Integrated exome and transcriptome analysis prioritizes MAP4K4 de novo frameshift variants in autism spectrum disorder as a novel disease-gene association.

The application of next-generation sequencing (NGS) to clinical practice is still hampered by the ability to interpret the clinical relevance of novel variants and the difficulty of evaluating their effect in specific tissues. Here, we applied integrated genomic approaches for interrogating blood samples of two unrelated individuals with neurodevelopmental disorders and identified a novel neuro-pathogenic role for the Mitogen-Activated Protein Kinase 4 gene (MAP4K4). In particular, we identified two novel frameshift variants in coding exons expressed in the blood and neuronal isoforms. Both variants were predicted to generate non-sense-mediated decay. By transcriptome analysis, we simultaneously demonstrated the deleterious effect of the identified variants on the splicing activity and stability of MAP4K4 mRNA. Therefore, we propose MAP4K4 as a novel causative gene for non-syndromic and syndromic neurodevelopmental disorders. Altogether, we prove the efficacy of an integrated approach of exome and transcriptome sequencing in the resolution of undiagnosed cases by leveraging the analysis of variants in genes expressed in peripheral blood.

A "spoon full of sugar" helps the medicine go down: How a participant friendly version of a psychophysics task significantly improves task engagement, performance and data quality in a typical adult sample.

Few would argue that the unique insights brought by studying the typical and atypical development of psychological processes are essential to building a comprehensive understanding of the brain. Often, however, the associated challenges of working with non-standard adult populations results in the more complex psychophysical paradigms being rejected as too complex. Recently we created a child- (and clinical group) friendly implementation of one such technique - the reverse-correlation Bubbles approach - and noted an associated performance boost in adult participants. Here, we compare the administration of three different versions of this participant-friendly task in the same adult participants to empirically confirm that introducing elements in the experiment with the sole purpose of improving the participant experience, not only boosts the participant's engagement and motivation for the task but results in a significantly improved objective task performance and stronger statistical results.

Penetration of rufloxacin into the cerebrospinal fluid in patients with inflamed and uninflamed meninges.

Forty-four patients scheduled for lumbar puncture (LP) were recruited to determine the level of penetration of orally administered rufloxacin into cerebrospinal fluid (CSF). The patients were divided into three clinical groups: those with normal CSF (groups A(1d) and A(7d)), those with aseptic meningitis (group B), and those with bacterial meningitis (group C). Members of group A(1d) received a single 400-mg rufloxacin dose, while group A(7d), B, and C constituents had a multiple-dose regimen (one 400-mg dose, followed by one 200-mg dose daily for 6 days). LP was performed on group A(1d) members 5 h after they had received treatment, while for group A(7d) it was undertaken 5 h after administration of the last dose. For group B, LP was performed 5 h after the first and the last doses, whereas for group C it was undertaken after the first, fourth, and last doses. Concentrations of rufloxacin in simultaneously collected CSF and plasma samples were determined. Mean CSF/plasma rufloxacin concentration ratios ranged from 0.57 to 0.84, depending on the study group. A higher, but not statistically significant, degree of penetration into CSF was observed in patients with bacterial meningitis than in those with normal CSF or aseptic meningitis. These data indicate that rufloxacin diffuses efficiently into the CSF of patients with either inflamed or uninflamed meninges.

Absence of effect of rufloxacin on theophylline pharmacokinetics in steady state.

