Association of NOD2 Mutations with Aggressive Periodontitis.

Aggressive periodontitis (AgP) is characterized by rapid alveolar bone destruction and tooth loss early in life, and its etiology remains unclear. To explore the genetic risk factors of AgP, we performed genome-wide single-nucleotide polymorphism genotyping for identity-by-descent mapping and identified 32 distinct candidate loci, followed by whole exome sequencing with 2 pedigrees of AgP consisting of 3 cases and 1 control in 1 family and 2 sibling cases in the other. After variant filtering procedures and validation by targeted Sanger sequencing, we identified 2 missense mutations at 16q12 in NOD2 (p.Ala110Thr and p.Arg311Trp), which encodes nucleotide-binding oligomerization domain protein 2. We further examined 94 genetically unrelated AgP patients by targeted sequencing of NOD2 and found that 2 patients among them also carried the p.Arg311Trp variant. Furthermore, we found 3 additional missense mutations in this gene (p.His370Tyr, p.Arg459Cys, and p.Ala868Thr). These mutations either had not been previously observed or are extremely rare (frequency <0.001) in Asian populations. NOD2 plays a crucial role in innate immunity as an intracellular receptor initiating nuclear factor κB-dependent and mitogen-activated protein kinase-dependent gene transcription. These results demonstrated NOD2 as a novel gene involved in AgP.

Human immunodeficiency virus DNA copies as a virologic marker in a clinical trial with beta-interferon.

The number of human immunodeficiency virus (HIV) DNA copies in peripheral blood mononuclear cells (PBMC) was quantitated by the polymerase chain reaction (PCR) and used as a virologic marker in a clinical trial with beta-interferon (beta-IFN) (6 x 10(6) IU/day administered intravenously for 4 weeks). In 11 HIV-infected patients who were clinically stable, the number ranged from 10 to 1,063 copies per 10(5) PBMC. However, percent change of the number in the individual untreated patients stayed between -46.2% and 203.0% of the basal level by one month interval. In six patients who were treated with beta-IFN, changes in the number were not significant before and after the trial.

Reoxygenation injury in a cultured corneal epithelial cell line protected by the uptake of lactoferrin.

PURPOSE: To investigate whether reoxygenation after extended hypoxia causes cellular damage in cultured corneal epithelial cells and to demonstrate the protective effects of lactoferrin. METHODS: Immortalized human corneal epithelial cells (T-HCECs) were cultured to confluence in 96-well culture plates, subjected to stringent hypoxia (1% O2, 5% CO2, 94% N2 at 37 degrees C) for 24 hours, and returned to normoxic conditions (5% CO2, 95% air at 37 degrees C). Cell viability was observed by 1 microM propidium iodide staining 0, 2, 4, and 6 hours after reoxygenation. Inhibition studies were performed after 2 hours' reoxygenation, using 2 mM iron chelator desferrioxamine and 0.2 mg/ml lactoferrin. Confocal immunocytochemistry for human lactoferrin and western blot analysis for lactoferrin-induced ferritin were performed in cultured T-HCECs to demonstrate the internalization of lactoferrin after application. RESULTS: After 2 hours, reoxygenation of T-HCECs after hypoxia produced an increase in cell death that was significantly greater than that observed in normoxic control cells or in cells subjected to hypoxia for the same time span without reoxygenation. The addition of desferrioxamine and lactoferrin at the time of reoxygenation significantly attenuated cellular damage. Confocal immunocytochemistry revealed that lactoferrin is taken into the cytoplasm of T-HCECs as early as 30 minutes after application. This was also demonstrated in western blot analysis by the upregulation of intracellular ferritin at 18 hours by the addition of iron-bound lactoferrin but not by iron-free lactoferrin. CONCLUSION: Reoxygenation is responsible for increased cellular damage after extensive hypoxia, which is attenuated by chelators of free iron in the cytosol, including the major tear protein lactoferrin.

Excimer laser-induced hydroxyl radical formation and keratocyte death in vitro.

PURPOSE: To characterize the type of reactive oxygen species (ROS) produced by excimer photoablation of aqueous solutions and to show the effects of ROS and antioxidants on corneal stromal cells in vitro. METHODS: Electron spin-resonance spectroscopy was performed using the spin-trapping agent 5,5-dimethyl-1-pyrroline N-oxide (DMPO) for the detection of the superoxide anion and the hydroxyl radical in an acellular DMPO solution irradiated with the excimer laser. Hydroxyl radicals were produced by the Fenton reaction in vitro by the mixture of hydrogen peroxide and ferrous iron (Fe2+), and the effects on cultured corneal fibroblasts were observed by fluorescent microscopy using the cell death marker, propidium iodide (PI) and TdT-mediated dUTP nick-end labeling (TUNEL). RESULTS: Excimer photoablation of a 1% DMPO solution produced a species-specific spin-trapping adduct for the hydroxyl radical ('OH), but not for the superoxide anion or other unidentified free radical. The signals were inhibited dose dependently by the hydroxyl radical scavenger dimethylsulfoxide (DMSO) and an L-ascorbic acid analogue, EPCK-1. The production of *OH in the supernatant of cultured rabbit corneal fibroblasts by the Fenton reaction caused an increase in PI (+) and TUNEL (+) cells by 90 minutes, which was significantly inhibited by the addition of DMSO. CONCLUSIONS: Hydroxyl radicals may be partly responsible for stromal fibroblast cell apoptosis after excimer photoablation.

