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PubMed1995

Thai Buddhist accounts of male homosexuality and AIDS in the 1980s.

In this paper the author considers the scriptural background to homophobic diatribes that emerged in the mid-1980s as the HIV epidemic slowly gained recognition in Thailand. By scrutiny of references to homosexuality in the Pali canon, he concludes that the scriptures are open to widely divergent interpretations. This doctrinal malleability is expressed in the contrasting views of contemporary Thai Buddhist intellectuals. The 'traditional' view claims that homosexuality is the kammic consequence of past sexual misconduct but in itself is not sinful and does not have future kammic consequences. The other, less tolerant view is that homosexuality derives from sexual excesses and, moreover, is the cause of the contemporary AIDS epidemic. Paradoxically it is the reformist Buddhist thinkers who have adopted the latter anti-homosexual position in that they tend to place less emphasis on kamma and more on individual responsibility for life circumstances.

Asia↗
PubMed1990

New approach to analysis of rotational panoramic radiography by computer simulation.

Computer simulation is an effective method for mathematical analysis of the image of rotational panoramic radiography. Conventional simulation methods have been used only for analysis of imaging factors and prediction of the characteristics of the obtained image. This paper describes a new computer simulation method that analyzes the imaging factors and produces the simulated image itself. The simulation is carried out by reproducing the actual image formation process in the computer. The head phantom for simulation, which represents the human dentomaxillofacial structures as a numerical value from 0 to 4, was constructed by scanning a real head phantom from neck to head with an X-ray CT scanner at a slice thickness of 1 mm and at 1 mm slice intervals. The diverging X-ray beam was expressed as a set of straight lines and was moved according to various parameters. The simulated image obtained was satisfactory for image analysis of rotational panoramic radiography.

Computer Simulation↗
PubMed1989

Ca2+ and Na+ currents in developing skeletal myoblasts are expressed in a sequential program: reversible suppression by transforming growth factor beta-1, an inhibitor of the myogenic pathway.

We have analyzed the biophysical and developmental properties of Ca2+ and Na+ currents in C2 muscle cells, whose morphological and biochemical phenotype closely resembles differentiated skeletal muscle. Both fused and unfused C2 myocytes possessed: (1) membrane capacitance consistent with the presence of complex sarcotubular invaginations, (2) tetrodotoxin-sensitive Na+ channels, and (3) "fast" and "slow" Ca2+ channels that inactivated at holding potentials of -40 and -20 mV, respectively. Thus, the passive electrical properties, Na+ currents, and Ca2+ currents expressed in C2 cells each differed from those found in the nonfusing muscle cell line, BC3H1, and corresponded more precisely to characteristic findings observed in skeletal muscle fibers. In further contrast to BC3H1 cells, C2 muscle also expressed "transient" Ca2+ channels similar to those reported in embryonic or neonatal skeletal muscle, which were detected within 12-24 hr of mitogen withdrawal, up to 60 hr before appearance of "fast" and "slow" currents. Na+ channels also were induced 12-24 hr after mitogen withdrawal. Unlike the "fast" and "slow" Ca2+ currents, which were maximally expressed at 8-14 d of serum withdrawal, "transient" Ca2+ channels became down-regulated upon prolonged differentiation (as found in postnatal skeletal muscle in vivo) and were no longer expressed at 14 d. Despite their divergent kinetic and developmental properties, all components of Ca2+ and Na+ current in C2 myocytes were suppressed reversibly in the presence of transforming growth factor beta-1, a purified growth factor that inhibits the myogenic phenotype. The results indicate that fusion is not essential for skeletal myoblasts to produce developmentally regulated voltage-gated channels that resemble those of intact muscle and demonstrate that the formation of diverse Ca2+ and Na+ channels can be mediated by a single peptide that affects the myogenic pathway.

Calcium↗
PubMed1989

Isolation and characterization of cDNA clones derived from the divergently transcribed gene in the region upstream from the human dihydrofolate reductase gene.

Transcripts initiated in the region immediately upstream from the human dihydrofolate reductase gene but transcribed from the opposite strand have been identified in human cells. These divergent upstream transcripts are polyadenylated, start at 89 base pairs upstream from the dihydrofolate reductase major initiation site, and are composed of two species of 5.0 and 3.8 kilobases in length. We have isolated complementary DNA clones derived from the divergent transcripts and identified a 3.5-kilobase open reading frame in one of these clones. Computer-assisted sequence analyses have shown that there is no significant sequence homology between the putative amino acid sequence and known protein sequences. This arrangement of two divergent transcriptional units suggests that the expression of these two genes may be regulated by a bidirectional promoter with common sequence elements.

