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Biomedical subjects

R Menon

Publications and source records attributed to R Menon.

At least 55 records · Page 3Linked to original sources

Interleukin-10 inhibition of interleukin-6 in human amniochorionic membrane: transcriptional regulation.

OBJECTIVE: Our purpose was to study the regulatory effects of recombinant interleukin-10 on interleukin-6 messenger ribonucleic acid and protein production by human fetal membranes. STUDY DESIGN: Amniochorionic membranes were collected from women undergoing elective cesarean section. Membranes were maintained in an organ explant system and stimulated with media containing lipopolysaccharide (50 ng/ml) and various amounts of recombinant interleukin-10 (10, 50, 100 ng/ml). Experiments were conducted in a dose- and time-dependent manner. Transcription and translation of interleukin-6 were monitored with quantitative reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assay. RESULTS: Interleukin-10 stimulation of amniochorionic membranes in culture produced a dose-dependent decrease in the production of interleukin-6 messenger ribonucleic acid and protein. Quantitative polymerase chain reaction was used to document a decrease in interleukin-6 messenger ribonucleic acid, which paralleled the decrease in peptide levels as detected with enzyme-linked immunosorbent assay. The interleukin-10 effect was present only when tissue was concurrently stimulated with lipopolysaccharide. Interleukin-10 inhibition could not be produced in the absence of lipopolysaccharide stimulation. CONCLUSION: Addition of interleukin-10 to culture media leads to transcriptional regulation of interleukin-6, which results in decreased production of both messenger ribonucleic acid and protein by human amniochorionic membranes. The decrease in interleukin-6 is a dose-dependent effect of interleukin-10. This finding may have important implications with respect to a possible role for interleukin-10 or an interleukin-10 stimulatory factor in the management of preterm labor associated with the presence of inflammatory cytokines.

Amnion↗

Sequence-specific priming and exonuclease-released fluorescence detection of HLA-DQB1 alleles.

Molecular typing of HLA DQB1 alleles, employing sequence-specific primers (SSP) for PCR amplification, was used to test a novel method that eliminates the requirement for subsequent gel electrophoresis or additional hybridization steps by directly detecting positive reactions. We have evaluated the performance of this fluorescence-based oligonucleotide probe assay to assign the most common DQB1 alleles on DNA from 14 homozygous cell lines and in a blind study of 50 diabetic patient samples that had been previously typed at the DQB1 locus using SSOP and conventional SSP-based approaches. We used a panel of 14 DQB1 SSP primer pairs, internal control primers, and a combination of 4 fluorescent oligonucleotide probes to detect 14 alleles or groups of alleles and controls. We can reliably detect single-base allelic differences, observe 100% concordance with the results obtained using both of the standard methods, and are able to further subtype several alleles that are not easily distinguished using SSOP (e.g. DQB1 *0401/0402 and DQB1 *0302/ 0303). Sequence-specific priming and exonuclease-released fluorescence (SSPERF) detection is technically simple and can be performed in less than 2 hours, including DNA extraction, PCR amplification, data analysis and allele identification. This method is particularly useful for the analysis of large numbers of samples, for which high throughput is critical and for which gel-based approaches are difficult to perform. This technique may also be useful for small-scale class I and class II molecular typing in clinically oriented laboratories.

Alleles↗

Third molars in the establishment of adult status--a case report.

Teeth are the most durable structures in the human body. The timing and sequence of their development, as contained in dental development charts, have been used as valid criteria for age determination. The third molars however are the last teeth to erupt and are regarded as the most variable in the dentition. Age estimation in a legal context, using developing third molars must be carefully applied otherwise justice may miscarry. A case of wrongful use of the technique is presented here.

Adolescent↗

High contrast and fast three-dimensional magnetic resonance imaging at high fields.

A new three-dimensional imaging strategy based on magnetization prepared ultrafast gradient recalled echo technique that demonstrates pronounced T1 contrast at high fields is introduced. High-resolution three-dimensional image sets of human brain showing high contrast between white and gray matter areas are presented. The ratio of contrast-to-noise was examined as a function of the relevant parameters in the imaging sequence; calculations based on high-field T1 values as well as the experimental data demonstrated that maximal contrast-to-noise ratio is attained under the same magnetization preparation conditions both for cortical and subcortical gray matter relative to white matter, leading to approximately equivalent appearance of all gray matter areas in the same image. In addition, the images displayed clear visualization of subtle anatomical structures such as the subthalamic nuclei (ventral tier nuclei, dorsomedial nucleus, and pulvinar) and mammillothalamic tracts.

Brain↗

Expression of inflammatory cytokines (interleukin-1 beta and interleukin-6) in amniochorionic membranes.

