MicroRNAs and liver function.

MicroRNAs (miRNAs) are small, noncoding RNA molecules that regulate gene expression post-transcriptionally through complementary base pairing with thousands of messenger RNAs. Although the target genes and precise biological functions of individual miRNAs remain largely unknown, miRNAs are speculated to play important roles in diverse biological processes in both normal and pathological states. The liver is a vital organ that plays major roles in a number of physiological functions. Recent advances in the study of liver miRNAs using gene-modified mice or in vivo nucleic acid delivery to overexpress specific miRNAs or inhibit miRNA functions have revealed the crucial biological roles of individual miRNAs in physiologically essential liver functions in vivo. Because miRNA-based strategies are being applied to clinical therapeutics, the importance of precise knowledge of miRNA functions cannot be underestimated, not only from a scientific point of view, but also from a clinical perspective to make the most of such drugs and avoid unexpected harmful effects. The aims of this review are to describe current knowledge regarding both known and as-yet-undiscovered molecular aspects of the biological roles of miRNAs in the liver, with a special emphasis on lipid, glucose, drug, and iron metabolism as vital functions of the liver as well as important therapeutic targets.

Prevalence of human papillomavirus in oral gargles and tonsillar washings.

OBJECTIVES: Human papillomavirus (HPV) infection drives carcinogenesis in the oropharynx. No standard sampling or HPV detection methods for evaluating oropharyngeal HPV infection exist. The prevalence of oral HPV infection in Japan is unknown. MATERIALS AND METHODS: We examined 435 healthy Japanese individuals to address whether adding tonsillar washing to oral gargling would improve HPV detection. We compared HPV assessment using GENOSEARCH HPV31 versus nested PCR and direct sequencing. Associations between HPV infection and demographic and behavioral characteristics were examined. RESULTS: Most participants who were HPV-positive based on oral gargles were also HPV-positive based on tonsillar washings: 11 (64.7%) of 17 on nested PCR and 12 (70.6%) of 17 on GENOSEARCH HPV31. Although HPV infection was more prevalent in oral gargles followed by tonsillar washings than in oral gargles alone, the difference was not statistically significant (nested PCR, 4.8% vs. 3.9%, P = 0.46; GENOSEARCH HPV31, 5.3% vs. 3.9%, P = 0.33). The overall agreement between nested PCR and GENOSEARCH HPV31 was 98.6%, with 76.0% positive agreement. The overall prevalence of oral HPV infection in Japan was 5.7% (95% confidence interval, 3.9-8.3%). Men had a significantly higher prevalence of oral HPV infection than women (8.3% vs. 2.6%, P = 0.02). Infection increased with number of lifetime sexual partners (P < 0.001 for trend). CONCLUSION: The oropharynx is probably the major source of HPV-infected cells in oral gargles. Oral gargling could be a standard sampling method for evaluating oropharyngeal HPV infection. GENOSEARCH HPV31 could be an option for oral HPV detection.

Metabolic tumor volume of metastatic lymph nodes and survival after total laryngectomy in laryngeal and hypopharyngeal cancer.

OBJECTIVES: The prognostic value of metabolic tumor volume (MTV) in locally advanced laryngeal or hypopharyngeal cancer is established in the setting of chemoradiotherapy, while it remains unknown in the setting of upfront total laryngectomy. MATERIALS AND METHODS: We retrospectively analyzed 88 patients receiving total laryngectomy and neck dissection, using Cox regression models. RESULTS AND CONCLUSION: Variables related to metastatic lymph node were associated with overall survival, whereas those related to primary tumor were not. In multivariable models, MTV of metastatic lymph nodes (N-MTV) as a continuous variable (Akaike's information criterion (AIC), 277.5) was equivalent to pathological nodal status (AIC, 278.2; P = 0.40), and superior to pathological nodal classification as an ordinal variable (AIC, 281.4; P < 0.05) in ability of predicting death. The risk of death was increased by 1.2-fold (95% confidence interval (CI), 1.0-1.4; P = 0.03) every 10-ml increment of N-MTV, while patients with pN+ disease were at a higher risk of death by 2.9-fold (95% CI, 1.0-12.2; P < 0.05) compared with patients with pN0 disease. Using recursive partitioning analysis (RPA), we classified the patients as having a low, intermediate, or high risk of death on the basis of N-MTV and extranodal extension (ENE). This RPA classification system exhibited greater concordance with overall survival than the classification considering pathological nodal status and ENE (AIC, 275.8 versus 281.4; P = 0.02). In the setting of upfront total laryngectomy, N-MTV is a critical predictor of mortality. A staging system in which N-MTV is incorporated may better inform adjuvant treatment decisions.

