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Ca 2+ influx through Ca v 3.3 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 Ca v 3.3 and the related regulatory pathways remains largely limited. To address this quest, we employed the rat Ca v 3.3 C-terminus as bait in yeast-two-hybrid screenings of a cDNA library, identifying rat Gβ 2 as an interaction partner. Subsequent assays revealed that the interaction of Gβ 2 subunit was specific to the Ca v 3.3 C-terminus. Through systematic dissection of the C-terminus, we pinpointed a 22 amino acid sequence (amino acids 1789-1810) as the Gβ 2 interaction site. Coexpression studies of rat Ca v 3.3 with various Gβγ compositions were conducted in HEK-293 cells. Patch clamp recordings revealed that coexpression of Gβ 2 γ 2 reduced Ca v 3.3 current density and accelerated inactivation kinetics. Interestingly, the effects were not unique to Gβ 2 γ 2, but were mimicked by Gβ 2 alone as well as other Gβγ dimers, with similar potencies. Deletion of the Gβ 2 interaction site abolished the effects of Gβ 2 γ 2 . Importantly, these Gβ 2 effects were reproduced in human Ca v 3.3. Overall, our findings provide evidence that Gβ(γ) complexes inhibit Ca v 3.3 channel activity and accelerate the inactivation kinetics through the Gβ interaction with the Ca v 3.3 C-terminus., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Ca v 3.1 T-type Ca 2+ 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 Ca v 3.1 channels. However, systematic identification of phosphorylation sites (phosphosites) in Ca v 3.1 channel has been poorly investigated. In this work, we analyzed rat Ca v 3.1 protein expressed in HEK-293 cells by mass spectrometry, identified 30 phosphosites located at the cytoplasmic regions, and illustrated them as a Ca v 3.1 phosphorylation map which includes the reported mouse Ca v 3.1 phosphosites. Site-directed mutagenesis of the phosphosites to Ala residues and functional analysis of the phospho-silent Ca v 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 Ca v 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 Ca v 3.1 at the N- and C-terminal regions play crucial roles in the regulation of the channel activity and voltage-dependent gating properties., (© 2022 Wiley Periodicals LLC.)

Ca v 1.2 channel phosphorylation plays an important role in regulating neuronal plasticity by action potential-dependent Ca 2+ entry. Most studies of Ca v 1.2 regulation by phosphorylation have been reported in heart and muscles. Here, we identified phosphorylation sites of neuronal Ca v 1.2 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 Ca v 1.2 channel is regulated by phosphorylation in a complex mechanism involving multiple phosphorylation sites., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

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 E x and E y 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., (© 2022. The Author(s).)

Ca 2+ entry through Ca v 1.3 Ca 2+ channels plays essential roles in diverse physiological events. We employed yeast-two-hybrid (Y2H) assays to mine novel proteins interacting with Ca v 1.3 and found Snapin2, a synaptic protein, as a partner interacting with the long carboxyl terminus (CT L ) of rat Ca v 1.3 L variant. Co-expression of Snapin with Ca v 1.3 L /Ca v β 3 /α 2 δ 2 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 Ca v 1.3 S containing a short CT (CT S ) in which a Snapin interaction site in the CT L was deficient. Luminometry and electrophysiology studies uncovered that Snapin co-expression did not alter the membrane expression of HA tagged Ca v 1.3 L but increased the slope of tail current amplitudes plotted against ON-gating currents, indicating that Snapin increases the opening probability of Ca v 1.3 L . Taken together, our results strongly suggest that Snapin directly interacts with the CT L of Ca v 1.3 L , leading to up-regulation of Ca v 1.3 L channel activity via facilitating channel opening probability.

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.

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.

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.

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., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

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 S 21 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.

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.

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.

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., Competing Interests: The authors declare no conflict of interest.

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., Competing Interests: The authors declared no conflict of interest.

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.

Voltage-activated Ca 2+ channels (Ca v ) play critical roles in excitable cells including neurons. Unlike the well-defined roles of Ca v 2 for pre-synaptic neurotransmission, the post-synaptic function of Ca v 2 is unclear. Based on our previous study demonstrating the postsynaptic association of the Ca v 2 with the AMPA receptor (AMPA-R), in this study we sought to further analyse the Ca v 2-AMPA-R association. We used a step-by-step dissociation of partially purified native Ca v 2-AMPA-R complexes and co-immunoprecipitation of the Ca v 2-AMPA-R complexes expressed in HEK293T cells to demonstrate that the main subunit of Ca v , α 1 , formed a complex with the AMPA-R without the auxiliary subunits β, α 2 δ, γ 2/3 . The α 1 subunit increased the cell-surface localisation of the AMPA-R, which could be a post-synaptic function of the Ca v 2., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Calcium-dependent inactivation of high voltage-activated Ca 2+ channels plays a crucial role in limiting rises in intracellular calcium (Ca 2+ 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 Ca 2+ i can regulate low voltage-activated T-type Ca 2+ channels. Using whole cell patch clamp, we examined the biophysical properties of Ca 2+ current through the three T-type Ca 2+ channel isoforms, Ca v 3.1, Ca v 3.2, or Ca v 3.3, comparing internal solutions containing 27 nM and l μM free Ca 2+ Both activation and inactivation kinetics of Ca v 3.3 current in l μM Ca 2+ i solution were more rapid than those in 27 nM Ca 2+ i solution. In addition, both activation and steady-state inactivation curves of Ca v 3.3 were negatively shifted in the higher Ca 2+ i solution. In contrast, the biophysical properties of Ca v 3.1 and Ca v 3.2 isoforms were not significantly different between the two internal solutions. Overexpression of CaM 1234 (a calmodulin mutant that doesn't bind Ca 2+ ) occluded the effects of l μM Ca 2+ i on Ca v 3.3, implying that CaM is involved in the Ca 2+ i regulation effects on Ca v 3.3. Yeast two-hybrid screening and co-immunoprecipitation experiments revealed a direct interaction of CaM with the carboxyl terminus of Ca v 3.3. Taken together, our results suggest that Ca v 3.3 T-type channel is potently regulated by Ca 2+ i via interaction of Ca 2+ /CaM with the carboxyl terminus of Ca v 3.3., (Copyright © 2017 by The American Society for Pharmacology and Experimental Therapeutics.)

