Behaviour of the black flying fox Pteropus alecto: 2. Territoriality and courtship

Flying foxes are commonly thought of as highly social mammals, yet little is known about the dynamics of their social interactions at a day roost. The aim of the present study was to examine the nature of the seasonal activities of territoriality and courtship amongst wild flying foxes in Australia. Focal observations were conducted at two permanent roosts of black flying foxes Pteropus alecto during periods of peak social interaction in the summers of 1999 and 2000 in urban Brisbane, Queensland. Observations of male territoriality were conducted at dawn and began eight weeks prior to the commencement of mating. The majority of defense bouts (87%) consisted of ritualised pursuit, while 13% of bouts involved physical contact expressed as either wrestling or hooking. One male with an unusually large territory took significantly longer to defend it than other males with less territory to defend. Observations of courtship revealed repetitive courtship sequences, including pre-copulatory approaches by the males, copulation attempts and grooming/resting periods. Thirty-four complete courtship sequences incorporating 135 copulation attempts were recorded over two seasons. Females actively resisted courtship approaches by males, forcing males to display a continuous determination to mate over time where determination can be considered an indicator of 'fitness'. The courtship bout length of females with suckling young was significantly longer ((x) over bar +/- SE; 230.9 +/- 22.16 s) than that of females unencumbered by large pups (158.5 +/- 9.69 s), although the length of copulations within those courtships was not (45.6 +/- 5.19 versus 36.2 +/- 3.43 s).

The 2000 Ogura Lecture: Olfactory neural cells: An untapped diagnostic and therapeutic resource

Objective. This is an over-view of the cellular biology of upper nasal mucosal cells that have special characteristics that enable them to be used to diagnose and study congenital neurological diseases and to aid neural repair. Study Design: After mapping the distribution of neural cells in the upper nose, the authors' investigations moved to the use of olfactory neurones to diagnose neurological diseases of development, especially schizophrenia. Olfactory-ensheating glial cells (OEGs) from the cranial cavity promote axonal penetration of the central nervous system and aid spinal cord repair in rodents. The authors sought to isolate these cells from the more accessible upper nasal cavity in rats and in humans and prove they could likewise promote neural regeneration, making these cells suitable for human spinal repair investigations. Methods: The schizophrenia-diagnosis aspect of the study entailed the biopsy of the olfactory areas of 10 schizophrenic patients and 10 control subjects. The tissue samples were sliced and grown in culture medium. The ease of cell attachment to fibronectin (artificial epithelial basement membrane), as well as the mitotic and apoptotic indices, was studied in the presence and absence of dopamine in those cell cultures. The neural repair part of the study entailed a harvesting and insertion of first rat olfactory lamina propria rich in OEGs between cut ends of the spinal cords and then later the microinjection of an OEG-rich suspension into rat spinal cords previously transected by open laminectomy. Further studies were done in which OEG insertion was performed up to 1 month after rat cord transection and also in monkeys. Results: Schizophrenic patients' olfactory tissues do not easily attach to basement membrane compared with control subjects, adding evidence to the theory that cell wall anomalies are part of the schizophrenic lesion of neurones. Schizophrenic patient cell cultures had higher mitotic and apoptotic indices compared with control subjects. The addition of dopamine altered these indices enough to allow accurate differentiation of schizophrenics from control patients, leading to, possibly for the first time, an early objective diagnosis of schizophrenia and possible assessment of preventive strategies. OEGs from the nose were shown to be as effective as those from the olfactory bulb in promoting axonal growth across transected spinal cords even when added I month after injury in the rat. These otherwise paraplegic rats grew motor and proprioceptive and fine touch fibers with corresponding behavioral improvement. Conclusions. The tissues of the olfactory mucosa are readily available to the otolaryngologist. Being surface cells, they must regenerate (called neurogenesis). Biopsy of this area and amplification of cells in culture gives the scientist a window to the developing brain, including early diagnosis of schizophrenia. The Holy Grail of neurological disease is the cure of traumatic paraplegia and OEGs from the nose promote that repair. The otolaryngologist may become the necessary partner of the neurophysiologist and spinal surgeon to take the laboratory potential of paraplegic cure into the day-to-day realm of clinical reality.