A role of mast cells for hepatic fibrosis in primary sclerosing cholangitis.

We encountered four patients with overt primary sclerosing cholangitis (PSC) which were histologically classified into stage 2 or 3. We examined the expression of stem cell factor (SCF), a ligand of c-kit, in injured bile ducts by immunohistochemistry, and mast cells were identified by immunohistochemistry using anti-HMCT (human mast cell tryptase) and anti-c-kit antibodies to clarify their relation with portal fibrosis coincident with destroyed bile ducts. SCF was detected in the epithelia of most bile ducts in PSC, and many HMCT- and c-kit-positive mast cells were found in portal tracts. Image analysis showed more significant numbers of c-kit-positive mast cells per area of portal tract in PSC than in chronic hepatitis C, and they might increase from stage 2 to 3. c-Kit-positive cells infiltrated into the portal tracts with SCF-positive destroyed bile ducts, and c-kit mast cells should be investigated in detail to make a role for portal fibrosis in PSC.

Characterization of DNA rearrangements of N-myc gene amplification in three neuroblastoma cell lines by pulsed-field gel electrophoresis.

We characterized N-myc gene amplification in three human neuroblastoma cell lines (IMR-32, TGW, GOTO). Rearrangements in long-range regions surrounding amplified N-myc genes were examined by pulsed-field gel electrophoresis. Since rare-cutting enzymes completely digested DNA at the middle of the N-myc gene, we were able to construct a physical map upstream and downstream of the germline N-myc gene, and to obtain information on restriction sites surrounding amplified N-myc genes. This method enables us to envisage the organization of amplified units over a long range. Digestion patterns differed considerably among the germline and the three cell lines, but were simple in each case. We estimated that the minimal distance between neighboring N-myc genes is at least several hundred kilobases. Our data suggest that amplification units contain several DNA fragments derived from different loci, but that they are homogeneous.

Effects of ritodrine, a beta 2-adrenoceptor agonist, on smooth muscle cells of the myometrium of pregnant rats.

1 Effects of ritodrine on the electrical and mechanical activities of the myometrium of pregnant rats were investigated in relation to cyclic adenosine 3',5'-monophosphate (cyclic AMP) production and the actions of isoprenaline. 2 In the longitudinal muscle cells of the 22nd day of gestation, ritodrine (greater than 10(-8) M) hyperpolarized the membrane, reduced the membrane resistance and suppressed the amplitude of contraction. Increased concentrations of ritodrine (greater than 10(-7) M) suppressed the generation of spikes and increased the amount of cyclic AMP produced. These actions of ritodrine were much the same as those of isoprenaline but the dose-response curves of all parameters were shifted to the right. 3 The effects of ritodrine on the circular muscle cells at the 22nd day of gestation were similar to those observed in the longitudinal muscle cells. 4 In the circular muscle cells at the 17-19th days of gestation, ritodrine produced neither a hyperpolarization nor an increase in cyclic AMP production (up to 10(-5) M) but did reduce the plateau potential and did relax the tissue (greater than 10(-7) M). 5 In the longitudinal muscle cells, during the 17-19th days of gestation, the responses of the tissues were similar to those observed in circular muscles on the 22nd day of gestation. These actions of ritodrine were suppressed by propranolol (10(-6) M). 6 These results indicate that ritodrine is a beta-adrenoceptor agonist and its mechanism of action similar to that of isoprenaline. However, the potency of ritodrine was approximately 100 times less than that of isoprenaline.

Moyamoya disease associated with renovascular hypertension.

Findings at the autopsy of a 7-year-old Japanese girl with moyamoya disease associated with renovascular hypertension are reported. There was disruption of a saccular aneurysm located at the junction of the left posterior cerebral and the posterior communicating arteries. Intimal thickening of carotid, coronary, and renal arteries, similar to findings in cases of intimal hyperplasia of fibromuscular dysplasia, was observed. These findings indicate that moyamoya disease may actually be an intracranial manifestation in a systemic arterial disorder.

