Maintenance of optogenetic channel rhodopsin (ChR2) function in aging mice: Implications for pharmacological studies of inhibitory synaptic transmission, quantal content, and calcium homeostasis.

Disruption of synaptic function is believed to represent a common pathway contributing to cognitive decline during aging. Optogenetics is a prodigious tool for studying relationships between function and synaptic circuitry but models utilizing viral vectors present limitations. Careful characterization of the functionality of channel rhodopsin in transgenic models is crucial for determining whether they can be used across aging. This includes verifying the light sensitivity of the protein and confirming its ability to generate action potentials in response to light stimulation. We combined in vitro optogenetic methodology and a reduced synaptic preparation of acutely isolated neurons to determine if the ChR2(H134R)-eYFP vGAT mouse model is well-suited for aging studies. We used neurons from young (2-6 mo), middle-aged (10-14 mo) and aged (17-25 mo) bacterial artificial chromosome (BAC) transgenic mouse line with stable expression of the channelrhodopsin-2 (ChR2) variant H134R in GABAergic cell populations. Cellular physiology and calcium dynamics were assessed in basal forebrain (BF) neurons using patch-clamp recording and fura-2 microfluorimetry, alongside 470 nm light stimulation of the transgenic ChR2 channel to characterize a wide array of physiological functions known to decline with age. We found ChR2 expression is functionally maintained across aging, while spontaneous and optically evoked inhibitory postsynaptic currents, and quantal content were decreased. Aged mice also showed an increase in intracellular calcium buffering. These results, which are on par with previous observations, demonstrate that the optogenetic vGAT BAC mouse model is well-suited for investigating age-related changes in calcium signaling and synaptic transmission.

Late-Onset, Short-Term Intermittent Fasting Reverses Age-Related Changes in Calcium Buffering and Inhibitory Synaptic Transmission in Mouse Basal Forebrain Neurons.

Aging is often associated with cognitive decline and recurrent cellular and molecular impairments. While life-long caloric restriction (CR) may delay age-related cognitive deterioration as well as the onset of neurologic disease, recent studies suggest that late-onset, short-term intermittent fasting (IF), may show comparable beneficial effects as those of life-long CR to improve brain health. We used a new optogenetic aging model to study the effects of late-onset (>18 months), short-term (four to six weeks) IF on age-related changes in GABAergic synaptic transmission, intracellular calcium (Ca2+) buffering, and cognitive status. We used male mice from a bacterial artificial chromosome (BAC) transgenic mouse line with stable expression of the channelrhodopsin-2 (ChR2) variant H134R [VGAT-ChR2(H134R)-EYFP] in a reduced synaptic preparation that allows for specific optogenetic light stimulation on GABAergic synaptic terminals across aging. We performed quantal analysis using the method of failures in this model and show that short-term IF reverses the age-related decrease in quantal content of GABAergic synapses. Likewise, short-term IF also reversed age-related changes in Ca2+ buffering and spontaneous GABAergic synaptic transmission in basal forebrain (BF) neurons of aged mice. Our findings suggest that late-onset short-term IF can reverse age-related physiological impairments in mouse BF neurons but that four weeks IF is not sufficient to reverse age-related cognitive decline.SIGNIFICANCE STATEMENT Here, we demonstrate plasticity of the aging brain and reversal of well-defined hallmarks of brain aging using short-term intermittent fasting (IF) initiated later in life. Few therapeutics are currently available to treat age-related neurologic dysfunction although synaptic dysfunction occurs during aging and neurologic disease is a topic of intense research. Using a new reduced synaptic preparation and optogenetic stimulation we are able to study age-related synaptic mechanisms in greater detail. Several neurophysiological parameters including quantal content were altered during aging and were reversed with short-term IF. These methods can be used to identify potential therapies to reverse physiological dysfunction during aging.

Amitriptyline Decreases GABAergic Transmission in Basal Forebrain Neurons Using an Optogenetic Model of Aging.

