TRPM8 Mediates Hyperosmotic Stimuli-Induced Nociception in Dental Afferents.

Hyperosmolar sweet foods onto exposed tooth dentin evoke sudden and intense dental pain, called dentin hypersensitivity. However, it remains unclear how hyperosmolar stimuli excite dental primary afferent (DPA) neurons and thereby lead to dentin hypersensitivity. This study elucidated whether TRPM8, which is well known as a cold temperature- or menthol-activated receptor, additionally mediates nociception in response to hyperosmolar stimuli in adult mouse DPA neurons, which are identified by a fluorescent retrograde tracer: DiI. Single-cell reverse transcription polymerase chain reaction revealed that TRPM8 was expressed in subsets of DPA neurons and that TRPM8 was highly colocalized with TRPV1 and Piezo2. Immunohistochemical analysis also confirmed TRPM8 expression in DPA neurons. By using Fura-2-based calcium imaging, application of hyperosmolar sucrose solutions elicited calcium transients in subsets of the trigeminal ganglion neurons, which was significantly abolished by a selective TRPM8 antagonist: N-(3-Aminopropyl)-2-[(3-methylphenyl)methoxy]-N-(2-thienylmethyl)benzamide (AMTB) hydrochloride. When we further examined changes of c-fos expression (a neuronal activation marker) in the spinal trigeminal nucleus after hyperosmolar stimulation onto exposed tooth dentin, c-fos mRNA and protein expression were increased and were also significantly reduced by AMTB, especially in the spinal trigeminal interpolaris-caudalis transition zone (Vi/Vc). Taken together, our results provide strong evidence that TRPM8 expressed in DPA neurons might mediate dental pain as a hyperosmosensor in adult mice.

TRPM7 Mediates Mechanosensitivity in Adult Rat Odontoblasts.

Odontoblasts, with their strategic arrangement along the outermost compartment of the dentin-pulp complex, have been suggested to have sensory function. In addition to their primary role in dentin formation, growing evidence shows that odontoblasts are capable of sensing mechanical stimulation. Previously, we found that most odontoblasts express TRPM7, the nonselective mechanosensitive ion channel reported to be critical in Mg2+ homeostasis and dentin mineralization. In line with this finding, we sought to elucidate the functional expression of TRPM7 in odontoblasts by pharmacological approaches and mechanical stimulation. Naltriben, a TRPM7-specific agonist, induced calcium transient in the majority of odontoblasts, which was blocked by TRPM7 blockers such as extracellular Mg2+ and FTY720 in a dose-dependent manner. Mechanical stretch of the odontoblastic membrane with hypotonic solution also induced calcium transient, which was blocked by Gd3+, a nonselective mechanosensitive channel blocker. Calcium transient induced by hypotonic solution was also blocked by high extracellular Mg2+ or FTY720. When TRPM7-mediated calcium transients in odontoblasts were analyzed on the subcellular level, remarkably larger transients were detected in the distal odontoblastic process compared with the soma, which was further verified with comparable immunocytochemical analysis. Our results demonstrate that TRPM7 in odontoblasts can serve as a mechanical sensor, with its distribution to facilitate intracellular Ca2+ signaling in the odontoblastic process. These findings suggest TRPM7 as a mechanical transducer in odontoblasts to mediate intracellular calcium dynamics under diverse pathophysiological conditions of the dentin.

Piezo2 Expression in Mechanosensitive Dental Primary Afferent Neurons.

Mechanosensitive ion channels have been suggested to be expressed in dental primary afferent (DPA) neurons to transduce the movement of dentinal fluid since the proposal of hydrodynamic theory. Piezo2, a mechanosensitive, rapidly inactivating (RI) ion channel, has been recently identified in dorsal root ganglion (DRG) neurons to mediate tactile transduction. Here, we examined the expression of Piezo2 in DPA neurons by in situ hybridization, single-cell reverse transcriptase polymerase chain reaction, and whole-cell patch-clamp recordings. DPA neurons with Piezo2 messenger RNA (mRNA) or Piezo2-like currents were further characterized based on their neurochemical and electrophysiological properties. Piezo2 mRNA was found mostly in medium- to large-sized DPA neurons, with the majority of these neurons also positive for Nav1.8, CGRP, and NF200, whereas only a minor population was positive for IB4 and peripherin. Whole-cell patch-clamp recordings revealed Piezo2-like, RI currents evoked by mechanical stimulation in a subpopulation of DPA neurons. RI currents were pharmacologically blocked by ruthenium red, a compound known to block Piezo2, and were also reduced by small interfering RNA-mediated Piezo2 knockdown. Piezo2-like currents were observed almost exclusively in IB4-negative DPA neurons, with the current amplitude larger in capsaicin-insensitive DPA neurons than the capsaicin-sensitive population. Our findings show that subpopulation of DPA neurons is indeed mechanically sensitive. Within this subpopulation of mechanosensitive DPA neurons, we have identified the Piezo2 ion channel as a potential transducer for mechanical stimuli, contributing to RI inward currents. Piezo2-positive DPA neurons were characterized as medium- to large-sized neurons with myelinated A-fibers, containing nociceptive peptidergic neurotransmitters.

