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Biomedical subjects

A Chaudhuri

Publications and source records attributed to A Chaudhuri.

At least 109 records · Page 6Linked to original sources
PubMed1998

A maximum motion technique for assessment of color vision defects.

Several luminance-matching methods, such as flicker fusion and the minimum motion technique (MMT), are capable of detecting certain forms of abnormal color vision. We present evidence that the heterochromatic fusion nystagmus (HFN) luminance matching technique can discriminate among normal trichromats, protanopes, and deuteranomals. The HFN luminance matching technique has the advantage that it provides a positive indication of isoluminance (maximization of motion) as opposed to the MMT and flicker fusion methods, which indicate isoluminance by the minimization of motion and flicker, respectively. We tested 16 normal trichromats, 6 protanopes, and 4 deuteranomals with the HFN technique. Results indicate that HFN is a useful tool for examining color vision. Because the HFN stimulus elicits reflexive eye movements (optokinetic nystagmus) that follow the apparent motion of the stimulus, HFN luminance matching will be particularly useful in animal and infant research, where other color vision tests are difficult to implement.

Adolescent↗
PubMed1997

Neural activity mapping with inducible transcription factors.

Neurons respond to extracellular stimulation by modulating the expression of certain immediate-early genes. Inducible transcription factors (ITFs), such as c-Fos and Zif268, are coded by this class of gene and are among the first proteins to appear. The rapid accumulation of these products combined with histological methods that offer detection at the cellular level are key features that have led to their wide use in visualizing activated neurons. However, neuroscientists have long recognized two major drawbacks of ITFs that limit their use in the CNS: cell-type expression specificity and stimulus-transcription coupling uncertainty. In this review, I discuss recent advances in the field that broaden our understanding of the molecular constraints on ITF expression as well as in techniques that may help to extend their utility in functional mapping.

Animals↗
PubMed1997

CD40-mediated signals inhibit the binding of TNF receptor-associated factor 2 to the CD40 cytoplasmic domain.

TNF receptor-associated factor 2 (TRAF2) is a signal-transducing protein associated with the CD40 cytoplasmic domain. It has been hypothesized that during signal transduction, TRAF2 must be released from CD40 in order for it to interact with downstream signaling molecules. We found that CD40 and TRAF2 were constitutively associated with each other in a human B cell line. Following stimulation with an anti-CD40 Ab, a decrease in the amount of CD40-associated TRAF2 was observed that could not be explained by a change in total level of either of the proteins. These results, as well as similar findings obtained with 293 cells overexpressing CD40 and TRAF2, suggested that CD40-mediated signals inhibited the CD40-TRAF2 interaction. We then conducted binding studies using CD40 cytoplasmic domain fusion proteins and TRAF2 derived from either control or CD40-stimulated cell lines. These in vitro studies also indicated that the binding of TRAF2 to the CD40 cytoplasmic domain was inhibited by CD40 stimulation. The results of these experiments, as well as differences between the in vitro and in vivo findings, indicated that multiple mechanisms were involved in the inhibition of the CD40-TRAF2 interaction by CD40 signals. Possible mechanisms of inhibition are discussed based on mapping of the TRAF2 binding site on the CD40 cytoplasmic domain.

B-Lymphocytes↗
PubMed1997

Neural activity mapping with inducible transcription factors.

Neurons respond to extracellular stimulation by modulating the expression of certain immediate-early genes. Inducible transcription factors (ITFs), such as c-Fos and Zif268, are coded by this class of gene and are among the first proteins to appear. The rapid accumulation of these products combined with histological methods that offer detection at the cellular level are key features that have led to their wide use in visualizing activated neurons. However, neuroscientists have long recognized two major drawbacks of ITFs that limit their use in the CNS: cell-type expression specificity and stimulus-transcription coupling uncertainty. In this review, I discuss recent advances in the field that broaden our understanding of the molecular constraints on ITF expression as well as in techniques that may help to extend their utility in functional mapping.

Animals↗
PubMed1997

Dual activity maps in primate visual cortex produced by different temporal patterns of zif268 mRNA and protein expression.

The inducible nature of the immediate-early genes (IEGs) c-fos and zif268 allows their products to be used as activity markers in the brain. The utility of such markers in general is restricted because they can resolve only neurons activated by a single stimulus. To overcome this limitation, we have developed a double-label technique that exploits the dissimilar time course of zif268 mRNA and protein induction, allowing them to be separately induced by two different stimuli and independently stained to provide a visual display of neurons that are responsive to each stimulus. Two powerful features of this new imaging technique-the possibility of staining separate populations of activated neurons and the ability to visualize them at the cellular level-should extend IEG applications in biological activity mapping.