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.

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., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Mammalian T-type Ca(2+) channels are encoded by three separate genes (Cav3.1, 3.2, 3.3). These channels are reported to be sleep stabilizers important in the generation of the delta rhythms of deep sleep, but controversy remains. The identification of precise physiological functions for the T-type channels has been hindered, at least in part, by the potential for compensation between the products of these three genes and a lack of specific pharmacological inhibitors. Invertebrates have only one T-type channel gene, but its functions are even less well-studied. We cloned Ca-α1T, the only Cav3 channel gene in Drosophila melanogaster, expressed it in Xenopus oocytes and HEK-293 cells, and confirmed it passes typical T-type currents. Voltage-clamp analysis revealed the biophysical properties of Ca-α1T show mixed similarity, sometimes falling closer to Cav3.1, sometimes to Cav3.2, and sometimes to Cav3.3. We found Ca-α1T is broadly expressed across the adult fly brain in a pattern vaguely reminiscent of mammalian T-type channels. In addition, flies lacking Ca-α1T show an abnormal increase in sleep duration most pronounced during subjective day under continuous dark conditions despite normal oscillations of the circadian clock. Thus, our study suggests invertebrate T-type Ca(2+) channels promote wakefulness rather than stabilizing sleep.

Certain voltage-activated Ca2+ channels have been reported to act as potential zinc entry routes. However, it remains to be determined whether zinc can permeate individual Ca2+ channel isoforms. We expressed recombinant Ca2+ channel isoforms in Xenopus oocytes and attempted to record zinc currents from them using a two-electrode voltage clamp method. We found that, in an extracellular zinc solution, inward currents arising from zinc permeation could be recorded from Xenopus oocytes expressing L-type Cav1.2 or Cav1.3 isoforms, but not from oocytes expressing Cav2.2, Cav2.3, Cav3.1, or Cav3.2. Zinc currents through Cav1.2 and Cav1.3 were blocked by nimodipine, but enhanced by (±)Bay K8644, supporting the finding that zinc can permeate both L-type Cav1.2 and Cav1.3 channel isoforms. We also examined the blocking effects of low concentrations of zinc on Ca2+ currents through the L-type channel isoforms. Low micro-molar zinc potently blocked Ca2+ currents through Cav1.2 and Cav1.3 with different sensitivities (IC50 for Cav1.2 and Cav1.3=18.4 and 34.1 μM) and de-accelerated the activation and inactivation kinetics in a concentration-dependent manner. Notably, mild acidifications of the external zinc solution increased zinc currents through Cav1.2 and Cav1.3, with the increment level for Cav1.3 being greater than that for Cav1.2. In overall, we provide evidence that Cav1.2 and Cav1.3 isoforms are capable of potentially functioning as zinc permeation routes, through which zinc entry can be differentially augmented by mild acidifications., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background: Since algal rhodopsins, the eukaryotic seven-transmembrane proteins, are generally difficult to express in Escherichia coli, eukaryotic cells have been used for heterologous expression. Mistic, a membrane-associated protein that was originally discovered in Bacillus subtilis, has been shown to improve the expression levels of many foreign integral membrane proteins in E. coli when used as a fusion partner linked to the N-terminus of cargo proteins., Methods: Here, we expressed two algal rhodopsins with N- and C-terminal Mistic domains in E. coli-Acetabularia rhodopsin I (ARI) and Chlamydomonas sensory rhodopsin B (CSRB, channel rhodopsin 2). UV/VIS spectroscopy, pH titration of proton acceptor residue, laser-induced photolysis and electrophysiological measurement were used for investigating important residues in proton transport and spectroscopic characters of the proteins., Results: Protein yield of two algal rhodopsins was enhanced, obtaining 0.12mg of Mistic-ARI and 0.04mg of Mistic-CSRB per liter of culture. Spheroplast expression Mistic-ARI had outward proton-pumping activity, indicating protein functionality. Asp89 of ARI changed its protonation state by light absorption, and Asp100 was important for O(600) formation. Electrophysiology revealed that both residues took part in proton transport. The spectroscopic analyses of Mistic-CSRB revealed its characteristics., Conclusions: Fusion to the membrane-integrating protein Mistic can enhance overexpression of eukaryotic type I rhodopsins in E. coli., General Significance: These findings indicate that Mistic fusion and E. coli expression method could be an effective, low cost technique for studying eukaryotic membrane proteins. This may have useful implications, for example, in studying structural characteristics and optogenetics for rhodopsins., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The Cav1.2 Ca2+ channel is essential for cardiac and smooth muscle contractility and many physiological functions. We mutated single, double, and quadruple sites of the four potential Asn (N)-glycosylation sites in the rabbit Cav1.2 into Gln (Q) to explore the effects of Nglycosylation. When a single mutant (N124Q, N299Q, N1359Q, or N1410Q) or Cav1.2/WT was expressed in Xenopus oocytes, the biophysical properties of single mutants were not significantly different from Cav1.2/WT. In comparison, the double mutant N124,299Q showed a positive shift in voltage-dependent gating. Furthermore, the quadruple mutant (QM; N124,299,1359,1410Q) showed a positive shift in voltage-dependent gating as well as a reduction of current. We tagged EGFP to the QM, double mutants, and Cav1.2/WT to chase the mechanisms underlying the reduced currents of QM. The surface fluorescence intensity of QM was weaker than that of Cav1.2/WT, suggesting that the reduced current of QM arises from its lower surface expression than Cav1.2/WT. Tunicamycin treatment of oocytes expressing Cav1.2/WT mimicked the effects of the quadruple mutations. These findings suggest that Nglycosylation contributes to the surface expression and voltage-dependent gating of Cav1.2.

