Cross-cultural validity of the Dutch sleep-related breathing disorder scale of the Pediatric Sleep Questionnaire in a general population.

BACKGROUND: Pediatric sleep-disordered breathing is associated with multiple health problems. Polysomnography is the reference standard for identifying this disorder, but availability is limited. Therefore, an alternative screening tool is needed. Globally, the Sleep-Related Breathing Disorder scale of the Pediatric Sleep Questionnaire (PSQ) has proven to be a feasible tool. Consequently, this study aimed to translate and culturally adapt the PSQ into Dutch and then to examine the cultural validity, internal consistency, and test-retest reliability of the Dutch version among a general population visiting oral healthcare centers. METHODS: The translation, review, adaptation, pretest, and documentation approach was used to ensure cross-cultural adaptation of the PSQ. Then, 220 children (2.4-18 years) were sampled for clinimetric evaluation. We estimated the cross-cultural validity by comparing the factor analyses of the original PSQ and the Dutch version. Reliability was assessed using Cronbach's alpha, Spearman's correlation, the intraclass correlation coefficient, the standard error of measurement, and a Bland-Altman plot. RESULTS: The factor loading patterns of the Dutch version matched with the original study around the four predetermined factors: breathing, sleepiness, behavior, and other. The internal consistency, with a Cronbach's α of 0.77, was acceptable. The test-retest reliability with an intraclass correlation coefficient and Spearman's correlation of 0.89 and 0.93, respectively, was good to excellent. CONCLUSIONS: Cultural adaptation was ensured and the results support cross-cultural validity, internal consistency, and test-retest reliability of the Dutch Sleep-Related Breathing Disorder scale of the PSQ. This questionnaire could therefore be a valuable tool for screening disordered breathing in Dutch children.

The accuracy of photographic soft-tissue profile analysis to determine Class II and vertical skeletal relationships in children.

BACKGROUND: Lateral cephalometric analysis (LCA) is the reference standard for identifying common skeletal relationships in orthodontics, such as Cl II and hyperdivergent skeletal discrepancies, but it entails radiation exposure. Therefore, photographic soft-tissue profile analysis (PSPA) could be a useful alternative for these diagnoses, particularly for paediatric patients. This study aims to estimate the accuracy of PSPA for determining common skeletal discrepancies in children. METHODS: Cephalometric radiographs and profile photographs of a consecutive series of 100 children (8.0-17.6 years old) made on the same day were included. The validity of PSPA was verified against comparable LCA. First, by assessing the Pearson correlation and then estimating the sensitivity, specificity, receiver operating characteristic (ROC) curves and area under the curve (AUC) in sample A (n = 50). After external validation in a new sample B (n = 50), the ROC-AUC, diagnostic odds ratio, best cut-off points and discriminative validity were assessed in the total sample. Interrater reliability was estimated using the intraclass correlation coefficient, the standard error of measurement and Bland-Altman plots. RESULTS: The measurement properties of the PSPA angles A'N'B', Gl'-Sn-Pog, N'-Sn-Pog', and N'-Tra-Me' were valid (ROC-AUC > 0.7) and reliable (ICCs > 0.92). The angles A'N'B', Gl'-Sn-Pog', and N'-Sn-Pog', with the respective cut-off points ≥7.7', ≥12.8', and ≤163.5', were accurate values for determining Cl II discrepancy. The N'-Tra-Me'-angle (≥63') was an accurate estimate for a hyperdivergent discrepancy. CONCLUSIONS: These validated PSPA angles could be used in clinical settings as a minimally invasive diagnostic tool to screen children suspected of having skeletal Cl II and hyperdivergent discrepancies.

Impact of adenotonsillectomy on the dentofacial development of obstructed children: a systematic review and meta-analysis.