Amino Acid Sequence↗
PubMed1988

Breast cancer--a woman's perspective.

This paper is bound to arouse controversy. It already has. No article in this special issue has been more thoroughly reviewed or had more widely divergent opinions. Some reviewers thought it expressed unrelieved anger; others felt it described the experience, perceptions, and feelings of many women with breast cancer. The paper seemed important to publish, however, because breast cancer is the second most common tumor occurring in American women. It is important to note that the author is a successful teacher and health professional who has a full and active family and social life. She describes encounters with breast cancer in an uncommonly fervent and articulate way. These encounters have clearly left deep scars. The essay is not scientific and not balanced in this issue by a paper by a women who had a wonderful experience with breast cancer. Such a paper may never be written. Read on!

Adaptation, Psychological↗
PubMed1985

On the use of dreams as a means of diagnosis of brain-injured patients.

In our psychotherapeutic work with brain injured patients it has become apparent that there exists a divergence between the two modes of expression with which the patient reflects his inner truth. In contrast to the patient's conscious self-perception, which is usually distorted by his cognitive deficits and by his defense mechanisms, the dream--the unconscious, uncovers another reality. The present paper cites a repeated pattern of initial dreams and an attempt is made to understand its diagnostic and prognostic meaning.

Brain Injuries↗
PubMed1994

Identification and mutation of primary and secondary proteolytic cleavage sites in murine stem cell factor cDNA yields biologically active, cell-associated protein.

Phenotypic abnormalities of melanocytes, germ cells, and hematopoietic cells of Steel mice demonstrate the critical role of stem cell factor (SCF) in development. Production of SCF in the hematopoietic microenvironment as either a membrane-associated or soluble factor leads to pleiotropic effects on hematopoietic stem and progenitor cells and significant effects on the production of erythroid cells. Although the production of these two forms of SCF is highly regulated, the physiologic role(s) of membrane-associated and soluble SCF remain unclear. We have demonstrated that the generation of soluble murine SCF by murine stromal cells derived from the fetal hematopoietic microenvironment is dependent on two distinct proteolytic cleavage sites. The primary site in exon 6 is preferentially utilized in these cells. The secondary site located in exon 7 is utilized only in the absence of the primary site. Proteolytic processing at this secondary site appears to be species-specific, since the human protein sequence diverges at this site, and protein expressed from the human cDNA encoding this site in murine stromal cells remains largely membrane-associated. Site-directed mutagenesis of the murine SCF cDNA encoding both proteolytic cleavage sites leads to the generation of membrane-associated and biologically active SCF on murine stromal cells. These results suggest that the regulation of processing of the secondary proteolytic cleavage site could play a critical role in the function of membrane-associated SCF protein.

Amino Acid Sequence↗
PubMed1996

Evidence for progenitors of chronic lymphocytic leukemia B cells that undergo intraclonal differentiation and diversification.

Peripheral blood mononuclear cells from five patients with IgG+ B-type chronic lymphocytic leukemia (B-CLL) were analyzed for the presence of clone-specific Ig H chain variable region gene mRNA transcripts linked to C mu and/or C alpha. This was assessed by (1) comparing the lengths of portions of the VHDJH of the IgG+ CLL clones with those of the mu and alpha isotype-expressing B cells, (2) performing clone-specific endonuclease digestion studies, and (3) determining the DNA sequences of the mu and alpha isotype-expressing cDNA. Thus, when B-cell mRNA from these five patients were reverse transcribed with C gamma-specific primers and then amplified by polymerase chain reaction, dominant cDNA were found with lengths corresponding to those of the IgG+ CLL B cell. In addition, in four cases, cDNA of lengths identical to those of the CLL B cell were detected when mRNA was reverse transcribed and amplified using c mu- and/or C alpha-specific primers, strongly suggesting clonal relatedness. These CLL-related mu- and alpha-expressing cDNA were present in greater amounts that unrelated (non-CLL) mu- and alpha-expressing cDNA from normal B cells that used genes of the same VH family. When the sequences of these CLL-related C mu- and C alpha-expressing cDNA were compared with those of the IgG+ CLL clones, it was clear that they were derived from the same ancestral gene as the IgG-expressing CLL B cell, thus documenting their common origin. Finally, nucleotide point mutations were observed in the mu- and alpha-expressing cDNA of certain patients, indicating divergence with the CLL. These data suggest that IgM+ B cells, which are precursors of the leukemic B cells, exist in increased numbers in the blood of most patients with IgG+ B-CELL and that these cells may differentiate, accumulate V genes mutations, and undergo isotype switching in vivo. In addition, the data are consistent with a sequential-hit model for the evolution of CLL.

