G protein ß subunits regulate Cav3.3 T-type channel activity and current kinetics via interaction with the Cav3.3 C-terminus.

Ca2+ influx through Cav3.3 T-type channel plays crucial roles in neuronal excitability and is subject to regulation by various signaling molecules. However, our understanding of the partners of Cav3.3 and the related regulatory pathways remains largely limited. To address this quest, we employed the rat Cav3.3 C-terminus as bait in yeast-two-hybrid screenings of a cDNA library, identifying rat Gß2 as an interaction partner. Subsequent assays revealed that the interaction of Gß2 subunit was specific to the Cav3.3 C-terminus. Through systematic dissection of the C-terminus, we pinpointed a 22 amino acid sequence (amino acids 1789-1810) as the Gß2 interaction site. Coexpression studies of rat Cav3.3 with various Gßγ compositions were conducted in HEK-293 cells. Patch clamp recordings revealed that coexpression of Gß2γ2 reduced Cav3.3 current density and accelerated inactivation kinetics. Interestingly, the effects were not unique to Gß2γ2, but were mimicked by Gß2 alone as well as other Gßγ dimers, with similar potencies. Deletion of the Gß2 interaction site abolished the effects of Gß2γ2. Importantly, these Gß2 effects were reproduced in human Cav3.3. Overall, our findings provide evidence that Gß(γ) complexes inhibit Cav3.3 channel activity and accelerate the inactivation kinetics through the Gß interaction with the Cav3.3 C-terminus.

Case report: Generalized lymphatic anomaly of multiple abdominal organs in a young dog.

A 9-month-old, female Pomeranian dog presented with vomiting and lethargy. Ultrasonography revealed multilobulated anechoic round shape structures at the ovarian and uterine locations. Through computed tomography scan, an extensive non-contrast multilobulated fluid-filled mass suspected of originating from the walls of the ovary, uterus, urinary bladder and rectum was observed. Ovariohysterectomy and urinary bladder biopsy were performed. Histopathological examination revealed numerous cystic lesions lined by plump cuboidal cells believed to be of epithelial origin. Immunohistochemical staining showed that the cyst-like lesions lining cells were strongly positive for lymphatic vessel endothelial hyaluronan receptor 1. Based on these results, lesions were identified as generalized lymphatic anomaly (GLA), in which lymphangiomas develop in multiple organs. After 6 months follow-up, the size of the cysts remaining in the region of the bladder did not undergo much change. GLA should be included in the differential diagnosis when multiple cystic lesions are interspersed in multiple organs.

Phosphorylation states greatly regulate the activity and gating properties of Cav 3.1 T-type Ca2+ channels.

Cav 3.1 T-type Ca2+ channels play pivotal roles in neuronal low-threshold spikes, visceral pain, and pacemaker activity. Phosphorylation has been reported to potently regulate the activity and gating properties of Cav 3.1 channels. However, systematic identification of phosphorylation sites (phosphosites) in Cav 3.1 channel has been poorly investigated. In this work, we analyzed rat Cav 3.1 protein expressed in HEK-293 cells by mass spectrometry, identified 30 phosphosites located at the cytoplasmic regions, and illustrated them as a Cav 3.1 phosphorylation map which includes the reported mouse Cav 3.1 phosphosites. Site-directed mutagenesis of the phosphosites to Ala residues and functional analysis of the phospho-silent Cav 3.1 mutants expressed in Xenopus oocytes showed that the phospho-silent mutation of the N-terminal Ser18 reduced its current amplitude with accelerated current kinetics and negatively shifted channel availability. Remarkably, the phospho-silent mutations of the C-terminal Ser residues (Ser1924, Ser2001, Ser2163, Ser2166, or Ser2189) greatly reduced their current amplitude without altering the voltage-dependent gating properties. In contrast, the phosphomimetic Asp mutations of Cav 3.1 on the N- and C-terminal Ser residues reversed the effects of the phospho-silent mutations. Collectively, these findings demonstrate that the multiple phosphosites of Cav 3.1 at the N- and C-terminal regions play crucial roles in the regulation of the channel activity and voltage-dependent gating properties.

3D visualization of microwave electric and magnetic fields by using a metasurface-based indicator.

