Practices in prescribing protein substitutes for PKU in Europe: No uniformity of approach.

BACKGROUND: There appears little consensus concerning protein requirements in phenylketonuria (PKU). METHODS: A questionnaire completed by 63 European and Turkish IMD centres from 18 countries collected data on prescribed total protein intake (natural/intact protein and phenylalanine-free protein substitute [PS]) by age, administration frequency and method, monitoring, and type of protein substitute. Data were analysed by European region using descriptive statistics. RESULTS: The amount of total protein (from PS and natural/intact protein) varied according to the European region. Higher median amounts of total protein were prescribed in infants and children in Northern Europe (n=24 centres) (infants <1 year, >2-3g/kg/day; 1-3 years of age, >2-3 g/kg/day; 4-10 years of age, >1.5-2.5 g/kg/day) and Southern Europe (n=10 centres) (infants <1 year, 2.5 g/kg/day, 1-3 years of age, 2 g/kg/day; 4-10 years of age, 1.5-2 g/kg/day), than by Eastern Europe (n=4 centres) (infants <1 year, 2.5 g/kg/day, 1-3 years of age, >2-2.5 g/kg/day; 4-10 years of age, >1.5-2 g/kg/day) and with Western Europe (n=25 centres) giving the least (infants <1 year, >2-2.5 g/kg/day, 1-3 years of age, 1.5-2 g/kg/day; 4-10 years of age, 1-1.5 g/kg/day). Total protein prescription was similar in patients aged >10 years (1-1.5 g/kg/day) and maternal patients (1-1.5 g/kg/day). CONCLUSIONS: The amounts of total protein prescribed varied between European countries and appeared to be influenced by geographical region. In PKU, all gave higher than the recommended 2007 WHO/FAO/UNU safe levels of protein intake for the general population.

How strict is galactose restriction in adults with galactosaemia? International practice.

Dietary management of 418 adult patients with galactosaemia (from 39 centres/12 countries) was compared. All centres advised lactose restriction, 6 restricted galactose from galactosides ± fruits and vegetables and 12 offal. 38% (n=15) relaxed diet by: 1) allowing traces of lactose in manufactured foods (n=13) or 2) giving fruits, vegetables and galactosides (n=2). Only 15% (n=6) calculated dietary galactose. 32% of patients were lost to dietetic follow-up. In adult galactosaemia, there is limited diet relaxation.

[A sensitive, specific and predictive isolation medium developed for Stenotrophomonas maltophilia study in healthcare settings]. / La mise au point d'un milieu sensible, spécifique et prédictif de recherche de Stenotrophomonas maltophilia dans l'environnement des soins.

BACKGROUND: Stenotrophomonas maltophilia (Smalto) is a prominent nosocomial pathogen, commonly isolated in the hospital environment. Multiple Smalto nosocomial outbreaks have been linked to contaminated water sources. This study aimed to develop a medium able to ease healthcare environment Smalto isolation. METHODS: Financed, from March 2007 to June 2008, by a university hospital of Amiens' clinical research program, this study allowed Stenotrophomonas maltophilia selective medium with coloured indicator (SM2i) development. SM2i is constituted of Mueller Hinton agar (MH), maltose, DL-methionine, bromothymol blue. The mixture sterilized is refreshed at 50 degrees C, its pH adjusted to 7.1, and render selective by addition of vancomycin, imipenem and amphotericin B. Then, SM2i agar is sunk into 90 cm diameter Petri dish dated and stored at 4 degrees C for 4 weeks. SM2i is developed using Pasteur Institute culture type collection (CIP) strains of Smalto, Burkholderia cepacia, Pseudomonas aeruginosa (Psa) and a Smalto strain of our hygiene laboratory collection. It was validate on Psa imipenem-resistant and Enterococcus faecium vancomycin-resistant strains, then, tested on cold water first jet and faucet cotton-swabs samples. SM2i tests were made in comparison with the MH agar, MH agar plus four paper disks loaded 10 microg of imipenem and Cetrimed agar. Its sensitivity, specificity, positive (PPV) and negative (NPV) predictive values, accuracy, likehood-ratio (LR) and Youden index have been determined. RESULTS: SM2i agar is better in culturing Smalto test-strains. On SM2i, Smalto colonies are smooth, round, greeny, olive or lime green, have a green olive centre with a peripheral lighter or a dark green centre with an olive green suburb surrounded by a blue halo. SM2i is a selective, specific, predictive, accurate medium to search for Smalto in healthcare environment. In 122 pairs of cold water first jet and taps cotton-swabs samples, Smalto was isolated from 14.8% of water samples, 10.7% of cotton-swabs samples. It was isolated alone in 6.6% of water samples and 2.5% of swab samples. Thus, smalto has biocontaminated 17.2% of cold water taps. Compared to MH agar, SM2i sensitivity, specificity, PPV, NPV, accuracy, LR were 100, 100, 100, 100, 100% and infinity, and 87.5, 100, 100, 98.1, 98.4% and infinity for water and cotton-swabs samples respectively. CONCLUSION: SM2i is a selective, specific, predictive medium which can allow easily isolating and identifying accurately Smalto in environmental samples. Its evaluation on clinical samples is on going.

