miRNA-regulated transcription associated with mouse strains predisposed to hypnotic effects of ethanol.

INTRODUCTION: Studying innate sensitivity to ethanol can be an important first step toward understanding alcohol use disorders. In brain, we investigated transcripts, with evidence of miRNA modulation related to a predisposition to the hypnotic effect of ethanol, as measured by loss of righting reflex (LORR). METHODS: Expression of miRNAs (12 samples) and expression of mRNAs (353 samples) in brain were independently analyzed for an association with LORR in mice from the LXS recombinant inbred panel gathered across several small studies. These results were then integrated via a meta-analysis of miRNA-mRNA target pairs identified in miRNA-target interaction databases. RESULTS: We found 112 significant miRNA-mRNA pairs where a large majority of miRNAs and mRNAs were highly interconnected. Most pairs indicated a pattern of increased levels of miRNAs and reduced levels of mRNAs being associated with more alcohol-sensitive strains. For example, CaMKIIn1 was targeted by multiple miRNAs associated with LORR. CAMK2N1 is an inhibitor of CAMK2, which among other functions, phosphorylates, or binds to GABAA and NMDA receptors. CONCLUSIONS: Our results suggest a novel role of miRNA-mediated regulation of an inhibitor of CAMK2 and its downstream targets including the GABAA and NMDA receptors, which have been previously implicated to have a role in ethanol-induced sedation and sensitivity.

Increased hepatocellular protein carbonylation in human end-stage alcoholic cirrhosis.

OBJECTIVE: Oxidative stress is a significant contributing factor in the pathogenesis of alcoholic liver disease (ALD). In the murine models of chronic alcohol consumption, induction of oxidative stress results in increased peroxidation of polyunsaturated fatty acids to form highly reactive electrophilic α/ß unsaturated aldehydes that post-translationally modify proteins altering activity. Data are presented here suggesting that oxidative stress and the resulting carbonylation of hepatic proteins is an ongoing process involved in alcohol-induced cirrhosis. METHODS: Using age-matched pooled hepatic tissue obtained from healthy humans and patients with end stage cirrhotic ALD, overall carbonylation was assessed by immunohistochemistry and LC-MS/MS of streptavidin purified hepatic whole cell extracts treated with biotin hydrazide. Identified carbonylated proteins were further evaluated using bioinformatics analyses. RESULTS: Using immunohistochemistry and Western blotting, protein carbonylation was increased in end stage ALD occurring primarily in hepatocytes. Mass spectrometric analysis revealed a total of 1224 carbonylated proteins in normal hepatic and end-stage alcoholic cirrhosis tissue. Of these, 411 were unique to cirrhotic ALD, 261 unique to normal hepatic tissue and 552 common to both groups. Bioinformatic pathway analysis of hepatic carbonylated proteins revealed a propensity of long term EtOH consumption to increase post-translational carbonylation of proteins involved in glutathione homeostatic, glycolytic and cytoskeletal pathways. Western analysis revealed increased expression of GSTA4 and GSTπ in human ALD. Using LC-MS/MS analysis, a nonenaldehyde post-translational modification was identified on Lysine 235 of the cytoskeletal protein vimentin in whole cell extracts prepared from human end stage ALD hepatic tissue. CONCLUSIONS: These studies are the first to use LC-MS/MS analysis of carbonylated proteins in human ALD and begin exploring possible mechanistic links with end-stage alcoholic cirrhosis and oxidative stress.

Genetics of gene expression characterizes response to selective breeding for alcohol preference.

Numerous selective breeding experiments have been performed with rodents, in an attempt to understand the genetic basis for innate differences in preference for alcohol consumption. Quantitative trait locus (QTL) analysis has been used to determine regions of the genome that are associated with the behavioral difference in alcohol preference/consumption. Recent work suggests that differences in gene expression represent a major genetic basis for complex traits. Therefore, the QTLs are likely to harbor regulatory regions (eQTLs) for the differentially expressed genes that are associated with the trait. In this study, we examined brain gene expression differences over generations of selection of the third replicate lines of high and low alcohol-preferring (HAP3 and LAP3) mice, and determined regions of the genome that control the expression of these differentially expressed genes (de eQTLs). We also determined eQTL regions (rv eQTLs) for genes that showed a decrease in variance of expression levels over the course of selection. We postulated that de eQTLs that overlap with rv eQTLs, and also with phenotypic QTLs, represent genomic regions that are affected by the process of selection. These overlapping regions controlled the expression of candidate genes (that displayed differential expression and reduced variance of expression) for the predisposition to differences in alcohol consumption by the HAP3/LAP3 mice.