NPS-2143 - Treatment of osteoporosis - Calcilytic

Efforts to discover new treatments for osteoporosis led to the identification of the potent and selective, small-molecule calcium receptor antagonist NPS-2143. NPS-2143 is the prototype calcilytic drug, designed to act on calcium receptors on the surface of parathyroid glands, stimulating the release of the body's own stores of native parathyroid hormone (PTH). In osteopenic ovariectomized rats, daily oral administration of NPS-2143 resulted in moderate but sustained increases in plasma PTH levels and marked increases in bone formation and resorption, with no net bone gain or loss. The combination of NPS-2143 and estrogen increases bone formation and density to a greater extent than either agent alone. These results suggest that NPS-2143 may be useful in the treatment of established osteoporosis.

Effect of tamoxifen on the enzymatic activity of human cytochrome CYP2B6

Tamoxifen is primarily used in the treatment of breast cancer. It has been approved as a chemopreventive agent for individuals at high risk for this disease. Tamoxifen is metabolized to a number of different products by cytochrome P450 enzymes. The effect of tamoxifen on the enzymatic activity of bacterially expressed human cytochrome CYP2B6 in a reconstituted system has been investigated. The 7-ethoxy-4-(trifluoromethyl) coumarin O-deethylation activity of purified CYP2B6 was inactivated by tamoxifen in a time- and concentration-dependent manner. Enzymatic activity was lost only in samples that were incubated with both tamoxifen and NADPH. The inactivation was characterized by a K-l of 0.9 muM, a k(inact) of 0.02 min(-1), and a t(1/2) of 34 min. The loss in the 7-ethoxy-4-(trifluoromethyl) coumarin O-deethylation activity did not result in a similar percentage loss in the reduced carbon monoxide spectrum, suggesting that the heme moiety was not the major site of modification. The activity of CYP2B6 was not recovered after removal of free tamoxifen using spin column gel filtration. The loss in activity seemed to be due to a modification of the CYP2B6 and not reductase because adding fresh reductase back to the inactivated samples did not restore enzymatic activity. A reconstituted system containing purified CYP2B6, NADPH-reductase, and NADPH-generating system was found to catalyze tamoxifen metabolism to 4-OH-tamoxifen, 4'-OH-tamoxifen, and N-desmethyl-tamoxifen as analyzed by high-performance liquid chromatography analysis. Preliminary studies showed that tamoxifen had no effect on the activities of CYP1B1 and CYP3A4, whereas CYP2D6 and CYP2C9 exhibited a 25% loss in enzymatic activity.

Amino Acids involved in differences in the pharmacological profiles of the rat and human noradrenaline transporters

It has been suggested from a previous study in our laboratory that differences in the pharmacology of the species variants of the noradrenaline transporter (NET) are the result of four non-conservative amino acid exchanges from the total of 26 amino acids that are divergent between the rat NET (rNET) and human NET (hNET). The aim of this study was to examine the effects of changing the rNET at each of these four amino acid residues, which markedly alter local charge distribution, to the amino acid found in hNET. Site-directed mutagenesis was used to create mutant cDNAs from rNET cDNA. The mutant NETs (rK71), rE62K, rK375N and rR612Q), rNET and hNET were expressed in transiently transfected COS-7 cells to determine the effects of the mutations on the differing pharmacological properties of the species variants. The ratios of V-max for noradrenaline uptake and B-max for nisoxetine binding (which are a measure of the turnover number of the transporter, i.e. the number of transport cycles per min) were greater for rNET and rR612Q than for hNET, rK71), rE62K and rK375N. The K-m of noradrenaline was lower for hNET, rK713, rE62K and rK375N than for rNET or rR612Q. There were no differences between the K-i values for inhibition of noradrenaline uptake by nisoxetine for rNET, hNET or the mutants, but the K-i values of cocaine were lower for hNET, rE62K and rR612Q than rNET or rK375N. Hence, the study showed that: (1) the aspartate 7. lysine 62 and asparagine 375 amino acid residues are important in determining the lower substrate translocation by hNET than rNET; (2) the aspartate 7 and lysine 62 residues in the N-terminus of hNET determine the higher affinities of substrates for the hNET than the rNET; and (3) the lysine 62 and glutamine 612 residues in the N- and C-termini, respectively, of hNET Lire determinants of the higher cocaine affinity for the hNET than rNET.