Several quinolone antibacterial agents are known to inhibit the metabolism of theophylline, with the potential to cause adverse events due to raised theophylline concentrations during coadministration. A randomized crossover study was therefore conducted with 12 healthy male volunteers (ages, 23 to 34 years; body weight, 64 to 101 kg) to evaluate a possible interaction between rufloxacin and theophylline. Both drugs were administered at steady state. Following the administration of an oral loading dose of 400 mg on day 1, rufloxacin was given orally at 200 mg once daily on days 2 to 7 during one period only. During both periods, 146 mg of theophylline was administered orally twice daily for 3 days (which were days 4 to 6 of the rufloxacin coadministration period) and intravenously once the next morning to test for an interaction. Theophylline and rufloxacin concentrations were measured by reversed-phase high-pressure liquid chromatography, the pharmacokinetics of theophylline at steady state following administration of the last dose were calculated by compartment-model-independent methods. To compare the treatments, analysis of variance-based point estimates and 90% confidence intervals (given in parentheses) were calculated for the mean ratios of the pharmacokinetic parameters from the test (rufloxacin coadministration) over those from the reference (theophylline without rufloxacin) period. These were as follows: maximum concentration at steady state, 1.01 (0.96 to 1.07); area under the concentration-time curve from 0 to 12 h, 0.98 (0.94 to 1.02); half-life, 0.99 (0.95 to 1.03); total clearance at steady state, 1. 02 (0.99 to 1.06); and volume of distribution in the elimination phase, 1.01 (0.97 to 1.05). In conclusion, rufloxacin did not affect theophylline pharmacokinetics at steady state. Therefore, therapeutic coadministration of rufloxacin and theophylline is not expected to cause an increased incidence of theophylline-related adverse events.

Acute liver injury related to the use of niperotidine.

BACKGROUND/AIMS: H2-receptor antagonists are widely used for the therapy of peptic disease, since they ensure a protracted and intense inhibition of gastric acidity. Niperotidine (piperonyl-ranitidine) is a new H2 blocking agent recently proposed for clinical use. METHODS: Twenty-five cases of acute hepatitis associated with the use of niperotidine were reported in Italy between March and August 1995. Intercurrent viral infections, recent drug and alcohol consumption and blood transfusions were excluded as causes. RESULTS: All patients showed an increase in the parameters of liver cell injury and the clinical symptoms of acute hepatitis. After withdrawal of the drug, all patients showed a good outcome, except one who developed a fulminant hepatitis and died from digestive tract bleeding. CONCLUSIONS: The absence of other causes of acute liver injury suggests that the observed liver injury may be a niperotidine-adverse reaction. Moreover, the lack of a relationship between the dose of the drug and the degree of liver damage, the variable latent period and the rarity and unpredictability of the injury are suggestive of an idiosyncratic reaction.

Rufloxacin once daily versus ciprofloxacin twice daily in the treatment of patients with acute uncomplicated pyelonephritis.

PURPOSE: We compare the bacteriological and clinical efficacy of rufloxacin and ciprofloxacin in patients with acute uncomplicated pyelonephritis. MATERIALS AND METHODS: A total of 110 outpatients was enrolled in a randomized, double-blind multicenter study and treated for 10 days with 200 mg. rufloxacin daily (after a loading dose of 400 mg. on day 1) or 500 mg. ciprofloxacin twice daily. Bacteriological and clinical efficacy was based on the accumulated outcomes assessed at the end of treatment, and at 2 and 4 to 6 weeks. RESULTS: The bacteriological and clinical success rates of rufloxacin and ciprofloxacin were comparable: 55.6% versus 58.8% and 74% versus 71%, respectively (95% confidence interval -28% to +22% and -20% to +25%, respectively). Both study medications were well tolerated. CONCLUSIONS: Rufloxacin once daily is a good alternative in the outpatient treatment of acute uncomplicated pyelonephritis.

Impact of rufloxacin and ciprofloxacin on the intestinal microflora in a germ-free mice model.

The impact of rufloxacin and ciprofloxacin on faecal microflora was investigated in an experimental germ-free mice model into which human gastrointestinal flora was introduced and stabilized. Animals received oral doses of 10, 40 or 80 mg rufloxacin/kg, 30 or 60 mg ciprofloxacin/kg once daily for 10 days, or no treatment. Microflora was studied before, during and 1 week after treatment. No significant effect ( < 2 log reduction, p > 0.05) on aerobic microflora was observed after rufloxacin as compared to controls, except for a marked decrease in enterobacteria and a slight decrease in enterococci, staphylococci and anaerobes at the highest dose. Both doses of ciprofloxacin significantly reduced enterobacteria and clostridia, while the lower dose slightly reduced peptococci, bifidobacteria and eubacteria. The depression of the intestinal ecosystem was reversible, except for enterobacteria at the higher ciprofloxacin dose. No bacterial resistance was detected after either treatment.

