The liposoluble proteome of <Emphasis Type="Italic">Mycoplasma agalactiae</Emphasis>: an insight into the minimal protein complement of a bacterial membrane

Background  

Mycoplasmas are the simplest bacteria capable of autonomous replication. Their evolution proceeded from gram-positive bacteria, with the loss of many biosynthetic pathways and of the cell wall. In this work, the liposoluble protein complement of Mycoplasma agalactiae, a minimal bacterial pathogen causing mastitis, polyarthritis, keratoconjunctivitis, and abortion in small ruminants, was subjected to systematic characterization in order to gain insights into its membrane proteome composition.     

Time of Progression to Osteopenia/Osteoporosis in Chronically HIV-Infected Patients: Screening DXA Scan

Background

Algorithms for bone mineral density (BMD) management in HIV-infected patients are lacking. Our objective was to assess how often a dual-energy x-ray absorptiometry (DXA) scan should be performed by assessing time of progression to osteopenia/osteoporosis.

Methods

All DXA scans performed between 2000 and 2009 from HIV-infected patients with at least two DXA were included. Time to an event (osteopenia and osteoporosis) was assessed using the Kaplan–Meier method. Strata (tertiles) were defined using baseline minimum T scores. Differences between strata in time to an event were compared with the log-rank test.

Results

Of 391 patients (1,639 DXAs), 49.6% had osteopenia and 21.7% osteoporosis at their first DXA scan. Of the 112 (28.6%) with normal BMD, 35.7% progressed to osteopenia; median progression time was 6.7 years. These patients were stratified: “low-risk" (baseline minimum T score >−0.2 SD), “middle-risk" (between −0.2 and −0.6 SD), and “high-risk" (from −0.6 to −1 SD); median progression time to osteopenia was 8.7, >7.2, and 1.7 years, respectively (p<0.0001). Of patients with osteopenia, 23.7% progressed to osteoporosis; median progression time was >8.5 years. Progression time was >8.2 years in “low-risk" tertile (T score between −1.1 and −1.6 SD), >8.5 years in “middle-risk" (between −1.6 and −2), and 3.2 years in “high-risk" (from −2 to −2.4) (p<0.0001).

Conclusions

Our results may help to define the BMD testing interval. The lowest T score tertiles would suggest recommending a subsequent DXA in 1–2 years; in the highest tertiles, ≥6 years. Early intervention in patients with bone demineralization could reduce fracture–related morbidity/mortality.     

Capacity for protein synthesis following heat stimulus of Drosophila associates with heat tolerance but does not underlie the latitudinal tolerance cline

Across populations of Drosophila melanogaster along the Australian eastern coastline latitudinal clines occur in both heat-knockdown tolerance and hardened heat-knockdown tolerance – low latitude tropical populations being more tolerant. A latitudinal cline also occurs for rates of total protein synthesis following a mild heat stress, with tropical populations having higher rates. Since the control of protein synthesis following heat stress is an important component of the cellular heat-shock response, we hypothesised that the higher rates of synthesis that follow a heat stimulus lead to higher knockdown tolerance and underpins the cline. However, levels of heat-stimulated total protein synthesis have been negatively related to heat-hardening capacity, a somewhat conflicting result. Here we examine the relationship between these physiological and adaptive traits in a set of 40 family lines derived from a hybrid laboratory population established by crossing populations from either end of the latitudinal transect. Among these lines high levels of heat-stimulated total protein synthesis were associated with both low basal and low heat-hardened adult knockdown time, confirming the importance of a negative relationship between protein synthesis and thermal tolerance. This result, when considered along with the directions of the latitudinal clines in protein synthesis and tolerance, suggests that variation in rates of heat-stimulated total protein synthesis is not a factor contributing to the latitudinal cline in heat tolerance. Given the robustness of this negative relationship we discuss possible explanations and future experiments to elucidate how the cellular heat stress response might facilitate increased knockdown tolerance.     