The role of conserved CXXXRXG motif in the expression and function of the human norepinephrine transporter

Highly conserved motifs in the monoamine transporters, e.g. the human norepinephrine transporter (hNET) GXXXRXG motif which was the focus of the present study, are likely to be important structural features in determining function. This motif was investigated by mutating the glycines to glutamate (causing loss of function) and alanine, and the arginine to glycine. The effects of hG117A, hR121G and hG123A mutations on function were examined in COS-7 cells and compared to hNET. Substrate K-m values were decreased for hG117A and hG123A, and their K values for inhibition of [3 H]nisoxetine binding were decreased 3-4-fold and 4-6-fold, respectively. Transporter turnover was reduced to 65% of hNET for hG117A and hR121G and to 28% for hG123A, suggesting that substrate translocation is impaired. K values of nisoxetine and desipramine for inhibition of [H-3]norepinephrine uptake were increased by 5-fold for hG117A, with no change for cocaine. The K-i value of cocaine was increased by 3-fold for hG123A, with no change for nisoxetine and desipramine. However, there were no effects of the mutations on the K-d of [H-3]nisoxetine binding or K-i values of desipramine or cocaine for inhibition of [H-3]nisoxetine binding. Hence, glycine residues of the GXXXRXG motif are important determinants of NET expression and function, while the arginine residue does not have a major role. This study also showed that antidepressants and psychostimulants have different NET binding sites and provided the first evidence that different sites on the NET are involved in the binding of inhibitors and their competitive inhibition of substrate uptake. (C) 2002 Elsevier Science B.V. All rights reserved.

(-)-CGP 12177 increases contractile force and hastens relaxation of human myocardial preparations through a propranolol-resistant state of the beta1-adrenoceptor

Two forms of the activated beta(1)-adrenoceptor exist, one that is stabilized by (-)-noradrenaline and is sensitive to blockade by (-)-propranolol and another which is stabilized by partial agonists such as (-)-pindolol and (-)-CGP 12177 but is relatively insensitive to (-)-propranolol. We investigated the effects of stimulation of the propranolol-resistant PI-adrenoceptor in the human heart. Myocardium from non-failing and failing human hearts were set up to contract at 1 Hz. In right atrium from non-ailing hearts in the presence of 200 nM (-)-propranolol, (-)-CGP 12177 caused concentration-dependent increases in contractile force (-logEC(50)[M] 7.3+/-0.1, E-max 23+/-1% relative to maximal (-)-isoprenaline stimulation of beta(1)- and beta(2)-adrenoceptors, n=86 patients), shortening of the time to reach peak force (-logEC(50)[M] 7.4+/-0.1, E-max 37+/-5%, n=61 patients) and shortening of the time to reach 50% relaxation (t(50%), -logEC(50)[M] 7.3+/-0.1, E-max 33+/-2%, n=61 patients). The potency and maxima of the positive inotropic effects were independent of Ser49Gly- and Gly389Arg-beta(1)-adrenoceptor polymorphisms but were potentiated by the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (-logEC(50)[M] 7.7+/-0.1, E-max 68+/-6%, n=6 patients, P

Effects of herbicides diuron and atrazine on corals of the Great Barrier Reef, Australia