BACKGROUND: Dentofacial deformities frequently require orthodontic treatment. Understanding of preventable risk factors is essential for reducing treatment need. Upper airway obstruction (for example due to hypertrophic adenoids and/or tonsils) has been hypothesized to be a risk factor. OBJECTIVES: This systematic review aimed to reflect the contemporary evidence on the risk of obstruction by hypertrophic adenoids and/or tonsils, by assessing the dentofacial changes after adeno- and/or tonsillectomy. SEARCH METHODS: A systematic search of electronic databases and manual searches of grey literature and reference lists of relevant studies was performed. SELECTION CRITERIA: No restrictions were placed on publication language. Experimental, cohort, and case-control studies were eligible for inclusion. Studies reporting associations between treatment of adenoid and/or tonsil hypertrophy and dentofacial deformities in children were included. Adenoidectomy and/or tonsillectomy were performed in all patients; outcomes were assessed before and after surgery. DATA COLLECTION AND ANALYSIS: Data were extracted by two independent reviewers in duplicate. The Cochrane Risk of Bias tool was used to assess the methodological quality of the included papers. RESULTS: The initial search yielded 1196 papers, of which 16 articles could be included. All papers described controlled prospective cohort studies, reporting on a total of 461 patients and controls (mean age, 4.1-13.9 years). A descriptive and quantitative synthesis of dentofacial change postoperatively is presented. Consistent findings across studies were the normalisation towards labial inclination of the upper and lower incisors and towards a more horizontal mandibular growth pattern. No change in vertical or sagittal maxillary growth was reported after surgical treatment. Post-surgical increase in maxillary archwidth and decrease in lateral crossbite-frequency were consistently reported. Findings on overjet, overbite and angle from S to N to B (SNB-angle), mandibular arch width, and gonial angle were inconsistent. CONCLUSION: The available literature suggests that treatment of hypertrophic adenoids and/or tonsils affects dentofacial deformity. This could indicate a relationship between nasopharyngeal obstruction (i.e. upper airway obstruction) and the dentofacial growth pattern. However, the high risk of bias and considerable diversity between studies impedes a clear conclusion regarding this effect. REGISTRATION: None.

A parent-focused intervention to reduce infant obesity risk behaviors: a randomized trial.

OBJECTIVE: To assess the effectiveness of a parent-focused intervention on infants' obesity-risk behaviors and BMI. METHODS: This cluster randomized controlled trial recruited 542 parents and their infants (mean age 3.8 months at baseline) from 62 first-time parent groups. Parents were offered six 2-hour dietitian-delivered sessions over 15 months focusing on parental knowledge, skills, and social support around infant feeding, diet, physical activity, and television viewing. Control group parents received 6 newsletters on nonobesity-focused themes; all parents received usual care from child health nurses. The primary outcomes of interest were child diet (3 × 24-hour diet recalls), child physical activity (accelerometry), and child TV viewing (parent report). Secondary outcomes included BMI z-scores (measured). Data were collected when children were 4, 9, and 20 months of age. RESULTS: Unadjusted analyses showed that, compared with controls, intervention group children consumed fewer grams of noncore drinks (mean difference = -4.45; 95% confidence interval [CI]: -7.92 to -0.99; P = .01) and were less likely to consume any noncore drinks (odds ratio = 0.48; 95% CI: 0.24 to 0.95; P = .034) midintervention (mean age 9 months). At intervention conclusion (mean age 19.8 months), intervention group children consumed fewer grams of sweet snacks (mean difference = -3.69; 95% CI: -6.41 to -0.96; P = .008) and viewed fewer daily minutes of television (mean difference = -15.97: 95% CI: -25.97 to -5.96; P = .002). There was little statistical evidence of differences in fruit, vegetable, savory snack, or water consumption or in BMI z-scores or physical activity. CONCLUSIONS: This intervention resulted in reductions in sweet snack consumption and television viewing in 20-month-old children.

Niche specialization of reef-building corals in the mesophotic zone: metabolic trade-offs between divergent Symbiodinium types.