The antidepressant drug amitriptyline is used in the treatment of clinical depression and a variety of neurological conditions such as anxiety, neuropathic pain disorders and migraine. Antidepressants are associated with both therapeutic and untoward effects, and their use in the elderly has tripled since the mid-1990s. Because of this widespread use, we are interested in testing the acute effects of amitriptyline on synaptic transmission at therapeutic concentrations well below those that block voltage-gated calcium channels. We found that 3 µM amitriptyline reduced the frequency of spontaneous GABAergic inhibitory postsynaptic currents (IPSCs) and reduced quantal content in mice at ages of 7-10 mo. and 23-25 mo., suggesting a presynaptic mechanism of action that does not diminish with age. We employed a reduced synaptic preparation of the basal forebrain (BF) and a new optogenetic aging model utilizing a bacterial artificial chromosome (BAC) transgenic mouse line with stable expression of the channelrhodopsin-2 (ChR2) variant H134R specific for GABAergic neurons [VGAT-ChR2(H134R)-EYFP]. This model enables optogenetic light stimulation of specific GABAergic synaptic terminals across aging. Age-related impairment of circadian behavior was used to confirm predictable age-related changes associated with this model. Our results suggest that low concentrations of amitriptyline act presynaptically to reduce neurotransmitter release and that this action is maintained during aging.

Flexural strength of denture base acrylic resins processed by conventional and CAD-CAM methods.

STATEMENT OF PROBLEM: High flexural strength is one of the desirable properties for denture base resins, yet only few studies have evaluated the physical properties of newer denture bases such as computer-aided design and computer aided manufacturing (CAD-CAM) milled products. PURPOSE: The purpose of this in vitro study was to compare the flexural strength of 3 different types of denture base resins: compression molded, injection molded, and CAD-CAM milled. MATERIAL AND METHODS: Three groups (n=10) of acrylic denture base resins were tested: injection molded, compression molded, and CAD-CAM milled resin. ISO-compliant, rectangular specimens were fabricated (64×10×3.3 mm) (n=30). Specimens were stored in water for 1 week, and flexural strength was measured by using a 3-point bend test until failure. The Student t test was used to evaluate differences in the flexural strength and modulus of elasticity among specimen groups. The Bonferroni formula was used to set significance at α=.017 to account for multiple comparisons among the 3 groups. RESULTS: The flexural strength of the CAD-CAM milled group was significantly higher than that of the other 2 groups (P<.001), while the strength of the compression molded group was significantly greater than that of the injection molded group (P<.001). The flexural modulus of the CAD-CAM group was significantly higher than that of the other 2 groups (P<.001). CONCLUSIONS: CAD-CAM milled denture bases may be a useful alternative to conventionally processed denture bases in situations where increased resistance to flexural strength is needed.

Effects of ethanol and varenicline on female Sprague-Dawley rats in a third trimester model of fetal alcohol syndrome.

Perinatal ethanol exposure disrupts a variety of developmental processes in neurons important for establishing a healthy brain. These ethanol-induced impairments known as fetal alcohol spectrum disorder (FASD) are not fully understood, and currently, there is no effective treatment. Further, growing evidence suggests that adult females are more susceptible to ethanol, with the effects of perinatal ethanol exposure also being sexually divergent. Female models have been historically underutilized in neurophysiological investigations, but here, we used a third-trimester binge-ethanol model of FASD to examine changes to basal forebrain (BF) physiology and behavior in female Sprague-Dawley rats. We also tested varenicline as a potential cholinomimetic therapeutic. Rat pups were gavage-treated with binge-like ethanol, varenicline and ethanol, and varenicline alone. Using patch-clamp electrophysiology in BF slices, we observed that binge-ethanol exposure increased spontaneous post-synaptic current (sPSC) frequency. Varenicline exposure alone also enhanced sPSC frequency. Varenicline plus ethanol co-treatment prevented the sPSC frequency increase. Changes in BF synaptic transmission persisted into adolescence after binge-ethanol treatment. Behaviorally, binge-ethanol treated females displayed increased anxiety (thigmotaxis) and demonstrated learning deficits in the water maze. Varenicline/ethanol co-treatment was effective at reducing these behavioral deficits. In the open field, ethanol-treated rats displayed longer distances traveled and spent less time in the center of the open field box. Co-treated rats displayed less anxiety, demonstrating a possible effect of varenicline on this measure. In conclusion, ethanol-induced changes in both BF synaptic transmission and behavior were reduced by varenicline in female rats, supporting a role for cholinergic therapeutics in FASD treatment.

