Involvement of AMPA Receptor GluR2 and GluR3 Trafficking in Trigeminal Spinal Subnucleus Caudalis and C1/C2 Neurons in Acute-Facial Inflammatory Pain

To evaluate the involvement of trafficking of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) GluR2 and GluR3 subunits in an acute inflammatory orofacial pain, we analyzed nocifensive behavior, phosphorylated extracellular signal-regulated kinase (pERK) and Fos expression in Vi/Vc, Vc and C1/C2 in GluR2 delta7 knock-in (KI), GluR3 delta7 KI mice and wild-type mice. We also studied Vc neuronal activity to address the hypothesis that trafficking of GluR2 and GluR3 subunits plays an important role in Vi/Vc, Vc and C1/C2 neuronal activity associated with orofacial inflammation in these mice. Late nocifensive behavior was significantly depressed in GluR2 delta7 KI and GluR3 delta7 KI mice. In addition, the number of pERK-immunoreactive (IR) cells was significantly decreased bilaterally in the Vi/Vc, Vc and C1/C2 in GluR2 delta7 KI and GluR3 delta7 KI mice compared to wild-type mice at 40 min after formalin injection, and was also significantly smaller in GluR3 delta7 KI compared to GluR2 delta7 KI mice. The number of Fos protein-IR cells in the ipsilateral Vi/Vc, Vc and C1/C2 was also significantly smaller in GluR2 delta7 KI and GluR3 delta7 KI mice compared to wild-type mice 40 min after formalin injection. Nociceptive neurons functionally identified as wide dynamic range neurons in the Vc, where pERK- and Fos protein-IR cell expression was prominent, showed significantly lower spontaneous activity in GluR2 delta7 KI and GluR3 delta7 KI mice than wild-type mice following formalin injection. These findings suggest that GluR2 and GluR3 trafficking is involved in the enhancement of Vi/Vc, Vc and C1/C2 nociceptive neuronal excitabilities at 16-60 min following formalin injection, resulting in orofacial inflammatory pain.     

Cytological and Reproductive Studies of Japanese Diplazium (Woodsiaceae; Pteridophyta). III. The Cytological Complexity of Species Groups with Simply Pinnate to Bipinnatifid Leaves

Diplazium with simply pinnate or bipinnatifid leaves. Diplazium wichurae var. wichurae, D. wichurae var. amabile, D. okudairae, and D. pin-faense are sexual diploids (2n=82; n=41II); D.× kidoi and D. × okudairaeoides are sterile diploids (2n= 82; meiosis irregular); D. donianum var. donianum is an apomictic triploid (2n=123; n=123II); D. donianum var. aphanoneuron is a sterile triploid (2n=123; meiosis irregular); D. crassiusculum, D. cavalerianum, D. incomptum, D. longicarpum, and D. pullingeri are sexual tetraploids (2n= 164; n=82II); and D. lobatum is an apomictic tetraploid (2n=164; n=164II). This is the first report of the chromosome numbers of D. lobatum, D. crassiusculum, D. incomptum, D. longicarpum, D. pullingeri, and D. × okudairaeoides, as well as the mitotic chromosome numbers of D. wichurae var. amabile, D. okudairae, D. pinfaense, and D. ×kidoi. The mitotic chromosome number, meiotic behavior, sterility, and allozyme analysis confirm that D. × kidoi and D. × okudairaeoides are hybrids between D. pin-faense and D. wichurae var. wichurae and D. okudairae and D. wichurae var. wichurae, respectively. Diplazium with simply pinnate to bipinnatifid leaves displayed an extraordinary cytological and reproductive complexity: a polyploidal series with diploids to hexaploids, sexual and apomictic reproduction, and natural hybridization. Received 14 August 2001/ Accepted in revised form 1 October 2001     

Diverse Expression of β1,4-N-Acetylgalactosaminyltransferase Gene in the Adult Mouse Brain

Abstract: Among various tissues of mouse, β1,4- N -acetylgalactosaminyltransferase (GM2/GD2 synthase) gene is expressed predominantly in the brain. Further analysis of the gene expression in the mouse CNS was performed by northern blotting and by enzyme assays using extracts from various parts of the CNS. In situ hybridization was also done to investigate the distribution of cells generating GM2/GD2 synthase. In northern blots, diverse levels of the gene expression were observed, depending on the regions examined. By in situ hybridization, pyramidal cells in the hippocampus, granular cells in dentate gyrus and cerebral cortex, Purkinje cells in cerebellum, and mitral cells in the olfactory bulb expressed high levels of the mRNA; these results corresponded to the results obtained by northern blot. Enzyme levels in these sites were accordingly high. However, enzyme levels in certain areas with low mRNA intensities, such as thalamus and pons medulla, were higher than expected from the results of northern blotting. The significance of the high gene expression in certain areas for brain function and the reason for the discrepancy between mRNA level and enzyme activity in some regions are discussed.     