A 30-year-old male was involved in a car accident. Radiographs revealed a depressed marginal fracture of the medial tibial plateau and an avulsion fracture of the fibular head. Magnetic resonance imaging showed avulsion fracture of Gerdy's tubercle, injury to the posterior cruciate ligament (PCL), posterior horn of the medial meniscus, and the attachments of the lateral collateral ligament and the biceps femoris tendon. The depressed fracture of the medial tibial plateau was elevated and stabilized using a cannulated screw and washer. The injured lateral and posterolateral corner (PLC) structures were repaired and augmented by PLC reconstruction. However, the avulsion fracture of Gerdy's tubercle was not fixed because it was minimally displaced and the torn PCL was also not repaired or reconstructed. We present a unique case of pure varus injury to the knee joint. This case contributes to our understanding of the mechanism of knee injury and provides insight regarding appropriate treatment plans for this type of injury.

The patellar ligament angle (PLA) was assessed in 105 normal stifle joints of 79 dogs and 33 stifle joints of 26 dogs with a ruptured cranial cruciate ligament (CrCL). The PLA of stifles with complete CrCL rupture was significantly lower than that of normal stifles, particularly at a flexion angle of 60 ˜ 80° in both plain and stress views. If the PLA was < 90.55° on the stress view with a 60 ˜ 80° flexion angle, the dog was diagnosed with a complete rupture of the CrCL with a sensitivity of 83.9% and specificity of 100%. In conclusion, measuring the PLA is a quantitative method for diagnosing complete CrCL rupture in canines.

Purpose: Despite the documented blood-saving effects of tranexamic acid (TNA) in total knee arthroplasty (TKA), the question whether clinical values of TNA are identical in unilateral and bilateral TKAs remains unclear. This study was undertaken to determine the clinical values of TNA in unilateral and simultaneous bilateral TKAs under a contemporary blood-saving protocol in terms of efficacy (total blood loss and transfusion rate) and safety (the incidences of symptomatic deep vein thrombosis and pulmonary embolism)., Methods: One hundred and eighty unilateral and 146 bilateral TKA patients were randomized into TNA group or control group. In unilateral TKA patients, TNA (10 mg/kg) was administered intravenously 20 min before tourniquet deflation and repeated 3 h after surgery. In bilateral TKA patients, one more dose (10 mg/kg) was given before tourniquet deflation in the second TKA. A contemporary blood-saving protocol was applied to all patients. The TNA and control groups were compared separately in unilateral and bilateral TKA patients for the efficacy and safety variables., Results: In unilateral TKA patients, the TNA group had less total blood loss (905 vs. 1,018 mL, p = 0.018) than the control group, but there was no difference in the allogenic transfusion rate (1 vs. 7 %, n.s.). In bilateral TKA patients, the TNA group showed no differences in total blood loss (1,282 vs. 1,379 mL, n.s.), but a significant reduction in the allogenic transfusion rate (7 vs. 27 %, p = 0.002). No symptomatic deep vein thrombosis or pulmonary embolism was found in all patients., Conclusion: This study demonstrates that the use of TNA reduces total blood loss, but the effects on the transfusion rate can differ depending on the type of TKAs (unilateral vs. bilateral) and the blood-saving protocols.

Synthetic molecules which selectively transport sodium or potassium chloride across a lipid membrane have been prepared. The salt carriers consist of two heteroditopic binding sites, an anion-binding cavity with three hydrogen bond donors and an azacrown ether for binding an alkali metal cation. The association constants between the carriers and chloride ion have been enhanced by 1 order of the magnitude in the presence of sodium or potassium ion in 10% (v/v) CD3OH/CD3CN, due to the formation of a contact ion-pair between the bound cation and chloride as demonstrated by the single-crystal X-ray structure of a sodium chloride complex. A series of transport experiments have demonstrated that the synthetic molecule functions as a mobile carrier of transporting salts via M(+)/Cl(-) symport. Among alkali metal chlorides, the carrier with an 18-azacrown-6 exhibits a strong selectivity toward potassium chloride, while the carrier with a 15-azacrown-5 displays a moderate selectivity for sodium chloride.