In response to recent reports of contamination of the nearshore marine environment along the Queensland coast by herbicides (including areas inside the Great Barrier Reef Marine Park), an ecotoxicological assessment was conducted of the impact of the herbicides diuron and atrazine on scleractinian corals. Pulse-amplitude modulated (PAM) chlorophyll fluorescence techniques were used to assess the herbicide effects on the symbiotic dinoflagellates within the tissues (in hospite) of 4 species of coral (Acropora formosa, Montipora digitata, Porites cylindrica, Seriatopora hystrix) in static toxicity tests, and in freshly isolated symbiotic dinoflagellates from Stylophora pistillata. Using change in the effective quantum yield (DeltaF/F-m') as an effect criterion, diuron (no observable effect concentration, NOEC = 0.3 mug 1(-1); lowest observable effect concentration, LOEC = 1 mug 1(-1); median effective concentration, EC50 4 to 6 mug 1(-1)) was found to be more toxic than atrazine (NOEC = 1 mug 1(-1), LOEC = 3 mug 1(-1), EC50 40 to 90 mug 1(-1)) in short-term (10 h) toxicity tests. In the tests with isolated algae, significant reductions in DeltaF/F-m' were recorded as low as 0.25 mug 1(-1) diuron (LOEC, EC50 = 5 mug 1(-1)). Time-course experiments indicated that the effects of diuron were rapid and reversible. At 10 mug 1(-1) diuron, DeltaF/F-m' was reduced by 25% in 20 to 30 min, and by 50% in 60 to 90 min. Recovery of DeltaF/F-m' in corals exposed to 10 mug 1(-1) diuron and then transferred to running seawater was slower, returning to within 10% of control values inside 1 to 7 h. The effect of a reduction in salinity (35 to 27%) on diuron toxicity (at 1 and 3 mug 1(-1) diuron) was tested to examine the potential consequences of contaminated coastal flood plumes inundating inshore reefs. DeltaF/F-m' was reduced in the diuron-exposed corals, but there was no significant interaction between diuron and reduced salinity seawater within the 10 h duration of the test. Exposure to higher (100 and 1000 mug 1(-1)) diuron concentrations for 96 h caused a reduction in DeltaF/F-m' the ratio variable to maximal fluorescence (F,1F.), significant loss of symbiotic dinoflagellates and pronounced tissue retraction, causing the corals to pale or bleach. The significance of the results in relation to diuron contamination of the coastal marine environment from terrestrial sources (mainly agricultural) and marine sources (antifouling paints) are discussed.

The Bambuí health and ageing study (BHAS): methodological approach and preliminary results of a population-based cohort study of the elderly in Brazil

OBJECTIVE: A cohort study has been designed to identify predictors of adverse health events in the elderly. The methodology of the study and preliminary descriptive results are presented. METHODS: The study population comprises all residents of Bambuí (Minas Gerais, Brazil), aged 60 or more years (n=1.742). From these, 92.2% were interviewed and 85.9% underwent clinical examination, consisting of haematological and biochemical tests, serology for Trypanosoma cruzi, anthropometric and blood pressure measures and electrocardiogram. Aliquots of serum, plasma and DNA were stored for future investigations. The baseline interview included sociodemographic characteristics, self-referred health condition and history of selected diseases, medication use, health service use, source of medical care, physical activities, smoking, drinking and eating habits, reproductive history, physical functioning, life events, social support and mental health. Individuals are being followed up annually. RESULTS: The following characteristics predominated among participants: women (60,0%), married (48.9%) or widowed (35.4%), people living in households with up to 2 residents (73.8%), heads of family (76.7%), people with monthly income between 1.00 and 2.99 Brazilian minimum wages (62.0%) and people with up to 4 years of schooling (89.1%). The median age was 68 years. Among the cohort members, only 1.7% were lost in the first follow-up. CONCLUSIONS: In general, the characteristics of the study population were very similar to those from other epidemiological studies of the elderly based on large Brazilian cities. The small number of losses to follow-up indicates that the choice of Bambuí was adequate, assuring the feasibility of a long term cohort study.

A new natural spiro heterocyclic compound and the cytotoxic activity of the secondary metabolites from Juniperus brevifolia leaves

Copyright © 2010 Elsevier B.V. All rights reserved.