Fracture Resistance of Ceramic Crowns Supported with Indirect Chair-side Composite Cores.

AIMS AND OBJECTIVES: To evaluate the influence of indirect chair-side polymerization of resin composite cores on the fracture resistance of overlaying IPS e.max Press crowns. MATERIALS AND METHODS: Root canals of 60 extracted premolars were prepared to receive #2 fiber posts after the crowns were sectioned 2 mm above the cervical line. In Groups 1-3 (n = 10 each), posts were luted to the prepared dowel spaces using self-adhesive resin cement. Resin composite cores were then bonded and incrementally built-up using Filtek Z250 XT, Filtek P60, and Filtek P90 resin composites. In Groups 4-6 (n = 10 each), the fabricated post-core systems were subjected to post-curing heat and pressure treatment before cementation to their respective teeth using self-adhesive resin cement. Another 10 sound premolars served as control. All teeth in the test and control groups were then subjected to standardized preparation to receive IPS e.max Press crowns before testing their fracture resistance and the mode of restorations' failure. The collected results were statistically analyzed using ANOVA, Kruskal-Wallis, and Tukey's tests on the past software used at α = 0.05 to stand on the significance of the detected differences. RESULTS: Significant differences were detected between the fracture resistance of teeth in different groups (ANOVA, P = 2.857E-35). Crowns in Groups 4-6 provided higher fracture resistance than those in Groups 1-3 (Tukey's test, P < 0.05). Crowns in Groups 4 and 6 provided higher fracture resistance than the control, while those in Groups 2 and 3 provided lower fracture resistance than the control (Tukey's test, P < 0.05). CONCLUSION: Indirect composite cores improved the fracture resistance of IPS e.max Press crowns when compared to directly fabricated post and cores. The directly and indirectly polymerized nanohybrid, methacrylate-based composite (Filtek Z250 XT) cores yielded the highest fracture resistance for the utilized all-ceramic crowns.

Characterization of age-related changes in synaptic transmission onto F344 rat basal forebrain cholinergic neurons using a reduced synaptic preparation.

Basal forebrain (BF) cholinergic neurons participate in a number of cognitive processes that become impaired during aging. We previously found that age-related enhancement of Ca(2+) buffering in rat cholinergic BF neurons was associated with impaired performance in the water maze spatial learning task (Murchison D, McDermott AN, Lasarge CL, Peebles KA, Bizon JL, and Griffith WH. J Neurophysiol 102: 2194-2207, 2009). One way that altered Ca(2+) buffering could contribute to cognitive impairment involves synaptic function. In this report we show that synaptic transmission in the BF is altered with age and cognitive status. We have examined the properties of spontaneous postsynaptic currents (sPSCs) in cholinergic BF neurons that have been mechanically dissociated without enzymes from behaviorally characterized F344 rats. These isolated neurons retain functional presynaptic terminals on their somata and proximal dendrites. Using whole cell patch-clamp recording, we show that sPSCs and miniature PSCs are predominately GABAergic (bicuculline sensitive) and in all ways closely resemble PSCs recorded in a BF in vitro slice preparation. Adult (4-7 mo) and aged (22-24 mo) male rats were cognitively assessed using the water maze. Neuronal phenotype was identified post hoc using single-cell RT-PCR. The frequency of sPSCs was reduced during aging, and this was most pronounced in cognitively impaired subjects. This is the same population that demonstrated increased intracellular Ca(2+) buffering. We also show that increasing Ca(2+) buffering in the synaptic terminals of young BF neurons can mimic the reduced frequency of sPSCs observed in aged BF neurons.