We review the ins and outs of T-channel structure, focusing on the extracellular high-affinity metal-binding site and intracellular loops. The high-affinity metal-binding site was localized to repeat I of Cav3.2. Interestingly, a similar binding site was found in the high voltage-activated Cav2.3 channel where it controls the channels' voltage dependence. Histidine at position 191 has a particularly interesting role in the high-affinity binding site, and its modification plays an important role in channel regulation by pharmacological agents that alter redox reactions. The intracellular loop connecting repeats I and II plays two important roles in Cav3.2 properties: one, its gating; and two, its surface expression. These studies have also identified a highly conserved intracellular gating brake that is predicted to form a helix-loop-helix structure. We conclude that the gating brake establishes important contacts with the gating machinery, thereby stabilizing a closed state of T-channels. This interaction is disrupted by depolarization, allowing the S6 segments to open and allowing Ca(2+) ions to flow through. Studies in cultured hippocampal neurons provided novel insights into how mutations found in idiopathic generalized epilepsy patients increase seizure susceptibility by both altering T-current pacemaker currents and by activating Ca-activated transcription factors that regulate dendritic arborization. These studies reveal novel roles for T-channels to control cellular physiology.

It has been established that Ca(V)3.2 T-type voltage-gated calcium channels (T-channels) play a key role in the sensitized (hyperexcitable) state of nociceptive sensory neurons (nociceptors) in response to hyperglycemia associated with diabetes, which in turn can be a basis for painful symptoms of peripheral diabetic neuropathy (PDN). Unfortunately, current treatment for painful PDN has been limited by nonspecific systemic drugs with significant side effects or potential for abuse. We studied in vitro and in vivo mechanisms of plasticity of Ca(V)3.2 T-channel in a leptin-deficient (ob/ob) mouse model of PDN. We demonstrate that posttranslational glycosylation of specific extracellular asparagine residues in Ca(V)3.2 channels accelerates current kinetics, increases current density, and augments channel membrane expression. Importantly, deglycosylation treatment with neuraminidase inhibits native T-currents in nociceptors and in so doing completely and selectively reverses hyperalgesia in diabetic ob/ob mice without altering baseline pain responses in healthy mice. Our study describes a new mechanism for the regulation of Ca(V)3.2 activity and suggests that modulating the glycosylation state of T-channels in nociceptors may provide a way to suppress peripheral sensitization. Understanding the details of this regulatory pathway could facilitate the development of novel specific therapies for the treatment of painful PDN.

Eugenol has been used as an analgesic in dentistry. Previous studies have demonstrated that voltage-gated Na(+) channels and high-voltage-activated Ca(2+) channels expressed in trigeminal ganglion (TG) neurons sensing dental pain are molecular targets of eugenol for its analgesic effects. However, it has not been investigated whether eugenol can affect T-type Ca(2+) channels, which are known to be detected in the afferent neurons. In this report, we investigate how eugenol can influence cloned T-type channel isoforms expressed in HEK293 cells, using whole-cell patch clamp. Application of eugenol inhibited Cav3.1, Cav3.2, and Cav3.3 currents in a concentration-dependent manner with IC50 values of 463, 486, and 708 μM, respectively. Eugenol was found to negatively shift the steady-state inactivation curves of the T-type channel isoforms, but it did not shift their activation curves. In addition, eugenol had little effect on the current kinetics of Cav3.1 and Cav3.2, but it accelerated the inactivation kinetics of Cav3.3 currents. Reduction of channel availability enhanced eugenol inhibition sensitivity for Cav3.1 and Cav3.2, but not for Cav3.3. Moreover, eugenol inhibition of T-type channel isoforms was found to be use dependent. Finally, we show that the T-type currents recorded from rat TG neurons were inhibited by eugenol with a similar potency to Cav3.1 and Cav3.2 isoforms. Taken together, our findings suggest that T-type Ca(2+) channels are additional molecular targets for the pain-relieving effects of eugenol.

