Modulation of cardiac troponin C-cardiac troponin I regulatory interactions by the amino-terminus of cardiac troponin I

Multidimensional heteronuclear magnetic resonance studies of the cardiac troponin C/troponin I(1-80)/troponin I(129-166) complex demonstrated that cardiac troponin I(129-166), corresponding to the adjacent inhibitory and regulatory regions, interacts with and induces an opening of the cardiac troponin C regulatory domain. Chemical shift perturbation mapping and (15)N transverse relaxation rates for intact cardiac troponin C bound to either cardiac troponin I(1-80)/troponin I(129-166) or troponin I(1-167) suggested that troponin I residues 81-128 do not interact strongly with troponin C but likely serve to modulate the interaction of troponin I(129-166) with the cardiac troponin C regulatory domain. Chemical shift perturbations due to troponin I(129-166) binding the cardiac troponin C/troponin I(1-80) complex correlate with partial opening of the cardiac troponin C regulatory domain previously demonstrated by distance measurements using fluorescence methodologies. Fluorescence emission from cardiac troponin C(F20W/N51C)(AEDANS) complexed to cardiac troponin I(1-80) was used to monitor binding of cardiac troponin I(129-166) to the regulatory domain of cardiac troponin C. The apparent K(d) for cardiac troponin I(129-166) binding to cardiac troponin C/troponin I(1-80) was 43.3 +/- 3.2 microM. After bisphosphorylation of cardiac troponin I(1-80) the apparent K(d) increased to 59.1 +/- 1.3 microM. Thus, phosphorylation of the cardiac-specific N-terminus of troponin I reduces the apparent binding affinity of the regulatory domain of cardiac troponin C for cardiac troponin I(129-166) and provides further evidence for beta-adrenergic modulation of troponin Ca(2+) sensitivity through a direct interaction between the cardiac-specific amino-terminus of troponin I and the cardiac troponin C regulatory domain.     

Interaction of bepridil with the cardiac troponin C/troponin I complex

Mammalian cells are characterized by an endomembrane system. Nevertheless, some cells lose these membranes during their terminal differentiation, e.g. red blood cells and lens fiber cells of the eye. 15-Lipoxygenase is believed to be critical for this membrane degradation. Here we use cultivated rabbit reticulocytes in the presence or absence of a lipoxygenase inhibitor to provide further evidence for the importance of 15-lipoxygenase for the in vivo degradation of mitochondria. We find that inhibitor treatment retarded mitochondrial degradation, as shown by persistence of marker proteins and by direct visualization of mitochondria by electron microscopy.     

Glomalean mycorrhizal fungi from tropical Australia

 A comparison of different methods for isolation of vesicular-arbuscular mycorrhizal (VAM) fungi into open-pot cultures was undertaken as part of a study of the diversity of these fungi. Four different isolation techniques using spores separated from soil, soil trap cultures, root samples, or transplanted seedlings grown in intact soil cores were used to obtain as many fungi as possible from each site. Isolation methods were compared using paired samples from the same locations within natural (savanna, rocky hill, wetland, rainforest) and disturbed (minesite) habitats in a seasonally dry tropical region in the Northern Territory of Australia. There were large differences in (i) the efficiency (rate of increase in mycorrhizal colonisation), (ii) the proportion of successful cultures, (iii) fungal diversity (number of fungal species in each culture) and (iv) specificity (identity of species isolated) between these four procedures. However, the less-efficient procedures generally resulted in a higher proportion of cultures of one fungus, which could be used without further isolation steps. Most species of Scutellospora, Acaulospora and Gigaspora were obtained primarily from field-collected spores, but only 50% of these culture attempts were successful. Spores from these initial cultures produced mycorrhizas much more rapidly and successfully when used to start second-generation cultures. Several species of fungi, rarely recovered as living spores from field soils, were dominant in many trap cultures started from soil or roots. Most of these fungi were Glomus species, that were first distinguished by colonisation patterns in roots and eventually identified after sporulation in second- or third-generation trap cultures. These experiments demonstrated that glomalean fungi in the habitats sampled belonged to two functional categories, based on whether or not spores were important propagules. The "non-sporulating" fungi were dominant in many trap cultures, which suggests that these fungi had higher total inoculum levels in soils than other fungi. Pot-culturing methods provided additional information on fungal diversity which complemented spore occurrence data obtained using the same soil samples and provided valuable new information about the biology of these fungi. Accepted: 26 December 1998     

Buffering capacity of whole corn mash alters concentrations of organic acids required to inhibit growth of Saccharomyces cerevisiae and ethanol production

Growth of Saccharomyces cerevisiae and fermentative ethanol production in the presence of acetic and lactic acids was measured in whole corn mash. In this industrial medium, as compared to glucose minimal medium, the yeast had increased tolerance to organic acid stress. It was concluded that the increased buffering capacity of whole corn mash, resulting in decreased concentration of undissociated acid, was responsible for this phenomenon.     

