Children who stutter show reduced action-related activity in the rostral cingulate zone.

Previous studies have indicated that children who stutter show not only speech-related problems, but also wider difficulties in self-control. In this study we test the novel hypothesis that children who stutter may experience difficulties with inhibitory control over voluntary actions. We used functional MRI to compare brain activity between children who stutter and children who do not stutter in a task that captures key cognitive aspects of voluntary action control. Participants performed a rolling marble task, in which they were instructed to press a key to stop a rolling marble from crashing on some of the trials (instructed action condition). They were also asked to choose voluntarily whether to execute or inhibit this prepotent response in other trials (volition condition). Children who stutter reported less motor and cognitive impulsivity and had shorter stop-signal reaction times when controlled for IQ, consistent with greater inhibition, compared to children who do not stutter. At the neural level, children who stutter showed decreased activation in the rostral cingulate zone during voluntary action selection compared to children who do not stutter. This effect was more pronounced for children who were rated as showing more stuttered syllables in the stutter screening, and was furthermore correlated with stop-signal reaction times and impulsivity ratings. These findings suggest that stuttering in childhood could reflect wider difficulties in self-control, also in the non-verbal domain. Understanding these neural mechanisms could potentially lead to more focused treatments of stuttering.

[Not every delirium protocol in Dutch hospitals is up-to-date: evaluation of the implementation of the new delirium guideline]. / Niet elk delierprotocol in ziekenhuizen is up-to-date.

OBJECTIVE: To describe the extent to which the essential changes in the new Dutch delirium guideline for adults and the elderly, published in April 2014, have been incorporated in local hospital protocols, so as to estimate the consequences this could have for patients. DESIGN: Quantitative study. METHOD: Dutch hospital protocols were collected for two periods: before (September to December 2012) and after publication of the guideline (March to July 2015). Protocols were compared with respect to basic delirium care (screening, diagnostic approach, therapy and follow-up care) and organisation of care. RESULTS: Of the 80 Dutch hospitals approached, we were able to include 57 (71%) protocols in this study. 16 hospitals (28%) had adapted their protocols to the new guideline. Screening for the risk of delirium using the questions from the Dutch safety management system (Veiligheidsmanagementsysteem) was described in 29 (51%) of the protocols. Use of the Delirium Observation Screening Scale was reported in 52 (91%) protocols. A policy of moderation regarding antipsychotic therapy was described in 12 of 53 (23%) protocols, but in 21/53 (40%) the haloperidol dosages were higher than advised by the guideline. Follow-up care is described in 40 (70%) of the protocols. Organisation of delirium care, for example restriction of consultation of an expert to complex cases, was advised in 33/57 (58%) protocols. CONCLUSION: 15 months after publication, only just over a quarter of the protocols incorporated the guideline. In terms of the treatment of patients with delirium, this may have led to unnecessary treatment with antipsychotics. Furthermore, basic delirium care is apparently still not considered as the responsibility of every medical specialist in the hospital.

Immune evasion cluster-positive bacteriophages are highly prevalent among human Staphylococcus aureus strains, but they are not essential in the first stages of nasal colonization.

The Staphylococcus aureus immune evasion cluster (IEC), located on ß-haemolysin-converting bacteriophages (ßC-Φs), encodes the immune-modulating proteins chemotaxis inhibitory protein, staphylococcal complement inhibitor (SCIN), staphylococcal enterotoxin A and staphylokinase. Its precise role in S. aureus colonization is unclear. We studied the presence of the IEC-carrying bacteriophages in human and animal S. aureus isolates, using PCR for the gene encoding SCIN (scn). Human isolates were obtained by collecting serial nasal swabs from 21 persistent carriers. S. aureus strains from 19 (90%) persistent carriers contained an IEC that was present and indistinguishable in 95% of cases at all five sampling moments over a 3-month period. Of the 77 infectious animal strains included in the study, only 26 strains (34%) were IEC-positive. Integration of these IEC-positive strains into an amplified fragment length polymorphism genotype database showed that 24 of 53 (45%) strains were human-associated and only two of 24 (8%) were 'true' animal isolates (p < 0.001). The high prevalence and stability of IEC-carrying ßC-Φs in human strains suggested a role for these ßC-Φs in human nasal colonization. To test this hypothesis, 23 volunteers were colonized artificially with S. aureus strain NCTC 8325-4 with or without the IEC type B-carrying ßC-Φ13. Intranasal survival was monitored for 28 days after inoculation. The strain harbouring ßC-Φ13 was eliminated significantly faster (median 4 days; range 1-14 days) than the strain without ßC-Φ13 (median 14 days; range 2-28 days; p 0.011). In conclusion, although IEC-carrying ßC-Φs are highly prevalent among human colonizing S. aureus strains, they are not essential in the first stages of S. aureus nasal colonization.