Spin-induced band modifications of graphene through intercalation of magnetic iron atoms.

Intercalation of magnetic iron atoms through graphene formed on the SiC(0001) surface is found to induce significant changes in the electronic properties of graphene due mainly to the Fe-induced asymmetries in charge as well as spin distribution. From our synchrotron-based photoelectron spectroscopy data together with ab initio calculations, we observe that the Fe-induced charge asymmetry results in the formation of a quasi-free-standing bilayer graphene while the spin asymmetry drives multiple spin-split bands. We find that Fe adatoms are best intercalated upon annealing at 600 °C, exhibiting split linear π-bands, characteristic of a bilayer graphene, but much diffused. Subsequent changes in the C 1s, Si 2p, and Fe 3p core levels are consistently described in terms of Fe-intercalation. Our calculations together with a spin-dependent tight binding model ascribe the diffuse nature of the π-bands to the multiple spin-split bands originated from the spin-injected carbon atoms residing only in the lower graphene layer.

Risk factors for hospital and long-term mortality of critically ill elderly patients admitted to an intensive care unit.

BACKGROUND: Data on long-term outcomes of elderly (≥65 years) patients in ICU are sparse. MATERIALS AND METHODS: Adult patients (n = 1563, 45.4% elderly) admitted over 28 months were analyzed by competing risks regression model to determine independent factors related to in-hospital and long-term mortality. RESULTS: 414 (26.5%) and 337 (21.6%) patients died in-hospital and during the 52 months following discharge, respectively; the elderly group had higher mortality during both periods. After discharge, elderly patients had 2.3 times higher mortality compared to the general population of the same age-group. In-hospital mortality was independently associated with mechanical ventilation (subdistribution hazard ratio (SHR) 2.74), vasopressors (SHR 2.56), neurological disease (SHR 1.77), and Mortality Prediction Model II score (SHR 1.01) regardless of age and with malignancy (SHR, hematological 3.65, nonhematological 3.4) and prior renal replacement therapy (RRT, SHR 2.21) only in the elderly. Long-term mortality was associated with low hemoglobin concentration (SHR 0.94), airway disease (SHR 2.23), and malignancy (SHR hematological 1.11, nonhematological 2.31) regardless of age and with comorbidities especially among the nonelderly. CONCLUSIONS: Following discharge, elderly ICU patients have higher mortality compared to the nonelderly and general population. In the elderly group, prior RRT and malignancy contribute additionally to in-hospital mortality risk. In the long-term, comorbidities (age-related), anemia, airway disease, and malignancy were significantly associated with mortality.

Memb-roller: an effective way of making membrane rolls for pathological examination and studies of human placenta.

For the pathological evaluation of the chorioamniotic membranes, a membrane roll is made to give an orientation to the sections and to cover more extended planes. We designed a new instrument, Annie's memb-roller, for easy preparation of membrane rolls. Annie's memb-roller has two non-toothed arms which are 2 mm wide, 1 mm thick, and 8 cm long. To test its effectiveness in making a membrane roll, the placentas obtained from pregnant women (n = 10) with various obstetrical conditions were used. Preparation of the membrane rolls using Annie's memb-roller was easy and successful in all cases tested. The lengths of the membranes measured from fresh placentas and paraffin sections showed a very good correlation (p < 0.001; r = 0.941). Application of the memb-roller would be very helpful in the semi-quantitative analysis of the chorioamniotic membranes and the standardization of placental pathology and research practice.

Distinct patterns of C4d immunoreactivity in placentas with villitis of unknown etiology, cytomegaloviral placentitis, and infarct.