[Stenotrophomonas maltophilia and Pseudomonas aeruginosa water-associated microbiologic risk assessment in Amiens' University Hospital Centre]. / Evaluation des risques microbiologiques hydriques associés à Stenotrophomonas maltophilia et Pseudomonas aeruginosa au CHU d'Amiens.

BACKGROUND: Pseudomonas aeruginosa (Psa) and Stenotrophomonas maltophilia (Smalto) are major opportunistic waterborne pathogens causing hospital-acquired infections. This study aimed to assess the biocontamination level of cold water used in Amiens' university hospital wards, from March to June 2008. METHODS: We cultivated 122 pairs of cold water first jet and taps cotton-swabs on Cetrimide agar for Psa, on Stenotrophomonas maltophilia selective medium with coloured indicator (SM2i) for Smalto, on Mueller Hinton agar used as isolation medium reference for both, 48h at 30 degrees C. Data analysed with Epi-Info 6.04dFr were compared with chi(2) test, significant at p<.05. RESULTS: Psa and Smalto were isolated in 26.2 and 14.8% of water samples and in 21.3 and 10.7% of swab samples respectively. They were associated in 11.5% of water samples and 5% of swab samples. Psa was alone in 13.1% of water samples and 7.4% of swab samples whereas Smalto was found in 6.6% of water and 2.5% of swabs. Psa and Smalto were isolated from 14.8% of water samples and 8.2% of swab samples of the same tap. Finally, respectively 35.2 and 17.2% of the cold water taps were biocontaminated by Psa and Smalto. In fact, microbiologic water taps contamination risk was two-fold higher for Psa than for Smalto, p<.001, without variation between wards. CONCLUSION: Sm2i and Cetrimide are suited and efficient medium respectively for Smalto and Psa isolation. Cold-water samples are sufficient for waterborne pathogens biocontamination risk appraisal. Our results urged healthcare workers on efficient water fittings microbiologic risk control to prevent healthcare associated waterborne infections, notably due to Psa and Smalto.

Weaning practices in phenylketonuria vary between health professionals in Europe.