Visualizations of the microwave electric and magnetic near-field distributions of radio-frequency (RF) filters were performed using the technique of thermoelastic optical indicator microscopy (TEOIM). New optical indicators based on periodic dielectric-metal structures were designed for electric field visualization. Depending on the structure orientation, such metasurface-based indicators allow separately visualization of the Ex and Ey components of the in-plane electric field. Numerical simulations were conducted to examine the working principle of the designed indicator structures, and the results were compared to the experimental, showing good agreement. In addition, the 3D visualization of the microwave near-field distribution was built, to show the field intensity and distribution dependencies on the distance from the RF filter.

Sites and Regulation of L-Type Ca2+ Channel Cav1.2 Phosphorylation in Brain.

Cav1.2 channel phosphorylation plays an important role in regulating neuronal plasticity by action potential-dependent Ca2+ entry. Most studies of Cav1.2 regulation by phosphorylation have been reported in heart and muscles. Here, we identified phosphorylation sites of neuronal Cav1.2 channel protein purified from rat brain using mass spectrometry. The functional characterization of these phosphorylation sites showed altered voltage-dependent biophysical properties of the channel, without affecting current density. These results show that neuronal Cav1.2 channel is regulated by phosphorylation in a complex mechanism involving multiple phosphorylation sites.

Snapin Specifically Up-Regulates Cav1.3 Ca2+ Channel Variant with a Long Carboxyl Terminus.

Ca2+ entry through Cav1.3 Ca2+ channels plays essential roles in diverse physiological events. We employed yeast-two-hybrid (Y2H) assays to mine novel proteins interacting with Cav1.3 and found Snapin2, a synaptic protein, as a partner interacting with the long carboxyl terminus (CTL) of rat Cav1.3L variant. Co-expression of Snapin with Cav1.3L/Cavß3/α2δ2 subunits increased the peak current density or amplitude by about 2-fold in HEK-293 cells and Xenopus oocytes, without affecting voltage-dependent gating properties and calcium-dependent inactivation. However, the Snapin up-regulation effect was not found for rat Cav1.3S containing a short CT (CTS) in which a Snapin interaction site in the CTL was deficient. Luminometry and electrophysiology studies uncovered that Snapin co-expression did not alter the membrane expression of HA tagged Cav1.3L but increased the slope of tail current amplitudes plotted against ON-gating currents, indicating that Snapin increases the opening probability of Cav1.3L. Taken together, our results strongly suggest that Snapin directly interacts with the CTL of Cav1.3L, leading to up-regulation of Cav1.3L channel activity via facilitating channel opening probability.

Prevalence and characteristics of isolated nocturnal hypertension in the general population.

BACKGROUND/AIMS: Ascertaining the prevalence of isolated nocturnal hypertension (INHT) in the general population and identifying the characteristics of patients with INHT may be important to determine patients who should receive 24- hour ambulatory blood pressure (BP) measurements. This study aimed to evaluate the prevalence and characteristics of INHT in the general population. METHODS: Of 1,128 participants (aged 20 to 70 years), we analyzed 823 who had valid 24-hour ambulatory BP measurements and were not on antihypertensive drug treatment. RESULTS: The prevalence of INHT in the study was 22.8%. Individuals with INHT had a higher office, 24-hour, and daytime and nighttime ambulatory systolic and diastolic BPs compared to individuals with sustained day-night normotension. INHT was more prevalent in individuals with masked hypertension (MH) than in those with sustained hypertension (59.8% vs. 15.6%, p < 0.001). Among individuals with INHT, 92.6% had MH. Among individuals with office BP-based prehypertension, 34.5% had both INHT and MH. The prevalence of INHT was highest in individuals with office BP-based prehypertension. INHT was an independent determinant of MH after adjustment for age, sex, body mass index, diabetes, low-density-lipoprotein cholesterol, 24-hour systolic and diastolic BP, systolic and diastolic BP dipping, and systolic and diastolic BP non-dipping. CONCLUSION: The present study showed that INHT is not uncommon and is a major determinant of MH. Our findings strongly suggest the use of 24-hour ambulatory BP measurement for individuals within the prehypertension range of office BP owing to the high prevalence of INHT and MH in this population.