C4d deposition is considered to be evidence of antibody-mediated rejection. This study was conducted to compare C4d immunoreactivity between villitis of unknown etiology (VUE) and cytomegaloviral placentitis. C4d immunohistochemistry was performed in cases with VUE (n = 16) and cytomegaloviral placentitis (n = 5). Distinct, linear C4d immunoreactivity along the syncytiotrophoblast was found in all VUE cases. In cytomegaloviral placentitis, the intensity of C4d immunoreactivity along the syncytiotrophoblast was not prominent, but cytoplasmic C4d immunoreactivity of villous cytotrophoblasts was frequently observed. Further screening of the cases with placental infarcts (n = 5) demonstrated prominent C4d immunoreactivity in the chorionic villi adjacent to the infarct. We report the characteristic co-localization of VUE and C4d immunoreactivity. The overall findings in this study strongly suggest that the complement activation is a common mechanism of diverse placental injuries associated with rejection, infection, and ischemia.

Molecular genetic, cardiac and neurodevelopmental findings in cases of prenatally diagnosed rhabdomyoma associated with tuberous sclerosis complex.

OBJECTIVE: Rhabdomyoma is the most common type of cardiac tumor in fetuses and is often associated with tuberous sclerosis complex (TSC) with neurologic sequelae. The purpose of this study was to investigate the cardiac and neurodevelopmental outcomes of fetal rhabdomyoma. METHODS: We reviewed the clinical characteristics of 23 cases of cardiac rhabdomyoma diagnosed prenatally by fetal echocardiography at the Asan Medical Center between January 1998 and December 2009. We also reviewed postnatal results of brain magnetic resonance imaging, echocardiography, renal ultrasound examination and molecular genetic analysis to confirm the presence of cardiac rhabdomyoma with or without TSC. RESULTS: Among 23 cases, outcome data were available for 17 (73.9%) and six cases (26.1%) were lost to follow-up. The survival rate was 100.0% (17/17). Among the 17 cases with outcome data, spontaneous tumor regression occurred in eight (47.1%), and no change in tumor size and number was observed in the remaining nine cases (52.9%). There was no evidence of long-term cardiac dysfunction caused by persisting rhabdomyomas, regardless of tumor size. TSC was found in nine patients (52.9%), of whom five (55.6%) showed neurodevelopmental morbidity. We identified mutations in one of the TSC1 or TSC2 genes in four of nine TSC infants whose parents allowed us to perform molecular genetic analysis. Three of these (75.0%) were found to have neurologic impairment. Seven (77.8%) of nine TSC cases were non-familial. CONCLUSIONS: The overall outcome of isolated cardiac rhabdomyoma appears to be favorable. We suggest that systematic postnatal evaluation of TSC be performed even in cases of cardiac rhabdomyoma without a family history of TSC. Molecular characterization of TSC1 and TSC2 might be helpful in predicting short- and long-term neurodevelopmental outcomes.

Protective effect of fetal pulmonary sequestration in two cases of postnatal manifestation of congenital diaphragmatic hernia.

We describe two cases of postnatally diagnosed congenital diaphragmatic hernia (CDH) combined with pulmonary sequestration, both of which were diagnosed as isolated pulmonary sequestration on prenatal ultrasound. In these cases, prenatal ultrasonography demonstrated only a hyperechoic mass on the left lower lung and the diaphragm seemed intact. In each case both lungs showed otherwise normal development throughout pregnancy. Pulmonary sequestration may serve as a 'protector', preventing herniation of abdominal contents into the thoracic cavity. The co-occurrence of CDH may be obscured by a lung mass, especially on the left lower lung, and therefore it is necessary to deliver these infants at a tertiary center and parents should be counseled about the possibility of postnatal CDH.

Profile of volatile compounds during papaya juice fermentation by a mixed culture of Saccharomyces cerevisiae and Williopsis saturnus.

This study investigated the formation and utilization of volatile compounds during papaya juice fermentation by a mixed culture of Saccharomyces cerevisiae and Williopsis saturnus. Time-course papaya juice fermentations were carried out using pure cultures of S. cerevisiae var. bayanus R2 and W. saturnus var. mrakii NCYC2251 and a mixed culture of the two yeasts at a ratio of 1:1000 (R2:NCYC2251). Changes in S. cerevisiae cell population, Brix, sugar consumption and pH were similar in the mixed culture and in the S. cerevisiae monoculture. There was an early growth arrest of W. saturnus in the mixed culture fermentation. A range of volatile compounds were produced during fermentation including fatty acids, alcohols, aldehydes and esters and some volatile compounds including those initially present in the juice were utilized. The mixed culture fermentation of S. cerevisiae and W. saturnus benefited from the presence of both yeasts, with more esters being produced than the S. cerevisiae monoculture and more alcohols being formed than the W. saturnus monoculture. The study suggests that papaya juice fermentation with a mixed culture of S. cerevisiae and W. saturnus may be able to result in the formation of more complex aroma compounds and higher ethanol level than those using single yeasts.

