Biomedical subjects
Publications and source records attributed to J B Murray.
Sexual dysfunction side effects have been associated with antidepressant medication, especially with serotonin reuptake inhibitors. Neurotransmitters appear to be involved, especially dopamine and serotonin, but the processes by which they influence sexual dysfunction are not clear.
Methadone maintenance programs have effectively reduced heroin dependency and are available in most countries affected by heroin addiction. Methadone, developed in Germany during World War II as a pain killer, does not have the euphoric effects of heroin and the goal of treatment is to substitute methadone for heroin use. Recidivism is probably a life-long risk. Methadone maintenance programs began in the 1960s in the United States in New York City. Once tolerance is developed, it may be used continually without harmful side effects. Dosage is important for effectiveness as are counseling, rehabilitation services, and employment support. Reduction in criminality and AIDS has been associated with methadone maintenance programs.
Crystal structures of two complexes between recombinant MS2 capsids and RNA operator fragments have been determined at 2.7 A resolution. The coat protein of the RNA bacteriophage MS2 is bifunctional; it forms the icosahedral virus shell to protect the viral nucleic acid and it acts as a translational repressor by binding with high specificity to a unique site on the RNA, a single stem-loop structure, containing the initiation codon of the gene for the viral replicase. In order to determine the structure of these protein-RNA complexes, we have used chemically synthesized variants of the stem-loop fragment and soaked them into crystals of recombinant capsids. The RNA stem-loop, as bound to the protein, forms a crescent-like structure and interacts with the surface of the beta-sheet of a coat protein dimer. It makes protein contacts with seven phosphate groups on the 5' side of the stem-loop, with a pyrimidine base at position -5, which stacks onto a tyrosine, and with two exposed adenine bases, one in the loop and one at a bulge in the stem. Replacement of the wild-type uridine with a cytosine at position -5 increases the affinity of the RNA to the dimer significantly. The complex with RNA stem-loop having cytosine at this position differs from that of the wild-type complex mainly by having one extra intramolecular RNA interaction and one extra water-mediated hydrogen bond.
Cardiovascular effects of gender-specific ability perceptions (ability perceptions linked to one's identity as being female or male) were examined under different task conditions. In Study I, participants were led to believe that either men (masculine task) or women (feminine task) tend to do well on a memory task and then were provided the chance to avoid noise by attaining a low or high performance standard. As expected, sex differences in systolic blood pressure response during performance depended not only on task type but also on the degree of challenge. In Study 2, high standard effects were strengthened and extended through the use of an appetitive procedure and the inclusion of conditions in which the performance standard was extreme. Findings are discussed in terms of an interactional analysis of ability percepts, task demand, and cardiovascular responsivity.
Munchausen syndrome (MS) is characterized by patients' chronic and relentless pursuit of medical treatment for combinations of symptoms of consciously self-inflicted injury and falsely reported symptomatology. MS patients are adults, as are perpetrators of Munchausen syndrome by proxy (MSBP). MSBP is an unusual form of child abuse in which a parent, usually the mother, brings a child for medical attention with symptoms falsified or directly induced by the mother. Most reports of MSBP have been of a few cases appearing in medical and pediatric literature. Motivation in both disorders is unclear, and diagnosis and treatment present difficult problems that require more research.
Tourette's syndrome (TS), once considered a rare disorder, has been investigated extensively in the last two decades. It is inherited, usually beginning in childhood, and waxes and wanes, usually decreasing in frequency and severity in adolescence and early adulthood. Pharmacotherapy is the usual treatment approach, reducing frequency and severity of symptoms, but it is not a cure and often has side effects. Psychological help for people with TS and their families may be needed for this complex disorder.
Motion sickness may occur during travel by sea, automobile, airplane, and space. Susceptibility changes with age and may be influenced by psychological factors. Susceptibility can be reduced in most people by medications that involve histamine or neurotransmitters acetylcholine and noradrenaline, and influence the vestibular system.
The crystal structure of an unmodified hammerhead RNA in the absence of divalent metal ions has been solved, and it was shown that this ribozyme can cleave itself in the crystal when divalent metal ions are added. This biologically active RNA fold is the same as that found previously for two modified hammerhead ribozymes. Addition of divalent cations at low pH makes it possible to capture the uncleaved RNA in metal-bound form. A conformational intermediate, having an additional Mg(II) bound to the cleavage-site phosphate, was captured by freeze-trapping the RNA at an active pH prior to cleavage. The most significant conformational changes were limited to the active site of the ribozyme, and the changed conformation requires only small additional movements to reach a proposed transition-state.
A screening of a range of common laboratory antibiotics for inhibition of the hammerhead ribozyme has shown that in addition to certain aminoglycosides (most notably neomycin B) the tetracyclines are also effective inhibitors, with chlorotetracycline being more effective than tetracycline. Inhibition by chlorotetracycline is not as strong as that by neomycin B but is more complicated, with at least two binding sites apparent. As with hammerhead inhibition by neomycin B, chlorotetracycline inhibition can be overcome by raising the concentration of the Mg2+ ion cofactor. We find that around six Mg2+ ions will displace neomycin B, compared with twelve for chlorotetracycline. Inhibition observed in the presence of mixtures of neomycin B and chlorotetracycline is consistent with separate binding sites on the hammerhead for these two classes of antibiotic. Under certain conditions of the mixing order and low concentration of chlorotetracycline, enhancement of single-turnover hammerhead cleavage by up to 20% is observed, with higher concentrations of antibiotic being inhibitory. We have also found that the presence of 2.5% (v/v) DMSO causes a 30% enhancement of the single-turnover cleavage. These results thus extend the range of known inhibitors of hammerhead cleavage, and also demonstrate how the cleavage can be accelerated.