The photobiology of two reef corals and the distribution of associated symbiont types were investigated over a depth gradient of 0-60 m at Scott Reef, Western Australia. Pachyseris speciosa hosted mainly the same Symbiodinium C type similar to C3 irrespective of sampling depth. By contrast, Seriatopora hystrix hosted predominantly Symbiodinium type D1a or D1a-like at shallow depths while those in deeper water were dominated by a Symbiodinium C type closely related to C1. The photosynthesis/respiration (P/R) ratio increased consistently with depth at the two sampling times (November 2008 and April 2009) for P. speciosa and in November 2008 only for S. hystrix, suggesting a reduction in metabolic energy expended for every unit of energy obtained from photosynthesis. However, in April 2009, shallow colonies of S. hystrix exhibited decreased P/R ratios down to depths of approximately 23 m, below which the ratio increased towards the maximum depth sampled. This pattern was mirrored by changes in tissue biomass determined as total protein content. The depth of change in the direction of the P/R ratio correlated with a shift from Symbiodinium D to C-dominated colonies. We conclude that while photobiological flexibility is vital for persistence in contrasting light regimes, a shift in Symbiodinium type may also confer a functional advantage albeit at a metabolic cost with increased depth.

Trends in work-related musculoskeletal disorder reports by year, type, and industrial sector: a capture-recapture analysis.

BACKGROUND: Musculoskeletal disorders (MSD) are thought to be declining based on Bureau of Labor Statistics survey data, but there is also evidence of MSD under-reporting, raising the possibility of contrary trends. The magnitude of MSD under-reporting over time, and its industry distribution have not been adequately described. METHODS: Capture-recapture analysis of 7 years of Connecticut MSD (1995-2001), utilizing Workers' Compensation and physician reporting data was performed. RESULTS: Only 5.5%-7.9% of MSD cases appear to be reported to Workers Compensation annually. The capture-recapture estimated average annual rate for upper-extremity MSD was 133.1 per 10,000 employed persons, far above BLS rates. By industry, Manufacturing, State Government, and the Finance/Insurance/Real Estate sectors all had significantly higher MSD rates than Wholesale/Retail Trade. CONCLUSIONS: Upper-extremity MSD appears to be significantly under-reported, and rates are not decreasing over time. Capture-recapture methods provide an improved surveillance method for monitoring temporal trends in injury rates.

Evidence for coupling of membrane targeting and function of the signal recognition particle (SRP) receptor FtsY.

Recent studies have indicated that FtsY, the signal recognition particle receptor of Escherichia coli, plays a central role in membrane protein biogenesis. For proper function, FtsY must be targeted to the membrane, but its membrane-targeting pathway is unknown. We investigated the relationship between targeting and function of FtsY in vivo, by separating its catalytic domain (NG) from its putative targeting domain (A) by three means: expression of split ftsY, insertion of various spacers between A and NG, and separation of A and NG by in vivo proteolysis. Proteolytic separation of A and NG does not abolish function, whereas separation by long linkers or expression of split ftsY is detrimental. We propose that proteolytic cleavage of FtsY occurs after completion of co-translational targeting and assembly of NG. In contrast, separation by other means may interrupt proper synchronization of co-translational targeting and membrane assembly of NG. The co-translational interaction of FtsY with the membrane was confirmed by in vitro experiments.

Evidence for simultaneous binding of dissimilar substrates by the Escherichia coli multidrug transporter MdfA.

The mechanism by which multidrug transporters interact with structurally unrelated substrates remains enigmatic. Based on transport competition experiments, photoaffinity labeling, and effects on enzymatic activities, it was proposed in the past that multidrug transporters can interact simultaneously with a number of dissimilar substrate molecules. To study this phenomenon, we applied a direct binding approach and transport assays using the Escherichia coli multidrug transporter MdfA, which exports both positively charged (e.g., tetraphenylphosphonium, TPP(+)), zwitterionic (e.g., ciprofloxacin), and neutral (e.g., chloramphenicol) drugs. The interaction of MdfA with various substrates was examined by direct binding assays with the purified transporter. The immobilized MdfA binds TPP(+) in a specific manner, and all the tested positively charged substrates inhibit TPP(+) binding. Surprisingly, although TPP(+) binding is not affected by zwitterionic substrates, the neutral substrate chloramphenicol stimulates TPP(+) binding by enhancing its affinity to MdfA. In contrast, transport competition assays show inhibition of TPP(+) transport by chloramphenicol. We suggest that MdfA binds TPP(+) and chloramphenicol simultaneously to distinct but interacting binding sites, and the interaction between these two substrates during transport is discussed.