Evaluation of the usefulness of the fetal femur length with respect to gestational age to detect Down syndrome in Korean subjects.

OBJECTIVE: We investigated the usefulness of shortening of the fetal femur length (FL) to predict Down syndrome at different gestational ages in Korean subjects. METHODS: This study involved 110 Korean Down syndrome fetuses and 602 randomly selected euploid controls. The expected FL for any biparietal diameter (BPD) was calculated based on the control group data. Subjects were divided into four groups according to gestational age periods: 14-18 weeks; 19-23 weeks; 24-28 weeks and 29-36 weeks. The value of measured/expected FL ratio to predict Down syndrome was analyzed for each group. RESULTS: The values of FL for any BPD in Down syndrome patients were significantly different from those in the control group (p < 0.001). A low ratio of measured/expected FL increased the risk of fetal Down syndrome (p < 0.001) with a mean measured/expected FL in Down syndrome of 0.907 (SD 0.075). At a fixed false positive rate of 5%, the sensitivities of FL were lower than 32.8% (95% CI 0.705-0.915) in three second trimester groups, and 71.1% (95% CI 0.924-0.997) in the third trimester group. CONCLUSION: Short FL is a poor marker of Down syndrome in the second trimester in Korean subject. It may be used as a screening marker only in the third trimester.

Diagnostic value of the matrix metalloproteinase-8 rapid test for detecting microbial invasion of the amniotic cavity.

The purpose of this paper was to compare the MMP-8 PTD Check (MPC) test with other indirect tests for detecting microbial invasion of the amniotic cavity (MIAC). Amniotic fluid (AF) was analyzed in 155 women for white blood cell (WBC) count, glucose concentration, and an MPC test and evaluated for MIAC using cultures for aerobic/anaerobic bacteria and mycoplasmas and polymerase chain reaction (PCR) of chlamydia. The median AF glucose concentration was lower and the median AF WBC count was higher in women with MIAC than in women without MIAC (p < 0.01 and p < 0.001, respectively). Also, the positive rate of the MPC test was higher in women with MIAC than in women without MIAC (p < 0.001). The sensitivities of AF glucose concentration, AF WBC count, and the MPC test for the detection of MIAC were 58.6%, 75.9%, and 86.2%, respectively. The specificities for the detection of MIAC were 76.2%, 80.2%, and 74.6%, respectively. We conclude that the MPC test is a rapid, easily performed, and accurate indirect method for detecting MIAC.

Spontaneous resolution of prenatally diagnosed dural sinus thrombosis: a case report.

Fetal thrombosis of the dural sinus is an extremely rare congenital cerebrovascular condition that is potentially fatal. We report a case of dural sinus thrombosis diagnosed by prenatal ultrasonography and fetal magnetic resonance imaging (MRI) in the second trimester. The thrombosis showed partial resolution during pregnancy and resolved spontaneously after birth without neurological complications. This is the first report of spontaneous postnatal resolution, and may provide helpful information on the natural history and prenatal counseling of fetal thrombosis of the dural sinus.

Successful vesicoamniotic shunting of posterior urethral valves in the first trimester of pregnancy.

Prenatal ultrasonography early in gestation allows diagnosis of posterior urethral valves. We report on a fetus with posterior urethral valves treated using vesicoamniotic shunting at 13 + 5 weeks' gestation. A double-basket catheter was used for shunting. A 2582-g male neonate was delivered at 33 weeks' gestation, and the infant continued to show normal renal function at 3 years of age. To the best of our knowledge this is the first report of successful vesicoamniotic shunt placement using a double-basket catheter in the first trimester of pregnancy.

Four-dimensional ultrasound guidance of prenatal invasive procedures.