Bronchoscopic and angiographic comparison of bronchial arterial lesions in patients with hemoptysis.

In seven patients with nonmalignant disease, we bronchoscopically observed various types of bronchial arterial lesions that may have caused hemorrhage. Five of the seven showed a bulging lesion, and the other two demonstrated an intrabronchial mass. We also examined these seven patients using selective bronchial arteriography. Herein we report our comparative study of the bronchoscopic findings and the bronchial arteriograms of these bronchial arterial lesions. The bulge observed in bronchoscopy corresponded either to an aneurysm or to a hypervascular area in the bronchial arteriogram. The mass lesions corresponded to a hypervascular area or a focal dilatation in the bronchial arteriogram. The intrabronchial lesions observed bronchoscopically either disappeared or were significantly diminished by bronchial arterial embolization for management of the hemorrhage. A histologic examination in two patients who underwent surgery revealed vascular lesions corresponding to the intrabronchial lesions in bronchoscopy. The results of this comparative study have important application in the bronchoscopic examination of bronchial arterial lesions in patients with hemoptysis.

Anomalous unilateral single pulmonary vein mimicking pulmonary varices.

Anomalous single pulmonary veins (AUSPV) is a rare anomaly of the pulmonary venous system. Six cases have been documented in the literature, and two additional are reported here. Since AUSPV mimics pulmonary varices on pulmonary angiography, careful interpretation of the pulmonary venous anatomy by bidirectional and stereoscopic angiography is mandatory for diagnosing this anomaly.

Recurrent bleeding after arterial embolization in patients with hemoptysis.

Thirty-three patients with hemoptysis caused by nonmalignant diseases underwent arterial embolization. In seven (21.2 percent) of 33 patients, hemoptysis recurred after initial embolization in a follow-up period ranging from one day to four years. Although there was no difference in age and daily quantity of bleeding among patients with and without recurrences, those with mycetoma suffered the highest recurrence of bleeding after initial embolization (three [75 percent] of four patients). In the initial arteriograms of the seven patients were found six cases of markedly increased vascularity, three of increased vascularity in the bilateral lung, and three of nonbronchial systemic arterial supply to the diseased lung. Of six patients who received reexamination at the time of recurrent bleeding, the arteriogram demonstrated recanalization of previously embolized artery in five and revascularization by collateral circulation in five. In addition, hemoptysis recurred in three (60 percent) of five patients after second embolization and in one (50 percent) of two after third embolization. Four patients underwent surgical therapy: two after the initial embolization, one after the second, and one after the third, and these patients had no recurrence. While arterial embolization as initial treatment of hemoptysis is a highly useful procedure, this is a palliative procedure and potential for recurrence of hemoptysis exists as the lesion that has initially caused hemoptysis is not cured by the embolization. We emphasize that a combination therapy of repeated embolization and surgery will probably improve the efficacy of treatment of recurrent bleeding after initial embolization.

A cellular uptake of cis-platinum-encapsulating liposome through endocytosis by human neuroblastoma cell.

To explore the mechanisms involved in the efficient intracellular delivery of an antitumor agent, the uptake mechanism of cis-diamminedichloroplatinum (CDDP)-encapsulating liposome (phosphatidylcholine:phosphatidylserine:cholesterol = 6:2:3 in molar ratio) (CDDP-liposome) by the human neuroblastoma cell, IMR-32, was investigated. Dansylcadaverine, chloroquine and antimycin A suppressed the growth-inhibitory activity of CDDP-liposome mostly due to the interference of the energy-dependent internalization process. Colchicine and vinblastine also neutralized the DNA synthesis-inhibitory activity of CDDP-liposomes. Stimulation with CDDP-liposomes at low temperature (4 degrees C), which induces the disruption of microtubules under immunofluorescent visualization, markedly lowered the growth-inhibitory activity of CDDP-liposomes. These findings suggested that the action of CDDP-liposome was temperature-dependent and microtubules participated in the intracellular drug delivery. A23187 (0.4 ?M), further, promoted the action of CDDP-liposome, suggesting that an increase of intracellular Ca(2+) levels promotes the growth inhibitory activities of CDDP-liposomes. Taking the uptake-mechanisms and intracellular delivery processes of CDDP-liposomes into consideration, the use of CDDP-liposomes may present a promising tool for neuroblastoma chemotherapy.