Visualization of microwave near-field distribution in sodium chloride and glucose aqueous solutions by a thermo-elastic optical indicator microscope.

In this study, a new optical method is presented to determine the concentrations of NaCl and glucose aqueous solutions by using a thermo-elastic optical indicator microscope. By measuring the microwave near-field distribution intensity, concentration changes of NaCl and glucose aqueous solutions were detected in the 0-100 mg/ml range, when exposed to microwave irradiation at 12 GHz frequency. Microwave near-field distribution intensity decreased as the NaCl or glucose concentration increased due to the changes of the absorption properties of aqueous solution. This method provides a novel approach for monitoring NaCl and glucose in biological liquids by using a CCD sensor capable of visualizing NaCl and glucose concentrations without scanning.

Impacts of undernutrition and maternal oral health status on dental caries in Korean children aged 3-5 years.

INTRODUCTION: The association between nutrition and oral health has been studied in the elderly. This study aimed to examine the impacts of undernutrition and the mothers' socioeconomic and oral health statuses on the incidence of dental caries in Korean preschool children. METHODS: Data of 610 children aged 3-5 years and their mothers who underwent oral examinations and responded to the questionnaires in the 6th KNAHNES were used. Caries prevalence was measured by dft and dt among the primary teeth in children and DMFT among mothers. Dietary reference intake values were used to evaluate nutritional status, a nutritional quality index and the mean nutrient adequacy ratio. Complex sample correlation analysis was performed by using children's dft and dt statuses as dependent variables. Multilevel linear regression was applied to investigate the impacts of undernutrition and mothers' socioeconomic and oral health status on children's dft and dt statuses. Statistical significance was set as P < .05. RESULTS: Factors significantly related to dft in children were age and food. Food insecurity was the only factor significantly associated with dt in children. Children's sex and mother's DMFT were likely to be relevant to dft in children. Children's age and a nutritional quality index value less than 1 were also likely to be relevant to the dt of children. CONCLUSIONS: Children with an undernourished status had a higher caries experience. A comprehensive community dental health promotion programme should be developed to prevent the incidence of dental caries in vulnerable undernourished children.

Incidence of newly developed postoperative low back pain with median versus paramedian approach for spinal anesthesia.

Background: The effects of anesthetic techniques on postdural puncture backache (PDPB) have not been specifically evaluated. The purpose of this study was to compare the incidence and severity of PDPB between median and paramedian techniques. Methods: Patients were randomized to receive spinal anesthesia by either a median (Group M, n = 50) or paramedian (Group P, n = 50) approach. We recorded each patient's personal number of puncture attempts, surgical position, and operation duration. We investigated the incidence and intensity of back pain 1 day, 1 week, and 1, 2, and 3 months postoperatively. Results: The overall incidence of PDPB was higher in the Group M (18/50, 36%) than in the Group P (8/50, 16%) (P = 0.023). Twenty-four hours after surgery, 8 patients in Group M and 6 patients in Group P complained of back pain. Seven days after the surgery, 16 patients in the Group M and 5 patients in the Group P complained of pain (P = 0.007). After 1 month, 5 patients in the Group M and 1 patient in the Group P complained of pain. Only one patient in each group complained of pain after 3 months. No significant differences were noted in NRSs between the groups during study period. Conclusions: The results of this study suggest that spinal anesthesia using the paramedian approach reduces the incidence of PDPB during the early postoperative period.

Real-Time Noninvasive Measurement of Glucose Concentration Using a Modified Hilbert Shaped Microwave Sensor.

We developed a microwave glucose sensor based on the modified first-order Hilbert curve design and measured glucose concentration in aqueous solutions by using a real-time microwave near-field electromagnetic interaction technique. We observed S21 transmission parameters of the sensor at resonant frequencies depend on the glucose concentration. We could determine the glucose concentration in the 0-250 mg/dL concentration range at an operating frequency of near 6 GHz. The measured minimum detectable signal was 0.0156 dB/(mg/dL) and the measured minimum detectable concentration was 1.92 mg/dL. The simulation result for the minimum detectable signal and the minimum detectable concentration was 0.0182 dB/(mg/dL) and 1.65 mg/dL, respectively. The temperature instability of the sensor for human glycemia in situ measurement range (27-34 °C for fingers and 36-40 °C for body temperature ranges) can be improved by the integration of the temperature sensor in the microwave stripline platform and the obtained data can be corrected during signal processing. The microwave signal-temperature dependence is almost linear with the same slope for a glucose concentration range of 50-150 mg/dL. The temperature correlation coefficient is 0.05 dB/°C and 0.15 dB/°C in 27-34 °C and 36-40 °C temperature range, respectively. The presented system has a cheap, easy fabrication process and has great potential for non-invasive glucose monitoring.