Nonhuman primates contribute unique understanding to anovulatory infertility in women

Anovulatory infertility affects a large proportion of reproductive-aged women. Major improvements in successful clinical treatment of this prevalent disorder in women's health have been made possible because of biomedical research employing nonhuman primates. Experiments on female rhesus monkeys were the first to demonstrate that the key hypothalamic neurotransmitter, gonadotropin-releasing hormone, involved in stimulating pituitary gonadotropin synthesis, storage, and release was bioactive only when released in approximately hourly bursts. This breakthrough in understanding gonadotropin regulation enabled identification of hypogonadotropic, apparently normogonadotropic, and hypergonadotropic forms of anovulatory infertility, and development of appropriate stimulatory or inhibitory gonadotropin therapies. Treatments to overcome anovulatory infertility represent one of the major advances in clinical reproductive endocrinology during the last 25 yr. The future promise of nonhuman primate models for human ovulatory dysfunction, however, may be based on an increased understanding of molecular and physiological mechanisms responsible for fetal programming of adult metabolic and reproductive defects and for obesity-related, hyperinsulinemic impairment of oocyte development.     

Plant introductions,hybridization and gene flow

Many regional floras contain a high proportion of recently introduced plant species. Occasionally, hybridization between an introduced species and another species (introduced or native) can result in interspecific gene flow. This may occur even in instances where the F(1) hybrid shows very high sterility, but occasionally produces a few viable gametes. We provide examples of gene flow occurring between some rhododendrons recently introduced to the British flora, and between an introduced and native Senecio species. Neutral molecular markers have normally been employed to obtain evidence of interspecific gene flow, but the challenge now is to isolate and characterize functional introgressed genes and to determine how they affect the fitness of introgressants and whether they improve adaptation to novel habitats allowing introgressants to expand the range of a species. We outline a candidate gene approach for isolating and characterizing an allele of the RAY gene in Senecio vulgaris, which is believed to have introgressed from S. squalidus, and which causes the production of ray florets in flower heads. We discuss the effects of this introgressed allele on individual fitness, including those that originate directly from the production of ray florets plus those that may arise from pleiotropy and/or linkage.     

A linkage map for the hybridising toads Bombina bombina and B. variegata (Anura: Discoglossidae)

Stable hybrid zones in which ecologically divergent taxa give rise to a range of recombinants are natural laboratories in which the genetic basis of adaptation and reproductive isolation can be unraveled. One such hybrid zone is formed by the fire-bellied toads Bombina bombina and B. variegata (Anura: Discoglossidae). Adaptations to permanent and ephemeral breeding habitats, respectively, have shaped numerous phenotypic differences between the taxa. All of these are, in principle, candidates for a genetic dissection via QTL mapping. We present here a linkage map of 28 codominant and 10 dominant markers in the Bombina genome. In an F2 cross, markers that were mainly microsatellites, SSCPs or allozymes were mapped to 20 linkage groups. Among the 40 isolated CA microsatellites, we noted a preponderance of compound and frequently interleaved CA-TA repeats as well as a striking polarity at the 5' end of the repeats.     

Barriers to uptake of services for coronary heart disease: qualitative study

ObjectivesTo identify factors within the South Yorkshire coalfields that influence use of health services by people with angina.DesignQualitative study using semistructured individual and group interviews.SettingGeneral practice and community settings in Barnsley and Rotherham health authorities.Participants14 patients with stable angina and nine primary care staff had individual interviews plus five community groups and one group of general practitioners.ResultsA complex web of factors was identified that prevented, delayed, or facilitated referral to secondary care. Delay, denial, and self management by patients meant that the full extent of symptoms often remained hidden from general practitioners, resulting in a delayed or missed referral. Barriers identified fell into six categories: structural, personal, social and cultural, past experience and expectations, diagnostic confusion, and knowledge and awareness.ConclusionsMany of the factors influencing referral operate before general practitioners become involved. Community development could be one way of tackling inequalities and promoting sustainable change. Structural changes are needed to improve access and increase the acceptability of general practice services. Primary care staff should be educated to detect underreporting of symptoms and promote appropriate referral.

What is already known on this topic

An inverse correlation has been shown between deprivation and cardiac revascularisationFear of hospitals, denial of ill health, and low expectations may prevent people with angina accessing health services

What this study adds

Fear, denial, and low expectations were important barriers to accessing health services, reinforcing earlier findingsOther factors may be specific to the study population—for example, coping, independence, and attributing symptoms to industrially related lung diseaseMany of the barriers operate before general practitioners are involved, making it difficult to identify solutions     

Kerogen-bound and free hopanoic acids in the messel oil shale kerogen

The distribution of the free and bound hopanoic acids in both unheated and heated (350 degrees C for 50 h) kerogens, isolated from the Messel oil shale, were analyzed by GC-MS. The bound acids were released by subjecting the kerogen to three different treatments, namely, thermochemolysis in the presence of tetramethylammonium hydroxide (TMAH), as well as basic and acidic hydrolyses. All of these methods gave a series of hopanoic acids ranging from C(30) to C(34), in which the biological 17beta, 21beta(H) configuration is prominent. Both 22R and 22S epimers are present for the C(30) acid, whereas the others are dominated by the sidechain 22R-configuration. Thermochemolysis in the presence of TMAH was the most efficient in releasing kerogen-bound hopanoids. Following pyrolysis, the acids are generated and released into the free fraction with apparent epimerization occurring at C-17, C-21, and C-22. The bound hopanoic acids may be both chemically bonded as well as possibly being physically encapsulated within the macromolecular fraction of sedimentary organic matter. They are therefore either generated by breaking the bonds which bind them to the kerogen or they are released as a result of the macromolecular cage being broken apart.