Tmem2 Deficiency Leads to Enamel Hypoplasia and Soft Enamel in Mouse.

Teeth consist of 3 mineralized tissues: enamel, dentin, and cementum. Tooth malformation, the most common craniofacial anomaly, arises from complex genetic and environmental factors affecting enamel structure, size, shape, and tooth eruption. Hyaluronic acid (HA), a primary extracellular matrix component, contributes to structural and physiological functions in periodontal tissue. Transmembrane protein 2 (TMEM2), a novel cell surface hyaluronidase, has been shown to play a critical role during embryogenesis. In this study, we demonstrate Tmem2 messenger RNA expression in inner enamel epithelium and presecretory, secretory, and mature ameloblasts. Tmem2 knock-in reporter mice reveal TMEM2 protein localization at the apical and basal ends of secretory ameloblasts. Micro-computed tomography analysis of epithelial-specific Tmem2 conditional knockout (Tmem2-CKO) mice shows a significant reduction in enamel layer thickness and severe enamel deficiency. Enamel matrix protein expression was remarkably downregulated in Tmem2-CKO mice. Scanning electron microscopy of enamel from Tmem2-CKO mice revealed an irregular enamel prism structure, while the microhardness and density of enamel were significantly reduced, indicating impaired ameloblast differentiation and enamel matrix mineralization. Histological evaluation indicated weak adhesion between cells and the basement membrane in Tmem2-CKO mice. The reduced and irregular expressions of vinculin and integrin ß1 suggest that Tmem2 deficiency attenuated focal adhesion formation. In addition, abnormal HA accumulation in the ameloblast layer and weak claudin 1 immunoreactivity in Tmem2-CKO mice indicate impaired tight junction gate function. Irregular actin filament assembly was also observed at the apical and basal ends of secretory ameloblasts. Last, we demonstrated that Tmem2-deficient mHAT9d mouse ameloblasts exhibit defective adhesion to HA-containing substrates in vitro. Collectively, our data highlight the importance of TMEM2 in adhesion to HA-rich extracellular matrix, cell-to-cell adhesion, ameloblast differentiation, and enamel matrix mineralization.

Synthesis of water-soluble anthracene-appended benzoxaboroles and evaluation of their cis-1,2-diol recognition properties.

Three series of water-soluble anthracene-appended benzoxaboroles 1a-c were developed; their binding affinity toward cis-1,2-diols was explored by conventional fluorescence titrations to demonstrate the role of benzoxaborole as a general recognition motif of cis-1,2-diols for fluorescent probes. The complex structures of the tetra-coordinated boronate adducts between 1 and the cis-1,2-diols were revealed.

Questionable expression of unstable DQ heterodimer containing HLA-DQA1*01:07.

Human leukocyte antigens (HLA)-DQA1*01:07 was identified as an HLA-DQ blank specificity that segregated with the serological HLA-A2, -B7, -DR14, -DR52 haplotype, which carried DQB1*05:03. The blank specificity of DQA1*01:07-DQB1*05:03 may be because of lack of reactivity of available typing sera, or disruption of proper assembly of DQ heterodimer. The cDNA sequence of DQA1*01:07 is nearly identical to DQA1*01:04 except for a variant at position 304, which results in the replacement of an arginine with a cysteine at 79α. To determine whether the DQA1*01:07 product can be expressed on cell-surface, we co-expressed DQA1*01:07 with various DQB1*05 or *06 alleles in fibroblast cells. Cell-surface expression of DQ was detectable when DQA1*01:07 was co-expressed with DQB1*06:04 but undetectable with other DQB1*05 and DQB1*06 alleles, including DQB1*05:03, to which DQA1*01:07 was encoded in cis. These data suggest that DQA1*01:07 may act as a phenotypically null allele in the DQA1*01:07-DQB1*05:03 haplotype, while it can be expressed at a low level in the presences of certain DQB1*06 alleles, such as DQB1*06:04, in trans. Based on the null or low expression of DQA1*01:07 as shown in the previous and present studies, DQA1*01:07 has recently been renamed to DQA1*01:07Q, indicating its questionable expression.

