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Biomedical subjects

M Subramaniam

Publications and source records attributed to M Subramaniam.

At least 55 records · Page 3Linked to original sources
PubMed1994

The role of middle ear muscles in the development of resistance to noise induced hearing loss.

The role of middle ear muscles (MEMs) in the development of increased resistance to noise induced hearing loss (NIHL) was studied using monaural chinchillas. Animals with severed MEMs as well as those with intact MEMs were exposed to an octave band noise (OBN) centered at 0.5 kHz at 95 dB for six hours/day for ten consecutive days. Results indicated that animals with severed MEMs showed greater initial threshold shifts (TS) than the animals with intact MEMs. Both the groups showed a decrease in TS over the ten days of exposure. The subjects were given five days of recovery and then re-exposed to the same noise at 106 dB for 48 h. Permanent threshold shifts (PTS) in each group was compared against those in a control group exposed to the noise only at the higher level. Interestingly, both the 'conditioned' groups incurred substantially less PTS than the control group exposed only to the higher level.

Animals↗
PubMed1994

Changes in distortion product otoacoustic emissions and outer hair cells following interrupted noise exposures.

Changes in distortion product otoacoustic emissions (DPOAEs) were examined during and after interrupted noise exposures and compared to the condition of the outer hair cells (OHCs) and inner hair cells (IHCs) as assessed by scanning electron microscopy (SEM). Binaural, adult chinchillas were exposed to a 95 dB SPL, octave band noise centered at 0.5 kHz for 15 days using a 3 h on/9 h off schedule. DPOAEs were measured before, during and after the exposures. DPOAE amplitudes decreased significantly during the first few days of the interrupted noise exposures and then began to recover. At most frequencies, the emission amplitudes recovered completely to pre-exposure baseline values by five days after the last exposure. The results of the present study indicate that the changes in DPOAE amplitude paralleled the recovery in the amplitude and threshold of the compound action potentials as reported previously (Boettcher et al., 1992). Although the DPOAEs completely recovered, considerable OHC loss and stereocilia disarray was evident even four weeks after exposure.

Acoustic Stimulation↗
PubMed1994

The relationship among distortion-product otoacoustic emissions, evoked potential thresholds, and outer hair cells following interrupted noise exposures.

Distortion-product otoacoustic emissions (DPOAEs) are gaining popularity as indicators of the status of the cochlea. The efficacy of DPOAEs as indicators of changes in thresholds and the status of outer hair cells (OHCs) were examined using an animal model. Monaural chinchillas were exposed to an octave band noise (OBN) centered at 0.5 kHz at 95 dB SPL for 6 hr/day for 10 days. DPOAEs and evoked potential thresholds were recorded before, during, and after the exposures. The animals were sacrificed 5 days after the last exposure, and the status of OHCs was assessed using scanning electron microscopy. Results indicate that both evoked potential thresholds and DPOAEs effectively track the temporary changes associated with interrupted noise exposures. However, DPOAEs often recovered to their baseline even when there was a threshold shift of > 25 dB. Furthermore, at 5 days postexposure, both evoked potential thresholds and DPOAEs were normal despite considerable OHC pathology. The findings suggest that normal DPOAEs may not guarantee normal cochlear status and, therefore, results of DPOAE measurements should be interpreted cautiously.

Acoustic Stimulation↗
PubMed1993

The proto-oncogene c-fms is overexpressed in endometrial cancer.

Recent studies have shown that macrophage colony-stimulating factor and its receptor c-fms protein are significantly overexpressed in endometrial and ovarian cancers. In the present study, we analyzed the steady-state levels of c-fms mRNA in benign and malignant endometrial tissues by Northern and slot blot analyses. The relative levels of c-fms mRNA were quantified by using a hybridization signal for each sample on Northern blot analysis. Slot blot analysis was used to further quantitate the relative increase in c-fms mRNA in malignant specimens compared to benign specimens. Correlation of c-fms expression in the endometrial cancers was made with traditional prognostic indicators. Secretory endometrium had low levels of c-fms mRNA, whereas the endometrial cancers had the highest levels. Proliferative and hyperplastic endometrium values were intermediate. Comparative assessment of c-fms expression in endometrial cancer relative to other prognostic factors demonstrated greater expression of c-fms in specimens from patients with abnormal DNA ploidy, high-grade lesions, and possibly extrauterine metastases. Our study confirms the overexpression of c-fms in endometrial cancer and demonstrates a positive correlation between the steady-state mRNA levels of c-fms and other select adverse prognostic indicators.