Heterogeneity of the humoral immune response following Staphylococcus aureus bacteremia.

Expanding knowledge on the humoral immune response in Staphylococcus aureus-infected patients is a mandatory step in the development of vaccines and immunotherapies. Here, we present novel insights into the antibody responses following S. aureus bacteremia. Fifteen bacteremic patients were followed extensively from diagnosis onwards (median 29 days, range 9-74). S. aureus strains (median 3, range 1-6) and serial serum samples (median 16, range 6-27) were collected. Strains were genotyped by pulsed-field gel electrophoresis (PFGE) and genes encoding 19 staphylococcal proteins were detected by polymerase chain reaction (PCR). The levels of IgG, IgA, and IgM directed to these proteins were determined using bead-based flow cytometry. All strains isolated from individual patients were PFGE-identical. The genes encoding clumping factor (Clf) A, ClfB, and iron-responsive surface-determinant (Isd) A were detected in all isolates. Antigen-specific IgG levels increased more frequently than IgA or IgM levels. In individual patients, different proteins induced an immune response and the dynamics clearly differed. Anti-ClfB, anti-IsdH, and anti-fibronectin-binding protein A IgG levels increased in 7 of 13 adult patients (p < 0.05). The anti-IsdA IgG level increased in 12 patients (initial to peak level: 1.13-10.72 fold; p < 0.01). Peak level was reached 7-37 days after diagnosis. In a bacteremic 5-day-old newborn, antistaphylococcal IgG levels declined from diagnosis onwards. In conclusion, each bacteremic patient develops a unique immune response directed to different staphylococcal proteins. Therefore, vaccines should be based on multiple components. IsdA is immunogenic and, therefore, produced in nearly all bacteremic patients. This suggests that IsdA might be a useful component of a multivalent staphylococcal vaccine.

Simple assessment of homogeneity in pharmaceutical mixing processes using a near-infrared reflectance probe and control charts.

Determination of homogeneous mixing of the active pharmaceutical ingredient (API) is an important in-process control within the manufacturing of solid dosage forms. In this paper two new near-infrared (NIR) based methods were presented; a qualitative and a quantitative method. Both methods are based on the calculation of net analyte signal (NAS) models which were very easy to develop, specific with respect to the API and required no additional reference analysis. Using a well-mixed batch as a 'golden standard' batch, control charts were developed and used for monitoring the homogeneity of other batches with NIR. The methods were fast, easy to use, non-destructive and provided statistical tests of homogeneity. A mixing study was characterized with the two methods and the methods were validated by comparison with traditional HPLC analysis.

Net analyte signal based statistical quality control.

Net analyte signal statistical quality control (NAS-SQC) is a new methodology to perform multivariate product quality monitoring based on the net analyte signal approach. The main advantage of NAS-SQC is that the systematic variation in the product due to the analyte (or property) of interest is separated from the remaining systematic variation due to all other compounds in the matrix. This enhances the ability to flag products out of statistical control. Using control charts, the analyte content, variation of other compounds, and residual variation can be monitored. As an example, NAS-SQC is used to appreciate the control content uniformity of a commercially available pharmaceutical tablet product measured with near-infrared spectroscopy. Using the NAS chart, the active pharmaceutical ingredient (API) content is easily monitored for new tablets. However, since quality is a multivariate property, other quality parameters of the tablets are also monitored simultaneously. It will be demonstrated that, besides the API content, the water content of the tablets as well as the homogeneity of the other compounds is monitored.

Perianal fistulas in Crohn's disease are predominantly colonized by skin flora: implications for antibiotic treatment?

In this study we aimed to determine the microorganisms found in perianal fistulas in Crohn's disease and whether treatment with ciprofloxacin affects these microorganisms. Thirteen patients (males/females, 7/6; median age, 34 years; range, 18-61 years) with fistulas were treated with infliximab, 5 mg/kg intravenously, at weeks 6, 8, and 12 and randomized to double-blind treatment with ciprofloxacin, 500 mg bd (n = 6), or placebo (n = 7) for 12 weeks. Samples were taken at baseline and at weeks 6 and 18. In the ciprofloxacin group 10 different genera of microorganisms were identified, while 13 genera could be identified in the placebo group. Gram-negative enteric floras were present in a small minority. The genera found in patients with perianal fistulas were predominantly gram-positive microorganisms. Therefore, antimicrobial treatment should be directed toward these microorganisms.