BACKGROUND: In phenylketonuria (PKU), weaning is considered more challenging when compared to feeding healthy infants. The primary aim of weaning is to gradually replace natural protein from breast milk or standard infant formula with solids containing equivalent phenylalanine (Phe). In addition, a Phe-free second stage L-amino acid supplement is usually recommended from around 6 months to replace Phe-free infant formula. Our aim was to assess different weaning approaches used by health professionals across Europe. METHODS: A cross sectional questionnaire (survey monkey®) composed of 31 multiple and single choice questions was sent to European colleagues caring for inherited metabolic disorders (IMD). Centres were grouped into geographical regions for analysis. RESULTS: Weaning started at 17-26 weeks in 85% (n = 81/95) of centres, >26 weeks in 12% (n = 11/95) and < 17 weeks in 3% (n = 3/95). Infant's showing an interest in solid foods, and their age, were important determinant factors influencing weaning commencement. 51% (n = 48/95) of centres introduced Phe containing foods at 17-26 weeks and 48% (n = 46/95) at >26 weeks. First solids were mainly low Phe vegetables (59%, n = 56/95) and fruit (34%, n = 32/95).A Phe exchange system to allocate dietary Phe was used by 52% (n = 49/95) of centres predominantly from Northern and Southern Europe and 48% (n = 46/95) calculated most Phe containing food sources (all centres in Eastern Europe and the majority from Germany and Austria). Some centres used a combination of both methods.A second stage Phe-free L-amino acid supplement containing a higher protein equivalent was introduced by 41% (n = 39/95) of centres at infant age 26-36 weeks (mainly from Germany, Austria, Northern and Eastern Europe) and 37% (n = 35/95) at infant age > 1y mainly from Southern Europe. 53% (n = 50/95) of centres recommended a second stage Phe-free L-amino acid supplement in a spoonable or semi-solid form. CONCLUSIONS: Weaning strategies vary throughout European PKU centres. There is evidence to suggest that different infant weaning strategies may influence longer term adherence to the PKU diet or acceptance of Phe-free L-amino acid supplements; rendering prospective long-term studies important. It is essential to identify an effective weaning strategy that reduces caregiver burden but is associated with acceptable dietary adherence and optimal infant feeding development.

Early feeding practices in infants with phenylketonuria across Europe.

BACKGROUND: In infants with phenylketonuria (PKU), dietary management is based on lowering and titrating phenylalanine (Phe) intake from breast milk or standard infant formula in combination with a Phe-free infant formula in order to maintain blood Phe levels within target range. Professionals use different methods to feed infants with PKU and our survey aimed to document practices across Europe. METHODS: We sent a cross sectional, survey monkey® questionnaire to European health professionals working in IMD. It contained 31 open and multiple-choice questions. The results were analysed according to different geographical regions. RESULTS: Ninety-five centres from 21 countries responded. Over 60% of centres commenced diet in infants by age 10 days, with 58% of centres implementing newborn screening by day 3 post birth. At diagnosis, infant hospital admission occurred in 61% of metabolic centres, mainly in Eastern, Western and Southern Europe. Breastfeeding fell sharply following diagnosis with only 30% of women still breast feeding at 6 months.53% of centres gave pre-measured Phe-free infant formula before each breast feed and 23% alternated breast feeds with Phe-free infant formula. With standard infant formula feeds, measured amounts were followed by Phe-free infant formula to satiety in 37% of centres (n = 35/95), whereas 44% (n = 42/95) advised mixing both formulas together. Weaning commenced between 17 and 26 weeks in 85% centres, ≥26 weeks in 12% and < 17 weeks in 3%. DISCUSSION: This is the largest European survey completed on PKU infant feeding practices. It is evident that practices varied widely across Europe, and the practicalities of infant feeding in PKU received little focus in the PKU European Guidelines (2017). There are few reports comparing different feeding techniques with blood Phe control, Phe fluctuations and growth. Controlled prospective studies are necessary to assess how different infant feeding practices may influence longer term feeding development.

Relevance of mild ineffective oesophageal motility (IOM) and potential pharmacological reversibility of severe IOM in patients with gastro-oesophageal reflux disease.

BACKGROUND: Several studies showed high prevalence of ineffective oesophageal motility (IOM) in gastro-oesophageal reflux disease (GERD) and suggested an important role for ineffective oesophageal motility in increased acid exposure. However, impedance-manometric studies proposed that only severe ineffective oesophageal motility might affect oesophageal clearance. OBJECTIVES: (i) To re-assess the relevance of mild IOM in GERD and (ii) to test the reversibility of IOM. METHODS: Oesophageal motility, clearance and acid exposure were assessed in 191 GERD patients: 99 without IOM; 58 with mild IOM (30-80% ineffective contractions) and 34 with severe IOM (>80% ineffective contractions). In 30 patients with oesophagitis, the potential reversibility of IOM was evaluated with edrophonium intravenously. RESULTS: Patients with mild IOM had identical oesophageal clearance and acid exposure in comparison with those without IOM. Patients with severe IOM had a higher probability of prolonged supine clearance and acid exposure [odds ratio: 2.88 (1.16-7.17); 2.48 (0.99-6.17)]. This effect was independent of the presence of hiatal hernia and male sex. Severe IOM could be transiently reverted in 55% of patients. CONCLUSIONS: Mild IOM does not affect oesophageal clearance. Only severe IOM is associated with prolonged clearance and acid exposure, particularly in supine periods. The edrophonium test might be useful to predict severe IOM response to prokinetic medications.