Base Sequence↗
PubMed1998

[Analysis of a novel delta gene during the formation of cortical layers].

The preplate is composed of earliest-generated neurons, including Cajal-Retzius cells and prospected subplate neurons which together play a critical role in the formation of cortical layers. Additionally, Notch and Delta have been identified as key components that mediate the lateral specification and fate of numerous cells during development. Here, we demonstrate that a divergent type of Delta (D113) is expressed transiently and specifically in a set of earliest generated neurons in the neural tube. Moreover, in developing cerebrum, D113 is specifically and transiently expressed in prospected subplate neurons. Thus, our results suggest that D113 mediates the specification of cell fate in the earliest stage of neurogenesis including the neurons that compose preplate, leading to the mechanism of genesis of cortical layers.

Animals↗
PubMed2003

Gender differences and hormonal modulation of G proteins Galpha(q/11) expression in lymphoid organs.

Androgens and estrogens exert potent divergent feedback effects on gonadotropin-releasing hormone (GnRH) production at the level of the hypothalamus and GnRH action at the level of the pituitary. Androgens exert generally suppressive effects on GnRH production and action, whereas rising levels of estradiol increase both GnRH release and action. In addition to its known endocrine actions, GnRH possesses immunomodulatory effects. We have previously demonstrated gender differences in immune responsiveness to GnRH that parallel gender differences in endocrine responsiveness: females appear to be more immunologically responsive to GnRH than males. GnRH exerts its actions via the stimulatory G protein Galpha(q) and Galpha(11) (referred to collectively as Galpha(q/11)) as well as via Galpha(s). We have recently demonstrated that the heightened immune responsiveness to GnRH in lupus-prone female mice correlated with increased expression of Galpha(q/11) in lymphoid cells from females compared to males. We hypothesize that the hormonal milieu of females may contribute to increased expression of stimulatory G proteins and to the heightened immune and endocrine responsiveness to GnRH. In this report, we document gender differences in expression of Galpha(q/11) protein in lymphoid organs in non-autoimmune DBA/2 mice. In an effort to address the mechanisms for the gender differences in G-protein expression, we used competitive reverse transcription PCR to quantitate mRNA for stimulatory G proteins in immune cells under various hormonal conditions. We quantitated the expression of Galpha(q/11) mRNA and protein under physiologic hormonal alterations, i.e. throughout the estrous cycle in female mice. We demonstrate that expression of Galpha(q/11) mRNA and protein in lymphoid organs is significantly increased on the afternoon of proestrus compared to metestrus. Additional studies demonstrate that exposure to GnRH or to estrogens significantly increases the expression of Galpha(q/11) mRNA in immune cells. These findings support an active role for hormonal modulation of G proteins in the gender differences in endocrinologic and immunologic responsiveness to GnRH.

Animals↗
PubMed1989

Expression of T-cell receptor antigens in mycosis fungoides and inflammatory skin lesions.

Using immunohistologic methods, we studied the expression of the T-cell receptor (TCR)-associated antigens CD3, TCR-beta, and TCR-delta by cutaneous T cells in mycosis fungoides (MF) (36 patients) and a variety of inflammatory diseases (16 patients). Most T cells in the inflammatory diseases and patch/plaque mycosis fungoides expressed the immunophenotype characteristic of the vast majority of mature peripheral T cells: CD3+ TCR-beta+ TCR-delta-. In contrast, abnormal CD3/TCR-beta antigen expression was seen in 3 of 6 cases (50%) of tumor stage mycosis fungoides. Furthermore, we were able to document its evolution from the normal pattern present in earlier patch/plaque lesions of the two cases in which serial biopsies were available for study. Divergence of epidermal versus dermal CD3/TCR-beta antigen expression was seen in 2 of 34 (6%) of biopsies of patch/plaque mycosis fungoides but not in inflammatory controls. The TCR-delta+ cells were generally rare regardless of diagnosis. We conclude that inflammatory skin diseases and most patch/plaque mycosis fungoides are typically composed of T lymphocytes that resemble mature peripheral T cells in regard to their expression of TCR-associated antigens. In contrast, aberrant patterns of TCR-associated antigen expression can be seen in tumor stage MF, and, more rarely in patch/plaque MF.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1996

Paralogous Hox genes: function and regulation.