Factors Related to the Number of Existing Teeth among Korean Adults Aged 55-79 Years.

This study aimed to determine the association between the number of existing teeth (NET) and socioeconomic status (SES), oral health-related behaviours, and metabolic syndrome in Korean adults aged 55-79 years. The study included 3255 adults who underwent oral health examinations and answered questionnaires regarding SES, oral health-related behaviours, and metabolic diseases in the Sixth Korea National Health and Nutrition Examination Survey (2013-2015). The dependent variable was the binary status based on the median NET in each age group. The independent variables were based on SES, oral health-related behaviours, and the presence of metabolic syndrome. The study findings showed that the factors associated with the NET were sex, household income, education level, region of residence, daily toothbrushing frequency, dental visit within 1 year, smoking, and metabolic syndrome. NET was lower in males (adjusted OR: 0.74), in low household income group (adjusted OR: 0.77), in primary school graduates (adjusted OR: 0.53), in rural residents (adjusted OR: 0.78), and in medicaid beneficiaries (adjusted OR: 0.78). The interventions aimed at preserving existing teeth in elderly population should consider their SES, oral health-related behaviours, and metabolic syndrome and overhauling current oral healthcare system and redefining the roles of oral health professionals.

Association between Periodontitis and Metabolic Syndrome in a Korean Nationally Representative Sample of Adults Aged 35-79 Years.

This study aimed to evaluate the association between periodontitis and metabolic syndrome (MetS) and to investigate risk factors associated with MetS in Korean adults aged 35 to 79 years. Among individuals aged 35-79 years who participated in the Korea National Health and Nutrition Examination Survey 2013-2015, 8314 participants who completed the required examinations and questionnaires were included. Confounding variables related to demographic and socioeconomic status and systemic and oral health-related behaviors were age, gender, household income, education level, smoking, alcohol intake, physical activity, and frequency of daily toothbrushing. Of the 8314 participants, 32.2% were diagnosed with MetS. The prevalence of MetS was 26.6% and 41.6% in those without and with periodontitis, respectively. Among individuals with periodontitis, the prevalence of MetS was 44.3% in males and 36.9% in females. Compared to non-periodontitis, periodontitis was associated with MetS (adjusted OR = 1.422, 95% CI: 1.26-1.61). Age, frequency of daily toothbrushing, and periodontitis were associated with MetS in both males and females. While current smoking and alcohol intake more than twice a week were significantly associated with MetS in males, household income and education level were significantly associated with MetS in females. The findings suggest that periodontitis can be associated with MetS.

The potential use of bromelain as a natural oral medicine having anticarcinogenic activities.

Bromelain (BR), a protease extracted from Ananas comosus, reportedly possesses pharmacological activities including the reduction of thrombogenesis, and antihypertensive, and antimicrobial effects. This study aimed to investigate the potential effects of BR on oral cancer cells. The effect of BR on the viability of Ca9-22 and SCC25 cells was determined using the MTT assay. These cells were also treated with different doses of BR, and Western blotting was conducted to monitor apoptosis. Finally, flow cytometry analysis was performed to identify sub-G1 populations of oral cancer cells. After treatment, the viability of both Ca9-22 and SCC25 cells was markedly reduced, in a dose-dependent manner. BR induced poly (ADP-ribose) polymerase (PARP) and lamin A/C degradation, and generated cleavage products. Flow cytometry analysis showed that BR treatment significantly increased the sub-G1 population. Our findings therefore indicate that BR has potential as a novel, natural anticarcinogenic medicine.

Reduction of Dental Caries Among Children and Adolescents From a 15-Year Community Water Fluoridation Program in a Township Area, Korea.