Voltage-activated Ca2+ channels are membrane protein machinery performing selective permeation of external calcium ions. The main Ca2+ selective filters of all high-voltage-activated Ca2+ channel isoforms are commonly composed of four Glu residues (EEEE), while those of low-voltage-activated T-type Ca2+ channel isoforms are made up of two Glu and two Asp residues (EEDD). We here investigate how the Asp residues at the pore loops of domains III and IV affect biophysical properties of the Ca(v)3.2 channel. Electrophysiological characterization of the pore mutant channels in which the pore Asp residue(s) were replaced with Glu, showed that both Asp residues critically control the biophysical properties of Ca(v)3.2, including relative permeability between Ba2+ and Ca2+, anomalous mole fraction effect (AMFE), voltage dependency of channel activation, Cd2+ blocking sensitivity, and pH effects, in distinctive ways., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Background: Dexamethasone is a potent analgesic and antiemetic. However, the benefit of dexamethasone after TKA is unclear, as is the efficacy in a current multimodal regime., Questions/purposes: We determined (1) whether the addition of dexamethasone to a protocol including ramosetron further reduces postoperative emesis compared with ramosetron alone; (2) whether it reduces postoperative pain; and (3) whether it increases the risk for wound complications in a current multimodal regime after TKA., Methods: We randomized 269 patients undergoing TKAs to receive dexamethasone (10 mg) 1 hour before surgery and ramosetron immediately after surgery (Dexa-Ra group, n = 135), or ramosetron alone (Ra group, n = 134). We recorded the incidence of postoperative nausea and vomiting (PONV), severity of nausea, incidence of antiemetic requirement, complete response, pain level, and opioid consumption. Patients were assessed 0 to 6, 6 to 24, 24 to 48, and 48 to 72 hours postoperatively. In addition, patients were evaluated for wound complications and periprosthetic joint infections at a minimum of 1 year after surgery., Results: The Dexa-Ra group had a lower incidence of PONV during the entire 72-hour evaluation period and experienced less severe nausea for the first 6 hours after TKA, although not between 6 to 72 hours. Overall use of a rescue antiemetic was less frequent, and complete response was more frequent in the Dexa-Ra group. Patients in the Dexa-Ra group experienced lower pain and consumed less opioids during the 6- to 24-hour period and during the overall study period. No differences were found in wound complications between the groups, and each group had one case of periprosthetic joint infection., Conclusions: Patients who received prophylactic dexamethasone in addition to ramosetron had reduced postoperative emesis and pain without increased risks for wound complications, compared with patients who received ramosetron alone in patients managed using a multimodal regimen after TKA.

A near-field scanning microwave microscope (NSMM) is used to readout and visualize homemade 10-mer oligonucleotide microarrays and an Agilent 60-mer DNA microarray as a realistic test of NSMM applicability to multiplexed sequence analysis. Sensitive characterization of DNA coverage and high resolution mapping of DNA spots in the microarray were realized by measuring the change of microwave reflection coefficient (S₁₁) at about 4 GHz operating frequency. Hybridization between target (free) and capture (immobilized) sequences leads to changes in the microwave reflection coefficient, which were measured by the NSMM. These changes are caused by hybridization-induced modification of the dielectric permittivity profile of the DNA film. The dynamic range based on analysis of the 10-mer microarrays is over 3 orders of magnitude with the detection limit estimated below 0.01 strands/μm². The NSMM method should be readily capable of detecting target coverages down to 98% of probe coverage. We also directly image the patterned DNA microarray by NSMM with a 2 μm resolution. The complementary optical image of the DNA microarray visualized by using a relative fluorescent intensity metric agrees well with the NSMM results., (Copyright © 2012 Elsevier B.V. All rights reserved.)

We study the opto-electrical properties of Natronomonas pharaonis sensory rhodopsin II (NpSRII) by using a near-field microwave microprobe (NFMM) under external light illumination. To investigate the possibility of application of NFMM to biological macromolecules, we used time dependent properties of NPSRII before/after light activation which has three distinct states - ground-state, M-state, and O-state. The diagnostic ability of NFMM is demonstrated by measuring the microwave reflection coefficient (S(11)) spectrum of NpSRII under steady-state illumination in the wavelength range of 350-650 nm. Moreover, we present microwave reflection coefficient S(11) spectra in the same wavelength range for two fast-photocycling rhodopsins: green light-absorbing proteorhodopsin (GPR) and Gloeobacter rhodopsin (GR). In addition the frequency sweep shift can be detected completely even for tiny amounts of sample (∼10(-3) OD of rhodopsin). Based on these results NFMM shows both very high sensitivity for detecting conformational changes and produces a good time-resolved spectrum., (Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

We have developed an electromagnetic microwave cavity sensor based on the resonant frequency shift for real time measurement of the glycemia in pig blood. We could determine the concentration of d-glucose in pig blood in the range of 150-550mg/dl at the resonance frequency near 4.75GHz with a bandwidth of 300MHz. The change in the d-glucose concentration in blood brings microwave reflection coefficient S(11) changes of about 6.26dB and resonance frequency shifts of about 11.25MHz due to the electromagnetic interaction between the cavity resonator and the blood filled plastic tube inserted into the cavity. This proposed system provides a unique approach for real time noninvasive and contactless glucose monitoring., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

Fluorophore-assisted light inactivation (FALI) is an investigative tool to inactivate fluorescently labeled proteins by a mechanism of in situ photodestruction. We found that Ca(v)1.2 (L-type) and Ca(v)3.1 (T-type) calcium channels, labeled by genetic fusion with GFP derivatives, show differential sensitivity to FALI. Specifically, FALI silences Ca(v)1.2 calcium channels containing EYFP-labeled α(1C)subunits but does not affect the EYFP-α(1G) Ca(v)3.1 calcium channels or Ca(v)1.2 channels containing EYFP-labeled β subunits. Our findings limit the applicability of acceptor photobleaching for the measurements of FRET but open an opportunity to combine the fluorescent imaging of the live cell expressing labeled calcium channels with selective functional inactivation of their specific subsets.