OBJECTIVE: This study was designed to investigate the expression of inflammatory cytokines (interleukin-1 beta and interleukin-6) by fetal membranes in response to infection in vivo and to endotoxin in organ culture. STUDY DESIGN: Amniochorionic membranes were collected from infected and uninfected women and analyzed for cytokine messenger ribonucleic acid and protein. Normal membranes were cultured and exposed to endotoxin. Messenger ribonucleic acid expression was analyzed by reverse transcriptase-polymerase chain reaction. Cellular localization of messenger ribonucleic acid and protein was determined by in situ hybridization and immunocytochemical evaluation, respectively. RESULTS: Messenger ribonucleic acid for interleukin-1 beta appeared to be increased in infected or endotoxin-stimulated amniochorionic membranes, whereas interleukin-6 messenger ribonucleic acid was only observed in infected membranes or after endotoxin stimulation. Interleukin-1 beta messenger ribonucleic acid was localized exclusively to chorionic cells, whereas protein was observed in both chorion and amnion. Interleukin-6 messenger ribonucleic acid and protein were produced in both amniotic and chorionic cells. CONCLUSION: Amniochorionic membranes are a site of inflammatory cytokine production. These findings may have significance in preterm labor or premature rupture of membranes.

Amnion↗

Magnetic resonance imaging volumetric measurements of the superior temporal gyrus, hippocampus, parahippocampal gyrus, frontal and temporal lobes in late paraphrenia.

Quantified magnetic resonance measurements were made of superior temporal gyrus, parahippocampal gyrus, hippocampal, frontal and temporal lobe volumes and of the planar area of the thalamus and basal ganglia structures in 47 late paraphrenic patients and 33 healthy elderly controls. The late paraphrenics were divided into 31 schizophrenics and 16 patients with delusional disorder according to ICD-10 guidelines. Patients with delusional disorder tended to have smaller left temporal volumes compared with control subjects and patients with schizophrenia, but this difference failed to reach accepted levels of statistical significance after correction for the effects of multiple statistical comparisons, age and total brain size. Physiological right-left asymmetry, reported for temporal and frontal lobe volumes, was present in control, schizophrenic and delusional disorder subjects but delusional disorder patients had a significantly greater degree of temporal lobe asymmetry. The results add to the evidence for heterogeneity among late-onset psychoses and emphasize the subtle nature of any structural brain abnormalities in these patients.

Aged↗

Amniochorion: a source of interleukin-8.

PROBLEM: Preterm labor and premature rupture of the membranes are major complications of pregnancy. We have reported the possible role of amniochorionic membrane in the production of inflammatory cytokines and the early onset of labor. This study was conducted to detect the expression of IL-8 mRNA and peptide production in cultured fetal membranes. METHOD: Amniochorionic membranes were collected from women undergoing elective cesarean section at term. Membranes were cultured in an organ explant system and the expression of IL-8 was studied over a 10-day period by RT-PCR and in situ hybridization. IL-8 peptide localization was accomplished using immunocytochemistry. RESULTS: Constitutive expression of IL-8 mRNA in cultured fetal membranes was demonstrated in both amniotic and chorionic leave cells. mRNA and peptide for IL-8 was homogeneously distributed throughout the amniotic and chorionic cells. CONCLUSION: Human amniochorionic membrane is a source of IL-8 mRNA and peptide.

Amnion↗

Organ culture of amniochorionic membrane in vitro.

PROBLEM: The purpose of the study was to develop a novel method of amniochorionic membrane culture aimed at maintaining tissue integrity. METHOD: Amniochorionic membranes were collected from women prior to labor, undergoing elective cesarean section, with no history of infection or pregnancy related complication. Fetal membranes were maintained in culture for up to ten days. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a "house keeping" gene and inflammatory cytokine mRNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) at various periods of culture. mRNA localization was performed by in situ hybridization. RESULTS: GAPDH gene expression was seen throughout the culture period. Inflammatory cytokine mRNA (IL-1 alpha, IL-1 beta and IL-6) were also detected during culture. Cellular and tissue morphology appeared normal. CONCLUSIONS: The culture technique we propose is a simple organ explant system which maintains the morphology and autocrine/paracrine relationships within this tissue.

Amnion↗

Expression of TNF-alpha and TNFR p55 in cultured amniochorion.

PROBLEM: Preterm labor and PROM are major complications of pregnancy. We have reported the possible role of amniochorionic membrane as it relates to the production of cytokines and the early onset of labor. Amniochorion is capable of responding to an infectious process with the production of IL-6 and IL-1 beta. Here we examine the expression of TNF-alpha and TNFR in amniochorion. METHOD: Amniochorionic membranes were collected and maintained in an organ explant system. Samples were frozen and/or fixed for RT-PCR, in situ hybridization, and immunocytochemistry. RESULTS: RT-PCR demonstrated mRNA for TNF-alpha and in situ hybridization localized mRNA in chorion and amnion. Immunocytochemistry demonstrated TNF-alpha peptide in amnion but not in chorion. Immunocytochemical localization of TNFR indicates presence of that peptide in both amnion and chorion. CONCLUSIONS: We conclude that the fetal membranes are sources of TNF-alpha and TNFR, supporting our previous work indicating that fetal membranes are active participants in the response to intraamniotic infection.

Amnion↗

Imaging at high magnetic fields: initial experiences at 4 T.

