this plane was used to evaluate the orientation of the transition moment, which was presumed to lie precisely within the plane of the ring. We have tilted this plane systematically to evaluate the error in transition moment direction resulting from uncertainties in the atomic co-ordinates. When 2-methylaspartate is diffused into the crystals if forms a Schiff base with the co-enzyme in which the ring has tilted about 32 degrees from its original position and the polarization ratio of the 436 nm band drops from 1.6 in the free enzyme to about 0.38. On the assumption that the orientation of the transition moment within the co-enzyme does not change during this rotation, this value of the polarization ratio is within experimental error of that predicted from X-ray structures on the two forms. The 2-methylaspartate binds only to subunit 1.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effect of sustained hypertransfusion on Rauscher leukemia virus-variant A (RLV-A) infection in BALB/c mice.

Infection of BALB/c mice with the RLV-A virus normally induces an erythropoietic dysplasia characterized by hepatosplenomegaly, erythroblastosis, erythroblastemia and severe anemia without reticulocytosis. Time to death varies between 20-30 weeks. Mice were inoculated with RLV-A after being hypertransfused with 75% packed red cells for 42 days which has been shown to eliminate erythropoiesis and modify the microenvironment to favor granulopoiesis. Following RLV-A inoculation, one group did not receive further transfusion (short-term) and another group continued with hypertransfusion weekly (long-term). The pathogenesis of RLV-A in the short-term group paralleled the characteristic RLV-A response. In the long-term group however, the characteristic RLV-A response was never observed. Instead, a granulocytic leukemia was developed. Continued hypertransfusion presumably after establishment of an altered microenvironment resulted in a completely different viral pathogenesis and the development of a transplantable myeloid leukemia.

Animals↗

Naloxone enhances the response to hypercapnia of spinal and cranial respiratory nerves.

To assess the effects of endogenous opiates on respiratory muscle responses to CO2, naloxone was administered intravenously to paralyzed, vagotomized and artificially ventilated cats anesthetized with alpha-chloralose. Neural activity was recorded from the phrenic, hypoglossal (HG), glossopharyngeal (GP) and recurrent laryngeal (RL) nerves. Before naloxone, phasic activity began first in the phrenic at a PETCO2 of 30.0 +/- 1.8 Torr, followed by the RL at a PETCO2 of 33.5 +/- 1.7 Torr, the HG at a PETCO2 of 39.9 +/- 2.1 Torr and the GP at a PETCO2 of 42.5 +/- 2.2 Torr during CO2 rebreathing. Naloxone had no significant effect on the apneic threshold of any of the nerves studied. Naloxone did, however, increase respiratory frequency (P less than 0.01) mainly by causing a significant (P less than 0.01) shortening of TE as it had no significant effect on TI. Naloxone also significantly increased the rate at which peak nerve activity increased with CO2 in the HG (P less than 0.01) and the GP (P less than 0.01) nerves, but not in the phrenic and RL nerves. Instead, the maximum activity produced by hypercapnia and the PETCO2 level at which maximum activity occurred in the phrenic, but not the RL, increased after naloxone. The result of these effects was that naloxone extended the range over which the HG and GP behaved proportionally with the phrenic, but it did not change the curvilinear nature of these relationships.

Animals↗

Respiratory and vasomotor responses to focal cooling of the ventral medullary surface (VMS) of the rat.

Experiments were performed on anesthetized, paralyzed and artificially ventilated rats after denervation of the vagus and carotid sinus nerves. The electrical activity of the phrenic and cervical sympathetic nerves (CS) along with the arterial blood pressure (BP) were monitored. Graded unilateral cooling of the ventral lateral surface (VMS) from 37 degrees C to 10 degrees C between 6th and 12th nerve rootlets did not affect the phrenic activity. Whereas, a significant depression or apnea was seen with cooling of an area between 1st cervical and 12th nerve rootlets. Bilateral cooling also produced similar respiratory responses. Respiratory depression could also be obtained during higher respiratory drive (7% CO2 in O2). On the other hand, a significant fall in BP and reduction in CS activity were observed with unilateral cooling in any of these VMS areas. However, the magnitude of BP decrease was less with 7% CO2 in O2 compared to 100% O2 breathing.