Epidemiology of nasopharyngeal carriage of Neisseria meningitidis in healthy Dutch children.

We investigated the prevalence and determinants of nasopharyngeal carriage of Neisseria meningitidis in 3200 healthy children aged 1-19 years. The incidence of meningococcal carriage was, on average, 1.5%. Peak incidences were seen at age 1 year and after age 15 years. The independent determinants of meningococcal carriage included age, regular visits to youth clubs (odds ratio [OR], 2.2) and discotheques (OR, 4.3), and pneumococcal carriage (OR, 4.1).

In-line reaction monitoring of a methyl methacrylate and N, N-dimethylacrylamide copolymerization reaction using near-infrared spectroscopy.

Fast and accurate monitoring of monomer concentration during copolymerization reactions is of much interest. It is known that near-infrared spectroscopy (NIRS) can be used to monitor polymerization reactions. Here, a free radical solution copolymerization reaction between methyl methacrylate and N,N-dimethylacrylamide is considered. NIR spectra were measured in-line with a transflectance probe. The spectra of both involved monomers are very similar, making monitoring with NIRS challenging. It is shown that the NIRS calibration can be set up with only a few (5) off-line measured mixtures. Several validation methods for such a NIRS calibration model are discussed and tested. NIRS is used to follow conversion of the two monomers in a copolymerization reaction on-line.

Low prevalence of methicillin-resistant Staphylococcus aureus (MRSA) at hospital admission in the Netherlands: the value of search and destroy and restrictive antibiotic use.

In the Netherlands, less than 1% of clinical isolates of Staphylococcus aureus are methicillin-resistant (MRSA). A national search and destroy policy prevents MRSA from becoming endemic. Some MRSA outbreaks cannot be related to patients at risk for MRSA carriage. This study was designed to measure the prevalence of MRSA among patients without risk factors for MRSA carriage at the time of admission to the hospital. In four Dutch hospitals, patients admitted to non-surgical departments in the period 1999-2000 were screened for MRSA nasal carriage. Nasal swabs were streaked on 5% sheep blood agar (BA), submerged in a selective broth, and incubated for two to three days at 35 degrees C. Colonies suspected of being S. aureus were identified with an agglutination test. Susceptibility testing was performed by an automated system and additional oxacillin disk diffusion. Methicillin resistance was confirmed by a DNA hybridization test and mecA PCR. MRSA strains were genotyped by pulsed-field gel electrophoresis (PFGE). Twenty-four percent (2332/9859) of the patients were S. aureus nasal carriers. Only three (0.03%) patients were MRSA carriers. These patients were not repatriated, nor known to be MRSA carriers before screening. Genotyping revealed that the strains were not clonally related and were not related to MRSA outbreaks in the hospital where the patients were admitted. We conclude that at routine admission to a Dutch hospital (excluding high-risk foreign admissions) the MRSA prevalence is low (0.03%), due to the Dutch search and destroy policy and restrictive antibiotic prescribing.

New indicator for optimal preprocessing and wavelength selection of near-infrared spectra.

Preprocessing of near-infrared spectra to remove unwanted, i.e., non-related spectral variation and selection of informative wavelengths is considered to be a crucial step prior to the construction of a quantitative calibration model. The standard methodology when comparing various preprocessing techniques and selecting different wavelengths is to compare prediction statistics computed with an independent set of data not used to make the actual calibration model. When the errors of reference value are large, no such values are available at all, or only a limited number of samples are available, other methods exist to evaluate the preprocessing method and wavelength selection. In this work we present a new indicator (SE) that only requires blank sample spectra, i.e., spectra of samples that are mixtures of the interfering constituents (everything except the analyte), a pure analyte spectrum, or alternatively, a sample spectrum where the analyte is present. The indicator is based on computing the net analyte signal of the analyte and the total error, i.e., instrumental noise and bias. By comparing the indicator values when different preprocessing techniques and wavelength selections are applied to the spectra, the optimal preprocessing technique and the optimal wavelength selection can be determined without knowledge of reference values, i.e., it minimizes the non-related spectral variation. The SE indicator is compared to two other indicators that also use net analyte signal computations. To demonstrate the feasibility of the SE indicator, two near-infrared spectral data sets from the pharmaceutical industry were used, i.e., diffuse reflectance spectra of powder samples and transmission spectra of tablets. Especially in pharmaceutical spectroscopic applications, it is expected beforehand that the non-related spectral variation is rather large and it is important to remove it. The indicator gave excellent results with respect to wavelength selection and optimal preprocessing. The SE indicator performs better than the two other indicators, and it is also applicable to other situations where the Beer-Lambert law is valid.

