The heterotrimeric G-protein complex modulates light sensitivity in arabidopsis thaliana seed germination

Release of dormancy and induction of seed germination are complex traits finely regulated by hormonal signals and environmental cues such as temperature and light. The Red (R):Far-Red (FR) phytochrome photoreceptors mediate light regulation of seed germination. We investigated the possible involvement of heterotrimeric G-protein complex in the phytochrome signaling pathways of Arabidopsis thaliana seed germination. Germination rates of null mutants of the alpha (Gα) and beta (Gβ) subunits of the G-protein (Atgpa1-4 and agb1-2, respectively) and the double mutant (agb1-2/gpa1-4) are lower than the wildtype (WT) under continuous or pulsed light. The Gα and Gβ subunits play a role in seed germination under hourly pulses of R lower than 0.1 μmol m -2 s -1 whereas the Gβ subunit plays a role in higher R fluences. The germination of double mutants of G-protein subunits with phyA-211 and phyB-9 suggests that AtGPA1 seems to act as a positive regulator of phyA and probably phyB signaling pathways, while the role of AGB1 is ambiguous. The imbibition of seeds at 4°C and 35°C alters the R and FR light responsiveness of WT and G-protein mutants to a similar magnitude. Thus, Gα and Gβ subunits of the heterotrimeric G-protein complex modulate light induction of seed germination by phytochromes and are dispensable for the control of dormancy by low and high temperatures prior to irradiation. We discuss the possible indirect role of the G-protein complex on the phytochrome-regulated germination through hormonal signaling pathways.Fil: Botto, Javier Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; ArgentinaFil: Ibarra, Silvia Elizabeth. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; ArgentinaFil: Jones, Alan M.. University of North Carolina; Estados Unido

Susceptibility to Mycobacterium ulcerans Disease (Buruli ulcer) Is Associated with IFNG and iNOS Gene Polymorphisms.

Buruli ulcer (BU) is a chronic necrotizing disease of the skin and subcutaneous fat tissue. The causative agent, Mycobacterium ulcerans, produces mycolactone, a macrolide toxin, which causes apoptosis of mammalian cells. Only a small proportion of individuals exposed to M. ulcerans develop clinical disease, as surrounding macrophages may control the infection by bacterial killing at an early stage, while mycolactone concentration is still low. Otherwise, bacterial multiplication leads to in higher concentrations of mycolactone, with formation of necrotizing lesions that are no more accessible to immune cells. By typing a cohort of 96 Ghanaian BU patients and 384 endemic controls without BU, we show an association between BU and single nucleotide polymorphisms (SNPs) in iNOS (rs9282799) and IFNG (rs2069705). Both polymorphisms influence promoter activity in vitro. A previously reported SNP in SLC11A1 (NRAMP, rs17235409) tended to be associated with BU. Altogether, these data reflect the importance of IFNG signaling in early defense against M. ulcerans infection

Transcriptional Profiling of Patient Isolates Identifies a Novel TOR/Starvation Regulatory Pathway in Cryptococcal Virulence.

Human infection with Cryptococcus causes up to a quarter of a million AIDS-related deaths annually and is the most common cause of nonviral meningitis in the United States. As an opportunistic fungal pathogen, Cryptococcus neoformans is distinguished by its ability to adapt to diverse host environments, including plants, amoebae, and mammals. In the present study, comparative transcriptomics of the fungus within human cerebrospinal fluid identified expression profiles representative of low-nutrient adaptive responses. Transcriptomics of fungal isolates from a cohort of HIV/AIDS patients identified high expression levels of an alternative carbon nutrient transporter gene, STL1, to be associated with poor early fungicidal activity, an important clinical prognostic marker. Mouse modeling and pathway analysis demonstrated a role for STL1 in mammalian pathogenesis and revealed that STL1 expression is regulated by a novel multigene regulatory mechanism involving the CAC2 subunit of the chromatin assembly complex 1, CAF-1. In this pathway, the global regulator of virulence gene VAD1 was found to transcriptionally regulate a cryptococcal homolog of a cytosolic protein, Ecm15, in turn required for nuclear transport of the Cac2 protein. Derepression of STL1 by the CAC2-containing CAF-1 complex was mediated by Cac2 and modulated binding and suppression of the STL1 enhancer element. Derepression of STL1 resulted in enhanced survival and growth of the fungus in the presence of low-nutrient, alternative carbon sources, facilitating virulence in mice. This study underscores the utility of ex vivo expression profiling of fungal clinical isolates and provides fundamental genetic understanding of saprophyte adaption to the human host.IMPORTANCECryptococcus is a fungal pathogen that kills an estimated quarter of a million individuals yearly and is the most common cause of nonviral meningitis in the United States. The fungus is carried in about 10% of the adult population and, after reactivation, causes disease in a wide variety of immunosuppressed individuals, including the HIV infected and patients receiving transplant conditioning, cancer therapy, or corticosteroid therapy for autoimmune diseases. The fungus is widely carried in the soil but can also cause infections in plants and mammals. However, the mechanisms for this widespread ability to infect a variety of hosts are poorly understood. The present study identified adaptation to low nutrients as a key property that allows the fungus to inhabit these diverse environments. Further studies identified a nutrient transporter gene, STL1, to be upregulated under low nutrients and to be associated with early fungicidal activity, a marker of poor clinical outcome in a cohort of HIV/AIDS patients. Understanding molecular mechanisms involved in adaptation to the human host may help to design better methods of control and treatment of widely dispersed fungal pathogens such as Cryptococcus