The neurological, neurochemical, and neurotransmitter level differences as well as genetic influences associated with autism have been studied extensively in the last two decades. The varied findings from research offer hope for better understanding, effective treatment, and, perhaps, cure of this pervasive developmental disorder.
Depression occurs in 40% to 50% of Parkinson's disease (PD) patients. From research over the last decade, this co-occurrence appears to be more than an emotional reaction to disability. Replacement of the neurotransmitter, dopamine, benefits the PD motor functions, and the neurotransmitter, serotonin, in antidepressants, reduces depressive symptoms. However, identification of the physiological linkage between PD and depression continues to evade research efforts.
We report details of the synthesis and characterization of oligoribonucleotides containing 4-thiouridine or 2-pyrimidinone ribonucleoside (4HC). We have used these probes to examine the roles of the conserved pyrimidines in the central core of the hammerhead ribozyme. The effects on catalysis of singly-substituted hammerhead ribozyme and substrate strands were quantified in multiple-turnover reactions. Various effects were observed on kcat. and Km, with up to a 7-fold decrease and a 3-fold increase respectively. For substitutions with 4HC at positions 3 or 17, catalytic activity in single turnover reactions can be increased up to 8-fold equivalent to 40% of wild-type activity, by increasing the concentration of the Mg2+ cofactor, implying that these substitutions had a deleterious effect on Mg2+ binding. Calculations of the change in the apparent free energy of binding for variants at positions 3, 4 or 17 are each consistent with deletion of a single hydrogen-bond to an uncharged group in the ribozyme. The cytidine 5' to the scissile phosphate had not previously been thought to play a direct role in catalysis, however, removal of the exocyclic amino group decreased kcat. 4-fold. Recently, the crystal structures of a hammerhead ribozyme bound to either a non-cleavable 2'-deoxy substrate strand or a ribo-substrate strand have been reported. The kinetic properties of the variants described here are consistent with several key interactions seen in the crystals, in particular they provide experimental support for the assignment of the proposed catalytically active magnesium ion-binding site.
We present the results of in vitro binding studies aimed at defining the key recognition elements on the MS2 RNA translational operator (TR) essential for complex formation with coat protein. We have used chemically synthesized operators carrying modified functional groups at defined nucleotide positions, which are essential for recognition by the phage coat protein. These experiments have been complemented with modification-binding interference assays. The results confirm that the complexes which form between TR and RNA-free phage capsids, the X-ray structure of which has recently been reported at 3.0 A, are identical to those which form in solution between TR and a single coat protein dimer. There are also effects on operator affinity which cannot be explained simply by the alteration of direct RNA-protein contacts and may reflect changes in the conformational equilibrium of the unliganded operator. The results also provide support for the approach of using modified oligoribonucleotides to investigate the details of RNA-ligand interactions.
Explore the source record for details and available documents.
The RNA bacteriophage MS2 is a convenient model system for the study of protein-RNA interactions. The MS2 coat protein achieves control of two distinct processes--sequence-specific RNA encapsidation and repression of replicase translation--by binding to an RNA stem-loop structure of 19 nucleotides containing the initiation codon of the replicase gene. The binding of a coat protein dimer to this hairpin shuts off synthesis of the viral replicase, switching the viral replication cycle to virion assembly rather than continued replication. The operator fragment alone can trigger self-assembly of the phage capsid at low protein concentrations and a complex of about 90 RNA operator fragments per protein capsid has been described. We report here the crystal structure at 3.0 A resolution of a complex between recombinant MS2 capsids and the 19-nucleotide RNA fragment. It is the first example of a structure at this resolution for a sequence-specific protein-RNA complex apart from the transfer RNA synthetase complexes. The structure shows sequence-specific interactions between conserved residues on the protein and RNA bases essential for binding.
A general procedure for the purification of chemically synthesized oligoribonucleotides is reported. Purification based on the use of a single reverse-phase HPLC column with buffer systems of differing ion-pairing capacity is described. These methods have been applied to the preparation of a series of RNAs which range in size from 10 to 46 nucleotides. The yields obtained are high, up to 53% (based on isolated product compared to those obtained from final trityl assay). The purity of the isolated material is 96-99%. Thus with this general procedure, milligram quantities of extremely pure RNA can be efficiently obtained.
Research on multiple personality disorder (MPD) has burgeoned, and large-scale investigations indicate that a typical MPD patient is a woman, a victim of childhood abuse (especially sexual abuse), a person whose symptoms meet criteria for other psychiatric disorders, and a person who would employ many psychological defenses. Treatment approaches have frequently included hypnotherapy, which requires skill and caution.
Explore the source record for details and available documents.