Phase II study of a new combined primary chemotherapy regimen, intravenous methotrexate and vincristine and intraarterial adriamycin and cisplatin, for locally advanced urinary bladder cancer: preliminary results.

A phase II study of a new combination therapy was performed using intraarterial (i.a.) cisplatin and Adriamycin in combination with i.v. methotrexate and vincristine for 27 patients with invasive urinary bladder carcinoma of stages T2-3NOMO, and the therapeutic effects were assessed. Methotrexate (20 mg/m2) was given i.v. on days 1,15, and 22, and vincristine (0.7 mg/m2) was injected i.v. on day 2 before i.a. infusion therapy and on days 15 and 22. The i.a. chemotherapy was performed after both superior gluteal arteries had been embolized using 3- or 5-mm stainless-steel coils. A mixture of cisplatin (50-70 mg/m2) and Adriamycin (20 mg/m2) was infused i.a. via both internal iliac arteries over a period of 20-30 min. Angiotensin II (mean dose, 21 micrograms) was simultaneously infused i.a. in 15 of 27 patients. In 24 of the 27 patients, at least 2 cycles of full-dose chemotherapy were completed. The dose was decreased in the remaining 3 patients because of their poor health status and advanced age. Among the 27 patients, 9 and 14 had complete (CR) and partial responses (PR), respectively; 3 manifested no change (NC), and 1 had progressive disease (PD). The objective response rate (CR+PR) was 85.2%. Among the 27 patients staged T2-3 NOMO, 6 (CR, 1; PR, 5) underwent total cystectomies and 18 (CR, 8; PR, 8; NC, 2) had transurethral resection of a bladder tumor (TUR-Bt) or partial resections following chemotherapy. The remaining 3 diminished-dose patients had no surgery. Of the 27 patients, 22 were alive after a median follow-up period of 21+ (range, 7-48+) months. No significant side effect was observed except for lower extremity paresthesias in 5 patients (18.5%). These results point to the effectiveness of this therapy and to the possibility of urinary bladder preservation in patients with invasive, advanced urinary bladder cancers.

The value of CT in diagnosis and prognosis of different inborn neurodegenerative disorders in childhood.

Inborn errors of metabolism in 40 children have been investigated by computed tomography to obtain data on the degree of cerebral involvement in neurodegenerative and storage disorders: 20 children had various mucopolysaccharidoses, 8 sphingolipidoses , 3 mucolipidoses, 2 oligosaccharidoses , 3 ceroidlipofuscinoses and 4 had various leucodystrophies . Diagnosis in all patients except Alexander's disease was established by biochemical or histological means. The main findings on CT were cerebral atrophy with enlargement of the ventricles and the subarachnoid spaces and hypodensity of the white matter. The degree of cerebral atrophy seemed to develop according to the age of the patients, as could be seen from the patients with mucopolysaccharidosis III, metachromatic leucodystrophy and GM1-gangliosidosis. Hypodensity of the white matter was found in mucopolysaccharidosis I-H, II-B, VI, in mucolipidosis II and in patients with leucodystrophies . On the other hand, there was great variability in these CT findings even in siblings, as seen in four patients with mucopolysaccharidosis VI. Among the series there were several patients who did not show any abnormalities in CT, so that a negative CT did not exclude these disorders, even the leucodystrophies . CT features such as cerebral atrophy or hypodensity were helpful in the evaluation of these disorders, though a diagnosis could not be made by CT alone.

Cranial computed tomography in disorders of complex carbohydrate metabolism and related storage diseases.

Computed tomography (CT) was performed on 34 children with different disorders of complex carbohydrate metabolism and related storage diseases to obtain data on the degree of cerebral involvement. The main findings on CT were cerebral atrophy and hypodensity of the white matter. There was a great variability in these CT findings, even in siblings. Among the patients there were several in whom CT was normal, so a negative study does not exclude one of these disorders. These findings show that CT features such as cerebral atrophy or hypodensity are helpful in the evaluation of these disorders, though a diagnosis cannot be made on the basis of CT alone.