Epithelial salivary glands neoplasms in children and adolescents: a forty-four-year experience

Epithelial neoplasms of salivary gland origin are relatively uncommon in children and adolescents. Over a 44-year period, there were 38 cases affecting children under 19 years of age in our Pediatric Hospital-Based Tumor Registry. Medical charts of 38 patients with epithelial neoplasms of salivary glands were reviewed. Data collected included demographic, clinical, and histological characteristics. Statistical analysis included descriptive statistics, Student t-test, and Kaplan-Meier method was used for survival analysis. The mean age was 11.8 years. There was a female preponderance of 1.9:1. The parotid gland was affected in most cases (65.8%). Twenty-seven patients had malignant tumors and eleven patients presented benign neoplasms. Pleomorphic adenoma was the most frequent benign tumor (7 out of 11) and mucoepidermoid carcinoma was the most frequent malignancy (17 out of 27). Five-year overall survival rate was 81.6% for patients with malignant tumors. Grade of differentiation was the only significant prognostic factor for patients with mucoepidermoid carcinomas. Epithelial salivary gland tumors are very rare in children. Surgery is the best option to achieve high cure rates and radiotherapy must have precise indications because of their long-term side effects in young age.

Experience with six children with fetal rhabdomyomatous nephroblastoma: review of the clinical, biologic, and pathologic features.

BACKGROUND: Fetal rhabdomyomatous nephroblastoma (FRN) is a rare variant of Wilms tumor. MATERIALS AND RESULTS: One hundred and thirty two children with kidney tumors were seen at our hospital from 1985 to 1993. Among them were 6 (4.5%) who had FRNs. Five were boys aged 8 months to 3 years; the girl was 17 months old. Three of the four with unilateral disease had tumors so large that they were considered unresectable at diagnosis. Five received pre-operative chemotherapy and three also received pre-operative radiation therapy. None of the tumors responded. Both patients with bilateral tumors died of progressive disease. Three of the four patients with unilateral disease followed for at least one year are alive for 1 to 10 years after diagnosis. CONCLUSIONS: FRN should be in the differential diagnosis of huge kidney tumors in children, and preoperative therapies escalated with caution since FRN is not responsive to treatment used for classic Wilms tumor.

Experience with six children with fetal rhabdomyomatous nephroblastoma: review of clinical, biologic, and pathologic features

Fetal rhabdomyomatous nephroblastoma (FRN) is a rare variant of Wilms tumor. One hundred and thirty two children with kidney tumors were seen at our hospital from 1985 to 1993. Among them were 6 (4.5%) who had FRNs. Five were boys aged 8 months to 3 years; the girl was 17 months old. Three of the four with unilateral disease had tumors so large that they were considered unresectable at diagnosis. Five received pre-operative chemotherapy and three also received pre-operative radiation therapy. None of the tumors responded. Both patients with bilateral tumors died of progressive disease. Three of the four patients with unilateral disease followed for at least one year are alive for 1 to 10 years after diagnosis. FRN should be in the differential diagnosis of huge kidney tumors in children, and preoperative therapies escalated with caution since FRN is not responsive to treatment used for classic Wilms tumor.

Renal tumors in children under one year of age.

Among 202 primary renal tumors of childhood collected over 33 year period, 22 (10.8%) occurred in children under one year of age. Fifteen were of classic triphasic Wilms' tumor. Three were partially cytodifferentiated variants of Wilms' tumor. The remaining 4 cases were distributed among the mesenchymal renal tumors: congenital mesoblastic nephroma, cellular mesoblastic nephroma, clear cell sarcoma and malignant rhaboid tumor. The morphology of these tumors is presented. Our findings and the review of the literature show the wide morphologic spectrum of these neoplasms and the importance of recognizing them as distinct clinicopathologic entities.

Fetal rhabdomyomatous nephroblastoma. Report of a case with unusual clinical behavior.

The authors report a case of fetal rhabdomyomatous nephroblastoma that presented pulmonary metastases at diagnosis, a feature infrequently observed in this type of tumor.

Fetal rhabdomyomatous nephroblastoma: report of a case with unusual clinical behavior

The authors report a case of fetal rhabdomyomatous nephroblastoma that presented pulmonary metastases at diagnosis, a feature infrequently observed in this type of tumor.

Odontogenic adenomatoid tumor of the mandible (adenoameloblastoma).

Adenoameloblastoma may recur when treated incompletely by curettage. In the rare case of recurrence, an extensive and mutilating resection may become necessary. A correct diagnosis must be made in cases of atypical ameloblastomas in order to avoid unnecessary radical surgery. In doubtful cases, biopsy with histopathologic examination is indicated. Acknowledgment: We thank Mrs. Frieda Werebe for organizing this manuscript.