Animals↗
PubMed1997

Activity-dependent regulation of cytochrome b gene expression in monkey visual cortex.

The recent demonstration that certain mitochondrial subunits of cytochrome oxidase (CO) are regulated by neuronal activity has stimulated interest in the molecular processes that coordinate nuclear and mitochondrial gene expression following synaptic stimulation. We have studied the constitutive expression and activity-guided regulation of cytochrome b (cyt b), a gene that is encoded by mitochondrial DNA and that was cloned by subtractive hybridization from the lateral geniculate nucleus in the monkey. We have found cyt b mRNA expression in monkey striate cortex to be similar to that of CO activity with regard to the laminar profile and the presence of blobs in the supragranular layers. Layers 2/3, 4C, and 6 contained large numbers of stained cells, many of which were judged to be excitatory neurons, because they showed a Zif268-immunopositive nucleus. We have also found that removal of functional activity reduced cyt b mRNA content in area V1. Columns of reduced cyt b staining were visible after 3 days and were especially striking after 6 days of monocular deprivation. After 3 months of deprivation, the columns lost their contrast and became blurred. Our principal finding, that neuronal activity regulates cyt b levels, suggests that expression of a mitochondrial gene can be affected in a manner similar to that of several known nuclear genes. The differences in cyt b mRNA levels and CO activity after long-term deprivation suggests that some form of differential control is exerted on cyt b. Cyt b expression, therefore, may be used as a marker of altered mitochondrial transcription that is guided by the metabolic demands of active neurons.

Animals↗
PubMed1997

Detection of Duffy antigen in the plasma membranes and caveolae of vascular endothelial and epithelial cells of nonerythroid organs.

The nonerythroid expression of the Duffy blood group protein (gp-Fy) was confined to certain cell types. Immunocytochemistry studies of the kidney showed gp-Fy in the endothelium of glomeruli, peritubular capillaries, vasa recta, and the principal cells (epithelial) of collecting ducts. Gp-Fy was also produced in the endothelial cells of large venules and epithelial cells (type-I) of pulmonary alveoli. In the thyroid, only the endothelial cells of capillaries produced gp-Fy. In the spleen, the endothelial cells of capillaries, high endothelial venule, and sinusoids produced abundant gp-Fy. Ultrastructural studies showed that apical and basolateral plasma membrane domains, including caveolae, had gp-Fy. Immunoblot analysis showed substantially less gp-Fy in nonerythroid cells than in erythrocytes. Moreover, the analyzed nonerythroid organs of Duffy-negative individuals did not produce more gp-Fy to compensate for the lack of this protein in their erythrocytes. The nucleotide sequence and the size of kidney mRNA from a Duffy-positive individual were the same as that of bone marrow. It is assumed, therefore, that nonerythroid Duffy protein is the product of the same gene as that of bone marrow. This notion is reinforced by the fact that nonerythroid and erythroid gp-Fy have the same antigenic domains.

Cell Membrane↗
PubMed1997

Sensory regulation of immediate-early gene expression in mammalian visual cortex: implications for functional mapping and neural plasticity.

The expression of immediate-early genes that code for transcription factors has been extensively studied in the brain with regard to imaging functional activity. The components of the AP-1 transcription factor--in particular, c-Fos--and Zif268 have been widely used for this purpose. However, the precise details by which they are induced after synaptic stimulation remain unknown. Furthermore, the roles of these two proteins in neurons remains speculative and include such varied functions as short-term maintenance of cellular homeostasis to long-term changes that guide cortical plasticity. Current efforts at elucidating the physiological roles of AP-1 and Zif268 rely on assessing their expression in response to different conditions of sensory and pharmacological stimulation. In this review, we have examined the expression patterns of these transcription factors in the mammalian visual cortex under different conditions, with particular emphasis on the constitutive levels and how they change after visual deprivation and stimulation. A synthesis of this information offers further insight into their likely functions and the extent to which transcription factors may represent patterns of neural activity as a possible prelude to plastic events.

Animals↗
PubMed1997

Face recognition with multi-tone and two-tone photographic negatives.

The effects of photographic negatives on face recognition are often studied in two different ways--faces are learned and tested in photographic negatives (NN) or the contrast is reversed between learning and testing, i.e. they are learned in positives and tested in negatives (PN) or vice versa (NP). We have examined recognition performance for faces under these three conditions along with a control condition where faces were learned and tested in positives (PP). Using multi-tone face images, we found that the effect of photographic negatives was more pronounced in PN and NP than in NN. No differences were found between PN and NP or between NN and PP. When two-tone face images were used, recognition performance was worse in all conditions, except PP, when compared to the multi-tone counterparts. Our results show that contrast incongruency between learning and testing is the predominant factor affecting performance and that deficits in sensory coding or retention of negative face images are unlikely to be major factors. The advantage of multi-tone over two-tone negatives can be attributed to preserved facial information carried by the high-spatial-frequency components of the image.