Since 2000, a community water fluoridation program (CWFP) has been implemented in Hapcheon for over 15 years. We aimed to evaluate the caries-reducing effect on permanent teeth after this implementation. In 2015, evaluation surveys were conducted by our study group, 498 subjects aged 8, 10, 12, and 15 years. As the control, 952 similarly aged subjects were selected from the Sixth Korean National Health and Nutrition Examination Survey (2013-2015 KNHANES). Data of a prospective cohort of 671 8-,10- and 12-year-olds, collected when CWFP started, were used for the evaluation. Caries-reducing effects were estimated by decayed, missing and filled teeth (DMFT) scores between CWFP and control groups, pre- and post-program. Confounders including the mean number of sealant teeth and gender were adjusted for. The mean adjusted DMFT scores of 10-, 12- and 15-year-olds in Hapcheon were significantly lower compared to KNHANES DMFT scores; in addition, those of 8-, 10- and 12-year-olds after the 15-year CWFP were significantly lower than in 2000. The caries-reducing effect among 12-year-olds was 37.6% compared to those recorded in KNHANES, and 67.4% compared to those in 2000. In conclusion, the caries-reducing effect was so high that health policy makers should consider CWFP as a priority policy for caries-reducing in Korean children and adolescents.

Association of Self-Perceived Oral Health and Function with Clinically Determined Oral Health Status among Adults Aged 35⁻54 Years: A Cross-Sectional Study.

This study aimed to analyse the association of self-perceived oral health status (OHS) and functions with clinical OHS in Korean adults aged 35⁻54 years. The study was designed as a cross-sectional study using data from the Fourth Korea National Health and Nutrition Examination Survey (2007⁻2009). A total of 6605 subjects aged 35⁻54 years who completed the oral examination and questionnaires were included. An association of self-perceived OHS and functions with clinically determined OHS was confirmed by a complex-samples general linear model. Data on socioeconomic variables, i.e., household income and education level, self-perceived OHS and functions, such as chewing and speaking, were collected by trained interviewers. The clinical OHS was determined by trained dentists and included the number of untreated decayed teeth (DT); decayed, missing, and filled teeth (DMFT); prosthetic and periodontal status. The combined score was estimated as the sum of self-perceived OHS and functions. Based on the estimation coefficient, the clinical variables that were most strongly associated with self-perceived OHS and functions were, in order, periodontal status, prosthetic status, DT, and DMFT. In addition, the combined score for self-perceived OHS and functions was associated with household income, education, and clinically determined OHS.

Neuronal calcium channel α1 subunit interacts with AMPA receptor, increasing its cell surface localisation.

Voltage-activated Ca2+ channels (Cav) play critical roles in excitable cells including neurons. Unlike the well-defined roles of Cav2 for pre-synaptic neurotransmission, the post-synaptic function of Cav2 is unclear. Based on our previous study demonstrating the postsynaptic association of the Cav2 with the AMPA receptor (AMPA-R), in this study we sought to further analyse the Cav2-AMPA-R association. We used a step-by-step dissociation of partially purified native Cav2-AMPA-R complexes and co-immunoprecipitation of the Cav2-AMPA-R complexes expressed in HEK293T cells to demonstrate that the main subunit of Cav, α1, formed a complex with the AMPA-R without the auxiliary subunits ß, α2δ, γ2/3. The α1 subunit increased the cell-surface localisation of the AMPA-R, which could be a post-synaptic function of the Cav2.

Ca2+ Regulation of Cav3.3 T-type Ca2+ Channel Is Mediated by Calmodulin.