CT-measured bone attenuation in patients with chronic obstructive pulmonary disease: relation to clinical features and outcomes.

Osteoporosis is highly prevalent in chronic obstructive pulmonary disease (COPD) patients and has been related to several clinical features. However, most studies have been in relatively small COPD cohorts. Therefore, the objectives of this study were to compare bone attenuation measured on low-dose chest computed tomography (CT) between COPD subjects and smoker and nonsmoker controls, and to relate bone attenuation to clinical parameters, inflammatory biomarkers, and outcomes in a large, well-characterized COPD cohort. We studied 1634 COPD subjects, 259 smoker controls, and 186 nonsmoker controls who participated in a large longitudinal study (ECLIPSE). We measured bone attenuation, extent of emphysema, and coronary artery calcification (Agatston score) on baseline CT scans, and clinical parameters, inflammatory biomarkers, and outcomes. Bone attenuation was lower in COPD subjects compared with smoker and nonsmoker controls (164.9 ± 49.5 Hounsfield units [HU] versus 183.8 ± 46.1 HU versus 212.1 ± 54.4 HU, p < 0.001). Bone attenuation was not significantly different between COPD subjects and smoker controls after adjustment for age, sex, and pack-years of smoking. In the COPD subjects, bone attenuation correlated positively with forced expiratory volume in 1 second (FEV1, r = 0.062, p = 0.014), FEV1/forced vital capacity (FVC) ratio (r = 0.102, p < 0.001), body mass index (r = 0.243, p < 0.001), fat-free mass index (FFMI, r = 0.265, p < 0.001), and C-reactive protein (r = 0.104, p < 0.001), and correlated negatively with extent of emphysema (r = -0.090, p < 0.001), Agatston score (r = -0.177, p < 0.001), and interleukin-8 (r = -0.054, p = 0.035). In a multiple regression model, older age, lower FFMI and higher Agatston score were associated with lower bone attenuation. Lower bone attenuation was associated with higher exacerbation (r = -0.057, p = 0.022) and hospitalization (r = -0.078, p = 0.002) rates but was not associated with all-cause mortality. In conclusion, CT-measured bone attenuation was lower in COPD subjects compared with nonsmoker controls but not compared with smoker controls, after adjustment for age, sex, and pack-years of smoking. In the COPD subjects, bone attenuation was associated with age, body composition, and coronary artery calcification but was not associated with all-cause mortality.

Enhanced calcium buffering in F344 rat cholinergic basal forebrain neurons is associated with age-related cognitive impairment.

Alterations in neuronal Ca(2+) homeostasis are important determinants of age-related cognitive impairment. We examined the Ca(2+) influx, buffering, and electrophysiology of basal forebrain neurons in adult, middle-aged, and aged male F344 behaviorally assessed rats. Middle-aged and aged rats were characterized as cognitively impaired or unimpaired by water maze performance relative to young cohorts. Patch-clamp experiments were conducted on neurons acutely dissociated from medial septum/nucleus of the diagonal band with post hoc identification of phenotypic marker mRNA using single-cell RT-PCR. We measured whole cell calcium and barium currents and dissected these currents using pharmacological agents. We combined Ca(2+) current recording with Ca(2+)-sensitive ratiometric microfluorimetry to measure Ca(2+) buffering. Additionally, we sought changes in neuronal firing properties using current-clamp recording. There were no age- or cognition-related changes in the amplitudes or fractional compositions of the whole cell Ca(2+) channel currents. However, Ca(2+) buffering was significantly enhanced in cholinergic neurons from aged cognitively impaired rats. Moreover, increased Ca(2+) buffering was present in middle-aged rats that were not cognitively impaired. Firing properties were largely unchanged with age or cognitive status, except for an increase in the slow afterhyperpolarization in aged cholinergic neurons, independent of cognitive status. Furthermore, acutely dissociated basal forebrain neurons in which choline acetyltransferase mRNA was detected had the electrophysiological profiles of identified cholinergic neurons. We conclude that enhanced Ca(2+) buffering by cholinergic basal forebrain neurons may be important during aging.

