Incidental prostate cancer prevalence at radical cystoprostatectomy--importance of the histopathological work-up.

The reported incidental prostate cancer prevalence rates at radical cystoprostatectomy cover a range from 4 to 60 %. We investigated the influence of the histopathological work-up on prostate cancer prevalence rates. We identified 114 patients who had undergone cystoprostatectomy for bladder cancer between 2000 and 2012. Complete histopathological assessment was defined as follows: (i) complete embedding of the prostate gland, (ii) sectioning of 15 or more prostate sections, and (iii) processing as whole mount slides. Prostate cancer prevalence rates derived from complete and incomplete histopathological assessments were compared. The overall prostate cancer prevalence rate was 59.6 %. A mean of 14.4 macroscopic tissue sections (thickness 3-5 mm) were sectioned. Sectioning ≥15 sections resulted in a prostate cancer detection rate of 75 %, compared to 42.6 % when sectioning <15 sections (p < 0.001). Complete embedding yielded a prostate cancer detection rate of 72.3 and of 23.1 % for partly embedded prostates (p < 0.0001). Prostate cancer was detected in 68.8 % of the whole mounted samples and in 38.2 % of the samples sectioned as standard slides (p < 0.01); according to the criteria described by Epstein and Ohori, 44.1 % of the detected prostate cancers were clinically significant. The quality of the histopathological work-up significantly influences prostate cancer detection rates and might at least partially explain the highly variable reported incidental prostate cancer prevalence rates at cystoprostatectomy (CP). The high proportion of significant prostate cancer found in our series calls for a careful surgical approach to the prostate during CP.

[To start eating again could be dangerous]. / Wieder zu essen, kann gefährlich sein.

A 36-year-old woman with anorexia nervosa and resulting malnutrition decided to change her life and eat properly. One day after the refeeding start, instead of feeling better she felt much worse: fatigue, nausea and leg swelling occurred. She consulted our emergency department. A «refeeding syndrome¼ has been diagnosed. In this case report we discuss causes, clinical presentation and treatment of this illness. The aim is to make practitians aware about the existing of this disease even with oral refeeding. Warning signs have to be interpreted correctly in order to avoid dramatic consequences.

Assessing analytical variability of measurement of vitamin A in corn-soy blend.

Two multilaboratory investigations were conducted by SUSTAIN to assess variability in the measurement of vitamin A, the marker used to verify levels of vitamin premix addition to enriched/fortified food aid products, including the widely distributed corn-soy blend (CSB). CSB specifications identify AACC Approved Method 86-06 or equivalent methods for vitamin A analysis, however there is no requirement to demonstrate equivalency. CSB samples with known and blinded levels of vitamin A and a reference standard were analyzed by 16 laboratories using their respective methods. Calculated coefficients of variation across all laboratories and methods for unknown samples and reference standard were 35 and 7.1%, respectively, suggesting the largest source of variation is the vitamin extraction procedure. Laboratories generally overestimated low levels and underestimated high levels of vitamin A within the range of 6000 and 16 000 IU/lb. Only two laboratories demonstrated excellent internal precision (+/- 300 IU vitamin A/lb) and reported values within 95% confidence interval for all blinded samples. Results of this study have implications both for quality control in food aid products (due to the use of vitamin A as a marker) and for regulatory oversight of vitamin A content in commercial food products.

Impact of sample storage on detection of periodontal bacteria.

