MicroRNA profiling in complete and partial hydatidiform moles.

INTRODUCTION: Hydatidiform mole is one of the gestational trophoblastic disease and comprises complete (CM) and partial moles (PM), which carries a risk of developing persistence disease, invasive mole or choriocarcinoma. MicroRNAs (miRNAs) have been discovered in various tissues, including neoplastic tissues. Its role in the pathogenesis of molar pregnancy or as biomarkers are still largely uncertain. The aim of this study is to identify the differentially expressed miRNAs in CM and PM. MATERIALS AND METHODS: Using next-generation sequencing, the miRNAs profiles of CM (n=3) and PM (n=3) moles, including placenta of non-molar abortus (n=3) as control were determined. The differentially expressed miRNAs between each group were analysed. Subsequently, bioinformatics analysis using miRDB and Targetscan was utilised to predict target genes. RESULTS: We found 10 differentially expressed miRNAs in CMs and PMs, compared to NMAs, namely miR- 518a-5p, miR-423-3p, miR-503-5p, miR-302a-3p, and miR-1323. The other 5 miRNAs were novel, not listed in the known database. The 3 differentially expressed miRNAs in CMs were predicted to commonly target ZTBT46 and FAM73B mRNAs. DISCUSSION: miR-518 was consistently observed to be downregulated in CM versus PM, and CM versus NMA. Further bioinformatic analysis to provide insight into the possible role of these miRNAs in the pathogenesis of HMs, progression of disease and as potential diagnostic biomarkers as well as therapeutic targets for HMs is needed.

Impact of temozolomide chemotherapy on seizure frequency in patients with low-grade gliomas.

OBJECT: Seizures occur in approximately 80% of patients with low-grade gliomas (LGGs). The majority of patients are treated with anticonvulsant monotherapy; however, many patients require multidrug therapy, or their seizures are refractory to antiepileptic drugs altogether. The oral alkylating agent temozolomide has emerged as a potential initial treatment option for LGG. A few reports suggest an association between temozolomide and reduced seizure frequency in patients with intractable epilepsy. METHODS: Using their clinical database, the authors identified adult patients whose LGGs were treated using temozolomide as the initial antineoplastic therapy at the University of Virginia Health System. As a control group, the authors assessed patients whose LGGs were under observation. All patients had seizure disorders that were treated with anticonvulsants. Seizure frequency in patients with intractable epilepsy was analyzed before and after treatment with temozolomide. Age at diagnosis, sex, antiepileptic drugs, pathological subtype, surgical treatment, and follow-up until progression were also assessed. Interval seizure frequency was meticulously analyzed at each neurooncology clinic visit. A meaningful difference in seizures was defined as a reduction in seizure frequency of greater than 50% per month. RESULTS: Thirty-nine patients were identified in the temozolomide cohort and 30 patients in the control cohort. The median age at diagnosis was 46 years for the former cohort and 41.5 years for the latter. The median length of follow-up was 39 months for the temozolomide group and 37 months for the control group. There was a significant difference in reduced seizure frequency between patients receiving temozolomide (59%) and those who did not receive temozolomide (13%, p < 0.001). Seven patients (18%) in the temozolomide group displayed this improvement independent of antiepileptic drug adjustment compared with no patient in the control group (p < 0.001). CONCLUSIONS: The authors' data suggest that a subset of patients with LGGs experience improvement in seizure frequency during treatment with temozolomide independent of antiepileptic drug adjustment. This decrease in seizure frequency appears independent of the natural history of seizures in patients whose tumors are under observation. Consequently, seizures in patients with LGGs may be better controlled with the combination of AEDs and temozolomide.

An acid phosphatase from Manihot glaziovii as an alternative to alkaline Phosphatase for molecular cloning experiments.

An acid phosphatase, free of deoxyribonuclease activity, was isolated from Manihot glaziovii leaves. It had a Mr of 78 kDa and was optimally active at pH 4.3 and 52 degrees C. It was inactivated at 65 degrees C over 15 min. It had a broad substrate specificity with strongest activity towards p-nitrophenyl phosphate. The enzyme dephosphorylated linearized pUC18 DNA and preventing self-ligation under the same conditions used for calf intestine alkaline phosphatase.

Assessing cyanogen content in cassava-based food using the enzyme-dipstick method.