Carbon stocks in above‐ground biomass of monoculture plantations,mixed species plantations and environmental restoration plantings in north‐east Australia

Summary The emergence of carbon markets has provided a potential source of funding for reforestation projects. However, there is concern amongst ecologists that these markets will promote the establishment of monoculture plantations rather than more diverse restoration plantings, on the assumption that fast‐growing monocultures are likely to store more carbon than restoration plantings. We examined the validity of this assumption for three predominantly rainforest plantation types established in the moist tropical uplands of north‐east Australia: monoculture plantations of native rainforest conifers (n = 5, mean age 13 years); mixed species plantations of rainforest cabinet timber species, rainforest conifers and eucalypts (n = 5, mean age 13 years); and, environmental restoration plantings comprised mostly of a diverse range of rainforest trees (n = 10, mean age 14 years). We found that restoration plantings stored significantly more carbon in above‐ground biomass than monoculture plantations of native conifers (on average, 106 t vs 62 t carbon per ha); and tended to store more carbon than mixed species timber plantations which were intermediate in value (86 t carbon per ha). Carbon stocks were higher in restoration plantings than in monoculture and mixed species plantations for three reasons. First, and most importantly, restoration plantings were more densely stocked than monoculture and mixed species plantations. Second, there were more large diameter trees in restoration plantings than monoculture plantations. Third, the trees used in restoration plantings had a higher average wood density than the conifers used in monoculture plantations. While, on average, wood density was higher in mixed species plantations than restoration plantings, the much higher stocking rate of restoration plantings meant they stored more carbon than mixed species plantations. We conclude that restoration plantings in the moist tropics of north‐east Australia can accumulate relatively high amounts of carbon within two decades of establishment. Comparison with reference rainforest sites suggests that restoration plantings could maintain their high stocking rates (and therefore high biomass) as they develop in future decades. However, because restoration plantings are currently much more expensive to establish than monoculture plantations, restoration plantings are unlikely to be favoured by carbon markets. Novel reforestation techniques and designs are required if restoration plantings are to both provide habitat for rainforest biota and store carbon in biomass at a cost comparable to monoculture plantations.     

Induction of methotrexate release from rat hepatocytes in suspension by alpha-adrenergic agents: involvement of calcium and metabolic energy

Hepatocytes in suspension which have accumulated [3H]methotrexate release the antifolate compound into the medium upon exposure to alpha-adrenergic agents. In the presence of metabolic poisons, such as sodium azide, dinitrophenol, or dicumarol, the release of methotrexate is attenuated, indicating that integrity of the cellular metabolic apparatus is required for response to the hormonal stimulus. In the presence of millimolar concentrations of the organic acid, probenecid, release of cellular methotrexate may be reduced (1 mM probenecid) or eliminated (2 mM probenecid), suggesting the involvement of a "membrane carrier." Microtubule poisons such as vincristine, vinblastine, and griseofulvin do not modify epinephrine + isobutyl methyl xanthine (IBMX)-induced release of methotrexate. The involvement of calcium in release of methotrexate from the hepatocyte is substantiated by a dose-dependent response to the calcium ionophore, A23187, in the presence of calcium, with a lack of response in the absence of calcium. These effects of A23187 are not related to inhibition of methotrexate influx. Other putative "calcium antagonists" such as tetracaine, neomycin sulfate, and 3,4,5-trimethoxybenzoic acid [8-(diethylamino)octyl ester], do not interfere with epinephrine + IBMX-induced release of [3H]methotrexate, suggesting that these agents may not be effective probes of calcium flux in the liver cell.     

Laccase immobilization on enzymatically functionalized polyamide 6,6 fibres

Polyamide matrices, such as membranes, gels and non-wovens, have been applied as supports for enzyme immobilization, although in literature the enzyme immobilization on woven nylon matrices is rarely reported. In this work, a protocol for a Trametes hirsuta laccase immobilization using woven polyamide 6,6 (nylon) was developed. A 2⁴ full factorial design was used to study the influence of pH, spacer (1,6-hexanediamine), enzyme and crosslinker concentration on the efficiency of immobilization. The factors enzyme dosage and spacer seem to have played a critical role in the immobilization of laccase onto nylon support. Under optimized working conditions (29 U mL⁻¹ of laccase, 10% of glutaraldehyde, pH = 5.5, with the presence of the spacer), the half-life time attained was about 78 h (18% higher than that of free enzyme), the protein retention was 30% and the immobilization yield was 2%. The immobilized laccase has potential for application in the continuous decolourization of textile effluents, where it can be applied into a membrane reactor.     