BACKGROUND/AIMS: Information on the impact of sample storage prior to analysis by DNA methods is limited. The aim of this study was to investigate the effect of subgingival sample storage on bacterial detection and enumeration. MATERIAL AND METHODS: Subgingival plaque samples were studied by a) checkerboard DNA-DNA hybridization by immediate processing, b) storage at + 4 degrees C for 6 weeks, c) storage at - 20 degrees C for 6 months or d) storage at - 20 degrees C for 12 months. RESULTS: No differences in total DNA were found between protocol 1 and 2, or between protocol 3 and 4. Protocol 1 yielded 2.4 times more total bacterial DNA than did protocol 3 (P < 0.001). Actinobacillus actinomycetemcomitans and Campylobacter gracilis were detected in 21.1% of the immediately processed samples but only in 6.6% of the samples after 12 months of storage. Similar changes were noticed for Treponema denticola, which was detected in 22.3% and 9.2%, respectively. Streptococci spp., Fusobacterium nucleatum and Tannerella forsythia did not seem to be affected by storage. In contrast, the level of Campylobacter rectus detection frequency changed from 2.6% if processed immediately to 15.8% if samples were stored for 12 months. CONCLUSIONS: In longitudinal clinical studies including microbiological samples and processed with DNA-DNA hybridization methods, samples should be stored for the same period of time before processing to avoid loss of microbiological information.

Recombinant AAV vectors containing the foot and mouth disease virus 2A sequence confer efficient bicistronic gene expression in cultured cells and rat substantia nigra neurons.

Recombinant adeno-associated viruses (rAAVs) are promising vectors for gene therapy since they efficiently and stably transduce a variety of tissues of immunocompetent animals. The major disadvantage of rAAVs is their limited capacity to package foreign DNA (< or =5 kb). Often, co-expression of two or more genes from a single viral vector is desirable to achieve maximal therapeutic efficacy or to track transduced cells in vivo by suitable reporter genes. The internal ribosome entry site (IRES) sequence of encephalomyocarditis virus has been widely used to construct bicistronic viral vectors. However, the IRES is rather long and IRES-mediated translation can be relatively inefficient when compared with cap-dependent translation. As an alternative to the IRES for in vivo gene expression, we studied the 16 amino-acid long 2A peptide of foot and mouth disease virus (FMDV). The 2A peptide mediates the primary cis-'cleavage' of the FMDV polyprotein in a cascade of processing events that ultimately generate the mature FMDV proteins. We have generated several different rAAV genomes in which two coding regions are fused in-frame via the FMDV 2A sequence. We show that FMDV 2A efficiently mediates the generation of the expected cleavage products from the artificial fusion proteins in cells. Furthermore, we find that both EGFP and alpha- synuclein are expressed at substantially higher levels from 2A vectors than from the corresponding IRES-based vectors, while SOD-1 is expressed at comparable or slightly higher levels. Finally, we demonstrate for the first time, that the 2A sequence results in effective bicistronic gene expression in vivo after injection of 2A-dependent rAAVs into the rat substantia nigra. We conclude that 2A-containing rAAVs may represent an attractive alternative to IRES-dependent vectors for ex vivo and in vivo gene expression and gene therapy.

Characterization of VX on concrete using ion trap secondary ionization mass spectrometry.

The nerve agent VX (O-ethyl S-2-diisopropylaminoethyl methyl phosphonothiolate) was analyzed on the surface of concrete samples using an ion trap secondary ion mass spectrometer (IT-SIMS). It was found that VX could be detected down to an absolute quantity of 5 ng on a concrete chip, or to a surface coverage of 0.0004 monolayers on crushed concrete. To achieve these levels of detection, the m/z 268-->128 ion fragmentation was measured using MS2, where m/z 268 corresponds to [VX + H]+, and 128 corresponds to a diisopropylvinylammonium isomer, that is formed by the elimination of the phosphonothiolate moiety. Detection at these levels was accomplished by analyzing samples that had been recently exposed to VX, i.e., within an hour. When the VX-exposed concrete samples were aged, the SIMS signature for intact VX had disappeared, which signaled the degradation of the compound on the concrete surface. The VX signature was replaced by ions which are interpreted in terms of VX degradation products, which appear to be somewhat long lived on the concrete surface. These compounds include ethylmethylphosphonic acid (EMPA), diisopropyl taurine (DIPT), diisopropylaminoethanethiol (DESH), bis(diisopropylaminoethane) disulfide [(DES)2], and a particularly tenacious compound that may correspond to diisopropylvinylamine (DIVA), or an isomer thereof. It was found that the thiolamine-derived degradation products DIPT, DESH, and (DES)2 were removed with isopropyl alcohol extraction. However, the DIVA-related degradation product was observed to strongly adhere to the concrete surface for longer than one week. Although quantitation was not possible in this set of experiments, the results clearly show the rapid degradation of VX on concrete, as well as the surface sensitivity of the IT-SIMS for intact VX and its adsorptive degradation products.