The cyanogen content of 35 cassava-based foods was determined using the enzyme-dipstick method. The analyses showed presence of residual cyanogens in these products, and they ranged from 2 to 88 mg HCN equivalent/kg. Foodstuff prepared from grated cassava roots exhibited a lower level of cyanogen content (2-31 mg HCN equivalent/kg) compared with those prepared as cassava root slices or from cassava flour (28-88 mg HCN equivalent/kg). These results showed that current commercial processing methods did not remove all the cyanogen present from cassava roots and it might be worthwhile re-examining these processing practices. In this respect, the report showed that an overnight treatment of cassava root slices with linamarase and cellulase might facilitate the removal of cyanogens. This study also demonstrated the usefulness of enzyme-dipsticks for assessing cyanogen content in cassava food products.

Somatic embryogenesis, plant regeneration and cyanogenesis in Manihot glaziovii Muell. Arg. (ceara rubber).

The report describes a system for somatic embryogenesis and direct plant regeneration from the embryos of Manihot glaziovii. Somatic embryos were obtained by culturing young leaf lobes (3-6 mm long) adjacent to the apex in Murashige and Skoog medium containing 18 µM 2,4-dichlorophenoxy acetic acid for 20 days and then transferring them to a maturation medium with 0.5 µM 6-benzylaminopurine. Secondary embryogenesis was induced from cotyledonary segments of somatic embryos by using the same protocol as that for primary embryogenesis. For regeneration, somatic embryos were cultured in medium supplemented with 10-4 M kinetin and 53.4% of them developed into plantlets. Linamarin and linamarase were not detected in calli or in somatic embryos. Linamarin content was found to be highest in leaves of regenerated plantlets, followed by stem and root tissues. Levels of linamarase activity were almost the same in leaves and stem tissues and very low in roots.

Customisation of AFLP analysis for cassava varietal identification.

Amplified fragment length polymorphism (AFLP) markers were used in the characterization of eight cassava varieties. This nonradioactive AFLP system was customized in terms of the choice of restriction enzymes used and the selection of nucleotides added to the 3' end of primers. EcoRI/MseI and HindIII/MseI fragments generally gave monomorphic profiles while ApaI/TaqI fragments produced polymorphic profiles suggesting a genome with high G + C content. It was possible to identify the eight cassava varieties used in this study using CTG as selective bases at the TaqI primer. For cassava, the AFLP system provided a higher number of loci detected per run when compared to RAPD. The reliability accompanying AFLP analysis would thus make it suitable for the characterization of cassava varieties.

Applying disease management strategies to Medicare.

Medicare coverage begins for many when they have already developed one or more chronic diseases, and it often pays for the latest and costliest phases. Population-based disease modeling, patient screening, and monitoring would be appropriate interventions for chronic renal disease. Patients who have not yet advanced to end-stage renal disease would benefit from management of diabetes and hypertension, avoidance of nephrotoxic substances, and better preparation for dialysis. Administrative support could take the form of clinical guidelines, physician-led multidisciplinary teams, integrated delivery systems, provider and patient education, and new information technologies. Medicare reflects the long-term public perspective, and thus should further this new direction by supporting education, reimbursing for prevention efforts and allied health services, encouraging efficiency, and monitoring cost and quality outcomes.

Migration of a central venous catheter into pulmonary vein complicated with lung edema in a premature infant.

In this paper, we present a preterm infant with acute focal lung edema due to central venous catheter migration into the right pulmonary vein. The substances aspirated from the endotracheal tube contained the parenteral nutrition fluid. Thus, parenteral nutrition fluid extravasation into the lung or a vasculopulmonary fistula was suspected. The course of complication was smooth after the catheter was withdrawn into the right atrium. This unusual presentation during TPN hyperalimentation was described in order to prevent a potentially dangerous event.

Maintenance of employment on dialysis.

This article describes the components of a multidisciplinary effort focused on promoting, among other goals, continued employment during end-stage renal disease (ESRD) treatment. The education and guidance of the patient begin during the pre-ESRD period, intensify through dialysis treatment, and continue even through posttransplantation follow-up. Such focused programs support patients in retaining their usual lifestyle, staying in their current jobs where possible, and maximizing self-esteem and quality of life.

Benefits of a multidisciplinary predialysis program in maintaining employment among patients on home dialysis.

