Increased frequency of anti-retina antibodies in asymptomatic patients with chronic t. gondii infection

PURPOSE:

To search for anti-retina antibodies that serve as markers for eye disease in uveitis.

MATERIALS AND METHODS:

Stored sera from patients with uveitis, ocular toxoplasmosis (n = 30) and non-infectious, immune-mediated uveitis (n = 50) and from asymptomatic individuals who were positive (n = 250) and negative (n = 250) for anti-Toxoplasma antibodies were tested. Serum anti-retina IgG was detected by an optimized ELISA using a solid-phase whole human retina extract, bovine S-antigen or interphotoreceptor retinoid-binding protein.

RESULTS:

Uveitis patients showed a higher mean reactivity to whole human retina extract, interphotoreceptor retinoid-binding protein and S-antigen in comparison to the asymptomatic population. These findings were independent of the uveitis origin and allowed the determination of the lower anti-retina antibody cut-off for the three antigens. Asymptomatic anti-Toxoplasma serum-positive individuals showed a higher frequency of anti-human whole retina extract antibodies in comparison to asymptomatic anti-Toxoplasma serum-negative patients. The bovine S-antigen and interphotoreceptor retinoid-binding protein ELISAs also showed a higher mean reactivity in the uveitis groups compared to the asymptomatic group, but the observed reactivities were lower and overlapped without discrimination.

CONCLUSION:

We detected higher levels of anti-retina antibodies in uveitis patients and in a small fraction of asymptomatic patients with chronic toxoplasmosis. The presence of anti-retina antibodies in sera might be a marker of eye disease in asymptomatic patients, especially when whole human retina extract is used in a solid-phase ELISA.     

Influence of hepatitis B virus genotypes on the development of preS deletions and advanced liver disease

Hepatitis B virus (HBV) mutants with deletions in the preS region have not been evaluated for association with viral genotypes. In a case-control study, HBV DNA samples collected from 80 each of carriers infected with HBV genotype B or C were examined for preS deletions. PreS deletion mutants were found in a total of 37 of 160 (23%) HBV carriers. Carriers with preS deletion mutants were older (56.0 +/- 12.7 vs 49.3 +/- 16.9 years, P < 0.05), were infected more frequently with HBV genotype C (84% vs 40%, P < 0.05), and had more advanced disease, such as liver cirrhosis and hepatocellular carcinoma (54% vs 31%; P < 0.05), than did those without such mutants. In a multivariate analysis, genotype C (odds ratio [OR] = 9.3, P < 0.001) and advanced liver disease (OR = 3.1, P < 0.01) were the most significant variables in association with preS deletions. A direct repeat sequence (TCAGG) was found at the start or at the end of preS1 deletions in 6 of the 20 (30%) cases examined, and preS2 deletions in these cases were clustered over the 5'-terminal half of this region. These results indicate that the development of preS deletion mutants depends on HBV genotypes and that it may be associated with progressive liver disease.     

Identification of 16 novel mutations in the argininosuccinate synthetase gene and genotype-phenotype correlation in 38 classical citrullinemia patients