Potentiating effect of (2-[2-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl)methyl]-phenyl]ethenyl) -phosphonic acid (MDL 74,428), a potent inhibitor of purine nucleoside phosphorylase, on the antiretroviral activities of 2',3'-dideoxyinosine combined with ribavirin in mice.

2',3'dideoxyinosine (ddI) has potent activity against human immunodeficiency virus (HIV) but is rapidly metabolized by erythrocytic purine nucleoside phosphorylase (PNP), and therefore has a very short plasma half-life in rodents, monkeys and in patients with acquired immunodeficiency syndrome. It is now reported that 100 microM (2-[2-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl)methyl]-phenyl]ethenyl) - phosphonic acid (MDL 74,428), a very potent inhibitor of PNP blocks the intracellular phosphorolysis of ddI in cultured human red blood cells, in T leukemic CEM lymphoblasts and prolongs ddI plasma effective concentration in mice at a dose of 250 mg/kg body weight given i.p. In MDL 74,428-treated CEM cells, despite marked reduction of ddI catabolism, neither further accumulation of ddATP, the active antiviral metabolite of ddI, nor potentiation of the activity of ddI against HIV cytopathogenicity is observed. MDL 74,428 does not also affect the inhibitory effect of ddI combined with ribavirin on the transformation in vitro of C3H/3T3 cells by Moloney murine sarcoma virus (MSV). In mice, on the contrary, MDL 74,428 (200 mg/kg body weight, i.p.) is effective at potentiating the effect of ribavirin used either alone, or combined with ddI on MSV-induced tumour formation and associated mortality. However, in the absence of ribavirin, co-administration of MDL 74,428 with ddI affords, no chemotherapeutic advantage.

Congenital intracranial cysts: clinical findings, diagnosis, treatment and follow-up. A multicenter, retrospective long-term evaluation of 72 children.

A multicenter, retrospective evaluation of 72 children born between 1973 and 1987 with intracranial cysts was performed: 37 had an arachnoid cyst, 25 a Dandy-Walker malformation and 10 a porencephalic cyst. The following criteria were evaluated: history, clinical findings, diagnostic work-up (neuroradiology, CSF and intracystic pressures), surgical therapy and follow-up (outcome and cyst volumes).

Spermidine nuclear acetylation in rat hepatocytes and in logarithmically growing rat hepatoma cells: comparison with histone acetylation.

Spermidine acetylation has been studied in nuclear homogenates and in entire nuclei from rat hepatocytes and rat hepatoma tissue culture (HTC) cells, isolated at different stages of logarithmic growth, and compared to histone acetylation. Under all experimental conditions, N8-acetylspermidine was the predominant product of the reaction (90%). Unlike histone, spermidine acetylation in HTC cell and hepatocyte entire nuclei was almost absent or strikingly reduced relative to acetylation using nuclear homogenates as the enzyme sources. This was due to the lack of a free minor pool of spermidine, most likely lost during the purification of entire nuclei. Thus, preincubation of intact nuclei in the presence of spermidine restored activities to values observed using nuclear sonicates. Spermidine acetylation in HTC cell nuclei fluctuated moderately during cell growth, being stimulated immediately after initiation of proliferation and decreasing progressively as cultures reached high cell density. This pattern corroborated that of N8-acetylspermidine intracellular accumulation induced by culturing cells in the presence of 1 mM 7-amino-2-heptanone, a competitive inhibitor of N8-acetylspermidine deacetylase. Histone acetylation during HTC cell growth was not markedly different qualitatively from that of spermidine. Moreover, spermidine and histone acetylations in hepatocyte nuclei were of the same order of magnitude as those seen in rat hepatoma cell nuclei. Finally, inhibition of deacetylation of N8-acetylspermidine had no apparent deleterious effects on cell and growth. It remains to be determined whether the acetylation step is of higher physiological importance, in particular, and as discussed in nuclear spermidine turnover.