Learning styles of orthodontic residents.

Significant challenges face many orthodontic residency programs, particularly a shortage of full-time experienced faculty members. Due to this shortage, it is critical that program directors design comprehensive curricula that incorporate the most effective and efficient teaching methods. It is theorized that teaching effectiveness and efficiency are optimized when the course design and content closely match students' learning preferences. This survey study was designed to distinguish the learning preferences of orthodontic residents utilizing Felder and Soloman's Index of Learning Styles, which assesses student learning preferences in four dimensions using dichotomous scales, thereby providing insight into how teaching strategies can best be structured. As a secondary focus, additional questions on the survey were asked to gain information about residents' access to the Internet and comfort level with online learning so as to address acceptance of web-based courses in response to the shortage of full-time faculty members. Orthodontic residents, contacted via email, were requested to complete an online survey; 261 responses were collected. The results indicate that orthodontic residents are highly visual learners and show a preference for sensing and sequential learning strategies. In terms of information technology, the residents are comfortable with and have adequate access to current technological assets; therefore, they may be well suited for inclusion of computer-based teaching modules and other multimedia devices in their residency curriculum.

Calcium buffering systems and calcium signaling in aged rat basal forebrain neurons.

Disturbances of neuronal Ca2+ homeostasis are considered to be important determinants of age-related cognitive impairment. Cholinergic neurons of the basal forebrain (BF) are principal targets of decline associated with aging and dementia. During the last several years, we have attempted to link these concepts in a rat model of 'normal' aging. In this review, we will describe some changes that we have observed in Ca2+ signaling of aged BF neurons and the reversal of one of these changes by dietary caloric restriction. Our evidence supports a scenario in which subtle changes in the properties of voltage-gated Ca2+ channels result in increased Ca2+ influx during aging. This increased Ca2+, in turn, triggers an increase in rapid Ca2+ buffering in the somatic compartment of aged BF neurons. However, this nominal 'compensation', along with other changes in Ca2+ handling machinery (notably mitochondria) alters the Ca2+ signal with age in a way that is dependent on the magnitude of the Ca2+ load. By combining whole-cell patch clamp electrophysiology, ratiometric Ca2+-sensitive microfluorimetry and single-cell reverse transcription-polymerase chain reaction, we have determined that age-related rapid buffering changes are present in identified cholinergic BF neurons and that these changes can be prevented by a caloric restriction dietary regimen. Because caloric restriction extends lifespan and retards the progression of age-related dysfunction, these findings suggest that increased Ca2+ buffering in cholinergic neurons may be relevant to cognitive decline during normal aging. Importantly, calcium homeostatic mechanisms of BF cholinergic neurons are amenable to dietary interventions that could promote cognitive health during aging.

Functional compensation by other voltage-gated Ca2+ channels in mouse basal forebrain neurons with Ca(V)2.1 mutations.