Classical citrullinemia (CTLN1), a rare autosomal recessive disorder, is caused by mutations of the argininosuccinate synthetase (ASS) gene, localized on chromosome 9q34.1. ASS functions as a rate-limiting enzyme in the urea cycle. Previously, we identified 32 mutations in the ASS gene of CTLN1 patients mainly in Japan and the United States, and to date 34 different mutations have been described in 50 families worldwide. In the present study, we report ASS mutations detected in 35 additional CTLN1 families from 11 countries. By analyzing the entire coding sequence and the intron-exon boundaries of the ASS gene using RT-PCR and/or genomic DNA-PCR, we have identified 16 novel mutations (two different 1-bp deletions, a 67-bp insertion, and 13 missense) and have detected 12 known mutations. Altogether, 50 different mutations (seven deletion, three splice site, one duplication, two nonsense, and 37 missense) in 85 CTLN1 families were identified. On the basis of primary sequence comparisons with the crystal structure of E. coli ASS protein, it may be concluded that any of the 37 missense mutations found at 30 different positions led to structural and functional impairments of the human ASS protein. It has been found that three mutations are particularly frequent: IVS6-2A>G in 23 families (Japan: 20 and Korea: three), G390R in 18 families (Turkey: six, U.S.: five, Spain: three, Israel: one, Austria: one, Canada: one, and Bolivia: one), and R304W in 10 families (Japan: nine and Turkey: one). Most mutations of the ASS gene are "private" and are distributed throughout the gene, except for exons 5 and 12-14. It seems that the clinical course of the patients with truncated mutations or the G390R mutation is early-onset/severe. The phenotype of the patients with certain missense mutations (G362V or W179R) is more late-onset/mild. Eight patients with R86H, A118T, R265H, or K310R mutations were adult/late-onset and four of them showed severe symptoms during pregnancy or postpartum. However, it is still difficult to prove the genotype-phenotype correlation, because many patients were compound heterozygotes (with two different mutations), lived in different environments at the time of diagnosis, and/or had several treatment regimes or various knowledge of the disease.     

Three-Dimensional Computed Tomographic Findings of Bilateral Tracheal Bronchus

A newborn male was admitted with cyanosis and respiratory distress. Echocardiography showed a right heart isomerism associated with a single right ventricle, a double-outlet right ventricle, and pulmonary atresia. Chest X-ray demonstrated severe left upper lobe emphysema and a shift of the mediastinal structures to the right. Two-dimensional computed tomography (CT) exhibited left upper lobe emphysema and right upper lobe atelectasis. Three-dimensional (3D) spiral CT angiography showed a bilateral tracheal bronchus. The left tracheal bronchus branch was compressed between the descending aorta and the ductus arteriosus. After a right arteriopulmonary shunt operation, the patient’s respiratory condition improved dramatically, with spontaneous closure of the ductus arteriosus. Subsequently, 3D-CT clearly exhibited the disappearance of tracheal compression. This combination of bilateral tracheal bronchus and congenital heart anomaly is extremely rare. The 3D-CT is a powerful noninvasive means for dynamically demonstrating the special relationships of arterial and tracheal anomalies.     

The efficiency of interference of Potato virus X infection depends on the target gene

RNA silencing is a natural defense response against viral infection. This phenomenon has been used to interfere with viral infections by exploiting fragments of viral genomes as sources of RNA silencing. Agrobacterium-mediated transient expression of a hairpin RNA derived from the TGBp1 gene of Potato virus X (PVX) induced RNA silencing of the TGBp1 gene and resulted in interference of PVX infection. The interference was induced in the infiltrated leaves but not in the upper non-infiltrated leaves. Transient expression of a CP hairpin RNA also induced interference of PVX. The TGBp1 hairpin RNA showed more efficient interference of PVX infection than the CP hairpin RNA.     

Readiness potential and movement initiation in the rat

Cortical field potentials were recorded by electrodes implanted chronically on the surface and at a 2.0 mm depth in various cortical areas in the left hemisphere in the rat during self-paced movements of the right forelimb. A surface-negative (s-N), depth-positive (d-P) cortical field potential appeared about 1.0 s (range: 0.5-1.5 s) before movement onset in the rostral (RFA) and caudal (CFA) forelimb areas of the motor cortex, and the somatosensory cortex, but not in the occipital cortex. Bipolar recording of electromyographic activities induced by the electrical stimulation of various cortical loci was also performed by pairs of steel electrodes inserted in the face, trunk, forelimb and hindlimb muscles on both sides. The stimulation of the forelimb motor cortex activated the face and/or forelimb muscles, while that of the somatosensory cortex generally activated several body part muscles including the forelimb muscle. Stronger stimulus intensity was requested to elicit the activities of most of the ipsilateral muscles to the cortex stimulated than the contralateral ones. The minimum intensity for inducing the forelimb muscle activity was lowest in the CFA among cortical areas producing the activity. The stimulation of cortical loci in which the s-N, d-P potential was recorded could induce muscle activities in the forelimb contralateral to the stimulation. It is suggested that the s-N, d-P potential is the readiness potential for activating muscles to initiate movement in the rat forelimb.     