Elevated immunoglobulin G antibodies to the proline-rich amino-terminal region of Epstein-Barr virus nuclear antigen-2 in sera from patients with systemic connective tissue diseases and from a subgroup of Sjögren's syndrome patients with pulmonary involvements.

Associations of Epstein-Barr virus (EBV) and autoimmune diseases have been hypothesized. We have analysed IgG antibodies to EBV nuclear antigen (EBNA)-2 in sera from Japanese patients with autoimmune systemic connective tissue diseases (CTD), exemplified by systemic lupus erythematosus (SLE), primary Sjogren's syndrome (SS), rheumatoid arthritis (RA), systemic sclerosis (SSc) and secondary SS (classical CTDs complicated with SS). An enzyme-linked immunosorbent assay (ELISA) which uses glutathione-S-transferase polypeptides fused to EBV nuclear antigen (EBNA)-2 and EBNA-1 was developed. Ratios of IgG antibody reactivity to whole IgG concentrations of sera were calculated to normalize EBNA-2 and EBNA-1 antibody levels to the hypergammaglobulinaemia that occurs in CTD. The ELISA optical density OD(450) readings of IgG antibodies to both the amino-terminal aa 1-116 of EBNA-2 and carboxyl-terminal aa 451-641 of EBNA-1 were elevated significantly in patients with SLE, primary SS, RA, SSc and secondary SS when compared to EBNA-1. The OD readings were divided by serum IgG concentrations to normalize for the hypergammaglobulinaemia. The specific levels of IgG antibodies to the amino-terminal region of EBNA-2 were elevated in patients with SLE, primary SS or RA, as well as those with secondary SS complicated with SLE or RA. The EBNA-2 amino-terminal region contains a polyproline tract and a proline-rich sequence and has considerable amino acid sequence homology with many cellular proline-rich proteins. High ratios of EBNA-2 aa 1-116 to EBNA-1 aa 451-641 IgG antibody levels which probably suggest reactivation of EBV latent infection were associated significantly with pulmonary involvement in SS patients. These results are consistent with the hypothesis that the sequence similarity between the amino-terminal region of EBNA-2 and proline-rich cellular proteins is associated with pathogenesis in a subpopulation of CTD patients, possibly by the molecular mimicry-epitope shift mechanism.

Liver abscess formation after hepatic chemoembolization for metastatic pancreatic neuroendocrine tumor.

Hepatic chemoembolization is an effective treatment of unresectable hypervascular hepatic tumors, but history of bilioenteric anastomosis is a major risk factor for liver abscess due to the retrograde enteric bacterial contamination. We report a case of culture-negative liver abscess developed after hepatic chemoembolization for a metastatic pancreatic endocrine tumor, and discuss the pathogenesis of liver abscess formation. We also present a potential pitfall for the prophylactic antibiotic therapy in a patient with a history of bilioenteric anastomosis. This is the first report of a culture-negative liver abscess caused by hepatic chemoembolization for a metastatic pancreatic endocrine tumor.

Endovascular management of multiple arterial aneurysms in Behcet's disease.

We report a case of Behcet's disease complicated by four arterial aneurysms successfully treated by coil embolisation and stent placement. Percutaneous endovascular repair offers a safe alternative to surgical management of this serious condition.

Centro de Testagem e aconselhamento de Brasília - CTA-DF: análise da capacidade de atendimento da demanda vulnerável no Distrito Federal / Brasília Center for testing, and counseling Center - CTA-DF: analysis of its capacity to fulfill the vulnerable demand in the Federal District

Com a expansäo da epidemia HIV-AIDS no Brasil, observaram-se avanços terapêuticos com aumento da sobrevida e melhoria da qualidade de vida dos portadores do vírus. Como também modificaçäo no perfil epidemiológico dos indivíduos infectados e, visíveis deslocamentos dos casos de infecçäo para o interior do País e setores marginalizados da sociedade. Objetiva conhecer se o CTA do Distrito Federal atendeu a populaçäo mais vulnerável e de risco para a infecçäo HIV

Severe complications after hypofractionated high dose rate intracavitary brachytherapy following external beam irradiation for oesophageal carcinoma.