OBJECTIVE: Technological advances in ultrasonography have revolutionized prenatal diagnosis and treatment. Here we evaluate the effectiveness of using four-dimensional (4D) ultrasonography to guide prenatal invasive procedures. PATIENTS AND METHODS: Prenatal invasive procedures using 4D ultrasound were recorded prospectively in 93 cases: 10 amnioinfusions, 50 amniocenteses, 8 chorionic villus samplings (CVS) and 25 cordocenteses. The needle target site was first identified using the two-dimensional (2D) mode, and was then confirmed using the three-dimensional (3D) mode. The needle was inserted under 4D ultrasound guidance. After selecting the needle target site, the true position of the needle was determined in three planes ('real-time 3D targeting'). RESULTS: Using 4D ultrasound guidance, most procedures were performed within 5 min and with a 100% success rate, even in cases involving severe oligohydramnios (amniocentesis), thin placentas (CVS) or narrow umbilical veins (cordocentesis). Moreover, there were no serious complications during or after any procedure. CONCLUSIONS: 4D ultrasonography can be used to guide various prenatal invasive procedures offering clear information in all three planes. It is likely that such imaging will reduce the time taken to complete the procedures and reduce the risks associated with them. Published by John Wiley & Sons, Ltd.

The variables affecting nuchal skin-fold thickness in mid-trimester.

OBJECTIVES: To find out variables that have an effect on the measured values of nuchal skin-fold thickness (NT) and to formulate a regression equation on the basis of those variables. METHODS: The data on gestational age (GA), cephalic index (CI), presentation (Pr, vertex or breech), and the presence or absence of nuchal cord (NC) were collected prospectively on 548 normal singleton fetuses between 16 and 24 weeks gestation, and independent correlation of those variables with NT was calculated by multiple regression analysis and a regression equation was produced. RESULTS: GA had positive correlation and CI had negative correlation with NT. The nuchal skin-fold was thicker among fetuses with breech presentation rather than those with vertex presentation and increased in the presence of nuchal cord. We calculated the expected NT through these observations: for fetuses presenting vertex, NT = 5.608 + 0.243GA - 0.066CI + NC* and for breech, NT = 2.803 + 0.392GA - 0.066CI + NC*. CONCLUSIONS: This is the first report, which takes GA, CI, Pr, and NC for correlation factors with NT as a whole. These equations and further studies on determining the cutoff values of the nuchal skin-fold thickness may improve the sensitivity and specificity for the detection of Down syndrome and enable to minimize unnecessary invasive procedures such as amniocentesis or cordocentesis.

Thyroid hormone regulation of N-methyl-D-aspartic acid receptor subunit mRNA expression in adult brain.

Thyroid hormone is an essential modulator of brain development, but little is known about its actions in the adult brain. Hypothyroidism is associated with gene expression changes in both central and peripheral nervous tissue. Functional consequences of adult-onset hypothyroidism include an inability to produce long-term potentiation in rat hippocampus and impaired learning and memory in both rats and man. Long-term potentiation is a form of learning that is dependent on functional N-methyl-d-aspartic acid (NMDA)-preferring ionotropic glutamate receptors. This work examines the expression of ionotropic glutamate receptor subunit mRNA following surgical thyroidectomy with or without thyroid hormone replacement. In situ hybridization histochemistry was used to determine the mRNA levels of the NMDA receptor subunits NR1, NR2A, NR2B, the AMPA receptor subunit GluR1, and the kainate receptor subunit KA2. Reducing circulating concentrations of thyroid hormone by surgical removal of the thyroid gland 2 weeks before sacrifice decreased the expression of NR1 mRNA exclusively in the hippocampus. Conversely, hyperthyroidism selectively reduced NR2B mRNA expression in the dorsal hippocampus. Altering thyroid hormone status had no effect on the expression of KA2 or GluR1 subunit mRNA. The regulation of expression of NR1 and NR2B mRNA by thyroid hormone is a novel mechanism for explaining the relationship between thyroid hormone and cognitive function.

Prenatal detection of mesoblastic nephroma by sonography and magnetic resonance imaging.

Congenital mesoblastic nephroma is the most common neonatal kidney tumor and surgical excision is almost always curative. We report the prenatal detection of congenital mesoblastic nephroma by sonography and magnetic resonance imaging. After birth, a right nephrectomy was performed and the baby recovered well.

Efg1, a morphogenetic regulator in Candida albicans, is a sequence-specific DNA binding protein.

Efg1 is essential for hyphal development in the human pathogen Candida albicans under most conditions. Efg1 is related to basic helix-loop-helix regulators, and therefore most workers presume that Efg1 is a transcription factor. Here we confirm that Efg1 is a DNA binding protein that can interact specifically with the E box.