A six-year-old intact female Maltese dog weighing 3.8 kg presented with a history of mild lameness and swelling on both forelimbs. Radiographic and computed tomographic views revealed an extensive periosteal reaction in all four limbs and a large round mass on the right middle lung lobe. A total lobectomy was performed and pulmonary adenosquamous carcinoma was histologically confirmed. A diagnosis of hypertrophic osteopathy (HO) secondary to a lung tumor was made. Periosteal proliferation decreased significantly after surgery; however, there was evidence of dyspnea, mass recurrence, and periosteal reaction three months post-operatively. This is the first case report of pulmonary adenosquamous carcinoma with HO in a dog in which we describe clinical, imaging, surgical, and histological findings.

PHACE association is a rare neurocutaneous condition in which facial hemangiomas associate with a spectrum of posterior fossa malformations, arterial cerebrovascular anomalies, cardiovascular anomalies, and eye anomalies. We reported a case of PHACE association in a premature infant showing facial, intracranial, and oropharyngeal hemangiomas with evidence of the Dandy-Walker variant and complicated cardiovascular anomalies, including a right-sided aortic arch and an atypical patent ductus arteriosus arising from a tortuous left subclavian artery. To our knowledge, intracranial hemangiomas are rare in PHACE association, and a concomitant oropharyngeal hemangioma has not been previously reported in the PHACE association literature. In infants presenting with large, plaque-like facial hemangiomas, it is important to conduct active cardiovascular and neurological evaluations. Special attention should be given to the laryngoscopic examination to search for additional hemangiomas in the airway.

Background: Patient-centered care requires physicians to respond to patients' preferences, including their preferences regarding treatment decision-making. The authors surveyed patients to determine their preoperative preferences and their retrospectively perceived levels of involvement in decision-making for carpal tunnel release, and they attempted to identify factors that affect patient preferences and experiences., Methods: Seventy-eight patients who underwent carpal tunnel release for carpal tunnel syndrome were requested to indicate their preferred level of involvement preoperatively and to assess their actual levels of involvement postoperatively, using a Control Preferences Scale containing five levels that range from fully active to fully passive. Clinical and demographic factors that potentially affected patients' preoperative preferences and postoperative assessments of levels of involvement were analyzed., Results: Fifty-nine patients (76%) indicated preoperatively that they preferred shared decision-making, and sixty-six (85%) thought postoperatively that they had experienced this type of decision-making. The correlation between preoperative and postoperative Control Preferences Scale assessments was significant (r = 0.525, p &lt; 0.001). A history of a surgical procedure was independently associated with a preoperative preference for a more active role (odds ratio = 4.2), and patients with a caregiver (odds ratio = 4.0) or private insurance (odds ratio = 2.6) were more likely to experience an active role. Patients who preferred a collaborative role had lower scores on the Disabilities of the Arm, Shoulder and Hand questionnaire than those who preferred a fully active role (p = 0.002) or a fully passive role (p = 0.009)., Conclusions: The majority of patients with carpal tunnel syndrome preferred to share surgical decision-making with the surgeon, and those who preferred a collaborative role had less severe symptoms than those who preferred a fully active or a fully passive role. A history of a surgical procedure, having a caregiver, and having private insurance were associated with a more active role. This information may assist the establishment of patient-centered consultation in patients with carpal tunnel syndrome.

Individual pharmacokinetic differences for herb-drug interaction have been associated with genetic variations of the multidrug resistance (MDR) gene. A high level expression of MDR protein increases cellular efflux and might decrease drug sensitivity. This study investigated the drug efflux activity difference of human MDR1 triallelic variant 2677G/T/A (rs2032582), as a nonsynonymous 893Ala/Ser/Thr, using Xenopus laevis oocytes and MDR1 overexpressing LLC-PK1 cells. Two MDR1 variants (2667T/893Ser and 2667A/893Thr) were generated using human MDR1 cDNA (2677G/893Ala). No significant difference in the expression of MDR1 893Ala/Ser/Thr was found in X. laevis oocytes. However, the MDR1 2667A/893Thr variant interestingly showed a significant decrease of efflux activity for both digoxin and daunorubicin compared with those of 893Ala and 893Ser variants. In further investigation assessing the inhibitory effects of three herbal extracts on MDR1, 893Ala and 893Ser showed significant decreases of efflux activities in treatments with P. cocos (p = 0.005 for 893Ser) and D. dasycarpus (p = 0.0009 for 893Ala; p = 0.002 for 893Ser) in X. laevis oocytes. The results in this study suggest that herbal medicines could interact with other drugs and change the therapeutic effects depending on the genetic polymorphisms of individuals., (Copyright © 2011 John Wiley & Sons, Ltd.)