Spectacle side panels and moist inserts for the treatment of dry-eye patients.

To increase the moisture level around the eyes, small, wet, acute, triangular sponges, from which water steadily evaporates, were attached to special side panels of modified eyeglasses. The moisture level in the region between the cornea and the spectacle lens was measured in 10 patients with moderately severe dry eyes and in 10 normal controls, by a miniature moisture sensor under ambient conditions of 22.3 degrees C and 29.8% humidity. The humidity was 32.0 +/- 4.2% with spectacles, frame only (no lenses); 32.3 +/- 6.0% with spectacles; 37.9 +/- 5.1% with spectacles equipped with side panels, and 42.7 +/- 5.7% with side panel moist inserts attached for dry eye patients. Humidity was the highest with moist inserts added to the spectacles of the control group, with or without moderate wind. All dry eye patients noticed symptomatic relief with the inserts, and the rose bengal score and fluorescein staining improved after 2 weeks of their use. Our findings indicate that the combination of side panels and periodically moistened sponge inserts offer a simple and noninvasive approach to increasing the humidity surrounding the eyes, thus decreasing the tear evaporation from the ocular surface and providing amelioration of frequently debilitating symptoms of dry eye.

Quantitative evaluation of atopic blepharitis by scoring of eyelid conditions and measuring the water content of the skin and evaporation from the eyelid surface.

PURPOSE: To quantitatively evaluate the condition of the eyelid skin of patients with atopic blepharitis, their symptoms were scored and the water content of the skin and evaporation from the skin were measured. METHODS: Forty patients with atopic blepharitis were examined. The condition of eyelid skin (erythema, edema/papulation/oozing/crust, excoriation/lichenification) was scored from 0 to 3 points. Water content and water evaporation were measured with a Moisture Checker and an evaporimeter, respectively. Eleven age-matched volunteers without atopic disorders were recruited as normal controls. RESULTS: The Moisture Checker values and water evaporation from lid skin were significantly correlated (r = -0.44, p = 0.006). The Moisture Checker values of the patients with atopic blepharitis was 35.5+/-8.2% (44.7 +/-10.6% in the normal controls, p = 0.009), and water evaporation from their lid skin was 3.6+/-0.9 g/cm2 per second (2.0+/-0.3 g/cm2 per second, p < 0.001); then, the patients were divided into four groups, from "asymptomatic" to "severe," according to the sum of their blepharitis scores. Patients with lower blepharitis scores tended to have higher Moisture Checker values and lower water evaporation values. CONCLUSION: Scoring of eyelid condition enabled us to objectively estimate the severity of atopic blepharitis. Measurements of the water content of lid skin and water evaporation from lid skin are useful in evaluation of the severity of this disease.

Origin of lumbar cerebrospinal fluid pulse wave.

System analysis was performed on 16 adult mongrel dogs to determine the origin of the lumbar cerebrospinal fluid pulse wave. The descending thoracic aorta was occluded to evaluate the effects of the spinal arterial pulsations, and the thoracic aorta and inferior vena cava were simultaneously occluded to evaluate the effects of the spinal venous pulsations. It was concluded that, in the first harmonic wave, the components of the lumbar cerebrospinal fluid pulse wave are as follows: spinal arterial pulsations, 39.4%; spinal vascular (arteries and veins) pulsations, 77%; venous pulsations in the spinal canal, 37.6%; and the intracranial pressure pulse wave transmitted through the spinal canal from the intracranial space to the lumbar level, 23%.

Apico-aortic conduit bypass operation in a patient with calcified ascending aorta due to Werner's syndrome.

A 53-year-old Japanese man with Werner's syndrome underwent successful apico-aortic conduit bypass operation. Preoperative examination showed severe calcification in the aortic root and ascending aorta. A composite graft was implanted from the left ventricular apex to the descending thoracic aorta with femoro-femoral bypass. To our knowledge, this case is the first surgical correction of a cardiac lesion in a patient with Werner's syndrome.

A meta-analysis of Hodgkin lymphoma reveals 19p13.3 TCF3 as a novel susceptibility locus.

Recent genome-wide association studies (GWAS) of Hodgkin lymphoma (HL) have identified associations with genetic variation at both HLA and non-HLA loci; however, much of heritable HL susceptibility remains unexplained. Here we perform a meta-analysis of three HL GWAS totaling 1,816 cases and 7,877 controls followed by replication in an independent set of 1,281 cases and 3,218 controls to find novel risk loci. We identify a novel variant at 19p13.3 associated with HL (rs1860661; odds ratio (OR)=0.81, 95% confidence interval (95% CI) = 0.76-0.86, P(combined) = 3.5 × 10(-10)), located in intron 2 of TCF3 (also known as E2A), a regulator of B- and T-cell lineage commitment known to be involved in HL pathogenesis. This meta-analysis also notes associations between previously published loci at 2p16, 5q31, 6p31, 8q24 and 10p14 and HL subtypes. We conclude that our data suggest a link between the 19p13.3 locus, including TCF3, and HL risk.