Calcium-dependent inactivation of high voltage-activated Ca2+ channels plays a crucial role in limiting rises in intracellular calcium (Ca2+i). A key mediator of these effects is calmodulin, which has been found to bind the C-terminus of the pore-forming α subunit. In contrast, little is known about how Ca2+i can regulate low voltage-activated T-type Ca2+ channels. Using whole cell patch clamp, we examined the biophysical properties of Ca2+ current through the three T-type Ca2+ channel isoforms, Cav3.1, Cav3.2, or Cav3.3, comparing internal solutions containing 27 nM and l µM free Ca2+ Both activation and inactivation kinetics of Cav3.3 current in l µM Ca2+i solution were more rapid than those in 27 nM Ca2+i solution. In addition, both activation and steady-state inactivation curves of Cav3.3 were negatively shifted in the higher Ca2+i solution. In contrast, the biophysical properties of Cav3.1 and Cav3.2 isoforms were not significantly different between the two internal solutions. Overexpression of CaM1234 (a calmodulin mutant that doesn't bind Ca2+) occluded the effects of l µM Ca2+i on Cav3.3, implying that CaM is involved in the Ca2+i regulation effects on Cav3.3. Yeast two-hybrid screening and co-immunoprecipitation experiments revealed a direct interaction of CaM with the carboxyl terminus of Cav3.3. Taken together, our results suggest that Cav3.3 T-type channel is potently regulated by Ca2+i via interaction of Ca2+/CaM with the carboxyl terminus of Cav3.3.

Association of stress, depression, and suicidal ideation with subjective oral health status and oral functions in Korean adults aged 35 years or more.

BACKGROUND: Oral health greatly affects well-being throughout the different stages of life from childhood to late adulthood. Loss of teeth due to poor oral health hinders mastication, leading to poor nutrition absorption, and affects pronunciation and aesthetics, leading to interpersonal difficulties. As social activities become limited, a sense of isolation and loneliness, stress, and depression grows while happiness decreases. This study aimed to examine the association of stress, depression, and suicidal ideation with oral health status and oral functions in a large nationwide sample of Korean adults aged 35 years or more. METHODS: The sample comprised 15,716 adults, selected using a rolling survey sampling method and data were extracted from the Fifth Korea National Health and Nutrition Examination Survey (KNHANES) (2010-2012). Participants were interviewed about their self-evaluation of health including oral health status and mental health, such as stress, depression, and suicidal ideation. Data from 11,347 adults were finally selected after excluding participants with missing answers. The dependent variables were stress, depression, and suicidal ideation. The independent variables were gender, age, household income, education, smoking, drinking, oral health perception, chewing, and speaking. Complex samples logistic regression analyses were used to estimate odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: Participants met the criteria for stress (25.4%), depression (13.0%), and suicidal ideation (13.9%). Subjective oral health status was not significantly associated with stress, depression, and suicidal ideation. However, the presence of very uncomfortable chewing problems was significantly associated with stress (OR = 2.294, 95% CI = 1.41, 3.72), depression (OR = 3.232, 95% CI = 1.97, 5.31), and suicidal ideation (OR = 2.727, 95% CI = 1.58, 4.72). The presence of very uncomfortable speaking problems was significantly associated with stress (OR = 1.592, 95% CI = 1.13, 2.24) but not significantly associated with depression and suicidal ideation. CONCLUSIONS: Oral functional problems including chewing and speaking difficulties can be associated with mental health. It is necessary to develop oral health promotion programs for adults and help them maintain a good quality of life and mental health.

Salivary bisphenol A levels and their association with composite resin restoration.

Composite resin has been increasingly used in an effort to remove minimal amount of tooth structure and are used for restoring not just carious cavities but also cervical abrasion. To synthesize composite resin, bisphenol A (BPA) is used. The aim of the study was to measure the changes in salivary BPA level related with composite resin restoration. ELISA was used to examine the BPA levels in the saliva collected from 30 volunteers whose teeth were filled with composite resin. Salivary samples were collected immediately before filling and 5 min and 7 d after filling. Wilcoxon signed-ranks test and linear regression were performed to test the significant differences of the changes in BPA levels in saliva. Before a new composite resin filling, there was no significant difference between with and without existing filling of composite resin and BPA level in the saliva was not correlated to the number of filled surfaces with composite resin. However, BPA level in the saliva increased to average 3.64 µg/L from average 0.15 µg/L after filling 5 min. BPA level increased in proportion with the number of filled surfaces. BPA level decreased to average 0.59 after filling 7 d. However it was higher than the BPA level before a new composite resin filling. Considering 50 µg/kg/day as the Tolerable Daily Intake of BPA suggested by European Food Safety Authority, the amount of BPA eluted in saliva after the composite resin filling is considered a safe level that is not a hazard to health at all.