Adolescent↗
PubMed1997

Cloning, characterization, and mapping of a murine promiscuous chemokine receptor gene: homolog of the human Duffy gene.

We report here the isolation and genomic organization of the orthologous mouse Duffy gene, named Dfy. It is a single copy gene located in chromosome 1 in a region homologous to the human Duffy gene (FY). Sequence analyses indicate that Dfy consists of two exons: exon 1 of 55 nucleotides, which encodes 7 amino acid residues; and exon 2 of 1038 nucleotides, which encodes 327 residues. The single intron consists of 462 nucleotides. The 5'-end promoter region contains motifs involved in vertebrate development in addition to potential binding sites of factors for globin transcription. The open reading frame (ORF) shows 60% homology with the human Duffy protein. However, mouse erythrocytes are serologically Duffy-negative and mouse erythrocyte membrane proteins do not cross-react with two Duffy-specific rabbit polyclonal antibodies. The deduced protein predicts a M(r) of 36,692 and carries three potential N-glycosylation sites to asparagine residues. Hydropathy analysis predicts an exocellular amino-terminal domain of 57 residues, seven transmembrane alpha-helices, and an endocellular carboxy-terminal domain of 29 residues. In bone marrow and spleen, Dfy expresses a major 1.4-kb and a minor 1.8-kb mRNA. Contrary to humans, Dfy is expressed in liver, synthesizing a 1.4-kb mRNA, and is repressed in kidney. Dfy is highly expressed in mouse brain and produces a major 8.5-kb and a minor 10.2-kb mRNA. The human erythroleukemia K562 cells, transfected with cDNA encoding the mouse Duffy-like protein and mouse erythrocytes, have the same chemokine binding profiles indicating that they contain the same protein.

Amino Acid Sequence↗
PubMed1996

The domain on the Duffy blood group antigen for binding Plasmodium vivax and P. knowlesi malarial parasites to erythrocytes.

Plasmodium vivax and the related simian malarial parasite P. knowlesi use the Duffy blood group antigen as a receptor to invade human erythrocytes and region II of the parasite ligands for binding to this erythrocyte receptor. Here, we identify the peptide within the Duffy blood group antigen of human and rhesus erythrocytes to which the P. vivax and P. knowlesi ligands bind. Peptides from the NH2-terminal extracellular region of the Duffy antigen were tested for their ability to block the binding of erythrocytes to transfected Cos cells expressing on their surface region II of the Duffy-binding ligands. The binding site on the human Duffy antigen used by both the P. vivax and P. knowlesi ligands maps to a 35-amino acid region. A 34-amino acid peptide from the equivalent region of the rhesus Duffy antigen blocked the binding of P. vivax to human erythrocytes, although the P. vivax ligand expressed on Cos cells does not bind rhesus erythrocytes. The binding of the rhesus peptide, but not the rhesus erythrocyte, to the P. vivax ligand was explained by interference of carbohydrate with the binding process. Rhesus erythrocytes, treated with N-glycanase, bound specifically to P. vivax region II. Thus, the interaction of P. vivax ligand with human and rhesus erythrocytes appears to be mediated by a peptide-peptide interaction. Glycosylation of the rhesus Duffy antigen appears to block binding of the P. vivax ligand to rhesus erythrocytes.

Amidohydrolases↗
PubMed1996

Visual stimulation regulates the expression of transcription factors and modulates the composition of AP-1 in visual cortex.

It is believed that long-term changes in neuronal function are orchestrated by transcription factors, such as AP-1 and ZIF 268, which are in turn regulated by synaptic stimulation. To further our understanding of the functional effects of such expression, we have examined the DNA-binding activities of both AP-1 and ZIF 268 by way of electrophoretic mobility shift assays (EMSA) on nuclear extracts from visual cortices of rats treated with selective light exposure. Visual stimulation after dark rearing increased the DNA-binding activities of both AP-1 and ZIF 268 to their highest levels within 2 hr. ZIF 268 thereafter dropped to levels similar to that observed in naive animals, whereas AP-1 DNA-binding activity continued to remain elevated even after 24 hr of stimulation. The components of the AP-1 complex, when assessed by EMSA-supershift analysis, showed considerable variability under different conditions of exposure. FosB and JunD were the major constituents of AP-1 in both naive and dark-reared animals. Brief visual stimulation (2 hr) added c-Fos, c-Jun, and JunB to this complex, whereas prolonged stimulation (6-24 hr) reduced c-Fos and c-Jun levels significantly, leaving only FosB, JunB, and JunD as the major components of AP-1. These results suggest that transcriptional control by AP-1 may be generated by selective combinatorial interactions of different members of the Fos and Jun families and that are guided by activity-dependent processes.