Heterogeneity among infecting strains of Pseudomonas aeruginosa in diverse departments of a large Tunisian hospital.

Clinical isolates of Pseudomonas aeruginosa were obtained during a half-year screening period of five different wards of the La Rabta Hospital (Tunis). Distinct clinical isolates (N= 82) were obtained from patients, 40 (48%) of which originated from the Department of Otolaryngology. In order to define the local epidemiology of this opportunistic organism, all strains were serotyped, analysed for pyocin production and genetically characterized with the help of pulsed-field gel electrophoresis (PFGE). The data show that, despite the frequent occurrence of identical serotypes, most of the isolates represent unique pyocin types (N= 53) and genotypes (N= 64). A combination of the pyocin and PFGE data showed that nearly all strains were of unique types, except for two pairs of strains. A limited number of strain clusters was observed on the basis of DNA typing data alone. This involved eight genotypes, some of which were clustered with respect to clinical environment or time. Genotype 22 occurred most frequently (6/83, 7%) and independently of time and locale, indicating that it may represent either a clonal type constituting a major fraction of all P. aeruginosa isolates in the region or a more prevalent organism. Despite a relatively high incidence of P. aeruginosa infections, the polyclonality of these strains shows that, in La Rabta Hospital, pseudomonal infections are not primarily due to excessive spread of a single bacterial genotype.

An epidemiological study on the occurrence of Staphylococcus aureus in superficial abscesses of patients presenting for surgery in a teaching hospital in Khartoum, Sudan.

A group of patients (n=86) suffering from superficial abscesses was recruited in the Khartoum Teaching Hospital, Sudan. Detailed clinical and socio-economic data were collected. It appeared that 83% of all patients were younger than 40. Labourers were most prevalent (28%), followed by students (23%) and housewives (16%). The head and neck were most often affected (22%), with hands being second (19%). In 92% of all pus cultures a microbial agents was identified, the large majority being Staphylococcus aureus (69%). Among patients, 47% were nasal carriers of S. aureus, similar to the carriage rate measured among controls, suggesting that nasal carriage is no risk factor for abscess development. Multivariate logistic regression analysis revealed that a history of abscess, recent traditional medical treatment, poor hygiene and low socio-economic status were significantly and independently associated with the occurrence of superficial abscesses.

Follow-up of Staphylococcus aureus nasal carriage after 8 years: redefining the persistent carrier state.

Studies of Staphylococcus aureus nasal carriage have distinguished three carriage patterns: persistent, intermittent, and noncarriage. The criteria used to identify these carriage patterns have been inconsistent. In 1988 the S. aureus nasal carrier index, i.e., the proportion of nasal swab specimen cultures yielding S. aureus, was determined for 91 staff members of various departments of a large university hospital by obtaining weekly nasal swab specimens for culture over a 12-week period. Thirty-three (36%) persons had carrier indices of 0.80 or higher, 15 (17%) had indices between 0.1 and 0.7, and 43 (47%) had indices of zero. In 1995, 17 individuals with carrier indices of 0.80 or higher in 1988 were available for reexamination. For 12 (71%) of these individuals, S. aureus was again isolated from a single nasal swab, i.e., from each individual with a 1988 carrier index of 1.0 but from only half of those with indices below 1.0. Genotyping (by randomly amplified polymorphic DNA analysis and pulsed-field gel electrophoresis) of all S. aureus strains showed that strains isolated from only three individuals, all with 1988 carrier indices of 1.0, in 1988 and 1995 showed genetic similarity. In conclusion, persistent S. aureus nasal carriage is a unique characteristic of a fraction of the population, and the attribute "persistent" should be confined to those individuals for whom serial nasal swab specimen cultures consistently yield S. aureus.

Glucose uptake kinetics and transcription of HXT genes in chemostat cultures of Saccharomyces cerevisiae.

The kinetics of glucose transport and the transcription of all 20 members of the HXT hexose transporter gene family were studied in relation to the steady state in situ carbon metabolism of Saccharomyces cerevisiae CEN.PK113-7D grown in chemostat cultures. Cells were cultivated at a dilution rate of 0.10 h-1 under various nutrient-limited conditions (anaerobically glucose- or nitrogen-limited or aerobically glucose-, galactose-, fructose-, ethanol-, or nitrogen-limited), or at dilution rates ranging between 0.05 and 0.38 h-1 in aerobic glucose-limited cultures. Transcription of HXT1-HXT7 was correlated with the extracellular glucose concentration in the cultures. Transcription of GAL2, encoding the galactose transporter, was only detected in galactose-limited cultures. SNF3 and RGT2, two members of the HXT family that encode glucose sensors, were transcribed at low levels. HXT8-HXT17 transcripts were detected at very low levels. A consistent relationship was observed between the expression of individual HXT genes and the glucose transport kinetics determined from zero-trans influx of 14C-glucose during 5 s. This relationship was in broad agreement with the transport kinetics of Hxt1-Hxt7 and Gal2 deduced in previous studies on single-HXT strains. At lower dilution rates the glucose transport capacity estimated from zero-trans influx experiments and the residual glucose concentration exceeded the measured in situ glucose consumption rate. At high dilution rates, however, the estimated glucose transport capacity was too low to account for the in situ glucose consumption rate.

