Evidence that Wra and Wrb are antithetical

Studies on 24 Wr(a+b+) and 23 Wr(a-b+) blood samples, using anti-Wrb in the enzyme-linked antiglobulin test (ELAT), have shown that Wr(a+b+) red cells bind, on average, a little over half the amount of anti-Wrb bound by Wr(a-b+) red cells. Similarly, ELAT studies using six different anti-Wra and 10 Wr(a+b+) samples, as well as red cells from the original Wr(a+b-) proposita, have shown that Wr(a+b+) red cells bind about half the amount of anti-Wra bound by Wr(a+b-) red cells. Various pitfalls that can arise when the ELAT is used to measure antigen ratios on red cells have been avoided but are described. This conclusive evidence that Wra and Wrb have an antithetical relationship is discussed in light of the knowledge that a ficin-resistant portion of MN sialoglycoprotein (SGP), when carried in liposomes, can inhibit anti-Wrb. It is possible that Wra, Wrb, or both may encode a post-translational change in MN SGP, or production of transferases that glycosylate membrane lipids that affect in situ orientation of MN SGP, or production of protein band 3 that then forms a complex with MN SGP at the red cell membrane surface.     

Heterogeneity of anti-U demonstrable by the use of papain-treated red cells

When red cells (RBCs) are treated with papain, one form of the U antigen, which we have named UPS (U papain-sensitive), is almost completely removed or denatured. A second form, UPR (U papain-resistant), remains unaltered on the treated RBCs. Tests on 42 examples of anti-U showed that two contained only anti-UPS, 19 contained only -UPR, and 21 contained separable -UPS and -UPR. In those sera containing both antibodies, anti-UPR was always the stronger of the two. These findings suggest 1) that UPS is located on the Ss sialoglycoprotein (glycophorin B) at a position distal to a papain-sensitive site or that the cleavage point is within the portion of the SGP that comprises UPS, and 2) that UPR is located between the papain-sensitive site and the RBC membrane. The UPS determinant was not denatured by neuraminidase, L-cysteine, trypsin, ficin, or alpha-chymotrypsin, and it was only partially denatured by pronase. The finding that RBCs treated with para-chloromercuribenzoic acid or para-chloromercuriphenyl sulfonic acid did not react with anti-UPR but did continue to react with anti-UPS suggests that the in situ configuration of UPR, but not UPS, is dependent on the presence of one or more disulfide bonds. RBCs of the S-s-U+(weak) phenotype were shown to carry markedly reduced amounts of both UPS and UPR.     

Contribution of the Vertebral Posterior Elements in Anterior–Posterior DXA Spine Scans in Young Subjects

Because DXA is a projection technique, anterior–posterior (AP) measurements of the spine include the posterior elements and the vertebral body. This may be a disadvantage because the posterior elements likely contribute little to vertebral fracture resistance. This study used QCT to quantify the impact of the posterior elements in DXA AP spine measures. We examined 574 subjects (294 females and 280 males), age 6–25 yr, with DXA and QCT. QCT measures were calculated for the cancellous bone region and for the vertebral body including and excluding the posterior elements. DXA data were analyzed for the entire L₃ vertebra and for a 10‐mm slice corresponding to the QCT scan region. BMC and BMD were determined and compared using Pearson's correlation. The posterior elements accounted for 51.4 ± 4.2% of the total BMC, with a significant difference between males (49.9 ± 4.0%) and females (52.8 ± 3.9%, p < 0.001). This percentage increased with age in younger subjects of both sexes (p < 0.001) but was relatively consistent after age 17 for males and 16 for females (p > 0.10). DXA areal BMD and QCT volumetric BMD correlated strongly for the whole vertebra including the posterior elements (R = 0.83), with BMC measures showing a stronger relationship (R = 0.93). Relationships were weaker when excluding the posterior elements. We conclude that DXA BMC provides a measure of bone that is most consistent with QCT and that the contribution of the posterior elements is consistent in young subjects after sexual maturity.     