The purpose of this study was to evaluate severe complications that developed after high dose rate (HDR) intracavitary brachytherapy for oesophageal carcinoma. Six consecutive patients with oesophageal carcinoma were treated by external beam irradiation (60 Gy in 30 fractions over 6 weeks) followed by hypofractionated intracavitary HDR brachytherapy (10 Gy in 2 fractions). Two of the six patients were alive and well for more than 2-3 years following therapy, but three of the six patients developed treatment-related oesophageal fistulae and died. HDR intracavitary brachytherapy following external beam irradiation is an effective method for radical treatment of oesophageal carcinoma. However, hypofractionated HDR brachytherapy should be used with care.

Primary Sjögren's syndrome initially manifested by optic neuritis: MRI findings.

We herein describe the MRI findings in a patient clinically diagnosed with primary Sjögren's syndrome (SjS) initially manifested by retrobulbar optic neuritis. A 63-year-old woman suddenly had left ocular pain and progressive visual disturbance. MR T2-weighted images revealed hyperintensity in the left optic nerve, with swelling. Contrast-enhanced T1-weighted images showed no abnormal enhancement. Follow-up MRI 6 months after admission revealed no significant changes in the affected optic nerve. To our knowledge, optic neuritis as a complication of SjS has been reported in ten patients [1, 2, 3, 4, 5, 6] and MRI findings in only one of them [6]. We thought MR images were useful for visualizing optic nerve involvement in SjS and observing its course.

Picture archiving and communication system: prospective study.

OBJECTIVES: To evaluate the use of a picture archiving and communication system and user satisfaction in order to further improve its quality. DESIGN: Prospective study. SETTING: Medical college hospital, Japan. MATERIALS AND METHODS: An automated computerised method was used to collect the data from March 1999 to February 2000. MAIN OUTCOME MEASURES: Each workstation automatically recorded data on the rank of the user, purpose of use, use of postprocessing tools, and user satisfaction. RESULTS: The number of resident users in the radiology reading room increased and those outside the reading room decreased, but the number of staff users changed little. The purpose of use and the use of postprocessing functions in the reading room were not significantly different from those outside it (P=0.179 and P=0.269, respectively). The average numbers of images accessed to per workstation monthly in the reading room, the general practice ward, and the gastroenterology ward were 1081, 970 and 741, respectively. Only 12 images in the orthopaedic surgery out-patient clinic and 70 images in the orthopaedic surgery ward, however, were accessed per month. The percentages of satisfied users decreased both inside and outside the reading room. The degree of satisfaction of users in the reading room was significantly different from that outside it (P=0.004). The most common reason for dissatisfaction was the length of time necessary to retrieve images. CONCLUSIONS: It is necessary to shorten response times of picture archiving and communication system workstations. Repeated data collection and evaluation, however, are also important.

Assessment of left ventricular volume by an ambulatory radionuclide monitoring system during head-up tilt in patients with unexplained syncope: relation to autonomic activity assessed by heart rate variability.

BACKGROUND: Decreased left ventricular volume during head-up tilt plays an important role in triggering syncope in patients with neurally mediated syncope. However, precise changes in left ventricular volume during head-up tilt have not been well investigated. This study was conducted to test the hypothesis that the decline in left ventricular volume during tilt could trigger ventricular mechanoreceptor activation. METHODS AND RESULTS: To investigate the mechanisms of tilt-induced syncope, we measured the temporal changes in left ventricular volume, ejection fraction, cardiac output, and heart rate variability indices during head-up tilt in 25 patients with syncope of undetermined etiology. Eleven patients had a cardioinhibitory response (CI group), 7 patients showed a vasodepressor response (VD group), and 7 patients demonstrated a negative response (NG group). Before syncope, ejection fraction increased most in the CI group, the left ventricular end-diastolic volume declined most in the VD group (VD group, -11.0% +/- 3.3%; CI group, -2.8% +/- 4.8%; NG group, -3.4% +/- 2.2%; P <.005), and the high-frequency spectra increased most in the CI group (CI group, 25.0% +/- 21.0%; VD group, -4.1% +/- 11.7%; NG group, -5.3% +/- 12.7%; P <.01). The vasodepressor response was dependent on left ventricular volume, whereas the cardioinhibitory response was related to the vagal activity reflected by high-frequency spectra. CONCLUSIONS: The precise evaluation of left ventricular volume by an ambulatory radionuclide monitoring system combined with a heart rate variability analysis is considered useful for clarifying the pathophysiology of neurally mediated syncope.

