Angiotensin I-converting enzyme transition state stabilization by HIS1089 : evidence for a catalytic mechanism distinct from other gluzincin metalloproteinases

Angiotensin (Ang) I-converting enzyme (ACE) is a member of the gluzincin family of zinc metalloproteinases that contains two homologous catalytic domains. Both the N- and C-terminal domains are peptidyl-dipeptidases that catalyze Ang II formation and bradykinin degradation. Multiple sequence alignment was used to predict His¹⁰⁸⁹ as the catalytic residue in human ACE C-domain that, by analogy with the prototypical gluzincin, thermolysin, stabilizes the scissile carbonyl bond through a hydrogen bond during transition state binding. Site-directed mutagenesis was used to change His¹⁰⁸⁹ to Ala or Leu. At pH 7.5, with Ang I as substrate, k_cat/K_m values for these Ala and Leu mutants were 430 and 4,000-fold lower, respectively, compared with wild-type enzyme and were mainly due to a decrease in catalytic rate (k_cat) with minor effects on ground state substrate binding (K_m). A 120,000-fold decrease in the binding of lisinopril, a proposed transition state mimic, was also observed with the His¹⁰⁸⁹ --> Ala mutation. ACE C-domain-dependent cleavage of AcAFAA showed a pH optimum of 8.2. H1089A has a pH optimum of 5.5 with no pH dependence of its catalytic activity in the range 6.5-10.5, indicating that the His¹⁰⁸⁹ side chain allows ACE to function as an alkaline peptidyl-dipeptidase. Since transition state mutants of other gluzincins show pH optima shifts toward the alkaline, this effect of His¹⁰⁸⁹ on the ACE pH optimum and its ability to influence transition state binding of the sulfhydryl inhibitor captopril indicate that the catalytic mechanism of ACE is distinct from that of other gluzincins.

Arg1098 is critical for the chloride dependence of human angiotensin I-converting enzyme C-domain catalytic activity

Angiotensin (Ang) I-converting enzyme (ACE) is a Zn²+ metalloprotease with two homologous catalytic domains. Both the N- and C-terminal domains are peptidyl dipeptidases. Hydrolysis by ACE of its decapeptide substrate Ang I is increased by Cl−, but the molecular mechanism of this regulation is unclear. A search for single substitutions to Gln among all conserved basic residues (Lys/Arg) in human ACE C-domain identified R1098Q as the sole mutant that lacked Cl− dependence. Cl−dependence is also lost when the equivalent Arg in the N-domain, Arg⁵⁰⁰, is substituted with Gln. The Arg¹⁰⁹⁸ to Lys substitution reduced Cl− binding affinity by ∼100-fold. In the absence of Cl−, substrate binding affinity (1/K m) of and catalytic efficiency (k cat/K m) for Ang I hydrolysis are increased 6.9- and 32-fold, respectively, by the Arg¹⁰⁹⁸ to Gln substitution, and are similar (<2-fold difference) to the respective wild-type C-domain catalytic constants in the presence of optimal [Cl−]. The Arg¹⁰⁹⁸ to Gln substitution also eliminates Cl− dependence for hydrolysis of tetrapeptide substrates, but activity toward these substrates is similar to that of the wild-type C-domain in the absence of Cl−. These findings indicate that: 1) Arg¹⁰⁹⁸ is a critical residue of the C-domain Cl−-binding site and 2) a basic side chain is necessary for Cl− dependence. For tetrapeptide substrates, the inability of R1098Q to recreate the high affinity state generated by the Cl−-C-domain interaction suggests that substrate interactions with the enzyme-bound Cl− are much more important for the hydrolysis of short substrates than for Ang I. Since Cl− concentrations are saturating under physiological conditions and Arg¹⁰⁹⁸ is not critical for Ang I hydrolysis, we speculate that the evolutionary pressure for the maintenance of the Cl−-binding site is its ability to allow cleavage of short cognate peptide substrates at high catalytic efficiencies.

Gyration

European Renaissance and Romantic landscape appeared in vistas. The conditions of the industrial revolution and, according to Patrick Maynard and Jonathon Crary, the film camera especially, led to a Modernist re-vision vividly recorded in Xavier Herbert’s contrary Modernist vision, prompted by seeing the Australian bush, its ‘... stunted trees, the mulga and the wilga and the gimlet gum, doing a kind of dance, spinning past, seeming to swing away from the train to the horizon and race ahead, to come back...the same set of trees in endless gyration’.