Endometrial Neoplasms↗
PubMed1993

Protection from noise induced hearing loss: is prolonged 'conditioning' necessary?

The effect of prior 'conditioning' noise exposures on the protection from subsequent higher level exposures was studied using four groups of chinchillas. The three experimental groups were 'conditioned' using a 0.5 kHz octave band noise at 95 dB SPL for 6 h a day. The first group was exposed to the noise once and allowed to recover for nine days prior to the second exposure. The second and third groups were exposed for ten and twenty days respectively. The first group showed only small reductions in threshold shift (TS) following the second exposure. The other two groups showed significant reductions in TS with repeated exposures. Following the last 'conditioning' exposure, all three experimental groups were allowed to recover for five days before exposing them to the same noise at 106 dB SPL for 48 h. Threshold shifts recorded following the 106 dB exposure were compared against those recorded in a control group exposed only to the higher level. Each of the three experimental groups developed significantly less permanent threshold shifts than the control group. However, there were no significant differences among the three experimental groups and the differences in hair cell losses were insignificant.

Acoustic Stimulation↗
PubMed1993

Divergent fates of P- and E-selectins after their expression on the plasma membrane.

P-selectin and E-selectin are related adhesion receptors for monocytes and neutrophils that are expressed by stimulated endothelial cells. P-selectin is stored in Weibel-Palade bodies, and it reaches the plasma membrane after exocytosis of these granules. E-selectin is not stored, and its synthesis is induced by cytokines. We studied the fate of the two proteins after their surface expression by following the intracellular routing of internalized antibodies to the selectins. By immunofluorescent staining, P-selectin antibody was first seen in endosomes, then in the Golgi region, and finally in Weibel-Palade bodies. In contrast, the E-selectin antibody was detected only in endosomes and lysosomes. Subcellular fractionation of cells after 4 h chase confirmed the localization of P-selectin antibody in storage granules and of the E-selectin antibody in lysosomes. In AtT-20 cells, a mouse pituitary cell line, transfected with P- or E-selectin, only P-selectin was delivered to the endogenous adrenocorticotrophic hormone storage granules after endocytosis. Deletion of the cytoplasmic domain abolished internalization. In summary, after a brief surface exposure, internalized E-selectin is degraded in the lysosomes, whereas P-selectin returns to the storage granules from where it can be reused.

Animals↗
PubMed1993

Individual susceptibility to noise-induced hearing loss: an old topic revisited.

The wide range in susceptibility to noise-induced hearing loss has intrigued researchers and hearing conservationists alike. Some of these differences in variability have been attributed to various intrinsic factors such as eye color, gender, age, etc. However, a review of controlled research shows that the influence of these intrinsic variables is relatively small and cannot explain the wide range of hearing loss observed in demographic studies. Furthermore, uncontrolled variables or unrecognized drug and noise interaction may obscure the relation between noise exposure and hearing loss. With the growing understanding of the physiology of the auditory system, new possibilities are emerging that may explain the range of susceptibility. A review of the role of acoustic reflex effectiveness, cochlear efferent function, and history of noise exposure provide a perspective for future strategies in predicting susceptibility to noise-induced hearing loss.

Age Factors↗
PubMed1993

Effect of low-frequency "conditioning" on hearing loss from high-frequency exposure.

Recent research has revealed that repeated exposures to a low-frequency noise results in a progressive reduction in threshold shifts (TS). This reduction in TS is not restricted to the exposure frequency, but can be observed at frequencies up to 3 or 4 octaves higher. Such "conditioning" exposures have also been observed to protect the auditory system against hearing loss from exposures to the same noise at higher levels. The aim of this study was to determine if "conditioning" using low-frequency exposures protects the auditory system against hearing loss from a high-frequency exposure. Monaural chinchillas were exposed to a 0.5-kHz octave band noise (OBN) at 95 dB SPL for 6 h a day. The animals were allowed to recover for 5 days, following which they were exposed to a 4-kHz OBN at 100 dB SPL for 48 h. Hearing thresholds determined using evoked potential recordings, indicated significantly greater permanent threshold shifts in this group of animals when compared to a control group exposed only to the 4-kHz OBN. These results were confirmed by histological examination which revealed greater hair cell loss in the experimental group.