MdfA, an interesting model protein for studying multidrug transport.

The resistance of cells to many drugs simultaneously (multidrug resistance) often involves the expression of membrane transporters (Mdrs); each can recognize and expel a broad spectrum of chemically unrelated drugs from the cell. Despite extensive research for many years, the actual mechanism of multidrug transport is still largely unknown. In addition to general questions dealing with energy coupling, the molecular view of substrate recognition by Mdrs is generally obscure. This mini-review describes structural and functional properties of the Escherichia coli Mdr, MdfA, and discusses the possibility that this transporter may serve as a model for studying the multidrug recognition phenomenon and the mechanism of multidrug transport.

New prospects in studying the bacterial signal recognition particle pathway.

In vivo and in vitro studies have suggested that the bacterial version of the mammalian signal recognition particle (SRP) system plays an essential and selective role in protein biogenesis. The bacterial SRP system consists of at least two proteins and an RNA molecule (termed Ffh, FtsY and 4.5S RNA, respectively, in Escherichia coli). Recent evidence suggests that other putative bacterial-specific SRP components may also exist. In vitro experiments confirmed the expected basic features of the bacterial SRP system by demonstrating interactions among the SRP components themselves, between them and ribosomes, ribosome-linked hydrophobic nascent polypeptides or inner membranes. The availability of a conserved (and essential) bacterial SRP version has facilitated the implementation of powerful genetic and biochemical approaches for studying the cascade of events during the SRP-mediated targeting process in vivo and in vitro as well as the three-dimensional structures and the properties of each SRP component and complex.

Association of Escherichia coli ribosomes with the inner membrane requires the signal recognition particle receptor but is independent of the signal recognition particle.

In mammalian cells, as well as Escherichia coli, ribosomes translating membrane proteins interact cotranslationally with translocons in the membrane, and this interaction is essential for proper insertion of nascent polypeptides into the membrane. Both the signal recognition particle (SRP) and its receptor (SR) are required for functional association of ribosomes translating integral membrane proteins with the translocon. Herein, we confirm that membrane targeting of E. coli ribosomes requires the prokaryotic SRalpha homolog FtsY in vivo. Surprisingly, however, depletion of the E. coli SRP54 homolog (Ffh) has no significant effect on binding of ribosomes to the membrane, although Ffh depletion is detrimental to growth. These and other observations suggest that, in E. coli, SRP may operate downstream of SR-mediated targeting of ribosomes to the plasma membrane.

A structure-based mechanism for drug binding by multidrug transporters.

Multidrug transporters bind chemically dissimilar, potentially cytotoxic compounds and remove them from the cell. How these transporters carry out either of these functions is unknown. On the basis of crystal structures of the multidrug-binding domain of the transcription activator BmrR and mutagenesis studies on the bacterial multidrug transporter MdfA, we propose a possible mechanism for the binding of cationic lipophilic drugs by multidrug transporters. The key element of this mechanism includes a conformational change in the transporter that exposes a buried charged residue in the substrate-binding pocket and allows access to this site by only those drugs that are its steric and electrostatic complements.

Resistance to bacitracin as modulated by an Escherichia coli homologue of the bacitracin ABC transporter BcrC subunit from Bacillus licheniformis.

A small open reading frame from the Escherichia coli chromosome, bcrC(EC), encodes a homologue to the BcrC subunit of the bacitracin permease from Bacillus licheniformis. We show that disruption of the chromosomal bcrC(EC) gene causes bacitracin sensitivity and, conversely, that BcrC(EC) confers bacitracin resistance when expressed from a multicopy plasmid.

A single membrane-embedded negative charge is critical for recognizing positively charged drugs by the Escherichia coli multidrug resistance protein MdfA.

