Search PubMed⌕ Search

Biomedical subjects

R Lal

Publications and source records attributed to R Lal.

At least 109 records · Page 6Linked to original sources

Gap junctional communication between vascular cells. Induction of connexin43 messenger RNA in macrophage foam cells of atherosclerotic lesions.

The structure and function of blood vessels depend on the ability of vascular cells to receive and transduce signals and to communicate with each other. One means by which vascular cells have been shown to communicate is via gap junctions, specifically connexin43. In atherosclerosis, the normal physical patterns of communication are disrupted by the subendothelial infiltration and accumulation of blood monocytes, which in turn can differentiate into resident foam cells. In this paper we report that neither freshly isolated human peripheral blood monocytes nor differentiated monocytes/macrophages exhibit functional gap junctional dye transfer in homo-cellular culture or in co-culture with endothelial cells or smooth muscle cells. By Northern analysis, neither freshly isolated blood monocytes nor pure cultures of differentiated monocyte/macrophages expressed gap junction messenger RNA. However, immunohistochemical staining followed by in situ hybridization on sections of human atherosclerotic carotid arteries revealed strong expression of gap junction connexin43 messenger RNA by macrophage foam cells. These results suggest that tissue-specific conditions present in atherosclerotic arteries induce expression of connexin43 messenger RNA in monocyte/macrophages.

Arteriosclerosis↗

Voltage-dependent gating and single-channel conductance of adult mammalian atrial gap junctions.

In the heart, the rapid propagation and synchronization of action potentials necessary for a normal heart rhythm and an effective cardiac output are mediated by specialized ionic channels that link adjacent cells and are known collectively as gap junctions. Cardiac gap junctions are gated by various physiological and pharmacological agents, but the role of voltage in their gating is unclear. Whereas embryonic or neonatal ventricular cells have voltage-gated gap junctions, adult cells are reported to have only voltage-independent gap junctions. We studied the voltage dependence of adult rat atrial gap junctions by individually voltage clamping each cell of a connected cell pair and controlling the transjunctional voltage (Vj), measuring transjunctional current (Ij), and calculating junctional conductance (gj). Two distinct populations of cell pairs were observed: highly coupled pairs with the peak gjs ranging from 3.4 to 40 nS and weakly coupled pairs with the peak gjs ranging from 0.3 to 2.0 nS. gj was dependent on Vj, and Ij decayed exponentially, with the time constants being voltage dependent. Voltage dependence was most apparent when cells were poorly coupled. The gj did not decrease to zero. The normalized conductance--Vj plot was fit with a two-state Boltzmann model as a first approximation, resulting in a half-inactivation potential and gating charge of 42.5 mV and 1.14 eV, respectively, for the weakly coupled cell pairs. For highly coupled cell pairs, the half-inactivation potential shifted to 53.3 mV. Single gap junctional channels had a gj of 36.2 +/- 7.6 pS (range, 27-49 pS), which was Vj independent.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effect of blockade of transdiaphragmatic absorption of bacteria on survival in peritonitis: an experimental study in rats.

An experimental rat model of established peritonitis was used to test the effect of intraperitoneal injection of platelet rich plasma (PRP) on blood and peritoneal fluid culture positivity and survival rates. Thirty animals divided into two groups of 15 each were studied. The first group served as control while animals in the second group received intraperitoneal injection of PRP. The use of PRP in established. Peritonitis was of no significant benefit.

Animals↗

Atomic force microscopy and dissection of gap junctions.

An atomic force microscope (AFM) was used to study the structure of isolated hepatic gap junctions in phosphate-buffered saline (PBS). The thickness of these gap junctions appears to be 14.4 nanometers, close to the dimensions reported by electron microscopy (EM). When an increasing force is applied to the microscope tip, the top membrane of the gap junction can be "dissected" away, leaving the extracellular domains of the bottom membrane exposed. When such "force dissection" is performed on samples both trypsinized and fixed with glutaraldehyde, the hexagonal array of gap junction hemichannels is revealed, with a center-to-center spacing of 9.1 nanometers.