This article reviews the preliminary experiences and the results obtained on the human brain at 4 T at the University of Minnesota. Anatomical and functional images are presented. Contrary to initial expectations and the early results, it is possible to obtain high-resolution images of the human brain with exquisite T1 contrast, delineating structures especially in the basal ganglia and thalamus, which were not observed clearly in 1.5-T images until now. These 4-T images are possible using a new approach that achieves maximal contrast for different T1 values at approximately the same repetition time and has built-in tolerance to variations in B1 magnitude. For functional images, the high field provides increased contribution from the venuoles and the capillary bed because the susceptibility-induced alterations in 1/T2* from these small-diameter vessels increase quadratically with the magnitude of the main field. Images obtained with short echo times at 4 T, and by implication at lower fields with correspondingly longer echo times, are expected to be dominated by contributions from large venous vessel or in-flow effects from the large arteries; such images are undesirable because of their poor spatial correspondence with actual sites of neuronal activity.

Brain↗

Intrinsic signal changes accompanying sensory stimulation: functional brain mapping with magnetic resonance imaging.

We report that visual stimulation produces an easily detectable (5-20%) transient increase in the intensity of water proton magnetic resonance signals in human primary visual cortex in gradient echo images at 4-T magnetic-field strength. The observed changes predominantly occur in areas containing gray matter and can be used to produce high-spatial-resolution functional brain maps in humans. Reducing the image-acquisition echo time from 40 msec to 8 msec reduces the amplitude of the fractional signal change, suggesting that it is produced by a change in apparent transverse relaxation time T*2. The amplitude, sign, and echo-time dependence of these intrinsic signal changes are consistent with the idea that neural activation increases regional cerebral blood flow and concomitantly increases venous-blood oxygenation.

Brain Chemistry↗

Influence of some dietary chemopreventive agents on the expression of functional differentiation of the mouse mammary gland in vitro.

The purpose of the present studies was to determine the influence of the chemopreventive agents selenium, 4-(hydroxyphenyl)-retinamide (4-HPR) and beta-carotene, on functional differentiation (lactogenesis) of the mouse mammary gland cells. The hormone-induced expression of the milk-protein genes, beta-casein, epsilon-casein and the whey acidic protein (WAP) was used as molecular marker of differentiation of the mammary cells in organ culture medium containing insulin, prolactin, aldosterone and hydrocortisone. Quantitative determination of the cellular concentration of the respective mRNA was ascertained by molecular hybridization of RNA to the specific cloned cDNA probes using both the solution and the filter hybridization methods. Selenium at 100 nm concentration caused a pronounced inhibition of accumulation of the respective mRNAs. The retinoid, 4-HPR, also caused a dose-dependent inhibition of expression of these mRNA sequences. In contrast, concentrations of the 3 mRNAs in beta-carotene-treated glands remained similar to those observed in glands cultured in medium containing the hormones and hexane (the latter being the solvent for beta-carotene). The significant antagonistic action of selenium and 4-HPR, however, was reversible after removal of the chemicals from the culture medium. Thus, among the 3 chemicals tested, selenium and retinoid can cause an adverse effect on functional differentiation (lactogenesis) of the mammary cells. This inhibitory effect, however, was reversible. beta-Carotene, on the other hand, caused no apparent antagonistic effect on expression of the milk-protein genes in the isolated whole mammary organ in culture.

Actins↗

Metabolism of 7,12-dimethylbenz[a]anthracene by mouse mammary cells in serum-free organ culture medium.

The murine mammary gland is a prime target organ for 7,12-dimethylbenz[a]anthracene, (DMBA)-induced carcinogenesis. We analyzed the metabolism of 3H-DMBA in a cell-free microsomal activation system derived from mouse mammary microsomes and in a whole mammary organs in culture. The in vitro microsomal activation system failed to show the more polar diol derivative of DMBA after HPLC. The metabolites obtained directly from the 3H-DMBA-treated whole mammary organs, however, revealed the presence of both the diol as well as the phenolic derivatives of DMBA. Analysis of the glands and the culture medium further showed that nearly 95% of the radioactivity added to the culture medium was associated with the adipose tissue and complete solubilization of the fat pad released substantial amounts of DMBA and its metabolites. It appears that a large portion of DMBA and its metabolites remain entrapped in the adipose tissue surrounding the parenchyma. Formic acid digestion of the gland releases the DMBA and the metabolites allowing their ethylacetate extraction, and HPLC characterization.

9,10-Dimethyl-1,2-benzanthracene↗

Job transfers: a neglected aspect of migration in Malaysia.

"This article examines the incidence of [job-related] transfers in Malaysia. The retrospective migration data from the Malaysia Family Life Survey [conducted in 1976-1977] are used to demonstrate that transfers comprise 18 percent of all migration in the country and that there has been a rise in the incidence of transfers over a 35 year period. Factors underlying this trend are outlined. Furthermore, significant differences in age, educational attainment and other characteristics between transferees and other types of migrants are identified and their implications discussed."

Age Factors↗