The impact of religious diversity and revitalization on inter-religious education for citizenship and human rights

The unanticipated rise of religious diversity and the re-entry of religion to the public sphere have radically increased the need and demand for education about religions – how they contribute to social and cultural capital – and about the management of religious diversity. The global movement of people and cultures has brought religious diversity to nearly every major city. With diversity has come a renewed interest in the religious identity of others and how to incorporate religious diversity in ways that produce social cohesion. Religious diversity has also raised interest in a values discourse where once atheistic secularity prevailed, made faith-based social and health service delivery both more appealing to governments and more difficult to deliver, and has challenged societies to accommodate a wider range of religious needs and lifestyles. Policies designed to promote social justice and peace have little chance of success without taking seriously the religious dimensions to the issues involved. This context makes clear the need for opportunities to learn about the religions in a society at all levels of education – opportunities that include direct experience of the ‘other’, curricula that appreciate the worlds of faith, spirituality and religion rather than demeaning them, education that provides both historical depth and local reality. Some of this education will be in school, some in remedial work required for a generation or two of leaders who have been raised in ignorance of religion, or trained to despise it.

A review of population-based prevalence studies of physical activity in adults in the Asia-Pacific region

Background: Physical activity (PA) surveillance is an important component of non-communicable disease risk factor monitoring, and occurs through national and international surveillance systems. This review identifies population PA estimates for adults in the Asia-Pacific region, and examines variation in trends and prevalence rates obtained using different PA measures.

Methods: Data were obtained from a MEDLINE search; World Health Organization’s Global Health Infobase; Government websites and reference lists of relevant papers. Inclusion criteria included: national studies or those reporting large scale population-level data; data published from 2000 to 2010 and trend data prior; sample sizes over n = 1000, or fewer subjects in small nations.

Results: In total, 56 population surveys from 29 Asia-Pacific countries were identified. Data on ‘sufficient physical activity’ amongst adults were available from 45 studies (80%), with estimates ranging from 7% to 93% (median 62%, inter-quartile range 40%-85%). For 14 countries, estimates of ‘sufficient activity’ were documented in multiple surveys using different methods, with the largest variation from 18% to 92% in Nepal. Median or mean METminutes/ day, reported in 20 studies, ranged from 6 to 1356. Serial trend data were available for 11 countries (22%), for periods spanning 2-10 years. Of these, five countries demonstrated increases in physical activity over time, four demonstrated decreases and three showed no changes.

Conclusions: Many countries in the Asia-Pacific region collect population-level PA data. This review highlights differences in estimates within and between countries. Some differences may be real, others due to variation in the PA questions asked and survey methods used. Use of standardized protocols and measures, and combined reporting of data are essential goals of improved international PA surveillance.

The effect of artificial night light on the abundance of nocturnal birds

Urbanisation is increasing rapidly, impacting on a broad range of species. The proliferation of electric light has transformed the night time environment; however, our understanding on the effects of artificial night lighting on fauna, including nocturnal birds, is extremely limited. The aim ofthis research was to determine whether artificial night light affected the abundance of nocturnal birds. Spotlighting surveys were undertaken in Research Park, Melbourne, Victoria, along three 300 m transects. Each transect was surveyed five times during three light treatments: when lights were on, 20 minutes after lights were turned off and when lights were absent, over a period often nights. A total of 123 nocturnal birds was detected during survey nights. Two species were recorded - the Southern Boobook Ninox novaeseelandiae and the Tawny Frogmouth Podargus strigoides. The Tawny Frogmouth was detected along all three transects (n=121); however, the Southern Boobook was detected along one transect only (n=2). None of the light treatments had a significant effect on bird abundance. Neither did location, habitat or the combined effects of light treatments, location and habitat. The results of this research will contribute to a growing body of knowledge and support future conservation activities for species in areas undergoing urbanisation. {The Victorian Naturalist 127 (5) 2010, 192-195).

Does self monitoring of blood blucose as opposed to urinalysis provide additional benefit in patients newly diagnosed with type 2 diabetes receiving structured education? The DESMOND SMBG randomised controlled trial protocol

Background
The benefit of self-monitoring of blood glucose (SMBG) in people with type 2 diabetes on diet or oral agents other than sulphonylureas remains uncertain. Trials of interventions incorporating education about self-monitoring of blood glucose have reported mixed results. A recent systematic review concluded that SMBG was not cost-effective. However, what was unclear was whether a cheaper method of self-monitoring (such as urine glucose monitoring) could produce comparable benefit and patient acceptability for less cost.

Methods/Design
The DESMOND SMBG trial is comparing two monitoring strategies (blood glucose monitoring and urine testing) over 18 months when incorporated into a comprehensive self-management structured education programme. It is a multi-site cluster randomised controlled trial, conducted across 8 sites (7 primary care trusts) in England, UK involving individuals with newly diagnosed Type 2 diabetes.