Acoustic Stimulation↗
PubMed1993

Rapid down-regulation of c-jun protooncogene transcription by progesterone in the avian oviduct.

Previous work in this and other laboratories has shown that steroids rapidly regulate the expression of nuclear protooncogenes. In this present study, we have investigated the effect of progesterone (Pg) on the expression of c-jun in the avian oviduct system and its promoter activity in avian liver cells. Pg treatment of estrogen-withdrawn chickens brings about a decrease in the steady state mRNA level of the protooncogene c-jun within 30 min. This decrease is steroid dose dependent and gene specific. Using nuclear run-off transcription analyses, this rapid regulation was shown to occur at the level of gene transcription, as the rate of c-jun transcription decreases by more than 80% within 15 min after progesterone treatment. As expected, ovalbumin gene transcription is increased only after a lag period of 4 h following Pg treatment. In other studies, we have linked the c-jun promoter sequences between -1000 and +192 to chloramphenicol acetyltransferase reporter gene and cotransfected them into transformed avian liver cells along with the expression vector for the Pg receptor. Pg treatment of these cells causes a decrease in chloramphenicol acetyltransferase gene expression, albeit to a lesser extent than Pg inhibition of c-jun gene transcription. These results suggest that the 5'-domain of the chicken c-jun gene contains sequence elements that negatively regulate c-jun promoter activity in response to Pg.

Animals↗
PubMed1992

High-performance capillary electrophoretic analysis of chloramphenicol acetyl transferase activity.

This study highlights the potential utility of high-performance capillary electrophoresis (HPCE) for monitoring enzyme activity. Free-zone capillary electrophoresis is used to rapidly and reproducibly analyze the activity of the bacterial enzyme chloramphenicol acetyl transferase (CAT) which converts the substrates acetyl coenzyme A (CoA) and chloramphenicol to acetyl chloramphenicol and CoA. The results of this study indicate that HPCE may be an excellent tool for studying enzyme activities since it has several advantages over standard single parameters assays, most notably, the ability to monitor both loss of substrate and appearance of products simultaneously. Conditions have been identified for optimal separation of the substrate (chloramphenicol) from the products (acetylated derivatives). This presents a unique potential of HPCE for the analysis of enzymatic reactions that may be applied to areas of analytical research presently utilizing enzymatic reactions. One such analytical method is the CAT assay used for analysis of gene promoter activity. In this study, HPCE is shown to yield similar quantitative results with nonradiolabelled substrate in a fraction of the time. HPCE has several advantages over standard techniques including speed of analysis, no need for radiolabelled substrate, small sample volumes, high sensitivity/resolution and excellent quantitative capabilities.

Chloramphenicol O-Acetyltransferase↗
PubMed1992

Glucocorticoid regulation of alkaline phosphatase, osteocalcin, and proto-oncogenes in normal human osteoblast-like cells.

In humans, glucocorticoids are known to have marked effects on bone metabolism and function, including the significant regulation of osteoblast cells. To aid in the understanding of the mechanism of glucocorticoid action on normal human osteoblasts (hOB), confluent cells were analyzed for the presence of glucocorticoid receptors (GR) as well as for the effects of the glucocorticoid dexamethasone (Dex) on the expression of both the rapid responding nuclear proto-oncogenes and the late responding structural genes for bone matrix proteins. The interactions between Dex and 1,25 dihydroxy vitamin D3 (1,25 D3) on the gene expression in these cells were also examined. Using a functional receptor assay, a mean of 11,600 functional nuclear bound glucocorticoid receptors (range 6,000-22,000) was measured in fifteen separate cell strains. Northern blot analysis with a cDNA probe to the human GR was used to demonstrate the presence of a 7Kb transcript which is a candidate mRNA for GR in these cells. In agreement with previous studies, treatment of the hOB cells with Dex increased the steady state mRNA levels for alkaline phosphatase (AP) but displayed little or no effect on the mRNA levels for osteocalcin (OC) and glyceraldehyde phosphate dehydrogenase (GAPDH). Interestingly, the 1,25 D3 inductions of mRNA levels for OC were blocked by Dex but enhanced for AP. The above effects of Dex on AP and OC gene expression, including the interaction with 1,25 D3, were also shown to occur at the level of protein. The effect of Dex on the mRNA levels of the nuclear proto-oncogenes c-myc, c-fos, and c-jun was also investigated, since the oncoproteins (Fos/Jun) appear to play a role in the delayed glucocorticoid regulation of structural genes. Interestingly, Dex increased the steady state levels of c-myc, c-fos, and c-jun mRNAs in nonproliferating (confluent) hOB cells by 3.5-, 10-, and 2.0-fold, respectively, over control (untreated cells) values within one h of steroid treatment. The Dex-induced mRNA levels were transient and returned to basal values within 24 h of the steroid treatment. A reduced but qualitatively similar pattern of response was found in proliferating hOB cells. The pattern of response of these genes to glucocorticoids in hOB cells mimics the response in avian liver cells but not in reproductive cells. These results support the theory that hOB cells are target cells for glucocorticoids, and that as a primary event glucocorticoids rapidly regulate the expression of the nuclear oncoproteins Fos/Jun in these cells.

