From childhood to adulthood: oral rehabilitation of a patient with ectodermal dysplasia.

Ectodermal dysplasia, in all its varieties, occurs in approximately 1 in every 100,000 live births. Depending on the variety, hypohidrosis, hypotrichosis, hypodontia, oligodontia, and/or anodontia may be exhibited. Few long-term clinical reports exist. This report follows the development, growth, and initial treatment of a 10-year-old patient with ectodermal dysplasia and his subsequent oral rehabilitation 20 years later.

The effect of various disinfectants on dental shade guides.

STATEMENT OF PROBLEM: Dental shade guides are used to evaluate tooth color before prosthodontic procedures and are subjected to disinfection after use. The effect of disinfection on shade guides has not been thoroughly investigated. PURPOSE: The purpose of this study was to evaluate the effect of disinfectants on the color of shade tabs. MATERIAL AND METHODS: Changes in the color (ΔE) of VITA Classical Shade Guide tabs were measured with a VITA Easyshade spectrophotometer in the CIELAB system and calculated after being subjected to Cavicide, Asepticare TB, Sporicidin, and distilled water (control) over a simulated period of 2 years. Statistical analysis was accomplished by a 2-way analysis of variance followed by the Tukey honestly significant difference (HSD) test (α=.05). RESULTS: A significant difference was noted in the degree of shade tab color change, depending on the type of disinfectant used (F=153.2, P<.001). No significant difference was noted in the amount of shade tab color change that occurred after disinfection among the different shade tabs used (F=0.611, P=.865), nor was a significant interaction noted between the type of disinfectant and the different shade tabs used (F=0.7, P=.919). Asepticare TB showed the least significant amount of change (ΔE=0.401), and Sporicidin (ΔE=0.889) and the control (ΔE=0.969) showed significantly more color change than Asepticare TB but less than Cavicide (ΔE=1.198). CONCLUSIONS: The average total CIELAB color difference for 50% human perceptibility is approximately 1 unit (under standardized laboratory conditions). In the oral cavity, however, an average change of 3.7 ΔE units could still allow teeth to be perceived as having the same color. Therefore, although the results are statistically significant, they may not be clinically important.

JNK1/2 regulate Bid by direct phosphorylation at Thr59 in response to ALDH1L1.

BH3 interacting-domain death agonist (Bid) is a BH3-only pro-apoptotic member of the Bcl-2 family of proteins. Its function in apoptosis is associated with the proteolytic cleavage to the truncated form tBid, mainly by caspase-8. tBid translocates to mitochondria and assists Bax and Bak in induction of apoptosis. c-Jun N-terminal kinase (JNK)-dependent alternative processing of Bid to jBid was also reported. We have previously shown that the folate stress enzyme 10-formyltetrahydrofolate dehydrogenase (ALDH1L1) activates JNK1 and JNK2 in cancer cells as a pro-apoptotic response. Here we report that in PC-3 prostate cancer cells, JNK1/2 phosphorylate Bid at Thr59 within the caspase cleavage site in response to ALDH1L1. In vitro, all three JNK isoforms, JNK 1-3, phosphorylated Thr59 of Bid with JNK1 being the least active. Thr59 phosphorylation protected Bid from cleavage by caspase-8, resulting in strong accumulation of the full-length protein and its translocation to mitochondria. Interestingly, although we did not observe jBid in response to ALDH1L1 in PC-3 cells, transient expression of Bid mutants lacking the caspase-8 cleavage site resulted in strong accumulation of jBid. Of note, a T59D mutant mimicking constitutive phosphorylation revealed more profound cleavage of Bid to jBid. JNK-driven Bid accumulation had a pro-apoptotic effect in our study: small interfering RNA silencing of either JNK1/2 or Bid prevented Bid phosphorylation and accumulation, and rescued ALDH1L1-expressing cells. As full-length Bid is a weaker apoptogen than tBid, we propose that the phosphorylation of Bid by JNKs, followed by the accumulation of the full-length protein, delays attainment of apoptosis, and allows the cell to evaluate the stress and make a decision regarding the response strategy. This mechanism perhaps can be modified by the alternative cleavage of phospho-T59 Bid to jBid at some conditions.

Bronchial reactivity and work-related symptoms in farmers.

Work-related respiratory symptoms and bronchial reactivity were studied in 76 never-smoking farmers and in a control group not exposed to organic dusts. The farmers were divided into those working with vegetables/grain crops, animals but not swine, and with swine. The extent of symptoms was evaluated using a specific organic dust questionnaire. Bronchial reactivity was assessed with the methacholine challenge test. An increased incidence of organic dust toxic syndrome (ODTS), mucous membrane irritation (MMI), and chronic bronchitis (CB) was found among farmers working with swine or other animals. Pulmonary function baseline values were normal. Bronchial reactivity was increased and related to subjective symptoms of MMI and CB. There was also a relation between fatigue at work and bronchial reactivity.

Environmental and health studies of farm workers in Swedish swine confinement buildings.

The relation between the health of workers and the environment in swine confinement buildings was investigated in a study of 57 workers on 30 swine farms in southern Sweden and 55 matched controls. Swine workers reported significantly higher frequencies of respiratory symptoms, more frequent colds and absence due to chest illness, and a history of pneumonia. The increased frequency of symptoms of respiratory disease was related to the number of years and percent of the day spent working with swine. Symptoms were also associated with respirable dust, total dust, endotoxin in total dust, and number of microbes in the air of the work environment. In a multiple regression analysis of the relation between 16 different environmental parameters to work period shifts of five pulmonary function parameters, endotoxin was found to be significantly related to the FEV1 in a dose dependent way.