The trial has 80% power to demonstrate equivalence in mean HbA1c (the primary end-point) at 18 months of within ± 0.5% assuming 20% drop out and 20% non-consent. Secondary end-points include blood pressure, lipids, body weight and psychosocial measures as well as a qualitative sub-study.

Practices were randomised to one of two arms: participants attend a DESMOND programme incorporating a module on self-monitoring of either urine or blood glucose. The programme is delivered by accredited educators who received specific training about equipoise. Biomedical data are collected and psychosocial scales completed at baseline, and 6, 12, and 18 months post programme. Qualitative research with participants and educators will explore views and experiences of the trial and preferences for methods of monitoring.

Discussion
The DESMOND SMBG trial is designed to provide evidence to inform the debate about the value of self-monitoring of blood glucose in people with newly diagnosed type 2 diabetes. Strengths include a setting in primary care, a cluster design, a health economic analysis, a comparison of different methods of monitoring while controlling for other components of training within the context of a quality assured structured education programme and a qualitative sub-study.

Inhibitory effect of antidepressant drugs on contact hypersensitivity reaction

Background: Contact hypersensitivity (CS) reaction in the skin is T-cell mediated immune reaction which plays a major role in the pathogenesis and chronicity of various inflammatory skin disorders and, like other delayed-type hypersensitivity (DTH) reactions, affords immunity against tumor cells and microbes. CS response is a self-limiting reaction, and interleukin (IL)-10 is considered to be a natural suppressant of cutaneous inflammatory response. Recently, it has been demonstrated that major depression is related to activation of the inflammatory response and elevation of some parameters of cell-mediated immunity. It has been suggested that such activation of the immune system may play a role in etiology of depression. If this immunoactivation is involved in etiology of depression, one would expect that antidepressant agents may have negative immunoregulatory effects. To the best of our knowledge, the effect of antidepressants on contact hypersensitivity has not been studied.

Methods: The aim of the present study was to establish the effect of prolonged desipramine or fluoxetine treatment on CS reaction to picryl chloride.

Results: Antidepressants significantly suppressed CS reaction, fluoxetine by 53% whereas desipramine by 47% compared to positive control. Moreover, desipramine and fluoxetine decreased relative weight of auxillary lymph nodes. Desipramine decreased also relative weight of inguinal lymph nodes and spleens whereas desipramine and fluoxetine increased production of IL-10 in comparison to positive control.

Conclusion: The observed effect of antidepressant drugs on CS reaction is consistent with the hypothesis that T-cell mediated immunity is targeted by antidepressants.

Conclusion : gender and politics and the changing status of women today

This book concerns a new body of knowledge and an emerging set of questions that has accompanied national, cross national and international global political movements aimed at trying to understand and to improve the situation of women by eliminating gender inequities and injustices.

Isolation and characterisation via 454 sequencing of microsatellites from the tawny frogmouth, Podargus strigoides (Class Aves, Family Podargidae)

We isolated 24 novel polymorphic microsatellite markers from the tawny frogmouth, a nocturnal bird endemic to Australia, which has successfully adapted to urban environments. Initially, 454 shotgun sequencing was used to identify 733 loci with primers designed. Of these, we trialled 30 in the target species of which all amplified a product of expected size. Subsequently, all 30 of these loci were screened for variation in 25 individuals, from a single population in Melbourne, Victoria, Australia. Twenty-eight loci were polymorphic with observed heterozygosity ranging from 0.03 to 0.96 (mean 0.58) and the number of alleles per locus ranged from 2 to 18 (average of 6.5); we confirmed that 24 loci conformed to Hardy–Weinberg expectations. The 24 loci identified here will be sufficient to unequivocally identify individuals and will be useful in understanding the reproductive ecology, population genetics and the gene flow amongst localities in urban environments where this bird thrives.

A population-based study investigating the association between sexual and relationship satisfaction and psychological distress among heterosexuals

This study examined whether sexual/relationship satisfaction are differentially associated with mental health issues. Using data from a population-based computer-assisted telephone survey, the authors included in this study 3,800 respondents who had a regular heterosexual partner. The authors used 2 methods of scoring the K6 to produce measures of moderate psychological distress and serious psychological distress. Overall, 8.8% of men and 12.1% of women were classified as having moderate psychological distress, whereas 1.6% of men and 3.2% of women were classified as currently experiencing serious psychological distress. The association between satisfaction and mental health was influenced by sex and the severity of the mental health issue but not by type of satisfaction. After adjusting for demographic differences in mental health, low ratings of sexual/relationship satisfaction were both consistently associated with higher levels of moderate psychological distress in men and women and higher proportions of serious psychological distress in men. Although women may be able to resolve their satisfaction issues during less severe stages of psychological distress, for men there was a strong association between low sexual/relationship satisfaction and serious psychological distress.