Alkaline Phosphatase↗
PubMed1992

The effect of 'conditioning' on hearing loss from a high frequency traumatic exposure.

The role of high frequency, low level 'conditioning' exposures as moderators of hearing loss from subsequent exposure to the same noise at a higher level was studied using monaural chinchillas. All the animals in the experimental groups were exposed to an octave band noise centered at 4 kHz at 85 dB SPL for 6 h a day for 10 days. One of the experimental groups was allowed to recover for 5 days and the other was allowed to recover for 18 h, prior to the higher level exposure at 100 dB for 48 h. A third group exposed only to the higher level constituted the control group. A comparison of threshold shifts and hair cell loss after 4 weeks of recovery across the three groups revealed: (a) the 5-day recovery group incurred greater threshold shifts than the other two groups; the hair cell loss in this group was greater than in the 18-h recovery group, but the same as in the control group and (b) the 18-h recovery group incurred considerably less threshold shift as well as hair cell loss than the other two groups. The results were also compared with the results from similar exposures using low frequency noise which indicated that the base vs. apex differences in the cochlea appear to extend to the effects of 'conditioning' exposures.

Animals↗
PubMed1991

Nuclear matrix localization and specific matrix DNA binding by receptor binding factor 1 of the avian oviduct progesterone receptor.

A chromatin acceptor protein for the avian oviduct progesterone receptor (PR), termed receptor binding factor 1 (RBF-1), has recently been shown to (1) be a component of the nuclear binding sites (acceptor sites) for PR and (2) generate high-affinity binding sites (termed the RBF-1 class of sites) on avian genomic DNA [Schuchard et al. (1991) Biochemistry 30, 4535-4542]. A second class of sites and its associated protein (termed RBF-2) were also identified. This paper demonstrates that RBF-1 and also the PR nuclear binding sites are localized in the oviduct nuclear matrix. RBF-1 is found in abundance in the nuclear matrix of liver but only in traces in the nuclear matrix of spleen. Extraction of the nuclear matrix with 4.0 M Gdn-HCl results in the complete removal of RBF-1 as occurs with whole chromatin. Interestingly, a second class of specific PR binding, termed RBF-2, remains on the nuclear matrix after the removal of all RBF-1. Southern blot analysis indicates that the nuclear matrix DNA contains sequences homologous with the 5'-flanking domains of the rapidly steroid regulated c-myc and c-jun protooncogenes and the beta-actin gene, but not genomic sequences of the late sex steroid regulated gene, ovalbumin, or the alpha-actin gene. A specific, small region in the 5'-flanking domain of the c-myc gene appears to be associated with the nuclear matrix. Southwestern blot analysis using partially purified RBF-1 shows a marked affinity and specificity of the RBF-1 for the nuclear matrix DNA.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Enrichment of a second class of native acceptor sites for the avian oviduct progesterone receptor as intact chromatin fragments.