Effect of polyamine deprivation on the survival of intracranial glioblastoma bearing rats.

It has previously been shown that systematic polyamine deprivation results in the almost complete inhibition of the growth of several solid tumors. The same polyamine deficient diet (containing antibiotics for the decontamination of the gastrointestinal tract, the ornithine decarboxylase inhibitor 2-(difluoromethyl)ornithine, and the polyamine oxidase inhibitor N1, N4-bis-(2,3-butadienyl)putrescine; "drug-containing polyamine deficient chow", DC-PDC) was applied for the first time to the treatment of rats with an intracranial tumor. Rats received intracortical grafts of C6 rat glioblastoma cells, and the length of their survival was determined. Treatment with DC-PDC, starting four days after tumor cell inoculation, significantly prolonged the median survival of the glioblastoma-bearing rats. The results underline the general growth inhibitory effect of systematic polyamine deprivation. Since the effect of polyamine restriction on tumor growth is reversible, combinations with cytotoxic drugs have to be found which exploit the changed functions of polyamine deficient tumor cells.

Comparative evaluation of the antitussive activity of butamirate citrate linctus versus clobutinol syrup.

In a double-blind randomized study 60 patients with either irritative cough due to seasonal respiratory disorders or chronic cough of any etiology were treated with either butamirate citrate linctus (Sinecod, Zyma) or with clobutinol syrup (Silomat, Boehringer, Ingelheim) for a period of 5 days at a dose regimen of 3 tablespoons daily. Efficacy was assessed based on the reduction of the severity as well as frequency of the cough and on the global opinion of the physician. Both groups showed highly significant improvements for the severity and frequency parameters (p less than 0.001), thus demonstrating the effectiveness of both treatments. No significant differences between groups were detected globally for the whole collective. For cough due to carcinomas (n = 14), however, a significantly better effect of butamirate on the frequency of cough (p = 0.026) was found which originated other significant differences in the global scores (p = 0.013) and in the physician's opinion (p = 0.026). Seven patients in both groups complained about side effects (mainly nausea and drowsiness).

[NAAGA versus disodium cromoglycate in perennial allergic rhinopathy: results of a multicenter study]. / Naaga versus Dinatrium-Cromoglykat bei perennialer allergischer Rhinopathie: Ergebnisse einer Multizenterstudie.

The efficacy and tolerability of Naaga (the magnesium salt of N-acetyl-aspartyl-glutamate) and of sodium cromoglycate after intranasal application were tested and compared in 100 patients suffering from chronic allergic rhinitis. The study was conducted according to a randomized, double-blind design in four centers. The intensity and frequency of typical allergic symptoms such as nasal obstruction, sneezing, runny and itching nose, conjunctivitis, ocular itching and tear flow were recorded on a 4-point scale (absent, light, moderate and severe). The intensity of nasal and ocular symptoms was found to be marked for patients in two centers. For the patients in the other two centers the drugs' efficacy was difficult to assess because symptoms were not marked (rated either as absent or light) at the beginning of the study. Thus the assessment of efficacy was based on 60 patients (centers 1 and 2), while the assessment of tolerability was based on the whole patient sample (n = 100). There was no statistically significant difference between the two compounds in terms of efficacy. A total of five patients in each of both groups complained of moderate untoward effects (all patients from the same center).

[Effectiveness and tolerance of the C3 convertase inhibitor, N-acetyl-aspartyl-magnesium glutamate with anti-allergic action. Results of a double-blind study]. / Wirksamkeit und Verträglichkeit des C3-Konvertasehemmers N-Acetyl-Aspartyl-Magnesiumglutamat mit antiallergischer Wirkung. Ergebnisse einer doppelblinden Phase-II-Studie.