Dietary practices in isovaleric acidemia: A European survey.

BACKGROUND: In Europe, dietary management of isovaleric acidemia (IVA) may vary widely. There is limited collective information about dietetic management. AIM: To describe European practice regarding the dietary management of IVA, prior to the availability of the E-IMD IVA guidelines (E-IMD 2014). METHODS: A cross-sectional questionnaire was sent to all European dietitians who were either members of the Society for the Study of Inborn Errors of Metabolism Dietitians Group (SSIEM-DG) or whom had responded to previous questionnaires on dietetic practice (n = 53). The questionnaire comprised 27 questions about the dietary management of IVA. RESULTS: Information on 140 patients with IVA from 39 centres was reported. 133 patients (38 centres) were given a protein restricted diet. Leucine-free amino acid supplements (LFAA) were routinely used to supplement protein intake in 58% of centres. The median total protein intake prescribed achieved the WHO/FAO/UNU [2007] safe levels of protein intake in all age groups. Centres that prescribed LFAA had lower natural protein intakes in most age groups except 1 to 10 y. In contrast, when centres were not using LFAA, the median natural protein intake met WHO/FAO/UNU [2007] safe levels of protein intake in all age groups. Enteral tube feeding was rarely prescribed. CONCLUSIONS: This survey demonstrates wide differences in dietary practice in the management of IVA across European centres. It provides unique dietary data collectively representing European practices in IVA which can be used as a foundation to compare dietary management changes as a consequence of the first E-IMD IVA guidelines availability.

Dietary practices in propionic acidemia: A European survey.

BACKGROUND: The definitive dietary management of propionic acidaemia (PA) is unknown although natural protein restriction with adequate energy provision is of key importance. AIM: To describe European dietary practices in the management of patients with PA prior to the publication of the European PA guidelines. METHODS: This was a cross-sectional survey consisting of 27 questions about the dietary practices in PA patients circulated to European IMD dietitians and health professionals in 2014. RESULTS: Information on protein restricted diets of 186 PA patients from 47 centres, representing 14 European countries was collected. Total protein intake [PA precursor-free L-amino acid supplements (PFAA) and natural protein] met WHO/FAO/UNU (2007) safe protein requirements for age in 36 centres (77%). PFAA were used to supplement natural protein intake in 81% (n = 38) of centres, providing a median of 44% (14-83%) of total protein requirement. Seventy-four per cent of patients were prescribed natural protein intakes below WHO/FAO/UNU (2007) safe levels in one or more of the following age groups: 0-6 m, 7-12 m, 1-10 y, 11-16 y and > 16 y. Sixty-three per cent (n = 117) of patients were tube fed (74% gastrostomy), but only 22% received nocturnal feeds. CONCLUSIONS: There was high use of PFAA with intakes of natural protein commonly below WHO/FAO/UNU (2007) safe levels. Optimal dietary management can only be determined by longitudinal, multi-centre, prospective case controlled studies. The metabolic instability of PA and small patient cohorts in each centre ensure that this is a challenging undertaking.

Construction by homologous recombination and phenotypic characterization of a DNA polymerase domain polA mutant of Mycobacterium smegmatis.

Gene replacement was achieved by homologous recombination in Mycobacterium smegmatis (Ms) using a cloned segment of the polA gene (encoding the DNA polymerase I) disrupted within the region encoding the C-terminal DNA polymerase domain by a kanamycin-resistance marker. The Ms polA755:aph mutant thus generated displayed a phenotype of hypersensitivity to DNA damage induced by UV irradiation and by hydrogen peroxide challenge.