Animals↗

Post-coital antifertility action of Ruta graveolens in female rats and hamsters.

Different preparations of Ruta graveolens were administered orally to female rats (Days 1-10 post coition) and female hamsters (Days 1-6 post coition). The powdered root (CDR), aerial parts (CDA) and the aerial parts aqueous extract (AEA) all showed potential anticonceptive activity in rats. Limited administration on selected days of CDA showed uniformly lesser activity than with 10-day treatment. Sequentially prepared petroleum ether and methanol extracts of CDA were as active as CDA itself. The benzene and chloroform extracts were toxic and inactive. Rutin, a known chemical constituent of the plant, was found to be inactive. None of the above preparations showed activity in hamsters.

Animals↗

Construction of a hybrid plasmid capable of replication in Amycolatopsis mediterranei.

A new plasmid, pA387, has been isolated from "Amycolatopsis sp." (DSM 43387). This plasmid could be isolated from liquid culture as well as mycelium from agar plates by a modified procedure. Plasmid pA387 is about 29.6 kb and can be cured at low frequency by protoplasting and ethidium bromide and heat treatment. Hybridization experiments showed that this plasmid is present in free form and does not integrate into the chromosome. A hybrid plasmid was constructed by cloning a 5.1-kb fragment of pA387 into the Escherichia coli vector pDM10. This hybrid plasmid, termed pRL1, could be transformed into Amycolatopsis mediterranei and A. orientalis by electroporation. A transformation frequency of 2.2 x 10(3) transformants per micrograms of DNA at 12.5 kV/cm and a pulse duration of 10.8 ms was obtained in A. mediterranei, whereas 1.1 x 10(5) transformants per microgram of DNA were obtained at a field strength of 7.5 kV/cm and a pulse duration of 7.6 ms in A. orientalis. Plasmid pRL1 is the first hybrid plasmid which could be used successfully for the transformation of A. mediterranei. The plasmid has a rather high copy number, is genetically stable, and can be easily reisolated from A. mediterranei. Plasmid pRL1 will be useful for further construction of a shuttle vector for E. coli and A. mediterranei and becomes the basis for the development of gene cloning techniques in Amycolatopsis spp.

Cloning, Molecular↗

Preparation and evaluation of 99mTc-t-butylisonitrile (99mTc-TBI) for myocardial imaging: a kit for hospital radiopharmacy.

A previous method was modified to obtain [99mTc(TBI)6]+ by reacting Zn(TBI)2Br2 directly with 99mTcO4- in the presence of Sn2+ ions. [Cu(TBI)4]Cl was next used as a source of TBI. On reaction with 99mTcO4- and Sn2+ ions for 3 min at 100 degrees C, [99mTc(TBI)6]+ product of radiochemical purity greater than 90% and yield greater than 70% was obtained. Data of biodistribution in rats (2-2.5% in heart) and biokinetics in rabbits were satisfactory. The kit formulation was found to be stable and also safe for administration.

Animals↗

Effect of passive eye position changes on retinogeniculate transmission in the cat.