Randomized, double-blind comparison of single-dose regimens of rufloxacin and pefloxacin for acute uncomplicated cystitis in women. French Multicenter Urinary Tract Infection-Rufloxacin Group.

In a double-blind, randomized, multicenter study, 463 adult women with symptomatic acute uncomplicated cystitis were treated orally with either a 400-mg single dose of rufloxacin (n = 226) or an 800-mg single dose of pefloxacin (n = 237). Escherichia coli (78%) and Proteus mirabilis (7%) were the most common isolates from 350 patients with significant pretreatment bacteriuria (uropathogens, > or = 10(5) CFU/ml). In the intention-to-treat analysis of patients with significant pretreatment bacteriuria, 343 patients were assessed for bacteriological outcome and 345 were assessed for clinical outcome. The bacteriological cure rate was 88% in the rufloxacin group and 84% in the pefloxacin group (95% confidence interval [CI] for difference in proportions, -4 to 12%), while the clinical resolution rate was 85 and 84%, respectively (95% CI, -8 to 9%). The per-protocol analysis demonstrated that among the 264 assessable patients, the bacteriological cure rate obtained with rufloxacin at 4 weeks of follow-up was comparable to that with pefloxacin (91 versus 85%; 95% CI, -3 to 15%). Among 295 clinically assessable patients, the clinical resolution rate at 4 weeks of follow-up was 89% in the rufloxacin group and 88% in the pefloxacin group (95% CI, -6 to 10%). Potentially drug-related adverse events occurred in 19% of the rufloxacin patients and in 18% of the pefloxacin patients. A single oral dose of 400 mg of rufloxacin is as effective and safe as a single oral dose of 800 mg of pefloxacin for the treatment of acute uncomplicated cystitis in women.

In vitro antimycoplasmal activities of rufloxacin and its metabolite MF 922.

The in vitro activities of rufloxacin and its metabolite, MF 922, were compared with those of ofloxacin, ciprofloxacin, erythromycin, and minocycline against Mycoplasma pneumoniae, Mycoplasma hominis, Mycoplasma fermentans, and Ureaplasma urealyticum. Rufloxacin, MF 922, and ciprofloxacin shared similar activities against all mycoplasmas tested. (MICs for 90% of isolates tested [MIC90s], 0.5 to 4 micrograms/ml. Ofloxacin had the lowest MIC90s for U. urealyticum, M. fermentans, and M. hominis (MIC90s, 0.25 to 1 micrograms/ml) and erythromycin had the lowest MIC90 for M. pneumoniae (MIC90, 0.004 micrograms/ml).

In vitro activity of rufloxacin against Listeria monocytogenes, Legionella pneumophila, and Chlamydia trachomatis.

The in vitro activity of a new fluoroquinolone, rufloxacin, was determined against both clinical isolates and standard strains of Legionella pneumophila, Chlamydia trachomatis, and Listeria monocytogenes and compared to that of ciprofloxacin, ofloxacin, and erythromycin. Among the antibacterials tested erythromycin was the most active. Rufloxacin inhibited the growth of L. pneumophila at concentrations in the range of 0.06-0.25 mg/l, that of C. trachomatis at concentrations in the range of 2-4, and that of L. monocytogenes at concentrations in the range of 1-2 mg/l. Rufloxacin demonstrated approximately the same activity as ciprofloxacin against C. trachomatis, but twice as less potency as ofloxacin and erythromycin against L. pneumophila, L. monocytogenes, and C. trachomatis, and twice as less potency as ciprofloxacin against L. pneumophila.

Biliary excretion of rufloxacin in humans.