N-Acetyl-aspartyl magnesium glutamate (Rhinaaxia, NAAGA) is a topically active antiallergic dipeptide. The compound acts in two different ways. On the one hand NAAGA inhibits the mast cell-degranulation, on the other hand this compound blocks the activation of the C3-convertase, subsequently followed by a blocked cleavage of the fragments C3a and C5a, respectively. 20 patients suffering from pollinosis were treated for 2 weeks according to a randomized double-blind placebo-controlled study. Besides subjective complaints nasal obstruction was objectively documented via rhinomanometria. 9 out of 10 patients under placebo had to use the rescue drug tritoqualine, a histidine decarboxylase inhibitor, compared to none in the verum group (p less than 0.01). After 14 days of treatment with NAAGA the nasal peak flow rate increased by 21.5 l/min and 21.8 l/min in the tritoqualine/placebo group, respectively (not significant). Nasal obstruction improved statistically significantly after 7 and 14 days of treatment in both groups. Tolerance was reported to be good in either group.

[Effectiveness and tolerance of the C3 convertase inhibitor, N-acetyl-aspartyl-magnesium glutamate in perennial rhinitis. Results of a double-blind, placebo-controlled study]. / Wirksamkeit und Verträglichkeit des C3-Konvertasehemmers N-Acetyl-Aspartyl-Magnesiumglutamat bei perennialer Rhinitis. Ergebnis einer doppelblinden, Plazebo-kontrollierten Untersuchung.

N-Acetyl-aspartyl magnesium glutamate (Rhinaaxia, NAAGA) is a new antiallergic substance for topical use. The compound covers a bilateral way of action. There is inhibition of the mast cell-degranulation and blocking of the C3-convertase, subsequently followed by a blocked cleavage of the fragments C3a and C5a, respectively. 20 patients suffering from perennial allergic rhinitis were treated according to a randomized double blind placebo-controlled study design. Apart from a nominal documentation of subjective complaints the degree of nasal obstruction was objectively rated via rhinomanometria. The nasal flow rate improved significantly in patients treated with NAAGA and subjective complaints decreased markedly. NAAGA showed to be well tolerated although 6 of 10 patients observed transient "nasal burning".

A comparison of the clinical utility of the radioimmunoassay, high-performance liquid chromatography, and TDx cyclosporine assays in outpatient renal transplant recipients.

Rapid and precise cyclosporine measurements are necessary to maximize immunosuppression and minimize toxicity. The new cyclosporine and metabolites TDx fluorescent polarization immunoassay was studied to determine its precision, sensitivity, and stability. Further, it was compared with the existing radioimmunoassay and high-performance liquid chromatography assays for clinical utility. The TDx procedure utilizes 50 microliter of serum or blood. The range of the assay is 0-1000 ng/ml for serum and 0-2000 ng/ml for whole blood. Precision studies of three control levels provided coefficients of variation of 2.1-5.8% for both assays. The sensitivities of the assays were less than 25 ng/ml for serum and less than 50 ng/ml for whole blood. In order to compare the TDx with the currently used RIA and HPLC methods, 362 simultaneous whole blood and serum samples were obtained from 122 renal transplant recipients over a 4-month period. The serum and whole blood TDx assays excluded fewer patients than either the RIA or HPLC assays with regard to sensitivity. The mean serum cyclosporine concentration for samples above the sensitivity was as follows: TDx 120 +/- 5 ng/ml, RIA 116 +/- 4 ng/ml, and HPLC 100 +/- 5 ng/ml. The mean whole-blood cyclosporine concentration for samples above the sensitivity was as follows: TDx 585 +/- 19 ng/ml, RIA 543 +/- 14 ng/ml, and HPLC 178 +/- 5 ng/ml. Serum and blood TDx levels correlated well with RIA levels, with regression coefficients of r = 0.813 and r = 0.897, respectively. Serum and blood TDx levels did not correlate strongly with HPLC levels, with regression coefficients of 0.332 and 0.781, respectively. Seventeen patients were diagnosed as having acute cyclosporine nephrotoxicity. The serum and whole-blood cyclosporine concentrations in these patients were at the upper end of the therapeutic range for all analytical methods. Five patients had acute cellular rejection; serum and whole-blood cyclosporine levels in these patients did not differ significantly from the stable patients when measured by each of the analytical methods. In conclusion, the TDx cyclosporine and metabolites assay provides reliable blood and serum concentrations that correlate well with RIA measurements in renal transplant recipients. This assay offers rapid sample turn-around times making possible same-day results for all patients without putting great demands upon the laboratory.