Animals↗

Comparison of the response of diaphragm and upper airway dilating muscle activity in sleeping cats.

The steady state and transient effects of hyperoxic hypercapnia on the electromyographic activities of the genioglossus (GG), posterior cricoarytenoid (PCA), and diaphragm (D) were studied in cats instrumented with chronically implanted electrodes during non-rapid eye movement sleep. Hypercapnia (inhalation of 3.4 and 7.4% CO2 in O2) increased the phasic electrical activity occurring during inspiration of all three muscles. Activities of the PCA and D increased in a parallel fashion with increasing CO2. The GG responded to steady state CO2 inhalation alinearly, with larger increases in activity occurring when CO2 was raised from 3.4 to 7.4% than when CO2 was increased from 0 to 3.4%. When gas mixtures containing CO2 were given, the GG reached its new steady state level more slowly than the D or PCA, and when CO2 was rapidly removed from the inspired gas mixture, the GG attained its steady state sooner than the D. These results suggest that in sleeping cats, hypercapnia does not affect either transient or steady state responses of the GG in the same way as the D or the PCA. These differences seem to be explained largely by different threshold characteristics of hypoglossal and phrenic neurons.

Animals↗

Respiratory and vasomotor effects of excitatory amino acid on ventral medullary surface.

Three glutamic acid analogues, N-methyl-D-aspartic (NMDA), quisqualic (QQ), and kainic (KAI) acids were applied topically to the ventral surface of the medulla (VMS) in paralyzed, vagotomized and carotid sinus denervated cats hyperventilated to apnea. Respiratory and vasomotor effects were assessed by changes in phrenic nerve activity and systemic arterial blood pressure. All three agents to varying degrees raised systemic blood pressure, but only NMDA consistently initiated phrenic nerve activity at pCO2 levels below that observed in control trials. KAI and QQ raised blood pressure even in those animals in which they had little effect on initiating phrenic nerve activity. Furthermore, respiratory responses were obtained from localized areas on VMS, namely the intermedio-caudal zone (I-C areas); whereas blood pressure elevations could be obtained from wider VMS areas including the rostral zone (R areas). In addition, the effects of the three amino acids on blood pressure were quantitatively different with KAI causing much greater increases in blood pressure than QQ or NMDA. The respiratory and vasomotor effects of NMDA and QQ were blocked by the use of 2-amino-5-phosphonovaleric acid and L-glutamic acid diethylester, their respective antagonists. The results suggest that neurons in the VMS which cause respiratory and vasomotor responses are not identical. Cells containing receptors stimulated by NMDA predominantly increase respiration, whereas cells containing receptors excited by KAI are more effective in eliciting vasomotor responses.

2-Amino-5-phosphonovalerate↗

Ventral medullary surface inputs to cervical sympathetic respiratory oscillations.

To examine the effects of focally cooling three areas (rostral, intermediate, and caudal) of the ventral medullary surface (VMS) on respiratory oscillations in cervical sympathetic and phrenic nerve activity, 12 cats were anesthetized, vagotomized, paralyzed, and artificially ventilated with 7% CO2 in O2. Cooling the intermediate area from 37 to 20 degrees C significantly reduced the magnitude of respiratory oscillations in both cervical sympathetic activity (P less than 0.001) and phrenic activity (P less than 0.001). Graded cooling of all three areas caused graded reductions in sympathetic respiratory-related activity that were comparable to the reductions in phrenic activity. The magnitude of the reductions in sympathetic respiratory oscillations was greatest for the intermediate area, followed in order by the caudal and rostral areas. Eight cats also underwent graded VMS cooling while ventilated with 3% CO2 in O2 and 100% O2. At each level of inspired CO2, graded cooling resulted in graded reductions in respiratory oscillations in sympathetic activity; conversely, at each medullary temperature, graded increases in inspired CO2 caused graded increases in cervical sympathetic respiratory activity. These results suggest that all three areas of the VMS influence respiratory oscillations in cervical sympathetic activity, although to different extents.

Animals↗