We have cloned a hamster Cav1.3 variant with a long carboxyl terminus. This differs from the first hamster Cav1.3 clone which has a short carboxyl terminus. When relative expression levels of the two variants were examined using quantitative RT-PCR, the long Cav1.3 transcripts were detected abundantly in the brain and testis, moderately in the heart, pancreas, and kidney, and weakly in the lung. Comparatively, the short Cav1.3 transcripts were detected less abundantly in most of the tissues. The two Cav1.3 variants were reconstituted in Xenopus oocytes and their electrophysiological properties were characterized using a two-electrode voltage clamping method. The long Cav1.3 variant was ~5-fold better expressed than the short Cav1.3 variant. When Ca2+ was used as a charge carrier, the long Cav1.3 variant containing an IQ (Ile-Gln) motif displayed strong calcium-dependent inactivation, while the short variant that was deficient of an IQ motif showed little calcium-dependent inactivation. Examination of other biophysical properties revealed that potentials for activation threshold, peak current, and half-activation and inactivation of the long Cav1.3 were significantly lower than those of the short Cav1.3. These findings suggest that the long carboxyl tail plays crucial roles in not only facilitating calcium-dependent inactivation, but also improving expression and negative shifting of the activation and inactivation properties., (Copyright © 2010 Elsevier B.V. All rights reserved.)

We cloned an intronless, nuclear-encoded opsin gene from an EST library of Acetabularia acetabulum. Acetabularia rhodopsin I (ARI) encodes a protein of 246 amino acids with molecular weight of 27 kDa. ARI was reconstituted in the Xenopus oocyte expression system to characterize its electrophysiological properties utilizing the two-electrode voltage-clamping technique. Oocytes where ARI cRNA was injected displayed outward directed currents in response to light. The maximum action spectrum of ARI was detected at 520 nm green light. Light-stimulated ARI current amplitude was altered by the protons, but not by the other ions in recording solutions, suggesting that the algal rhodopsin is a light-stimulated proton pump. Typical proton-mediated outward current elicited by 520 nm light was characterized with two phases of non-inactivating outward current following initial transient current. Taken together, we here reported cloning of a novel Acetabularia opsin gene which was characterized to be a proton-pump stimulated by light.

The Neer type II distal clavicle fracture is notorious for its high nonunion rate, and surgical treatment is usually recommended. We reviewed articles from January 1990 to September 2009, and among them, 425 cases from 21 studies were included. According to the 425 cases in the literature, sixty patients were treated nonsurgically and 365 surgically. From 365 patients who were treated surgically, 105 were identified as receiving the coracoclavicular stabilization, 162 hook plate, 42 intramedullary fixation, 16 interfragmentary fixation, and 40 K-wire plus tension band wiring. The nonsurgical treatment resulted in 20 (33.3%) nonunions and 4 (6.7%) other complications. The surgical treatment resulted in 6 (1.6%) nonunions, 81 (22.2%) complications other than nonunion. The nonunion rate was significantly high with nonsurgical treatment (p < 0.001), and the complication rate was statistically high with surgery (p = 0.002). With surgical treatment, the nonunion rate was not significantly different among the modalities (p = 0.391). The complication rate was significantly higher in cases of the hook plate (40.7%) and the K-wire plus tension band wiring (20.0%) than those of the coracoclavicular stabilization (4.8%), the intramedullary (2.4%) and the interfragmentary fixation (6.3%). For the nonsurgical treatment, the functional outcomes were generally acceptable despite the high nonunion rate. The nonsurgical treatment could be considered as the first line treatment after sufficient counsel with the patient. The nonunion rate is high, however, the functional outcome is acceptable in most of the cases with nonunion. If the surgical treatment is considered, the intramedullary screw fixation, CC stabilization and interfragmentary fixation would be preferred because of their low complication rate.

Activation of group I metabotropic glutamate receptors (subtypes mGluR1 and mGluR5) regulates neural activity in a variety of ways. In CA1 pyramidal neurons, activation of group I mGluRs eliminates the post-burst afterhyperpolarization (AHP) and produces an afterdepolarization (ADP) in its place. Here we show that upregulation of Ca(v)2.3 R-type calcium channels is responsible for a component of the ADP lasting several hundred milliseconds. This medium-duration ADP is rapidly and reversibly induced by activation of mGluR5 and requires activation of phospholipase C (PLC) and release of calcium from internal stores. Effects of mGluR activation on subthreshold membrane potential changes are negligible but are large following action potential firing. Furthermore, the medium ADP exhibits a biphasic activity dependence consisting of short-term facilitation and longer-term inhibition. These findings suggest that mGluRs may dramatically alter the firing of CA1 pyramidal neurons via a complex, activity-dependent modulation of Ca(v)2.3 R-type channels that are activated during spiking at physiologically relevant rates and patterns., Competing Interests: The authors have declared that no competing interests exist.

Experimental and clinical studies on the accuracy of the intramedullary alignment method have produced different results, and few have addressed accuracy in the sagittal plane. Reported deviations are not only attributable to the alignment method but also to radiological errors. The purpose of this study was to evaluate the accuracy of the intramedullary alignment method in the sagittal plane using computed tomography (CT) and 3-dimensional imaging software. Thirty-one TKAs were performed using an intramedullary alignment method involving the insertion of a long 8-mm diameter rod into the medullary canal to the distal metaphysis of the tibia. All alignment instruments were set to achieve an ideal varus/valgus angle of 0° in the coronal plane and a tibial slope of 0° in the sagittal plane. The accuracy of the intramedullary alignment system was assessed by measuring the coronal tibial component angle and sagittal tibial slope angles, i.e., angles between the tibial anatomical axis and the tangent to the medial and lateral tibial plateau or the cut-surface. The mean coronal tibial component angle was 88.5° ± 1.2° and the mean tibial component slope in the sagittal plane was 1.6° ± 1.2° without anterior slope. Our intramedullary tibial alignment method, which involves passing an 8-mm diameter long rod through the tibial shaft isthmus, showed good accuracy (less than 3 degrees of variation and no anterior slope) in the sagittal plane in neutral or varus knees.