Biosynthesis of vitamin B12 by Propionibacterium freudenreichii subsp. shermanii ATCC 13673 using liquid acid protein residue of soybean as culture medium

Vitamin B12 deficiency still persists, mainly caused by low intake of animal food products affecting vegetarians, vegans, and populations of underdeveloped countries. In this study, we investigate the biosynthesis of vitamin B12 by potential probiotic bacterium using an agroindustry residue, the liquid acid protein residue of soybean (LAPRS), as a low-cost, animal derivate-free alternative culture medium. Cultures of Propionibacterium freudenreichii subsp. shermanii ATCC 13673 growing in LAPRS for vitamin B12 biosynthesis were studied using the Plackett–Burman experimental approach, followed by a central composite design 2² to optimize the concentration of significant variables. We also performed a proteolytic treatment of LAPRS and evaluated the optimized–hydrolyzed medium influence on the microbial growth and metabolism in shaker flask and bioreactor experiments. In this all-plant source medium, P. freudenreichii subsp. shermanii produced high concentrations of cells and high amounts of vitamin B12 (0.6 mg/g cells) after process optimization. These results suggest the possibility of producing vitamin B12 by a potential probiotic bacterium in a very cheap, animal derivate-free medium to address the needs of specific population groups, at the same time reducing the production costs of this essential vitamin.     

Complex Bileaflet Mitral Valve Repair (Barlow's) Using the da Vinci Robotic Surgical System

Robotic mitral valve repair (RMVR) is less invasive and potentially more precise. However, RMVR lengthens both cardiopulmonary bypass and arrested heart times. In our initial experience, only posterior leaflet repair and/or annuloplasty were performed. With increasing experience, we have performed more complex bileaflet RMVR. A 50-year-old man presented with severe mitral regurgitation. Transesophageal echocardiography (TEE) demonstrated a complex bileaflet prolapse and preserved left ventricular function. Through a 4 cm working port and with the da Vinci Robotic Surgical System (Intuitive Surgical, Sunnyvale, CA) RMVR was performed. Details of the technique and patient's hospital course are described. The repair comprised closure of clefts between A3 and P3, quadrangular resection of P2, transfer of multiple chords from P2 to A2/A3 and a #38 Cosgrove-Edwards (Edwards Lifesciences, Irvine, CA) band annuloplasty. Nitinol U-Clips (Medtronic, Minneapolis, MN) were used to complete the annuloplasty. Postoperative TEE showed no mitral regurgitation. The patient was discharged on the third postoperative day. Cardiopulmonary bypass and arrested heart times were 3 hours and 29 minutes and 2 hours and 59 minutes, respectively. Complex bileaflet repair of mitral valve with Barlow's disease can be successfully performed with the da Vinci Robotic Surgical System. Long-term follow-up is needed to assess the durability of repair.     

Ligninolytic activity of tropical rainforest basidiomycetes

A total of 116 strains of Brazilian tropical rainforest basidiomycetes were evaluated in terms of their ability to oxidize the dye rhemazol brilliant blue R (RBBR) and guaiacol. Laccase and peroxidase activities were detected by the drop test using solutions of α-naphthol and pyrogallol, respectively. RBBR and guaiacol oxidation occurred in 96.6 and 87.1% of the strains tested, respectively. One hundred strains oxidized both substrates. In the drop test, most strains presented laccase (96.6%) and peroxidase (92.2%) activity. The quick screening method used here can be useful to identify ligninolytic fungal strains to be used in various biotechnological applications. This revised version was published online in November 2006 with corrections to the Cover Date.