An evaluation of the cyclosporine and metabolites whole blood TDx assay.

The new Abbott TDx cyclosporine and metabolites fluorescent polarization immunoassay procedure provides a 20-min sample turn-around time, using 50 microliters of sample for the analysis of cyclosporine in whole blood. A precipitation agent and a lysing agent are utilized as a pretreatment step. The range of the whole blood assay is from 0 to 2,000 ng/ml, with a sensitivity of 50 ng/ml. Precision studies at 3 control levels provided coefficients of variation of 2.1-4.8% for both assays. In order to compare this assay with the currently used Sandoz polyclonal radioimmunoassay (RIA) method. 200 whole blood samples were obtained from 20 renal, cardiac, and hepatic transplant recipients. The mean whole blood cyclosporine concentrations for samples above the sensitivity level were as follows: TDx 754 ng/ml (+/- 31) and RIA 619 ng/ml (+/- 22). Blood TDx levels correlated strongly with RIA levels, with a regression coefficient of r = 0.915. This new assay provides reliable blood cyclosporine concentrations that correlate well with RIA measurements. This assay offers rapid sample turn-around times, making same-day results for outpatient drug monitoring possible.

Chronopharmacodynamics and kinetics after symmetric and asymmetric multiple theophylline doses in patients with chronic obstructive pulmonary disease.

In a randomized double-blind crossover study, the effects of 4 days of symmetric or asymmetric dosing with sustained-release theophylline (Sabidal-SR) were compared with placebo in ten patients with chronic obstructive pulmonary disease in a stable state. The doses were 450 mg b.i.d. and 270 mg plus 630 mg, respectively. Special attention was paid to circadian rhythms in pharmacokinetics and pharmacodynamic effects, during 24 h of measuring. The peak plasma theophylline concentrations at night with a symmetric dosage were lower than with an asymmetric dosage: 11.5 mg/l (+/- 3.8) and 13.6 mg/l (+/- 4.4), respectively. Both theophylline regimens caused a significant increase in vital capacity (8%) and in forced expiratory volume in one second (15%), and also a decrease of specific viscous work of breathing (49%). There were no significant differences in pharmacodynamic effects between the symmetric and asymmetric dosing regimen. An exception was the heart rate, that increased significantly only with an asymmetric dose. We found no correlation between the theophylline plasma concentration and the changes in lung function and heart rate. Theophylline had no influence on subjective findings of the patients, nor on the oxygen saturation of the blood during sleep.

[NAAGA and cromoglycate. A comparison in the treatment of seasonal allergic rhinitis]. / NAAGA und Cromoglycat. Ein Vergleich in der Behandlung der saisonalen allergischen Rhinitis.

A multicentre double-blind clinical trial comparing two nasal solutions of naaga (magnesium salt of N-acetyl aspartyl glutamic acid-rhinaaxia) and of disodium cromoglycate (LOMUPREN) has been performed on 91 patients suffering from an acute symptomatic seasonal rhinitis treated for 28 days. Nasal, ocular and bronchial symptoms as well as side effects were recorded by the physician at the control visits and daily by the patient in a diary. Haematological parameters, blood chemistry and serum levels of non specific IgE were also measured. The results show: On the whole, there was no significant difference between groups, neither for the global appreciation of the therapeutic effect as assessed by the physician and the patient, nor for the daily evolution of the symptoms based on the patient's daily notebook. Both treatments were very well tolerated. In a selected population with IgE greater than 150 U/ml, there was a significantly better activity of naaga (p less than 0.05) on the disappearance of the ocular symptoms. The nasal symptoms appeared to improve significantly better in the cromoglycate group (p less than 0.05).