Habit-reversal treatment for children's stuttering: assessment in three settings.

Two boys (9 and 14 years old) participated in an evaluation of treatment for stuttering. Habit reversal procedures (awareness training, regulated breathing, and social support) were combined with teaching positive attitudes to parents. During treatment, stuttering frequencies decreased, speech rates increased, and speech naturalness ratings increased. These changes occurred in the clinic, the child's home, and the child's school. After treatment, stuttering remained low in the clinic and at home, but increased stuttering was found at school.

Efficacy and safety of teicoplanin plus rifampicin in the treatment of bacteraemic infections caused by Staphylococcus aureus.

An open study was carried out on 16 patients with hospital-acquired, bacteraemic Staphylococcus aureus infections to evaluate the safety and efficacy of teicoplanin plus rifampicin. Patients received teicoplanin 400 mg bd for the first 24 h followed by 400 mg od thereafter, and rifampicin 600 mg bd. Both agents were given intravenously. Serum samples were collected to determine trough and peak antibiotic concentrations. The MIC of teicoplanin and rifampicin and the MBC of teicoplanin were determined for all S. aureus isolates. Time-kill curves were performed for the drugs individually and in combination. Clinical efficacy was assessed by the APACHE II scoring system. Bacteriological success was evaluated by elimination, persistence or recurrence of S. aureus. Safety was carefully monitored by regular biochemical and haematological testing and recording of adverse events. Fifteen patients were evaluable, of whom 13 (86.7%) were clinically cured with elimination of S. aureus. One patient died, but death was not attributed to the study drugs. Treatment failed in another patient who relapsed with a high fever. S. aureus was recovered from blood cultures from this patient, and resistance to rifampicin had developed. Time-kill curves all showed adequate killing of S. aureus at the drug concentrations measured in vivo. Neither synergy nor antagonism between teicoplanin and rifampicin was demonstrated. The combination of teicoplanin and rifampicin is an effective and well-tolerated treatment for bacteraemic S. aureus infections, but in deep-seated foci of infection resistance to rifampicin may develop.

Comparison of five methods for the study of drug-protein binding in affinity capillary electrophoresis.

The qualitative and quantitative aspects of capillary electrophoretic methods used to study drug-protein interactions, viz. the affinity capillary electrophoresis (ACE). Hummel-Dreyer (HD), frontal analysis (FA), vacancy peak (VP) and vacancy affinity capillary electrophoresis (VACE) methods have been investigated. In the ACE and the VACE methods the binding parameters can be calculated from the change in the electrophoretic mobility of the drug on complexation with a protein. In the frontal analysis and the vacancy peak method the free drug concentration is measured with UV detection. In the Hummel-Dreyer method the amount of drug bound is measured with UV detection. For the comparison of these five methods the warfarin-bovine serum albumin (BSA) system was used. Several factors that might influence the determination of association parameters were examined. With the FA, VP, HD and VACE methods the absolute numbers of the different binding sites involved in the complex formation can be determined, a major advantage in drug-binding studies.

Teaching students with mental retardation and other disabilities to make simple drawings through a computer system and special cards.

3 students with mental retardation and other disabilities were trained to make simple drawings on a computer screen (with a light-pen) following the color sequence used for the same drawings on special cards. The screen provided immediate feedback on the students' drawing accuracy. Teachers rated the students' posttraining drawings much higher than the pretraining ones.

Critical evaluation of the applicability of capillary zone electrophoresis for the study of hapten-antibody complex formation.

The applicability of capillary zone electrophoresis (CZE) for the determination of association constants of an hapten-antibody complex with values as high as 10(7) (mol/I)-1 was investigated (dissociation constant in the nmol range). As a reference method the well known, enzyme-linked immunosorbent assay (ELISA) was selected. The study describes the optimisation of the experimental conditions of the CZE technique. The CZE measurements were optimised according to an experimental design. The results of the CZE and ELISA methods are compared giving consideration to the reproducibility (repeatability) of the two methods.