Rufloxacin is a new once-daily antibacterial fluoroquinolone with a long half-life. The aim of the present study was to evaluate the plasma and biliary kinetics and biliary and urinary excretion of rufloxacin in patients with extrahepatic cholestasis. Twelve patients with total external percutaneous transhepatic biliary drainage were given a single oral dose of 400 mg of rufloxacin. Plasma, bile, and urine samples and fractions were collected over 72 h after drug administration. Rufloxacin and its major metabolite, the N-desmethyl derivative, were measured by high-performance liquid chromatography. Maximum rufloxacin concentrations in plasma and bile (means +/- standard deviations) were 4.05 +/- 1.38 micrograms/ml and 8.24 +/- 7.16 micrograms/ml, respectively, and were reached in 4.2 +/- 3.0 h and 4.2 +/- 3.5 h, respectively. The terminal elimination half-life of rufloxacin in plasma was 45.1 +/- 13.5 h. Apparent plasma clearance was 31.3 +/- 10.5 ml/min, while biliary clearance was 0.4 +/- 0.2 ml/min and renal clearance was 12.7 +/- 6.0 ml/min. In 72 h, 0.9% +/- 0.8% of the dose given was recovered in bile and 27.2% +/- 12.0% was recovered in urine. Biliary concentrations exceeded the MICs of most common biliary tract pathogens for at least 24 h after administration. The broad antibacterial spectrum of rufloxacin and its high and prolonged biliary concentrations suggest that this drug may be useful for treatment of biliary tract infections.

Double-blind, comparative study of rufloxacin once daily versus amoxicillin three times a day in treatment of outpatients with exacerbations of chronic bronchitis.

In a double-blind, randomized, multicenter study, the efficacy and safety of two dosage schedules of rufloxacin once daily were compared with those of amoxicillin three times a day in the treatment of 192 outpatients with exacerbations of chronic bronchitis. Rufloxacin was given as a single oral dose of 400 mg on day 1 and single daily doses of 200 mg on the subsequent 9 days (n = 64) or as 300 mg on day 1 and then 150 mg daily for 9 days (n = 63); amoxicillin was given as 500 mg orally three times a day for 10 days (n = 65). Clinical and bacteriological assessments were carried out before treatment, between study days 3 and 5, and at days 1 and 8 after treatment. Pretreatment cultures were positive for 139 patients, the most frequently isolated pathogens being Streptococcus pneumoniae, Moraxella catarrhalis, and Haemophilus influenzae. Clinical success rates were comparable in the three groups (94, 95, and 98%, respectively), as were bacteriological success rates at the end of treatment (93, 95, and 91%, respectively) and at follow-up (88, 95, and 98%, respectively). The power to detect a significant 15% difference in cure rates was 74.9%. Follow-up bacteriological failures from pneumococcal infection were 18% in both rufloxacin groups combined and 5% in the amoxicillin group. The 200-mg dose regimen achieved average steady-state concentrations in plasma higher than did the 150-mg dose regimen (3.75 versus 2.72 micrograms/ml). Adverse events occurred in 11 and 13 patients, respectively, on rufloxacin and 8 on amoxicillin. This study shows that rufloxacin once daily ay be a possible option for the treatment of acute exacerbations of chronic bronchitis. The 200-mg daily oral dose preceeded by a loading dose of 400 mg displays a better pharmacokinetic profile than the lower dose.

Rufloxacin once daily versus ofloxacin twice daily for treatment of complicated cystitis and upper urinary tract infections. Italian Multicentre UTI Rufloxacin Group.

The efficacy and safety of rufloxacin once daily was compared with that of ofloxacin b.i.d. for therapy of complicated cystitis and upper urinary tract infections. Eighty-three patients were randomly assigned to receive rufloxacin as a loading dose of 400 mg on the first day, followed by 200 mg s.i.d., and 80 received ofloxacin 300 mg b.i.d. Both agents were administered orally for a median duration of eight days. Bacterial elimination rates after treatment were 90% for rufloxacin and 81% for ofloxacin. Half of the treatment failures occurred in patients with infections caused by uropathogens that became either less sensitive or resistant to the quinolones being studied. At a two-week follow-up, recurrences had not occurred in any of the rufloxacin patients and had occurred in 17% of the ofloxacin patients. Minor adverse reactions were reported by 12 and 13 patients, respectively. Rufloxacin once daily is as effective as ofloxacin b.i.d. for the treatment of complicated cystitis and upper urinary tract infections.