Autopsy report of primary CNS B-cell lymphoma indistinguishable from multiple sclerosis: diagnosis with the immunoglobulin gene rearrangements analysis.

We report a case of primary CNS B-cell lymphoma indistinguishable from multiple sclerosis (MS). MRI of the head showed the spontaneous disappearance of the white matter lesions and the progressive cerebral atrophy. The brain biopsy failed to make a diagnosis of CNS lymphoma but rather suggested MS. Although the primary CNS lymphoma was suspected at autopsy, the immunohistochemical study showed the CNS-infiltrating lymphoid cells comprising both T-cells and B-cells. Analysis of the immunoglobulin and T-cell receptor gene rearrangements first provided evidence of primary CNS B-cell lymphoma.

Spontaneous resolution of multiple hemangiomas of the brain. Case report.

A case of multiple intracerebral hemangiomas is reported. The lesions were angiographically hypervascular and accompanied by marked perifocal brain edema. After removal of the main lesion and postoperative steroid administration, the rest of the lesions resolved spontaneously. Histologically, the lesion was composed of capillaries and fibromyxoid stroma. The distinctive clinical, radiological and histological features of this lesion are discussed in contrast to several other types of intracranial vascular tumors.

Effects of oral repetitive loading of disopyramide on acute-onset atrial fibrillation with concurrent monitoring of serum drug concentration.

We investigated the efficacy and safety of oral repetitive loading of disopyramide, for the termination of acute-onset (i.e., therapy started within 2 days after the onset of palpitations) atrial fibrillation (AF) in 96 consecutive patients, with concurrent monitoring of the serum concentration of this agent in fifteen of the patients. Outpatients with AF verified by standard electrocardiogram (ECG) were hospitalized and received disopyramide (200 mg) every 4-6 h, with a maximal dose of 800 mg daily, until the termination of AF under ECG monitoring was obtained. Conversion to sinus rhythm occurred within the first day of treatment in 88 patients (92%), on the second day of treatment in six patients (6%), and on the third and fifth days of treatment in the remaining two patients. No major adverse effects, such as hypotension, congestive heart failure, proarrythmic events or systemic embolism were noted. The serum levels of disopyramide evaluated in fifteen of the enrolled patients were found to be maintained within the therapeutic range throughout the treatment period. In spite of the absence of any placebo-controlled group in this study, these findings suggest that repetitive oral loading of disopyramide (200 mg) with an interval of 4-6 h is effective and safe for the termination of acute-onset AF under a stable therapeutic serum drug concentration, hence offering the possibility of self-medication for patients with episodic AF.

Intracranial venous angiomas.

Publications in the scientific literature are controversial in regard to the clinical significance of intracerebral venous angiomas. The present study of 11 patients with venous angiomas underscores the clinical importance of these lesions as potential causes of cerebral hemorrhage and obstructive hydrocephalus. The clinical and radiographic manifestations in 9 of these 11 patients correlated well. In cases of venous angiomas, the venous phase of angiography must be carefully scrutinized, particularly in patients with subarachnoid or intracerebral hemorrhage. Some diagnostic problems related to the angiography of venous angiomas are discussed.

Alterations in the properties of the rat myometrium during gestation and post partum.

The relationships among progesterone and oestradiol contents in the sera, membrane activities, catecholamine actions and contents of cyclic nucleotides were investigated in the rat myometrium during gestation and post partum, and under conditions of hormone treatment. Progesterone contents declined and oestradiol contents increased in the sera on the 21st day of gestation. During the late stage of gestation, oestradiol related to regularity, and progesterone to an irregularity, in the spike generation of myometrial cells. In the longitudinal muscle, catecholamines dominantly activated the beta-adrenoceptors during gestation, but activation of the alpha-adrenoceptors was seen during and after delivery, while in the circular muscle, alpha-adrenoceptor dominancy altered to beta-adrenoceptors by application of catecholamines appeared simultaneously. At the late stage of gestation, oestradiol treatment rather than progesterone increased the cyclic AMP contents, in both muscle layers. In the light of the results, alteration in the myometrial properties during the last stage of gestation and delivery cannot be explained solely by progesterone withdrawal phenomena.