We evaluated the benefits of a multidisciplinary predialysis intervention to help home dialysis patients maintain employment. This program was shown to be successful with in-center hemodialysis patients in a case-controlled study published in the American Journal of Kidney Disease in September, 1993. We looked at 30 patients on home dialysis (28 on continuous ambulatory peritoneal dialysis and 2 on home hemodialysis) as of March, 1995. The age of the patients ranged from 28 to 63 years, with a mean of 46.8 years. Forty-three percent of the patients were diabetic. The patients went through a multidisciplinary predialysis program which consisted of: (1) psychosocial assessment, (2) education about dialysis and choice of modalities, (3) orientation to dialysis unit, and (4) counseling sessions with patient, significant family members, and others. During this time, patients were referred to the program approximately six months before beginning dialysis. There was continued collaboration among members of the multidisciplinary team. Of the 30 patients, 11 were already disabled when beginning dialysis. Of the 19 who were working once dialysis began, 14 maintained employment (3 diabetic), with 1 retiring and 4 becoming disabled. Therefore, 74% of patients maintained employment. In conclusion, employment was maintained by this predialysis multidisciplinary program.

Linamarin sensors: interference-based sensing of linamarin using linamarase and peroxidase.

An interference-based linamarin sensor is developed. Horseradish peroxidase (HRP) is adsorbed on a pyrolytic graphite (PG) electrode, and then linamarase from cassava is cross-linked with glutaraldehyde on the electrode surface. The prepared bienzyme electrode is poised at -300 mV vs Ag/AgCl for 40 s to reduce dissolved O(2) to H(2)O(2) at the PG surface. The potential is then stepped to 0 mV, at which point the accumulated H(2)O(2) is reduced, though the O(2) reduction does not proceed. Since the H(2)O(2) reduction is catalyzed by HRP, the transient cathodic current is inhibited by cyanide, which is liberated from linamarin by linamarase. Therefore, the transient current is a function of the linamarin concentration. This sensor responds to 1 × 10(-5)-5 × 10(-3) M linamarin and can estimate a linamarin concentration of a cassava extract.

Hypodensity of platelet serotonin uptake sites in posttraumatic stress disorder: associated clinical features.

We have previously reported that binding to blood platelets of paroxetine, a selective serotonin (5-HT) reuptake inhibitor which binds to 5-HT uptake sites, is decreased in patients with posttraumatic stress disorder (PTSD). Specifically, we found a lower number of platelet 3H-paroxetine binding sites (Bmax) and a lower dissociation constant (Kd) for 3H-paroxetine binding in combat veterans with PTSD compared to normal control subjects. In the current study we assessed the relationship of platelet 3H-paroxetine binding to clinical features in 41 Vietnam combat veterans with SCID-diagnosed PTSD. The results indicated that Bmax of platelet 3H-paroxetine binding was negatively correlated with both state and trait anxiety, as well as with depressive and overall PTSD symptoms. However, there was no evidence that platelet 3H-paroxetine binding differed as a function of comorbid psychiatric diagnoses including major depression, other anxiety disorders, and substance abuse in these patients.

Sources of fungal linamarases.

Forty-four strains of Aspergillus, Penicillium, Fusarium, Trichoderma and Rhizopus were grown on a liquid medium containing glucose and cassava-root extract. All of the Aspergillus and Fusarium strains, eight out of 10 Penicillium strains and three of seven Trichoderma strains showed linamarase activity. No such activity was detected in any Rhizopus strain. The crude enzyme preparation from F. oxysporum had the highest affinity for linamarin whereas that from A. nidulans was the most heat-stable.

Increased risk of renal cell cancer among women using diuretics in the United States.

Use of prescription diuretics and incidence of renal cell cancer have increased in the United States in the past 25 years. Recent interview-based epidemiologic studies have reported an association between diuretic use and renal cell cancer risk. Our study evaluated this hypothesis using, for the first time, medical records as the source of the information on prescription diuretic use. Using medical records of women from a prepaid health plan, we identified 191 cases and 191 controls matched on age, membership duration, and membership at diagnosis. Diuretics use and history of potential confounding factors were ascertained by a standardized review of the medical records of each subject, without reference to case or control status. There was a strong and statistically significant association between renal cell cancer and prescription diuretics (odds ratio [OR] adjusted for hypertension, smoking, and obesity = 2.9, 95 percent confidence interval [CI] = 1.7-4.7). Risk tended to increase with dose, measured by number of prescriptions. Since renal cancer can induce hypertension, which is treated by diuretics, and thereby confound the association with diuretics, we examined diuretic use 10 or more years prior to diagnosis when secondary hypertension would be unlikely. The OR for prescriptions 10 or more years before diagnosis was 3.5 (CI = 1.7-7.4). Our results support earlier reports of an excess risk of renal cell cancer among users of prescription diuretics and indicate need for further study to evaluate this relationship, especially due to the extensive use of diuretics and the increasing incidence of this cancer.