Cellular uptake and activity of rufloxacin in different cell types and professional phagocytic cells.

The penetration of the new quinolene rufloxacin into human professional phagocytic cells and different cell types was determined. The intracellular distribution was demonstrated in all cells studied. At 4 degrees C the transport of rufloxacin was reduced. An intracellular dose-dependent activity was demonstrated for rufloxacin in monocytes and granulocytes infected with E. coli and S. aureus. In all the experiments rufloxacin was able to eliminate all intracellular bacteria.

Effect of single doses of rufloxacin on the disposition of theophylline and caffeine after single administration.

Several reports indicate that quinolone antibiotics affect the pharmacokinetics of theophylline and caffeine. This study investigated the effects of one single oral dose of rufloxacin, a new quinolone antibacterial, on the pharmacokinetics of theophylline and caffeine, given as single oral doses. Twelve healthy male volunteers were randomly given one of the following treatments: treatment T: 300 mg theophylline; treatment T + R: 300 mg theophylline followed by 400 mg rufloxacin after 30 min; treatment C: 200 mg caffeine; treatment C + R: 200 mg caffeine followed by 400 mg rufloxacin after 30 min. Blood samples for determination of xanthine plasma levels were taken immediately before and 1, 2, 4, 6, 8, 10, 12, 24 and 48 hours after xanthine administration. Urine were collected before treatment and at 0-6, 6-12, 12-24 and 24-48 h intervals. There were no significant differences in the pharmacokinetic parameters or in urinary excretion after administration of either theophylline or caffeine when combined with rufloxacin. Sleep disturbances were reported by two subjects after both treatment T + R and C + R, and by four subjects after treatment T + R. The results of this study indicate that there is no significant pharmacokinetic interaction between a single oral dose of rufloxacin and single doses of theophylline and caffeine.

Inter- and intrasubject variabilities in the pharmacokinetics of rufloxacin after single oral administration to healthy volunteers.

Rufloxacin is a new long-acting, once-daily quinolone antibacterial agent. We evaluated inter- and intrasubject variations in pharmacokinetics of rufloxacin following oral administration of 400 mg (two capsules) under controlled conditions, at an interval of 2 weeks (periods I and II), to 12 healthy male subjects. Plasma and urine samples were collected up to 48 h after drug administration. Plasma drug levels determined by bioassay were higher than those measured by high-performance liquid chromatography, indicating that one or more active metabolites were formed. Individual high-performance liquid chromatography plasma rufloxacin concentrations were fitted with a one-compartment open model with first-order input. There were considerable variations in the plasma concentration-time profiles among subjects; for example, the elimination half-life in plasma varied from 14.6 to 95.5 h. However, pharmacokinetic parameters calculated for the two periods did not differ significantly. These results suggest that the intrasubject variation in the pharmacokinetics of rufloxacin is usually small in spite of the considerable intersubject variation.

In vitro activity of a new quinolone, rufloxacin, against nosocomial isolates.

The in vitro activity of rufloxacin (MF 934), a new broad-spectrum fluoroquinolone, was tested against 1,032 gram-positive and gram-negative clinical isolates and compared to that of five other compounds of this class. All quinolones except for ciprofloxacin had limited activity against group A and B streptococci and pneumococci (MIC 90% of 4-64 mg/l) and no activity against enterococci. Most species of the enterobacteriaceae and staphylococci were found to be sensitive to rufloxacin (MIC 90% of 0.5-8 and 2-8 mg/l). Like the other quinolones except for ciprofloxacin, rufloxacin was not active against Pseudomonas aeruginosa. The antibacterial activity of rufloxacin was affected only minimally by an increase in the bacterial inoculum or by alterations in the pH of the medium. In spite of the relatively higher MICs of rufloxacin compared to those of the other quinolones, its favorable pharmacokinetic properties may account for its good clinical efficacy.

Pharmacokinetics of rufloxacin in healthy volunteers after repeated oral doses.