The Hox homeobox gene family plays a pivotal role in regulating patterning and axial morphogenesis in vertebrates. Molecular characterization of the four Hox clusters has shown that they are evolutionarily related with respect to sequence, organization, and expression, suggesting they arose by duplication and divergence. Transgenic analysis has clearly demonstrated the functional roles of individual genes in a broad range of embryonic tissues, and in compound mutants has addressed the issues of cooperativity and redundancy. There is an emerging picture of the cis-regulatory elements underlying Hox expression, and for the 3' members of the clusters there is a considerable degree of conservation between paralogous genes with respect to their functional roles and regulatory control.

Animals↗
PubMed1997

The TATA-less promoter of hepatitis B virus S gene contains a TBP binding site and an active initiator.

The surface antigen (S) gene promoter, one of the major hepatitis B virus (HBV) promoters, directs the synthesis of a 2.1 kb mRNA which encodes the preS2 and S polypeptides. The preS2/S promoter does not contain a classical TATA box, and transcription regulation of the preS2/S gene has not been fully elucidated. We analysed two regions involved in preS2/S gene transcription of the HBV adw subtype: the diverged TATA box and a putative initiator element. We demonstrated sequence specific promoter activity of the putative TATA-like sequences in the preS2/S gene promoter (-25 to -32 bp). Using end labeled synthetic oligonucleotides we observed specific binding of nuclear extracts to the diverged TATA sequence, that was significantly reduced using a mutated oligonucleotide. Specific binding of yeast TBP to the diverged TATA sequence was shown which was increased in the mutant containing a classical TATA box. We analysed the proposed initiator (Inr) sequence of the preS2/S promoter region (-13 to -16 bp). Deletion of the inr element markedly reduced promoter activity as assessed by CAT expression. Gel shift assays showed specific binding of nuclear extracts to wild type but not to mutant Inr. Expression studies with double mutants of the diverged TATA and the Inr element established that both elements are active in transcription regulation.

Base Sequence↗
PubMed2004

Regulation of Egr-1, SRF, and Sp1 mRNA expression in contracting skeletal muscle cells.

The early cellular signals associated with contractile activity initiate the activation and induction of transcription factors that regulate changes in skeletal muscle phenotype. The transcription factors Egr-1, Sp1, and serum response factor (SRF) are potentially important mediators of mitochondrial biogenesis based on the prevalence of binding sites for them in the promoter regions of genes encoding mitochondrial proteins, including PGC-1 alpha, the important regulator of mitochondrial biogenesis. Thus, to further define a role for transcription factors at the onset of contractile activity, we examined the time-dependent alterations in Egr-1, Sp1, and SRF mRNA and the levels in electrically stimulated mouse C(2)C(12) skeletal muscle cells. Early transient increases in Egr-1 mRNA levels within 30 min (P < 0.05) of contractile activity led to threefold increases (P < 0.05) in Egr-1 protein by 60 min. The increase in Egr-1 mRNA was not because of increased stability, as Egr-1 mRNA half-life after 30 min of stimulation showed only a 58% decline. Stimulation of muscle cells had no effect on Sp1 mRNA but led to progressive increases (P < 0.05) in SRF mRNA by 30 and 60 min. This was not matched by increases in SRF protein but occurred coincident with increases (P < 0.05) in SRF-serum response element DNA binding at 30 and 60 min as a result of SRF phosphorylation on serine-103. To assess the importance of the recovery period, 12 h of continuous contractile activity was compared with four successive 3-h bouts, with an intervening 21-h recovery period after each bout. Continuous contractile activity led to a twofold increase (P < 0.05) in Egr-1 mRNA, no change in SRF mRNA, and a 43% decrease in Sp1 mRNA expression. The recovery period prevented the decline in Sp1 mRNA, produced a decrease in Egr-1 mRNA, and had no effect on SRF mRNA. Thus continuous and intermittent contractile activity evoked different specific transcription factor expression patterns, which may ultimately contribute to divergent qualitative, or temporal patterns of, phenotypic adaptation in muscle.