Tottering (tg/tg) and leaner (tg(la)/tg(la)) mutant mice exhibit distinct mutations in the gene encoding the voltage-activated Ca(2+) channel alpha(1A) subunit (CACNA1A), the pore-forming subunit of the Ca(V)2.1 (P/Q type) Ca(2+) channels. These mice exhibit absence seizures and deficiencies in motor control and other functions. Previous work in cerebellar Purkinje neurons has shown that these mutations cause dramatic reductions in calcium channel function. Because Purkinje cell somata primarily express the Ca(V)2.1 channels, the general decrease in Ca(V)2.1 channel function is observed as a profound decrease in whole-cell current. In contrast to Purkinje cells, basal forebrain (BF) neurons express all of the Ca(2+) channel alpha(1) subunits, with Ca(V)2.1 contributing approximately 30% to the whole-cell current in wild-type (+/+) mice. Here, we show that whole-cell Ba(2+) current densities in BF neurons are not reduced in the mutant genotypes despite a reduction in the Ca(V)2.1 contribution. By blocking the different Ca(2+) channel subtypes with specific pharmacological agents, we found a significant increase in the proportion of Ca(V)1 Ca(2+) current in mutant phenotypes. There was no change in tissue mRNA expression of calcium channel subtypes Ca(V)2.1, Ca(V)2.2, Ca(V)1.2, Ca(V)1.3, and Ca(V)2.3 in the tottering and leaner mutant mice. These results suggest that Ca(V)1 channels may functionally upregulate to compensate for reduced Ca(V)2.1 function in the mutants without an increase in Ca(v)1 message. Single-cell reverse transcription polymerase chain reaction (RT-PCR) experiments in a subset of sampled neurons revealed that approximately 90% of the cells could be considered cholinergic based on choline acetyltransferase (ChAT) mRNA expression.

Oral surgical handpiece use time parameters.

PURPOSE: To evaluate the clinical usage time parameters of handpieces used in oral surgical procedures. METHOD AND MATERIALS: One hundred randomly selected clinical oral surgery exodontia procedures were timed to record lengths of continuous segments of both handpiece use and non-usage. Providers with experience ranging from general dentists to board certified oral surgeons were timed during surgical exodontia treatment involving 1 to 4 teeth of various complexities. Usage times were compared with manufacturers' recommendations that on times should not exceed 20 seconds in any 50-second interval (20/50 rule). RESULTS: Handpiece run time increased with the number of teeth and surgical case complexity (both P < .001) but was unrelated to operator experience (P = .763), in a 3-predictor model (R2 = 0.20; P < .001). Ninety-four of the 100 cases experienced at least 1 second in violation of the 20/50 rule and 42% of all run seconds were in violation. CONCLUSION: Clinicians should be aware of recommended handpiece duty use cycles. Manufacturers' recommendations about handpiece use time cycles do not reflect actual clinical usage. Under the conditions of this study, actual surgical handpiece use time was not correlated with user experience. Less experienced providers did require longer to complete treatment, but increased treatment times were due to time spent that did not require surgical handpiece use.

Low voltage-activated calcium and fast tetrodotoxin-resistant sodium currents define subtypes of cholinergic and noncholinergic neurons in rat basal forebrain.

Neurons of the basal forebrain (BF) possess unique combinations of voltage-gated membrane currents. Here, we describe subtypes of rat basal forebrain neurons based on patch-clamp analysis of low-voltage activated (LVA) calcium and tetrodotoxin-resistant (TTX-R) sodium currents combined with single-cell RT-PCR analysis. Neurons were identified by mRNA expression of choline acetyltransferase (ChAT+, cholinergic) and glutamate decarboxylase (GAD67, GABAergic). Four cell types were encountered: ChAT+, GAD+, ChAT+/GAD+ and ChAT-/GAD- cells. Both ChAT+ and ChAT+/GAD+ cells (71/75) displayed LVA currents and most (34/39) expressed mRNA for LVA Ca(2+) channel subunits. Ca(v)3.2 was detected in 31/34 cholinergic neurons and Ca(v)3.1 was expressed in 6/34 cells. Three cells expressed both subunits. No single neurons showed Ca(v)3.3 mRNA expression, although BF tissue expression was observed. In young rats (2-4 mo), ChAT+/GAD+ cells displayed larger LVA current densities compared to ChAT+ neurons, while these latter neurons displayed an age-related increase in current densities. Most (29/38) noncholinergic neurons (GAD+ and ChAT-/GAD-) possessed fast TTX-R sodium currents resembling those mediated by Na(+) channel subunit Na(v)1.5. This subunit was expressed predominately in noncholinergic neurons. No cholinergic cells (0/75) displayed fast TTX-R currents. The TTX-R currents were faster and larger in GAD+ neurons compared to ChAT-/GAD- neurons. The properties of ChAT+/GAD+ neurons resemble those of ChAT+ neurons, rather than of GAD+ neurons. These results suggest novel features of subtypes of cholinergic and noncholinergic neurons within the BF that may provide new insights for understanding normal BF function.