The role of regional research laboratories of the local government in administering regional EQA programs

In 1981, the Japanese Ministry of Health and Welfare revised the enforcement of regulations of the Medical Technologists' Act. The amendments stipulate that all independent laboratories are legally obliged to introduce laboratory quality assurance programs and are responsible for the quality of all test results. To ensure adherence to these regulations, regional research laboratories of local governments such as the Tokyo Metropolitan Research Laboratory of Public Health should conduct regional external quality assessment (EQA) programs. We did a survey, in the form of a questionnaire, of the regional research laboratories of public health across the country. We found that commitment to the regional EQA in almost all of these public laboratories is insufficient. The main problem is that restructuring of local governments has resulted in lower budgets and so they are short of human resources. Nationwide EQA programs are only able to detect gross errors and use invalid methods for evaluating routine performance. We conclude that the regional EQA should be further developed.     

Impact of hepatitis B virus X protein on the DNA damage response during hepatocarcinogenesis

Hepatocellular carcinoma (HCC) is one of the most prevalent and lethal cancers worldwide. The main HCC-associated diseases are chronic infections with hepatitis B virus (HBV) and hepatitis C virus (HCV), and HBV-associated HCC is still prevalent in Asia. Many studies have suggested that HBV X protein (HBX), which is the most common ORF integrated into the host genome, plays a crucial role in hepatocarcinogenesis. However, the accumulated evidence regarding HBX-mediated signaling pathways is not concordant, and it is difficult to understand the mechanistic nature of HBX-associated hepatocarcinogenesis. For example, HBX was reported to inactivate the early responses to DNA damage via p53-dependent and -independent pathways by interacting with several DNA damage-binding proteins and was also reported to sensitize cells to p53-mediated apoptosis via ataxia-telangiectasia and Rad3-related (ATR)-dependent signaling. HBX also interferes with the centrosome replication process, resulting in rearrangement of chromosomes with micronuclei. Moreover, HBX was found to sensitize protein kinases such as Ras/Raf/mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK), stress-activated protein kinase/NH2-terminal-Jun kinase (SAPK/JNK), protein kinase B (PKB/Akt), and Janus kinase/STAT (JAK/STAT), indicating that a variety of signaling pathways may be activated by HBX. In this review, we focus on the roles of HBX in DNA damage repair during HCC development, with a view to achieving a better understanding of the significance of HBX in the early steps of hepatocarcinogenesis.     

Immunohistochemical Mapping of TRK-Fused Gene Products in the Rat Brainstem

The TRK-fused gene (TFG in human, Tfg in rat) was originally identified in human papillary thyroid cancer as a chimeric form of the NTRK1 gene. It was since reported that the gene product (TFG) plays a role in regulating phosphotyrosine-specific phosphatase-1 activity. As shown in the accompanying paper, we produced an antibody to rat TFG and used it to localize TFG to selected neurons in specific regions. In the present study, we mapped the TFG-positive neurons in the brainstem, cerebellum, and spinal cord of rats. In the brainstem, neurons intensely positive for TFG were distributed in the raphe nuclei, the gigantocellular reticular nucleus, the reticulotegmental nucleus of the pons, and some cranial nerve nuclei such as the trigeminal nuclei, the vestibulocochlear nuclei, and the dorsal motor nucleus of the vagus. Purkinje cells in the cerebellum and motor neurons in the spinal anterior horn were also positive for TFG. These results provide fundamental data for studying the functions of TFG in the brain.