1. Extracellular recordings were made from single neurons in layer A of the left dorsal lateral geniculate nucleus (LGNd) of anesthetized and paralyzed adult cats. Responses to retinotopically identical visual stimuli (presented through the right eye) were recorded at several positions of the left eye in its orbit. Visual stimuli consisted of drifting sinusoidal gratings of optimal temporal and spatial frequencies at twice threshold contrast. Visual stimulation of the left eye was blocked by a variety of methods, including intravitreal injection of tetrodotoxin (TTX). The change in position of the left eye was achieved by passive movements in a randomized and interleaved fashion. Of 237 neurons studied, responses were obtained from 143 neurons on 20-100 trials of identical visual stimulation at each of six eye positions. Neurons were classified as X- or Y- on the basis of a standard battery of physiological tests (primarily linearity of spatial summation and response latency to electrical stimulation of the optic chiasm). 2. The effect of eye position on the visual response of the 143 neurons was analyzed with respect to the number of action potentials elicited and the peak firing rate. Fifty-seven (40%) neurons had a significant effect [by one-factor repeated-measure analysis of variance (ANOVA), P less than 0.05] of eye position on the visual response by either criterion (number of action potentials or peak firing rate). Of these 57 neurons, 47 had a significant effect (P less than 0.05) with respect to the number of action potentials and 23 had a significant effect (P less than 0.05) by both criteria. Thus the permissive measure by either criterion and the conservative measure by both criteria resulted in 40% and 16%, respectively, of all neurons' visual responses being significantly affected by eye position. 3. For the 47 neurons with a significant effect of eye position (number of action potentials criterion), a trend analysis of eye position versus visual response showed a linear trend (P less than 0.05) for 9 neurons, a quadratic trend (P less than 0.05) for 32 neurons, and no significant trend for the 6 remaining neurons. The trends were approximated with linear and nonlinear gain fields (range of eye position change over which the visual response was modulated). The gain fields of individual neurons were compared by measuring the normalized gain (change in neuronal response per degree change of eye position). The mean normalized gain for the 47 neurons was 4.3. 4. The nonlinear gain fields were generally symmetric with respect to nasal versus temporal changes in eye position.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

Effect of passive eye movement on retinogeniculate transmission in the cat.

1. The nature and time window of interaction between passive phasic eye movement signals and visual stimuli were studied for dorsal lateral geniculate nucleus (LGNd) neurons in the cat. Extracellular recordings were made from single neurons in layer A of the left LGNd of anesthetized paralyzed cats in response to a normalized visual stimulus presented to the right eye at each of several times of movement of the left eye. The left eye was moved passively at a fixed amplitude and velocity while varying the movement onset time with respect to the visual stimulus onset in a randomized and interleaved fashion. Visual stimuli consisted of square-wave modulated circular spots of appropriate contrast, sign, and size to elicit an optimal excitatory response when placed in the neurons' receptive-field (RF) center. 2. Interactions were analyzed for 78 neurons (33 X-neurons, 43 Y-neurons, and 2 physiologically unclassified neurons) on 25-65 trials of identical visual stimuli for each of eight times of eye movement. 3. Sixty percent (47/78) of the neurons tested had a significant eye movement effect (ANOVA, P less than 0.05) on some aspect of their visual response. Of these 47 neurons, 42 (89%) had a significant (P less than 0.05) effect of an appropriately timed eye movement on the number of action potentials, 36 (77%) had a significant effect on the mean peak firing rate, and 31 (66%) were significantly affected as evaluated by both criteria. 4. The eye movement effect on the neurons' visual responses was primarily facilitatory. Facilitation was observed for 37 (79%) of the affected neurons. For 25 of these 37 neurons (68%), the facilitation was significant (P less than 0.05) as evaluated by both criteria (number of action potentials and mean peak firing rate). Ten (21%) of the affected neurons had their visual response significantly inhibited (P less than 0.05). 5. Sixty percent (46/78) of the neurons were tested for the effect of eye movement on both visually elicited activity (visual stimulus contrast = 2 times threshold) and spontaneous activity (contrast = 0). Eye movement significantly affected the visual response of 23 (50%) of these neurons. However, spontaneous activity was significantly affected for only nine (20%) of these neurons. The interaction of the eye movement and visual signals was nonlinear. 6. Nine of 12 neurons (75%) tested had a directionally selective effect of eye movement on the visual response, with most (8/9) preferring the temporal ward direction.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

Gating of retinal transmission by afferent eye position and movement signals.

Vision in most vertebrates is an active process that requires the brain to combine retinal signals with information about eye movement. Eye movement information may feed forward from the motor control areas of the brain or feed back from the extrinsic eye muscles. Feedback signals elicited by passive eye movement selectively gate retinal outflow at the first relay, the dorsal lateral geniculate nucleus. The gating predominantly facilitates retinogeniculate transmission immediately after eye movement and inhibits transmission when a new steady-state eye position is achieved. These two gating effects are distributed in a complementary fashion across the dorsal lateral geniculate nucleus such that the spatiotemporal activity profile could contribute to object detection and localization.

Action Potentials↗