Computed tomography-guided frameless stereotactic radiotherapy for stage I non-small cell lung cancer: a 5-year experience.

PURPOSE: Stereotactic radiotherapy (SRT) is highly effective for brain metastases from non-small-cell lung cancers (NSCLCs). As such, primary lesions of NSCLC may also be treated effectively by similar focal high-dose SRT. METHODS AND MATERIALS: Between October 1994 and June 1999, 50 patients with pathologically proven T1-2N0 M0 NSCLC were treated by CT-guided frameless SRT. Of these, 21 patients were medically inoperable and the remainder were medically operable but refused surgery. In most patients, SRT was 50-60 Gy in 5-10 fractions for 1-2 weeks. Eighteen patients also received conventional radiotherapy of 40-60 Gy in 20-33 fractions before SRT. RESULTS: With a median follow-up period of 36 months (range 22-66), 30 patients were alive and disease free, 3 were alive with disease, 6 had died of disease, and 11 had died intercurrently. Local progression was not observed on follow-up CT scans in 47 (94%) of 50 patients. The 3-year overall survival rate was 66% in all 50 patients and 86% in the 29 medically operable patients. The 3-year cause-specific survival rate of all 50 patients was 88%. No definite adverse effects related to SRT were noted, except for 2 patients with a minor bone fracture and 6 patients with temporary pleural pain. CONCLUSIONS: SRT is a very safe and effective treatment for Stage I NSCLC. Additional studies involving a larger patient population and longer follow-up periods are warranted to assess this new treatment for early-stage lung cancer.

Amino acid substitution analyses of the DNA contact region, two amphipathic alpha-helices and a recognition-helix-like helix outside the dimeric beta-barrel of Epstein-Barr virus nuclear antigen 1.

OBJECTIVES AND METHODS: Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA-1), which is essential for EBV latency, homodimerizes and binds to the EBV replication origin, oriP. We analyzed the dimerization/DNA-binding domain of EBNA-1 by random and site-directed amino acid substitution. RESULTS: Random point mutations that resulted in reduced DNA binding clustered in the DNA contact region (a.a. 461-473) and at or near the termini of alpha-helix II (514-527). Three substitutions of Gly in the DNA contact region each greatly reduced binding to a single binding site oligonucleotide. Substitutions at and near the termini of alpha-helix II diminished DNA binding. A helix-deforming substitution in alpha-helix I (477-489) blocked DNA binding. A helix-deforming substitution in alpha-helix III (568-582) abolished dimerization and DNA binding. Similarities in surface electrostatic properties and conserved amino acids were found between alpha-helix II and recognition helices of papillomavirus E2 proteins. CONCLUSIONS: The basic DNA contact region is crucial for the specific interaction of EBNA-1 with a single binding site. Alpha-helix I477 is indispensable for oriP binding, and alpha-helix III568 contributes to the homodimeric structure of EBNA-1. Alpha-helix II514 contributes to oriP binding, perhaps changing its alignment with DNA.

Epstein-Barr virus nuclear antigen-1-dependent and -independent oriP-binding cellular proteins.

OBJECTIVE: Epstein-Barr virus (EBV) nuclear antigen-1 (EBNA-1) and the replication origin, oriP, are essential for the replication and maintenance of latent EBV DNA in cells, but no enzymatic activity has been associated with EBNA-1 protein alone. In this study, we have searched for host cellular proteins that interact with EBNA-1 protein in various B cell lines latently infected with EBV, including a recently EBV growth-transformed cell line. METHODS: By using gel shift analysis, we investigated the interactions of an oligonucleotide containing a single EBNA-1 recognition site, derived from the family of repeats (FR) element of oriP, with protein from cell extracts. RESULTS: The FR oligonucleotide bound a (72-kD) cellular protein in the absence of EBNA-1 and without induction of the previously reported 'anti-EBNA-1 proteins'. The FR oligonucleotide formed complexes with additional proteins from EBNA-1-synthesizing cell lines; these complexes were abolished or supershifted by anti-EBNA-1 monoclonal antibodies. SDS-PAGE analyses of 35S-Met-labeled proteins that bound to a biotin- conjugated FR oligonucleotide, fractionated by a glycerol gradient centrifugation and affinity-purified with streptavidin, showed three major bands, a 72-kD protein, the FR binding of which seemed to be independent of EBNA-1, a 64-kD protein in both EBNA-1-transfected and latently EBV-infected cell lines, and a 45-kD protein in EBV-infected cell lines, which was most prominent in a recently EBV growth-transformed cell line. CONCLUSIONS: The FR element forms complexes with cellular proteins in the absence and presence of EBNA-1. These 72-, 64- and 45-kD cellular proteins might be involved in the function of the oriP and EBNA-1 system.