Mould and yeast identification in archival settings: preliminary results on the use of traditional methods and molecular biology options in Portuguese archives

This project was developed to fully assess the indoor air quality in archives and libraries from a fungal flora point of view. It uses classical methodologies such as traditional culture media – for the viable fungi – and modern molecular biology protocols, especially relevant to assess the non-viable fraction of the biological contaminants. Denaturing high-performance liquid chromatography (DHPLC) has emerged as an alternative to denaturing gradient gel electrophoresis (DGGE) and has already been applied to the study of a few bacterial communities. We propose the application of DHPLC to the study of fungal colonization on paper-based archive materials. This technology allows for the identification of each component of a mixture of fungi based on their genetic variation. In a highly complex mixture of microbial DNA this method can be used simply to study the population dynamics, and it also allows for sample fraction collection, which can, in many cases, be immediately sequenced, circumventing the need for cloning. Some examples of the methodological application are shown. Also applied is fragment length analysis for the study of mixed Candida samples. Both of these methods can later be applied in various fields, such as clinical and sand sample analysis. So far, the environmental analyses have been extremely useful to determine potentially pathogenic/toxinogenic fungi such as Stachybotrys sp., Aspergillus niger, Aspergillus fumigatus, and Fusarium sp. This work will hopefully lead to more accurate evaluation of environmental conditions for both human health and the preservation of documents.

Isolation and characterisation of metallothionein from the clam Ruditapes decussatus

Aquatic Toxicology 63 (2003) 307-318

Convergent validity between SF-36 and WHOQOL-BREF in older adults

OBJECTIVE : To compare the reliability and convergent validity of instruments assessing quality of life in Brazilian older adults. METHODS : Cross-sectional study of 278 literate, community-dwelling older adults attending a municipal university for the elderly in Sao Carlos, SP, Southeastern Brazil between 2006 and 2008. The Brazilian versions of the SF-36 and WHOQOL-BREF instruments to assess quality of life were compared. Cronbach’s alpha coefficient was used to estimate reliability and Pearson’s correlation for comparison between the two scales. RESULTS : Most of participants were women (87.8%) with a mean age of 63.83±7.22 years. Both scales showed an acceptable internal consistency – WHOQOL-BREF Cronbach’s alpha was 0.832 and SF-36 was 0.868. There was a weak (r ≤ 0.6) correlation between the related fields in the two questionnaires. CONCLUSIONS : The SF-36 and WHOQOL-BREF are reliable instruments for clinical and research uses in Brazilian older women. To select one, researchers should consider which aspects of quality of life they aim to capture because of weak convergent validity signs. This study’s results indicate that WHOQOL-BREF may be more relevant to evaluate changes in the quality of life of older women because it prioritizes responses to the aging process and avoids focusing on impairment.

Self-reported diabetes in older people: comparison of prevalences and control measures

OBJECTIVE The objective of this study was to analyze the prevalence of diabetes in older people and the adopted control measures.METHODS Data regarding older diabetic individuals who participated in the Health Surveys conducted in the Municipality of Sao Paulo, SP, ISA-Capital, in 2003 and 2008, which were cross-sectional studies, were analyzed. Prevalences and confidence intervals were compared between 2003 and 2008, according to sociodemographic variables. The combination of the databases was performed when the confidence intervals overlapped. The Chi-square (level of significance of 5%) and the Pearson’s Chi-square (Rao-Scott) tests were performed. The variables without overlap between the confidence intervals were not tested.RESULTS The age of the older adults was 60-69 years. The majority were women, Caucasian, with an income of between > 0.5 and 2.5 times the minimum salary and low levels of schooling. The prevalence of diabetes was 17.6% (95%CI 14.9;20.6) in 2003 and 20.1% (95%CI 17.3;23.1) in 2008, which indicates a growth over this period (p at the limit of significance). The most prevalent measure adopted by the older adults to control diabetes was hypoglycemic agents, followed by diet. Physical activity was not frequent, despite the significant differences observed between 2003 and 2008 results. The use of public health services to control diabetes was significantly higher in older individuals with lower income and lower levels of education.CONCLUSIONS Diabetes is a complex and challenging disease for patients and the health systems. Measures that encourage health promotion practices are necessary because they presented a smaller proportion than the use of hypoglycemic agents. Public health policies should be implemented, and aimed mainly at older individuals with low income and schooling levels. These changes are essential to improve the health condition of older diabetic patients.