Reduced mitochondrial buffering of voltage-gated calcium influx in aged rat basal forebrain neurons.

Alterations of neuronal Ca(2+) homeostatic mechanisms could be responsible for many of the cognitive deficits associated with aging in mammals. Mitochondrial participation in Ca(2+) signaling is now recognized as a prominent feature in neuronal physiology. We combined voltage-clamp electrophysiology with Ca(2+)-sensitive ratiometric microfluorimetry and laser scanning confocal microscopy to investigate the participation in Ca(2+) buffering of in situ mitochondria in acutely dissociated basal forebrain neurons from young and aged F344 rats. By pharmacologically blocking mitochondrial Ca(2+) uptake, we determined that mitochondria were not involved in rapid buffering of small Ca(2+) influx through voltage-gated Ca(2+) channels (VGCCs) in the somatic compartment. For larger Ca(2+) influx, aged mitochondria showed a significant buffering deficit. Evidence obtained with the potentiometric indicator, JC-1, suggests a significantly reduced mitochondrial membrane potential in aged neurons. These results support the interpretation that there is a fundamental difference in the way young and aged neurons buffer Ca(2+), and a corresponding difference in the quality of the Ca(2+) signal experienced by young and aged neurons for different intensities of cytoplasmic Ca(2+) influx.

Surface roughness of various packable composites.

Packable composite restorations have become a popular alternative to dental amalgam restorations in posterior teeth. A drawback inherent to composites is their difficulty in polishing, which often results in a dull or rough surface. This study compared the surface roughness of a resin-based hybrid composite material and five packable resin-based composites polished with either aluminum oxide disks or a rubber polishing system. Sixteen specimens of each of the six composite materials were polished with either Sof-Lex XT disks or Enhance rubber polishers followed by fine and superfine polishing pastes. The specimens were evaluated for surface roughness using surface profilometry. Mean values were calculated for each material type and method of polishing. Data were subjected to a two-way ANOVA. Post hoc comparison was accomplished using Tukey's HSD. No significant difference in surface roughness was detected among polishing techniques (p=0.067); however, a strong trend-that aluminum oxide disks provided a smoother surface than rubber polishers in five out of six materials-was noted.

Fracture strength of amalgam crowns with repaired endodontic access.

Endodontic therapy is accessed occlusally in posterior teeth, many of which have large, pre-existing amalgam restorations. These teeth are also commonly restored with an occlusal amalgam to repair the access opening. This study determined the fracture resistance of complex amalgam restorations that have repaired endodontic access compared with original, unrepaired, complex amalgams on endodontically-treated teeth. Two groups of 30 molars were used in the study. The first group was decoronated and received an endodontic access preparation. These teeth were restored using chamber retention and four TMS pins. The second group was decoronated and restored using pin retention. Later, they received an endodontic access through the restoration. The access was then repaired with amalgam. The samples were loaded in an Instron Universal Testing Machine until failure. The Group 1 samples failed at a mean force of 2297.5 N. The mean failure load for the samples in Group 2 was 1586.1 N. Student's t-test found this difference to be statistically significant. Endodontic access through an amalgam crown significantly compromises the fracture strength of the original restoration.

The effect of resin composite pins on the retention of class IV restorations.

Standardized Class IV cavity preparations were made in 48 human incisors. They were then divided into three groups of 16 teeth each. Group 1 was prepared with no internal retentive features. Groups 2 and 3 included an internal pin channel prepared with a #330 and a #329 bur, respectively. All specimens were restored with a micro-hybrid resin-based composite restorative material and a fifth generation dentin bonding system. Specimens were loaded to failure in an Instron Universal Testing Machine perpendicular to the long axis of the teeth. There was no evidence to suggest a difference in force required to dislodge the restoration among the three groups tested (p=0.185). Resin composite pins do not increase the retention of Class IV resin composite restorations.