Functional analysis of isoforms of NADPH: protochlorophyllide oxidoreductase (POR), PORB and PORC, in Arabidopsis thaliana

NADPH:protochlorophyllide oxidoreductase (POR) catalyzes the light-dependent reduction of protochlorophyllide. To elucidate the physiological function of three differentially regulated POR isoforms (PORA, PORB and PORC) in Arabidopsis thaliana, we isolated T-DNA tagged null mutants of porB and porC. The mature seedlings of the mutants had normal photosynthetic competencies, showing that PORB and PORC are interchangeable and functionally redundant in developed plants. In etiolated seedlings, only porB showed a reduction in the photoactive protochlorophyllide and the size of prolamellar bodies (PLBs), indicating that PORB, as well as PORA, functioned in PLB assembly and photoactive protochlorophyllide formation in etiolated seedlings. When illuminated, the etiolated porB seedling was able to green to a similar extent as the wild type, whereas the greening was significantly reduced under low light conditions. During greening, high light irradiation increased the level of PORC protein, and the greening of porC was repressed under high light conditions. The porB, but not porC, etiolated seedling was more sensitive to the far-red block of greening than the wild type, which is caused by depletion of endogenous POR proteins resulting in photo-oxidative damage. These results suggest that, at the onset of greening, PLBs are important for efficient capture of light energy for photoconversion under various light conditions, and PORC, which is induced by high light irradiation, contributes to photoprotection during greening of the etiolated seedlings.     

Complete denture hygiene and nocturnal wearing habits among patients attending the Prosthodontic Department in a Dental University in Brazil

Gerodontology 2010; doi: 10.1111/j.1741‐2358.2010.00369.x Complete denture hygiene and nocturnal wearing habits among patients attending the Prosthodontic Department in a Dental University in Brazil Objective: The aim of this study was to evaluate the overnight wearing and cleaning habits of complete denture wearers. Background: Successful complete denture treatment can be achieved when the patients are motivated and aware of appropriate denture wear and hygiene. Materials and methods: A sample of 224 complete denture wearers (162 women) aged 37–89 years was studied. Inclusion criteria comprised edentulous subjects who had received their new complete dentures between 2000 and 2005 in the Dental Clinic of the Araçatuba and Araraquara Dental School, São Paulo State University. Ethical approval was sought and granted. Subjects were interviewed using questions related to overnight denture wearing and denture cleaning habits. Possible statistical relationships among some of items were analysed by the chi‐square test at 5% significance level. Results: Of the patients, 55.8% removed their dentures during the overnight period and 88% did this every day. Among them, 66.4% removed both dentures. Most of the patients used brushing with toothpaste (105 patients – 46.87%) as a cleaning method. More than a half of the subjects (63.4%) showed biofilm and calculus on their dentures. Conclusion: The patients need instructions and motivation concerning denture hygienic and denture removal overnight.     

Prolamellar bodies formed by cyanobacterial protochlorophyllide oxidoreductase in Arabidopsis

In angiosperms, chlorophyll biosynthesis is light dependent. A key factor in this process is protochlorophyllide oxidoreductase (POR), which requires light to catalyze the reduction of protochlorophyllide to chlorophyllide. It is believed that this protein originated from an ancient cyanobacterial enzyme that was introduced into proto‐plant cells during the primary symbiosis. Here we report that PORs from the cyanobacteria Gloeobacter violaceus PCC7421 and Synechocystis sp. PCC6803 function in plastids. First, we found that the G. violaceus POR shows a higher affinity to its substrate protochlorophyllide than the Synechocystis POR but a similar affinity to plant PORs. Secondly, the reduced size of prolamellar bodies caused by a knockdown mutation of one of the POR genes, PORA, in Arabidopsis could be complemented by heterologous expression of the cyanobacterial PORs. Photoactive protochlorophyllide in the etioplasts of the complementing lines, however, was retained at a low level as in the parent PORA knockdown mutant, indicating that the observed formation of prolamellar bodies was irrelevant to the assembly of photoactive protochlorophyllide. This work reveals a new view on the formation of prolamellar bodies and provides new clues about the function of POR in the etioplast–chloroplast transition.