Does the presence of distant and local malignancy alter parenchymal perfusion in apparently disease-free areas of the liver?

Our objective was to investigate if hepatic arterial (HAP) and portal venous perfusion (PVP) in apparently normal areas of liver, as measured by functional CT, are affected by the presence of extra- and intrahepatic malignancy Three patient groups were compared: A, controls with no malignancy (N = 10); B, extrahepatic malignancy without liver involvement (N = 12); and C, subjects with metastases elsewhere in the liver (N = 13). HAP, PVP, and a CT hepatic perfusion index (CT-HPI) calculated as HAP/(HAP + PVP) were calculated on a section free of metastatic disease, using a previously published method. Figures for PVP were (median and interquartile range) in group A were 1.06 (0.9-1.30), in B 1.03 (0.81-1.09), and in C 0.75 (0.54-1.02) ml/min/ml; for HAP group A values were 0.07 (0.052-0.078), in B 0.07 (0.053-0.147), and in C 0.12 (0.091-0.146) ml/min/ml and for CT-HPI Group A values were 4.9% (4.8-6.6%), in B 5.6% (3.8-13.6%), and in C 14.3% (10.4-15.4%). Significant differences in all indices were seen between groups A and C. A significant difference (P = 0.017) was seen between groups B and C in the CT-HPI values. In conclusion, patients with liver metastases show abnormal blood flow in apparently normal liver compared to controls. This difference was not seen in subjects with malignancy without liver metastases. Possible explanations would be either the unmasking of occult metastatic disease or vasoactive or mechanical effects due to liver malignancy.

An experience of stereotactic radiation therapy for primary intracranial choriocarcinoma.

We report on a patient with choriocarcinoma in the pineal region who was successfully treated with stereotactic radiation therapy (SRT). The increased level of serum human chorionic gonadotropin (HCG) was lowered during chemotherapy with etoposide, cisplatin, and ifosfamide. However, HCG was not normalized and magnetic resonance images still showed an enhanced tumor mass with gadolinium. The patient underwent SRT of 40 Gy at an 80% isodose line per 10 fractions over two weeks, followed by conventional craniospinal irradiation of 32.4 Gy. The level of HCG dropped below the detection limit. The patient has been in good condition for more than four years after the completion of treatment, without any signs of recurrence. We propose SRT as a valid treatment option for malignant germ cell tumors in the pineal region.

Preassignment examination for personnel on Iwo Jima.

The Japan Maritime Self-Defense Force (JMSDF) requires all personnel assigned to Iwo Jima to undergo a preassignment medical examination. The efficacy and benefit of this mandatory examination has not been evaluated objectively in depth. Our purpose was to review the health status of JMSDF personnel assigned to Iwo Jima and to evaluate the correlation of the preassignment medical examination to their current health status for the study period. We divided the participants into two groups. Group Y was composed of JMSDF personnel receiving a preassignment medical examination, and group N was composed of those personnel who did not receive the examination. After the annual health examination, health status was evaluated using objective criteria. We found little statistically significant difference among group Y and group N participants. However, proportionally more patients currently receiving care for or diagnosed with hypertension, hyperuricemia, or severe obesity were identified as being members of group N than group Y. We have demonstrated that the preassignment medical examination may contribute to predicting the health status of potential Iwo Jima personnel and may contribute to controlling the cost of care associated with these specific diagnoses by limiting the assignment of at-risk personnel.