The nature of the broad substrate specificity phenomenon, as manifested by multidrug resistance proteins, is not yet understood. In the Escherichia coli multidrug transporter, MdfA, the hydrophobicity profile and PhoA fusion analysis have so far identified only one membrane-embedded charged amino acid residue (E26). In order to determine whether this negatively charged residue may play a role in multidrug recognition, we evaluated the expression and function of MdfA constructs mutated at this position. Replacing E26 with the positively charged residue lysine abolished the multidrug resistance activity against positively charged drugs, but retained chloramphenicol efflux and resistance. In contrast, when the negative charge was preserved in a mutant with aspartate instead of E26, chloramphenicol recognition and transport were drastically inhibited; however, the mutant exhibited almost wild-type multidrug resistance activity against lipophilic cations. These results suggest that although the negative charge at position 26 is not essential for active transport, it dictates the multidrug resistance character of MdfA. We show that such a negative charge is also found in other drug resistance transporters, and its possible significance regarding multidrug resistance is discussed.

The role of the ribosome-translocon complex in translation and assembly of polytopic membrane proteins.

Newly synthesized polytopic membrane proteins and secretory proteins often share the same target membrane as their primary destination, and in some cases, the cellular machinery that targets and transfers them into or across the membrane. Unlike secretory proteins, which are localized to the external compartment, each polytopic membrane protein molecule must be partitioned among the cytoplasm, the membrane and the external milieu. How does the ribosome-translocon complex cope with the different domains of polytopic membrane proteins?

The NG domain of the prokaryotic signal recognition particle receptor, FtsY, is fully functional when fused to an unrelated integral membrane polypeptide.

Recent studies have revealed that Escherichia coli possesses an essential targeting system for integral membrane proteins, similar to the mammalian signal recognition particle (SRP) machinery. One essential protein in this system is FtsY, a homologue of the alpha-subunit of the mammalian SRP-receptor (SR-alpha). However, E. coli does not possess a close homologue of the integral membrane protein SR-beta, which anchors SR-alpha to the membrane. Moreover, although FtsY can be found as a peripheral membrane protein, the majority is found soluble in the cytoplasm. In this study, we obtained genetic and biochemical evidence that FtsY must be targeted to the membrane for proper function. We demonstrate that the essential membrane targeting activity of FtsY is mediated by a 198-residue-long acidic N-terminal domain. This domain can be functionally replaced by unrelated integral membrane polypeptides, thus avoiding the need for specific FtsY membrane targeting factors. Therefore, the N terminus of FtsY constitutes an independent domain, which is required only for the targeting of the C-terminal NG domain of FtsY to the membrane.

MdfA, an Escherichia coli multidrug resistance protein with an extraordinarily broad spectrum of drug recognition.

Multidrug resistance (MDR) translocators recently identified in bacteria constitute an excellent model system for studying the MDR phenomenon and its clinical relevance. Here we describe the identification and characterization of an unusual MDR gene (mdfA) from Escherichia coli. mdfA encodes a putative membrane protein (MdfA) of 410 amino acid residues which belongs to the major facilitator superfamily of transport proteins. Cells expressing MdfA from a multicopy plasmid are substantially more resistant to a diverse group of cationic or zwitterionic lipophilic compounds such as ethidium bromide, tetraphenylphosphonium, rhodamine, daunomycin, benzalkonium, rifampin, tetracycline, and puromycin. Surprisingly, however, MdfA also confers resistance to chemically unrelated, clinically important antibiotics such as chloramphenicol, erythromycin, and certain aminoglycosides and fluoroquinolones. Transport experiments with an E. coli strain lacking F1-F0 proton ATPase activity indicate that MdfA is a multidrug transporter that is driven by the proton electrochemical gradient.

FtsY, the prokaryotic signal recognition particle receptor homologue, is essential for biogenesis of membrane proteins.

In mammalian cells, many secretory proteins are targeted to the endoplasmic reticulum co-translationally, by the signal recognition particle (SRP) and its receptor. In Escherichia coli, the targeting of secretory proteins to the inner membrane can be accomplished post-translationally. Unexpectedly, despite this variance, E. coli contains essential genes encoding Ffh and FtsY with a significant similarity to proteins of the eukaryotic SRP machinery. In this study, we investigated the possibility that the prokaryotic SRP-like machinery is involved in biogenesis of membrane proteins in E. coli. The data presented here demonstrate that the SRP-receptor homologue, FtsY, is indeed essential for expression of integral membrane proteins in E. coli, indicating that, in the case of this group of proteins, FtsY and the mammalian SRP receptor have similar functions.