Rufloxacin is a new broad-spectrum fluoroquinolone antibacterial agent. The pharmacokinetics and safety of rufloxacin were evaluated after repeated oral administration to healthy volunteers. The drug was administered once a day for 6 consecutive days following two different dose schedules. The first group of 11 subjects was given a loading dose of 300 mg on the first day and 150 mg on the subsequent 5 days. The second group of 12 subjects was given a loading dose of 400 mg and 200 mg for 5 days. Serum levels and urine concentrations of rufloxacin were determined by microbiological assay. A simultaneous fit of all data points for each subject was done according to a one-compartment open model. The drug was rapidly absorbed (absorption half-life 17 +/- 6 min in the 300 + 150 mg and 11 +/- 5 min in the 400 + 200 mg dose regimen group) and reached maximal serum concentrations (2.77 +/- 0.24 and 3.62 +/- 0.35 micrograms/ml) 4.2 +/- 0.4 and 4.0 +/- 0.9 h after the first administration. Steady-state serum concentrations (3.19 +/- 0.31 and 4.06 +/- 0.33 micrograms/ml) were reached in 3.7 +/- 0.7 and 4.5 +/- 0.4 days. Elimination half-lives were 29.5 +/- 2.4 and 36.0 +/- 2.8 h. Apparent volumes of distribution were 111 +/- 8 and 136 +/- 16 liters and apparent plasma clearances were 46 +/- 5 and 44 +/- 4 ml/min. Renal clearances were 18 +/- 3 and 17 +/- 2 ml/min.(ABSTRACT TRUNCATED AT 250 WORDS)

Bone involvement in primary hemochromatosis and alcoholic cirrhosis.

Biochemical indexes of bone metabolism, bone mineral density, and histomorphometry were evaluated in 14 male patients with noncholestatic cirrhosis due to primary hemochromatosis (six cases) or to chronic alcohol abuse (eight cases), and in 30 male controls of similar age. Alkaline phosphatase in alcoholic patients was significantly higher than in controls (mean +/- SD 50.4 +/- 33.7 vs 33.0 +/- 7.1 U/L, p less than 0.01), as was urinary hydroxyproline in both hemochromatotics and alcoholics (mean +/- SD, 44.3 +/- 8.4 and 40.4 +/- 16.8, respectively, vs 30.1 +/- 4.5 mg/g, p less than 0.001 and p less than 0.005). Bone mineral density was significantly lower in hemochromatotics than in alcoholics and controls (mean +/- SD, 591 +/- 90 vs 765 +/- 87 and 759 +/- 34 mg/cm2, respectively, p less than 0.005 and p less than 0.001). At bone biopsy, trabecular osteoporosis was observed in two hemochromatotics and four alcoholics, and osteomalacia was seen in another alcoholic. Overall densitometric and histomorphometric findings indicate a derangement of trabecular bone in both alcoholic and hemochromatotic cirrhosis, whereas cortical osteoporosis seems limited to hemochromatotic patients.

HLA typing in 67 Italian patients with idiopathic hemochromatosis and their relatives.

The frequency of HLA A3 and B7 antigens was significantly higher in 67 unrelated patients with idiopathic hemochromatosis (IH) than in 700 controls (62.7% vs 22.5%, p less than 10(-8) and 26.9% vs 9.3%, p less than 10(-3), respectively). A3 B7, A3 B35 and A3 B5 were significantly more frequent in 72 haplotypes linked to IH gene than in 278 control haplotypes. The prevalence of B35 and A3 B35 was significantly higher in IH patients from North-Eastern Italy than from other regions (60% vs 21%, p less than .05 and 54.5% vs 8.2%, p less than 0.0001, respectively). All 15 siblings HLA identical to the respective proband were homozygous for IH with variable expression of the disease, whereas minor abnormalities of iron-related indexes were present in 23% of heterozygous relatives. Homozygous-heterozygous mating probably occurred in three of 40 families, accounting for the overt disease in three offspring and in one HLA semi-identical sibling; however, in this last case the possibility of a recombination event cannot be excluded.