Ca(v)3.2 T-type channels contain a high affinity metal binding site for trace metals such as copper and zinc. This site is occupied at physiologically relevant concentrations of these metals, leading to decreased channel activity and pain transmission. A histidine at position 191 was recently identified as a critical determinant for both trace metal block of Ca(v)3.2 and modulation by redox agents. His(191) is found on the extracellular face of the Ca(v)3.2 channel on the IS3-S4 linker and is not conserved in other Ca(v)3 channels. Mutation of the corresponding residue in Ca(v)3.1 to histidine, Gln(172), significantly enhances trace metal inhibition, but not to the level observed in wild-type Ca(v)3.2, implying that other residues also contribute to the metal binding site. The goal of the present study is to identify these other residues using a series of chimeric channels. The key findings of the study are that the metal binding site is composed of a Asp-Gly-His motif in IS3-S4 and a second aspartate residue in IS2. These results suggest that metal binding stabilizes the closed conformation of the voltage-sensor paddle in repeat I, and thereby inhibits channel opening. These studies provide insight into the structure of T-type channels, and identify an extracellular motif that could be targeted for drug development.

We investigated the biophysical mechanism of inhibition of recombinant T-type calcium channels Ca(V)3.1 and Ca(V)3.2 by nitrous oxide (N(2)O). To identify functionally important channel structures, chimeras with reciprocal exchange of the N-terminal domains I and II and C-terminal domains III and IV were examined. In whole-cell recordings N(2)O significantly inhibited Ca(V)3.2, and - less pronounced - Ca(V)3.1. A Ca(V)3.2-prevalent inhibition of peak currents was also detected in cell-attached multi-channel patches. In cell-attached patches containing < or = 3 channels N(2)O reduced average peak current of Ca(V)3.2 by decreasing open probability and open time duration. Effects on Ca(V)3.1 were smaller and mediated by a reduced fraction of sweeps containing channel activity. Without drug, single Ca(V)3.1 channels were significantly less active than Ca(V)3.2. Chimeras revealed that domains III and IV control basal gating properties. Domains I and II, in particular a histidine residue within Ca(V)3.2 (H191), are responsible for the subtype-prevalent N(2)O inhibition. Our study demonstrates the biophysical (open times, open probability) and structural (domains I and II) basis of action of N(2)O on Ca(V)3.2. Such a fingerprint of single channels can help identifying the molecular nature of native channels. This is exemplified by a characterization of single channels expressed in human hMTC cells as functional homologues of recombinant Ca(V)3.1.

Electrophysiological characterization of T-type Ca2+ channel isoforms (Cav3.1, Cav3.2, and Cav3.3) has shown that all of the isoforms are low voltage-activated around resting membrane potential, but their current kinetics are distinctly different, with the activation and inactivation kinetics of the Cav3.1 and Cav3.2 channels being much faster than those of the Cav3.3 channel. We previously reported that multiple structural regions of the Cav3.3 T-type channel participate in determining its current kinetics. Here we have evaluated the relative contributions of individual cytoplasmic and trans-membrane regions to the current kinetics of the channel, by systematically replacing individual regions of Cav3.3 with the corresponding regions of Cav3.1. Introduction of the Cav3.1 III-IV loop into the Cav3.3 backbone accelerated both the activation and inactivation kinetics more prominently than any other intracellular loop or tail. Among the trans-membrane domains, introduction of the domain I of Cav3.1 into Cav3.3 accelerated both the activation and inactivation kinetics most effectively. These findings suggest that the current kinetics of the Cav3.3 channel are differentially controlled by several structural regions, among which the III-IV loop and domain I are the most prominent in governing both activation and inactivation kinetics.

Purpose: To correlate the incidence and rate of placental migration with the mode of delivery, pregnancy outcome, and maternal complication according to the location (anterior versus posterior) and type of placenta previa (PP)., Methods: We prospectively evaluated the placentas of 98 pregnant women with PP or low-lying placenta (LLP) at the prenatal sonographic examination performed between 20 and 27 gestational weeks. We divided the pregnant women into groups according to type and location of PP. Follow-up sonographic examination was performed between 32 and 37 weeks. We compared incidence of migration to the normal position and calculated the migration rate as the migrated distance divided by the weeks of interval between 2 sonographic examinations. We compared the incidences of cesarean section, fetal outcome, and maternal complications during the pregnancy., Results: The incidence of migration in the group of anterior placentas was significantly higher than that in the group of posterior placentas. The mean migration rate in the anterior group was 2.6 mm/week, whereas that in the posterior group was 1.6 mm/week. The migration rate of incomplete PP was significantly higher than that of LLP. Incidence of cesarean section for nonmigrated PP was significantly higher in the posterior group. The incidences of premature delivery and vaginal spotting were also significantly higher in the posterior group., Conclusion: Anterior PP and LLP may migrate more often and faster than posterior PP. Our results may be useful for planning of prenatal management and counseling patients with PP and LLP., ((c) 2007 Wiley Periodicals, Inc.)