Higher-concentration carbamide peroxide effects on surface roughness of composites.

BACKGROUND: Ten percent carbamide peroxide has been used extensively within the dental profession for the purpose of bleaching teeth. Although this method has been successful, the use of higher-concentration carbamide peroxides (20-35%) for home bleaching has increased substantially in the past few years. The purpose of this study was to evaluate changes in surface roughness of a hybrid and a microfilled composite after exposure to bleaching agents containing higher concentrations of carbamide peroxide. MATERIALS AND METHODS: Sixty-eight circular, resin-based composite specimens (8 mm x 2 mm) were prepared against a Mylar surface. Half of the specimens were fabricated from a hybrid composite material and the other half with a microfilled composite. Specimens were polished with aluminum oxide finishing disks, divided into four groups, and then exposed either to 20% carbamide peroxide for 3 hours per day for 14 days or to 35% carbamide peroxide for 1 hour per day for 14 days. The mean surface roughness (Ra) was determined for each specimen before and after exposure to bleaching agents, using mechanical profilometry. Data were analyzed by two-way analysis of variance (ANOVA) for differences between composite type, carbamide peroxide concentrations, and interaction of the two factors with respect to average surface roughness. All data analysis was performed at alpha = 0.05. RESULTS: Mean change from baseline of surface roughness (Ra) of microfilled composite and 20% carbamide peroxide was -0.03 microm +/- 0.10 microm; microfilled composite and 35% carbamide peroxide: 0.01 microm x 0.02 microm; hybrid composite and 20% carbamide peroxide: -0.43 microm x 0.91 microm; hybrid composite and 35% carbamide peroxide: -0.19 microm x 0.43 microm. Surface roughness of hybrid or microfilled composite did not change significantly from baseline with either concentration of carbamide peroxide (p = .300). CLINICAL SIGNIFICANCE: Higher-concentration carbamide peroxide bleaching agents, used as intended by the manufacturer, pose no significant risk to resin composite restoration surfaces.

Single-cell RT-PCR detects shifts in mRNA expression profiles of basal forebrain neurons during aging.

The medial septum and nucleus of the diagonal band (MS/nDB) contain cholinergic and GABAergic neuronal populations that have been identified based on immunohistochemical staining and/or electrophysiological properties. We explored the molecular diversity of MS/nDB neurons using single-cell reverse transcription-polymerase chain reaction (scRT-PCR) to assess gene expression profiles during aging in individual neurons acutely isolated from young (2-4 months) and aged (26-27 months) F344 rats. Neuronal gene expression profiles were characterized by detection of mRNAs for choline acetyltransferase (ChAT, cholinergic) and glutamate decarboxylase (GAD67, GABAergic), as well as mRNAs for calcium binding proteins (CaBPs) calbindin-D28k, calretinin and parvalbumin. Four major neuronal populations were identified: ChAT-positive (ChAT+) cells, GAD-positive (GAD+) cells, ChAT+/GAD+ cells and ChAT negative/GAD negative (ChAT-/GAD-) cells. With age, the percentage of cells expressing ChAT mRNA decreased from 53% in young to 40%, and the expression of GAD67 mRNA was reduced from 56 to 35% of the cells tested. The percentage of cells with detectable levels of both ChAT and GAD67 mRNA was reduced from 24% in young to 9% in aged. Concomitantly, the percentage of ChAT-/GAD- cells increased from 15 to 34% with age. Of the CaBPs, calretinin expression was observed most frequently in this study, and its detection decreased from 33 to 22% of the cells with age. Observations concerning the CaBPs were confirmed using in situ hybridization. These results suggest a shift in the mRNA expression profiles of MS/nDB neuronal populations during aging and exemplify the molecular diversity of cholinergic and GABAergic cells.