Selective interaction of AGS3 with G-proteins and the influence of AGS3 on the activation state of G-proteins.

AGS3 (activator of G-protein signaling 3) was isolated in a yeast-based functional screen for receptor-independent activators of heterotrimeric G-proteins. As an initial approach to define the role of AGS3 in mammalian signal processing, we defined the AGS3 subdomains involved in G-protein interaction, its selectivity for G-proteins, and its influence on the activation state of G-protein. Immunoblot analysis with AGS3 antisera indicated expression in rat brain, the neuronal-like cell lines PC12 and NG108-15, as well as the smooth muscle cell line DDT(1)-MF2. Immunofluorescence studies and confocal imaging indicated that AGS3 was predominantly cytoplasmic and enriched in microdomains of the cell. AGS3 coimmunoprecipitated with Galpha(i3) from cell and tissue lysates, indicating that a subpopulation of AGS3 and Galpha(i) exist as a complex in the cell. The coimmunoprecipitation of AGS3 and Galpha(i) was dependent upon the conformation of Galpha(i3) (GDP GTPgammaS (guanosine 5'-3-O-(thio)triphosphate)). The regions of AGS3 that bound Galpha(i) were localized to four amino acid repeats (G-protein regulatory motif (GPR)) in the carboxyl terminus (Pro(463)-Ser(650)), each of which were capable of binding Galpha(i). AGS3-GPR domains selectively interacted with Galpha(i) in tissue and cell lysates and with purified Galpha(i)/Galpha(t). Subsequent experiments with purified Galpha(i2) and Galpha(i3) indicated that the carboxyl-terminal region containing the four GPR motifs actually bound more than one Galpha(i) subunit at the same time. The AGS3-GPR domains effectively competed with Gbetagamma for binding to Galpha(t(GDP)) and blocked GTPgammaS binding to Galpha(i1). AGS3 and related proteins provide unexpected mechanisms for coordination of G-protein signaling pathways.

Stabilization of the GDP-bound conformation of Gialpha by a peptide derived from the G-protein regulatory motif of AGS3.

The G-protein regulatory (GPR) motif in AGS3 was recently identified as a region for protein binding to heterotrimeric G-protein alpha subunits. To define the properties of this approximately 20-amino acid motif, we designed a GPR consensus peptide and determined its influence on the activation state of G-protein and receptor coupling to G-protein. The GPR peptide sequence (28 amino acids) encompassed the consensus sequence defined by the four GPR motifs conserved in the family of AGS3 proteins. The GPR consensus peptide effectively prevented the binding of AGS3 to Gialpha1,2 in protein interaction assays, inhibited guanosine 5'-O-(3-thiotriphosphate) binding to Gialpha, and stabilized the GDP-bound conformation of Gialpha. The GPR peptide had little effect on nucleotide binding to Goalpha and brain G-protein indicating selective regulation of Gialpha. Thus, the GPR peptide functions as a guanine nucleotide dissociation inhibitor for Gialpha. The GPR consensus peptide also blocked receptor coupling to Gialphabetagamma indicating that although the AGS3-GPR peptide stabilized the GDP-bound conformation of Gialpha, this conformation of Gialpha(GDP) was not recognized by a G-protein coupled receptor. The AGS3-GPR motif presents an opportunity for selective control of Gialpha- and Gbetagamma-regulated effector systems, and the GPR motif allows for alternative modes of signal input to G-protein signaling systems.

AGS3 inhibits GDP dissociation from galpha subunits of the Gi family and rhodopsin-dependent activation of transducin.

A number of recently discovered proteins that interact with the alpha subunits of G(i)-like G proteins contain homologous repeated sequences named G protein regulatory (GPR) motifs. Activator of G protein signaling 3 (AGS3), identified as an activator of the yeast pheromone pathway in the absence of the pheromone receptor, has a domain with four such repeats. To elucidate the potential mechanisms of regulation of G protein signaling by proteins containing GPR motifs, we examined the effects of the AGS3 GPR domain on the kinetics of guanine nucleotide exchange and GTP hydrolysis by G(i)alpha(1) and transducin-alpha (G(t)alpha). The AGS3 GPR domain markedly inhibited the rates of spontaneous guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) binding to G(i)alpha and rhodopsin-stimulated GTPgammaS binding to G(t)alpha. The full-length AGS3 GPR domain, AGS3-(463-650), was approximately 30-fold more potent than AGS3-(572-629), containing two AGS3 GPR motifs. The IC(50) values for the AGS3-(463-650) inhibitory effects on G(i)alpha and transducin were 0.12 and 0.15 microm, respectively. Furthermore, AGS3-(463-650) and AGS3-(572-629) effectively blocked the GDP release from G(i)alpha and rhodopsin-induced dissociation of GDP from G(t)alpha. The potencies of AGS3-(572-629) and AGS3-(463-650) to suppress the GDP dissociation rates correlated with their ability to inhibit the rates of GTPgammaS binding. Consistent with the inhibition of nucleotide exchange, the AGS3 GPR domain slowed the rate of steady-state GTP hydrolysis by G(i)alpha. The catalytic rate of G(t)alpha GTP hydrolysis, measured under single turnover conditions, remained unchanged with the addition of AGS3-(463-650). Altogether, our results suggest that proteins containing GPR motifs, in addition to their potential role as G protein-coupled receptor-independent activators of Gbetagamma signaling pathways, act as GDP dissociation inhibitors and negatively regulate the activation of a G protein by a G protein-coupled receptor.