Activating Transcription Factor 2↗
PubMed1996

A novel invertase from a thermophilic fungus Thermomyces lanuginosus: its requirement of thiol and protein for activation.

Unlike the invertases from the mesophilic fungi and yeasts, invertase from a thermophilic fungi, Thermomyces lanuginosus, was unusually unstable both in vivo and in vitro. The following observations suggested that the unstable nature of the enzyme activity in the cell-free extracts was due to the oxidation of the cysteine residue(s) in the enzyme molecule: (a) the addition of dithiothreitol or reduced glutathione stabilized invertase activity during storage of the extracts and also revived enzyme activity in the extracts which had become inactive with time; (b) N-ethylmaleimide, iodoacetamide, oxidized glutathione, cystine, or oxidized coenzyme A-inactivated invertase; (c) invertase activity was low when the ratio reduced/oxidized glutathione was lower and high when this ratio was higher, suggesting regulation of the enzyme by thiol/disulfide exchange reaction. In contrast to the activation of invertase by the thiol compounds and its inactivation by the disulfides in the cell-free extracts, the purified enzyme did not respond to these compounds. Following its inactivation, the purified enzyme required a helper protein in addition to dithiothreitol for maximal activation. A cellular protein was identified that promoted activation of invertase by dithiothreitol and it was called "PRIA" for the protein which helps in restoring invertase activity. The revival of enzyme activity was due to the conversion of the inactive invertase molecules into an active form. A model is presented to explain the modulation of invertase activity by the thiol compounds and the disulfides, both in the crude cell-free extracts and in the purified preparations. The requirement of free sulfhydryl group(s) for the enzyme activity and, furthermore, the reciprocal effects of the thiols and the disulfides on invertase activity have not been reported for invertase from any other source. The finding of a novel invertase which shows a distinct mode of regulation demonstrates the diversity in an enzyme that has figured prominently in the development of biochemistry.

Cell-Free System↗
PubMed1996

Differential expression of neurofilament protein in the visual system of the vervet monkey.

It has been previously reported that the monoclonal antibody SMI-32 reveals a characteristic pattern of immunostaining which may be used to delineate various cortical modules in the monkey visual system. We wished to examine staining patterns with this antibody at both the lateral geniculate nucleus (LGN) and cortical levels with regard to magno- and parvocellular processing schemes in the vervet monkey. Using standard immunohistochemical procedures, we have found that the M-layers of the LGN were intensely stained in comparison to P-layers and that there were regional variations in staining within the visual cortex that reflected this input. The transition between areas V1 and V2 was especially prominent due to differences in the laminar staining profiles. Another striking result was found within the superior temporal sulcus where heavy SMI-32 immunostaining confined to the floor of the sulcus coincided with a similar zone of intense myelin staining. We have also found a number of other areas within the intraparietal and lateral sulci that show foci of heavy SMI-32 staining. As with Cat-301 immunostaining, the regional variabilities that are observed with SMI-32 in the visual cortex reflect molecular distinctions that may provide further criteria for functional segmentation.

Animals↗
PubMed1996

Influence of thyroxine on different ion-dependent ATPase activities in fat body of tasar silkworm, Antheraea mylitta D.

The activities of Na(+)-K(+)-, Ca(2+)-, and Mg(2+)-ATPase of tasar silkworm, Antheraea mylitta D. fat body were investigated from fifth larval stage to adult emergence after injection of various doses (0.5, 1.0, 2.0, and 5.0 micrograms/g) of mammalian thyroxine (T4) to 1-hr-old fifth instar larvae. In normal silkworms, both sexes exhibited maximum enzyme activity before spinning (Day 12). Na(+)-K(+)-, Ca(2+)-, and Mg(2+)-ATPase activities in fat body of silkworms markedly declined after pupation and more so on the 1-day-old adults. All doses of thyroxine treated on fifth instar larvae significantly altered ATPase activity in the larval, pupal, and adult stages in both sexes. ATPase activity was not altered by lower dose of T4 (0.5 micrograms/g) in 2-day-old fifth stage larvae, while the higher dose (5.0 micrograms/g) surprisingly caused a reduction in ATPase activity during the different developmental stages. The fat body ATPases were influenced by thyroxine in all the stages of silkworm in a dose-dependent manner. Our results thus indicate that thyroxine has a controlling influence on the ATPase system in silkworm fat body.

Aging↗