Animals↗
PubMed2002

Expression of myostatin is not altered in lines of poultry exhibiting myofiber hyper- and hypoplasia.

Decades of selective breeding have yielded lines of poultry with substantial myofiber hyperplasia, vet little is known about what genes have been altered during the course of selection. Myostatin is a strong negative regulator of muscle mass in mice and cattle and could have been one of many genetic factors contributing to increased myofiber deposition in growth-selected lines of poultry. To test this hypothesis, the sequence and expression patterns of myostatin were analyzed in growth-selected lines of chickens and quail. The sequence of broiler myostatin cDNA, amplified via reverse transcription (RT)-PCR from embryonic muscle RNA, contained no missense mutations in the coding sequence when compared to that of White Leghorn layers, although two silent single nucleotide polymorphisms (SNP) were found. Northern analysis of myostatin transcripts from embryonic pectoralis and quadriceps showed no significant differences in expression levels between broiler and layer muscle RNA. However, levels of myostatin transcripts were greatly reduced in muscles of posthatch chicks compared to embryonic muscle. Myostatin protein was also present in broiler and layer embryonic muscle at similar levels. No significant polymorphisms or differences in RNA expression levels were found in embryonic muscles of divergently selected lines of Japanese quail. These results indicate that intense artificial selection in these growth-selected lines of poultry has neither silenced the expression of myostatin nor created null alleles via mutation in the lines analyzed.

Animal Husbandry↗
PubMed2004

Prediction of site-specific amino acid distributions and limits of divergent evolutionary changes in protein sequences.

We derive an analytic expression for site-specific stationary distributions of amino acids from the structurally constrained neutral (SCN) model of protein evolution with conservation of folding stability. The stationary distributions that we obtain have a Boltzmann-like shape, and their effective temperature parameter, measuring the limit of divergent evolutionary changes at a given site, can be predicted from a site-specific topological property, the principal eigenvector of the contact matrix of the native conformation of the protein. These analytic results, obtained without free parameters, are compared with simulations of the SCN model and with the site-specific amino acid distributions obtained from the Protein Data Bank. These results also provide new insights into how the topology of a protein fold influences its designability, i.e., the number of sequences compatible with that fold. The dependence of the effective temperature on the principal eigenvector decreases for longer proteins, as a possible consequence of the fact that selection for thermodynamic stability becomes weaker in this case.

Amino Acids↗
PubMed1990

Two evolutionarily divergent genes encode a cytoplasmic ribosomal protein of Arabidopsis thaliana.

Two clones of Arabidopsis thaliana possessing high sequence identity to the yeast gene encoding ribosomal (r) protein L3 were isolated by heterologous DNA hybridization. The coding regions of these two clones have approx. 63% amino acid (aa) sequence identity to the yeast L3 r-protein and 85% aa sequence identity to each other. Both genes are expressed in shoots. The presence of two divergent genes in A. thaliana raises the possibility that the gene products participate in the formation of functionally distinct ribosomes.

Alleles↗
PubMed1996

The chromosomal distribution of mouse odorant receptor genes.

Odorant receptors (ORs) on nasal olfactory sensory neurons are encoded by a large multigene family. Each member of the family is expressed in a small percentage of neurons that are confined to one of several spatial zones in the nose but are randomly distributed throughout that zone. This pattern of expression suggests that when the sensory neuron selects which OR gene to express it may be confined to a particular zonal gene set of several hundred OR genes but select from among the members of that set via a stochastic mechanism. Both locus-dependent and locus-independent models of OR gene choice have been proposed. To investigate the feasibility of these models, we determined the chromosomal locations of 21 OR genes expressed in four different spatial zones. We found that OR genes are clustered within multiple loci that are broadly distributed in the genome. These loci lie within paralogous chromosomal regions that appear to have arisen by duplications of large chromosomal domains followed by extensive gene duplication and divergence. Our studies show that OR genes expressed in the same zone map to numerous loci; moreover, a single locus can contain genes expressed in different zones. These findings raise the possibility that OR gene choice may be locus-independent or involve consecutive stochastic choices.

Animals↗