Several classes of specific progesterone receptor (PR) nuclear binding sites (acceptor sites) have previously been identified in avian oviduct chromatin on the basis of different binding affinities. Recently, two classes of acceptor proteins (AP) that are associated with these binding sites in the avian oviduct have been identified. These APs were termed receptor binding factors (RBF-1 and -2), and one (RBF-1) has been purified [Schuchard et al. (1991) Biochemistry 30, 4535-4542]. The RBF-1 is associated with the highest affinity class of sites in the intact chromatin, and the RBF-2 is associated with the second highest affinity class of sites. The PR binding sites and their associated RBF-2 protein remain with the residual chromatin fraction following extraction by 4 M Gdn-HCl. This Gdn-HCl-treated chromatin has been termed nucleoacidic protein (NAP). This paper describes the 200-fold enrichment of the native RBF-2 class of PR acceptor sites beginning with the DNase I digestion of NAP to obtain DNase-resistant fragment (NAPf) containing approximately 150 bp of DNA. The PR binding sites are further enriched by high-performance or fast protein liquid chromatography and chromatofocusing. Anti-RBF-1/RBF-2 protein antibodies identify antigens that coelute with the PR binding activity. Hybridization analysis of the DNAf from the enriched NAPf demonstrates sequence homologies with the nuclear matrix DNA as well as with genomic sequences of the rapid steroid responding nuclear protooncogenes c-myc and c-jun. However, comparative analyses of the whole genomic DNA with the nuclear matrix DNA indicate that the RBF-2 (NAPf) is largely nonnuclear matrix.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Rapid induction of the c-jun protooncogene in the avian oviduct by the antiestrogen tamoxifen.

This report describes a rapid regulation of the expression of the c-jun protooncogene by the antiestrogen tamoxifen (Tam). The c-jun protooncogene codes for an important component of the AP-1 transcription factor complex, which regulates the expression of many unlinked genes. Repeated experiments have shown that Tam rapidly increases the steady-state c-jun mRNA levels in the avian oviduct but decreases the levels in the liver. The Tam effects are time- and dose-dependent. These results are supported by other studies that have demonstrated that 17 beta-estradiol decreases steady-state levels of c-jun protooncogene mRNA in oviducts of animals fully withdrawn from estradiol. The effect of Tam in the avian oviduct is in contrast to the reported effects of Tam on the expression of practically all other genes in the avian oviduct and other animal tissues. Transcription analyses using nuclear runoff experiments with oviduct nuclei demonstrate a decrease in the c-jun gene transcription within minutes after Tam treatment with a return to 75% of control values by 4 hr. The fact that Tam transiently decreases the transcription of the c-jun gene but increases the steady-state c-jun mRNA levels suggests that Tam must alter both transcriptional and post-transcriptional events. The results support a role of the c-jun protooncogene as a regulatory gene in the cascade model for steroid action whereby steroids rapidly regulate the regulatory genes, which in turn regulate many other structural genes.

Animals↗
PubMed1991

Cloning of murine tissue factor and regulation of gene expression by transforming growth factor type beta 1.

We have cloned a serum- and cycloheximide-inducible mRNA from AKR-2B murine fibroblasts which encodes a protein with significant sequence similarity to human tissue factor, a cellular initiator of the blood coagulation cascade. Information derived from this clone was used to establish the presence of a virtually identical sequence in mouse brain. Most importantly, cDNA-directed expression in a quail fibroblast cell line produced high levels of tissue factor procoagulant activity, confirming the identity of this protein as murine tissue factor. Additional studies demonstrate that transforming growth factor type beta 1 stimulates tissue factor gene transcription and is a potent inducer of tissue factor procoagulant activity in fibroblasts. Other tested mitogens such as platelet-derived growth factor, epidermal growth factor, and insulin were weak inducers. These results may reflect a role for transforming growth factor beta 1 in the maintenance of hemostasis or, alternatively, a role for tissue factor in cellular functions unrelated to blood coagulation.

Amino Acid Sequence↗
PubMed1991

The effect of 'conditioning' exposures on hearing loss from traumatic exposure.

The role of 'conditioning' exposures as moderators of hearing loss produced by exposure to a higher level noise was explored using chinchillas. Monaural chinchillas were exposed to an octave band of noise centered at 0.5 kHz at 95 dB for six hours/day for ten days. The subjects were allowed to recover to pre-exposure sensitivity and at five days after the last exposure they were re-exposed to the same noise at 106 dB. Thresholds recorded at various time intervals following the second exposure were compared with those recorded in a control group exposed only to the higher level noise. The experimental animals were found to have less threshold shift at all stages of recovery. Results are discussed in the light of results of other related studies and possible mechanisms involved are hypothesized.

Acoustic Stimulation↗