Campylobacter jejuni Glycosylation Island Important in Cell Charge,Legionaminic Acid Biosynthesis,and Colonization of Chickens

Previously, we identified five genes (Cj1321 to Cj1326, of which Cj1325 and Cj1326 are a single gene) in the O-linked flagellin glycosylation island that are highly prevalent in Campylobacter jejuni isolates from chickens. We report mutagenesis, functional, and structural data to confirm that this locus, and Cj1324 in particular, has a significant contributory role in the colonization of chickens by C. jejuni. A motile ΔCj1324 mutant with intact flagella was considerably less hydrophobic and less able to autoagglutinate and form biofilms than the parent strain, 11168H, suggesting that the surface charge of flagella of Cj1324-deficient strains was altered. The physical and functional attributes of the parent were restored upon complementation. Structural analysis of flagellin protein purified from the ΔCj1324 mutant revealed the absence of two legionaminic acid glycan modifications that were present in the parent strain, 11168H. These glycoform modifications were shown to be prevalent in chicken isolates and confirm that differences in the highly variable flagellin glycosylation locus can relate to the strain source. The discovery of molecular mechanisms influencing the persistence of C. jejuni in poultry aids the rational design of approaches to control this problematic pathogen in the food chain.Campylobacter jejuni is the leading bacterial cause of human gastroenteritis worldwide (7). Infection can cause symptoms including abdominal pain and fever with watery to bloody diarrhea (54). Occasionally, postinfectious sequelae follow C. jejuni infection and include reactive arthritis and Guillain-Barré syndrome (8). Recently, C. jejuni has been associated with immunoproliferative small intestine disease, which is a rare type of mucosa-associated lymphoid tissue lymphoma (31). The main source of transmission through the food chain is the consumption and handling of contaminated poultry, but the underlying reasons why chickens are particularly susceptible to colonization by C. jejuni are unknown (15). C. jejuni has also been recovered from nonavian livestock, unpasteurized milk, and contaminated water (7). The socioeconomic burden of this pathogen means that it is imperative that ways of reducing the levels of C. jejuni in the food chain, particularly poultry, are investigated.The glycosylation of flagellin in a number of gram-negative pathogenic bacteria, including Pseudomonas aeruginosa, Helicobacter pylori, and Aeromonas spp., is increasingly recognized as playing significant roles (2, 24, 32, 43, 49). Glycosylation modifications have been shown to influence the cell''s immunogenicity, interaction with eukaryotic cells, and host cell specificity. Aeromonads are waterborne bacteria that can cause disease in fish, reptiles, and amphibians. Mesophilic aeromonads are important human pathogens causing gastrointestinal infections and, in severe cases, wound disease and septicemia in healthy and immunocompromised patients (63). Flagella of the mesophilic aeromonad Aeromonas caviae have been shown to be glycosylated (43) with a derivative of pseudaminic acid (50). In the plant pathogen Pseudomonas syringae pv. glycinea, the mutation of three genes located in a flagellin glycosylation island results in alterations to host specificity (61). Mutants of P. syringae pv. glycinea fail to cause symptoms in the normal host, soybean plants, but can grow on nonhost tobacco leaves, causing symptom-like changes on leaves. Takeuchi et al. proposed that the posttranslational modification of flagellin may be an adaptation of the bacterium to avoid recognition by host defenses (61). In P. aeruginosa strain PAK, a flagellin glycosylation island comprising 14 genes was discovered and shown to cause glycosylation exclusively for P. aeruginosa isolates expressing a-type flagellin (2). Further studies have shown that there appears to be variation in the glycosylation islands of strains containing the a-type flagellin (4). A glycosylation island comprising four genes in the type b