A study on the fracture strength of collarless metal-ceramic fixed partial dentures

PURPOSE. The objective of this study was to evaluate fracture strength of collarless metal-ceramic FPDs according to their metal coping designs. MATERIALS AND METHODS. Four different facial margin design groups were investigated. Group A was a coping with a thin facial metal collar, group B was a collarless coping with its facial metal to the shoulder, group C was a collarless coping with its facial metal 1 mm short of the shoulder, and group D was a collarless coping with its facial metal 2 mm short of the shoulder. Fifteen 3-unit collarless metal-ceramic FPDs were fabricated in each group. Finished FPDs were cemented to PBT (Polybutylene terephthalate) dies with resin cement. The fracture strength test was carried out using universal testing machine (Instron 4465, Instron Co., Norwood MA, USA) at a cross head speed of 0.5 mm/min. Aluminum foil folded to about 1 mm of thickness was inserted between the plunger tip and the incisal edge of the pontic. Vertical load was applied until catastrophic porcelain fracture occurred. RESULTS. The greater the bulk of unsupported facial shoulder porcelain was, the lower the fracture strength became. However, there were no significant differences between experimental groups (P>.05). CONCLUSION. All groups of collarless metal-ceramic FPDs had higher fracture strength than maximum incisive biting force. Modified collarless metal-ceramic FPD can be an alternative to all-ceramic FPDs in clinical situations. [J Adv Prosthodont 2010;2:134-41]

Effect of Discontinuation of Fluoride Intake from Water and Toothpaste on Urinary Excretion in Young Children

As there is no homeostatic mechanism for maintaining circulating fluoride (F) in the human body, the concentration may decrease and increase again when intake is interrupted and re-started. The present study prospectively evaluated this process in children exposed to F intake from water and toothpaste, using F in urine as a biomarker. Eleven children from Ibiá, Brazil (with sub-optimally fluoridated water supply) aged two to four years who regularly used fluoridated toothpaste (1,100 ppm F) took part in the study. Twenty-four-hour urine was collected at baseline (Day 0, F exposure from water and toothpaste) as well as after the interruption of fluoride intake from water and dentifrice (Days 1 to 28) (F interruption) and after fluoride intake from these sources had been re-established (Days 29 to 34) (F re-exposure). Urinary volume was measured, fluoride concentration was determined and the amount of fluoride excreted was calculated and expressed in mg F/day. Urinary fluoride excretion (UFE) during the periods of fluoride exposure, interruption and re-exposure was analyzed using the Wilcoxon test. Mean UFE was 0.25 mg F/day (SD: 0.15) at baseline, dropped to a mean of 0.14 mg F/day during F interruption (SD: 0.07; range: 0.11 to 0.17 mg F/day) and rose to 0.21 (SD: 0.09) and 0.19 (SD: 0.08) following F re-exposure. The difference between baseline UFE and the period of F interruption was statistically significant (p < 0.05), while the difference between baseline and the period of F re-exposure was non-significant (p > 0.05). The findings suggest that circulating F in the body of young children rapidly decreases in the first 24 hours and again increases very fast after discontinuation and re-exposure of F from water and toothpaste

Fluoride Consumption and Its Impact on Oral Health

Objective. The purpose of this study was to evaluate caries and dental fluorosis among Mexican preschoolers and school-aged children in a non-endemic zone for fluorosis and to measure its biological indicators. Methods. DMFT, DMFS, dmft, dmfs, and CDI indexes were applied. Fluoride urinary excretion and fluoride concentrations in home water, table salt, bottled water, bottled drinks, and toothpaste were determined. Results. Schoolchildren presented fluorosis (CDI = 0.96) and dental caries (DMFT = 2.64 and DMFS = 3.97). Preschoolers presented dmft = 4.85 and dmfs = 8.80. DMFT and DMFS were lower in children with mild to moderate dental fluorosis (DF). Variable fluoride concentrations were found in the analyzed products (home water = 0.18–0.44 ppm F, table salt = 0–485 ppm F, bottled water = 0.18–0.47 ppm F, juices = 0.08–1.42 ppm F, nectars = 0.07–1.30 ppm F, bottled drinks = 0.10–1.70 ppm F, toothpaste = 0–2,053 ppm F). Mean daily fluoride excretion was 422 ± 176 µg/24 h for schoolchildren and 367 ± 150 µg/24 h for preschoolers. Conclusions. Data from our study show that, despite values of excretion within an optimal fluoride intake range, the prevalence of caries was significant in both groups, and 60% of the 11- to 12-year-old children presented with dental fluorosis. In addition, variable fluoride concentrations in products frequently consumed by children were found