Platelet serotonin studies in hyperserotonemic relatives of children with autistic disorder.

Platelet serotonin (5-HT) studies were conducted with 12 hyperserotonemic and 12 normoserotonemic age-, sex-, and relationship-matched relatives of autistic probands. Each group consisted of 7 mothers, 4 fathers, and 1 sister of autistic children and adolescents. The density (Bmax) of platelet 5-HT2 receptor binding sites, labelled with [3H]-lysergic acid diethylamide (LSD), was significantly lower in 11 hyperserotonemic subjects compared to 12 normoserotonemic subjects (40.9 +/- 13.5 fmol/mg protein, 59.6 +/- 13.2; p < 0.004). The affinity (Kd) for [3H]-LSD binding did not differ. Although the density (Bmax) of [3H]-paroxetine binding did not differ between groups, there was a small difference in the affinity (Kd) for [3H]-paroxetine binding (hyperserotonemic 47.6 +/- 9.0 pM, normoserotonemic 54.8 +/- 12.1; p < 0.05). There were no significant differences in platelet 5-HT uptake, or in thrombin-stimulated 5-HT release. Basal, 5-HT-stimulated, and arginine-vasopressin (AVP)-stimulated inositol phosphate production, as well as basal, prostaglandin E1 (PGE1)-, and forskolin-stimulated cAMP production did not differ. There were significant correlations between whole blood 5-HT levels and LSD Bmax (rs = -0.63, N = 23, p < 0.002) and whole blood 5-HT levels and 5-HT uptake Vmax (rs = 0.56, N = 18, p < 0.02). However, [3H]-LSD labelled 5-HT2 binding and 5-HT uptake were not correlated with each other. Hyperserotonemia of autism may be heterogeneous with one subgroup of subjects with increased 5-HT uptake and another subgroup with decreased 5-HT2 binding.

Isolation and characterization of ß-glucosidases from Aspergillus nidulans mutant USDB 1183.

Two extracellular ß-glucosidases (cellobiase, EC 3.2.1.21), I and II, from Aspergillus nidulans USDB 1183 were purified to homogeneity with molecular weights of 240,000 and 78,000, respectively. Both hydrolysed laminaribiose, ß-gentiobiose, cellobiose, p-nitrophenyl-ß-L-glucoside, phenyl-ß-L-glucoside, o-nitrophenyl-ß-L-glucoside, salicin and methyl-ß-L-glucoside but not α-linked disaccharides. Both were competitively inhibited by glucose and non-competitively (mixed) inhibited by glucono-1,5-lactone. ß-Glucosidase I was more susceptible to inhibition by Ag(+) and less inhibited by Fe(2+) and Fe(3+) than ß-glucosidase II.

Theophylline produces over-additive relaxation of canine tracheal smooth muscle when combined with beta-agonists: the dose-response relationship.

The interaction between theophylline (T) and the beta-agonists albuterol (A) and isoproterenol (I) was examined using canine cervical tracheal smooth muscle devoid of epithelium contracted with 0.1 or 0.3 microM methacholine. Greater functional antagonism with beta-agonists vs. T was confirmed and an ability of T to potentiate beta-agonist relaxation was demonstrated. The EC50 for T increased from 0.13 +/- 0.02 to 0.37 +/- 0.07 mM (mean +/- SEM) in preparations contracted with 0.1 or 0.3 microM methacholine, respectively, while that for I increased from 0.036 +/- 0.008 to 0.17 +/- 0.03 microM, a significantly larger change (P < 0.025). In tissues contracted with 0.3 microM methacholine and pretreated with 10 micrograms/ml of T IC50 values from composite concentration-response curves for I and A were displaced to the left and Emax was increased (56.6 to 71.5% for I, 44 to 61% for A, P < 0.0002). Addition of 10 micrograms/ml T resulted in relaxations which exceeded that calculated by the fractional product method for additive, independent action (P < 0.0001 for I, P < 0.0002 for A at 0.3 microM methacholine), suggesting that at least part of T's action was over-additive. Five, 10 and 20 micrograms/ml T enhanced the effectiveness of single concentrations of I by factors of 1.47 +/- 0.14 (P < 0.05), 2.72 +/- 0.26 (P < 0.01) and 5.34 +/- 0.55 (P < 0.01), respectively, in preparations contracted with 0.1 microM methacholine: I enhanced the effectiveness to a lesser degree. Using two approaches, positive interaction or over-additivity between T and beta-agonists has been demonstrated.