flagellin strain P. aeruginosa PAO has been found. When a mutant unable to glycosylate flagellin was tested in a murine model of burn wound infection, it exhibited a reduction in virulence compared to that of the wild type (3). Thus, it appears that in P. aeruginosa different glycoforms on the flagellin are required for the colonization of different hosts or environments and that these glycoforms may provide the bacterium with a specific survival advantage.We recently examined 111 strains of C. jejuni, including human, chicken, bovine, ovine, and environmental isolates, using comparative phylogenomics (whole-genome comparisons of microbes using DNA microarrays combined with Bayesian-based phylogenies) (10). Isolates fell into two distinct clades, which based on the origins of the isolates were defined as livestock-associated and non-livestock-associated clades. Over 40 genes were identified as being significantly prevalent in either of the clades. Among these was a set of six genes, Cj1321, Cj1322, Cj1323, Cj1324, Cj1325, and Cj1326 (as identified in the initial annotation by Parkhill et al. for the original sequenced C. jejuni strain, NCTC11168 [42]), that lie within a region of the genome encoding the flagellin O-linked glycosylation system. Thus, although genes Cj1321 to Cj1326 are located within a region of the genome which has variability, they are conserved among some C. jejuni strains that are often associated with livestock. Microarray data have shown that the six genes are all transcribed in the same orientation, but it is unknown if they are an operon (N. Dorrell and B. W. Wren, unpublished data). NCTC11168 has since been reannotated, and as a result, Cj1325 and Cj1326 are now considered to be one gene, hereinafter referred to as Cj1325/6 (22). Previous BLAST analyses have shown that the Cj1321 protein has amino acid similarity to many bacterial acetyl transferases, both Cj1322 and Cj1323 proteins are similar to hydroxyacyl dehydrogenases, and the product of Cj1324 is similar to WbpG, a protein involved in lipopolysaccharide synthesis in many bacteria.In C. jejuni NCTC11168 (the original sequenced strain, found in the livestock clade), the O-linked flagellar glycosylation system is thought to consist of a cluster of approximately 50 genes (Cj1293 to Cj1342) adjacent to flaA and flaB which encode the structural flagellin proteins (42). The full glycan structure(s) in NCTC11168 (and most other strains associated with livestock) is unknown, but given the considerably larger size of the O-linked glycosylation loci in the livestock-associated strains than in the non-livestock-associated strains, it is likely that the livestock-associated strains may have additional modifications to the pseudaminic acid basic structure, as well as other unique glycan moieties, compared to those of the non-livestock-associated strains. The flagellin O-linked glycosylation locus in C. jejuni 81-176 (a frequently studied human strain found in the non-livestock-associated clade) is far simpler than that in C. jejuni NCTC11168, comprising just 26 genes (21). Two modifications predominantly decorate FlaA and FlaB of strain 81-176, the nine-carbon sugar pseudaminic acid (5,7-diacetamido-3,5,7,9-tetradeoxy-l-glycero-α-l-manno-nonulosonic acid [Pse5Ac7Ac]) and an acetamidino form of pseudaminic acid, 5-acetamido-7-acetamidino-3,5,7,9-tetradeoxy-l-glycero-α-l-manno-nonulosonic acid (Pse5Ac7Am). Derivatives of Pse5Ac7Am also decorate the flagellin of 81-176 in minor quantities (34, 37, 62). Genetic analysis of 81-176 showed that pse genes are involved in the biosynthesis of pseudaminic acid and its derivatives (21, 37, 62). More recently, the full biosynthetic pathway for pseudaminic acid was determined; in a six-step reaction, UDP-N-acetylglucosamine (UDP-GlcNAc) is converted to pseudaminic acid through the actions of PseB/Cj1293, PseC/Cj1294, PseH/Cj1313, PseG/Cj1312, PseI/Cj1317, and PseF/Cj1311 proteins (The Cj designations refer to predicted coding sequences in C. jejuni NCTC11168) (11, 19, 21, 35, 52, 62).The most detailed analysis of the flagellin O-linked glycosylation locus has been undertaken with C. jejuni strain 81-176 and the related species Campylobacter coli (strain VC167) (34). Structural studies of the flagellum modifications of C. coli VC167 revealed that in addition to Pse5Ac7Ac, acetamidino and N-methylacetimidoyl derivatives of legionaminic acid [5-acetamidino-7-acetamido-3,5,7,9-tetradeoxy-d-glycero-d-galacto-nonulosonic acid (Leg5Am7Ac) and 5-E/Z-N-(N-methlyacetimidoyl)-7-acetamidino-3,5,7,9-tetradeoxy-d-galacto-nonulosonic acid (Leg5AmNMe7Ac), respectively] decorate the C. coli flagellin, the first demonstration of a legionaminic acid derivative modification of bacterial flagellin (36). Biosynthesis of these legionaminic acid derivatives involves a distinct pathway encoded by the posttranslational modification (ptm) genes (34, 36). Although the precise pathway for the production of legionaminic acid has yet to be determined, tentative functions have been assigned which have identified PtmA to PtmH to be required for biosynthesis (PtmA, PtmB, PtmC, PtmD, PtmE, PtmF, PtmG, and PtmH are equivalent to the Cj1332, Cj1331, Cj1327, Cj1328, Cj1329, Cj1330, Cj1324, and Cj1325/6 proteins, respectively) (36). This ptm pathway is absent in C. jejuni strain 81-176. PtmG and PtmH from C. coli VC167 show 86 and 76% amino acid sequence similarity, respectively, to two hypothetical proteins, the Cj1324 and Cj1325/6 proteins of C. jejuni NCTC11168. The enzyme(s) involved in the attachment of glycan(s) to the flagellin protein of Campylobacter strains and the consensus sequence for the O-linked glycosylation process have yet to be identified.In C. jejuni strain 81-176, glycosylation of flagellin has been shown to be necessary for the assembly of flagella and subsequent motility (19). There is extensive polymorphism in the C. jejuni O-linked glycosylation cluster, suggesting that selective pressure may cause the bacterium to alter surface antigens in attempts to evade the host immune defenses (59). Evidence supporting this possibility is demonstrated by comparing the glycan moieties of the flagella of C. jejuni 81-176 and C. coli VC167, as these strains produce unique modifications on their flagella which affect serospecificity (34).Given the diversity of the O-linked glycosylation system in C. jejuni and the prevalence of the locus of Cj1321 to Cj1325/6 in chicken isolates, we hypothesized that these genes may be important for the abilities of some C. jejuni strains to colonize poultry and that colonization may be mediated through structural and surface charge changes in the glycan that modifies the flagellin. In this study, we demonstrate that Cj1324 is involved in the biosynthesis of two novel legionaminic acid modifications found on the flagellin of strain 11168H. The presence of these modifications affects autoagglutination, cell charge, and the efficiency with which C. jejuni 11168H colonizes chickens.     

Risk of sleep apnea in orchestra members

BackgroundObstructive sleep apnea (OSA) is a common condition with substantial health consequences. A recent randomized trial found that playing the didgeridoo improved both subjective and objective sleep measures. We undertook a cross-sectional survey of professional orchestra players to test the hypothesis that playing a wind instrument would be associated with a lower risk of OSA.MethodsAn anonymous internet-based survey of professional orchestra members assessed risk of sleep apnea using the Berlin questionnaire. Multivariable logistic regression was used to test the association between playing a wind instrument and having a high risk score on the Berlin questionnaire, both unadjusted and adjusted for age, body mass index, and gender.ResultsA total of 1,111 orchestra members responded, including 369 (33%) wind instrument players. Wind players were more often male and had a higher body mass index than non-wind players. Of all musicians, 348 (31%) had a high risk of sleep apnea. Wind players were more likely than non-wind players to be at high risk in unadjusted analysis (Odds ratio = 1.47, 95% CI 1.13, 1.91), though this association was not significant in adjusted analysis (Odds ratio = 1.12 (0.82, 1.54)).ConclusionPlaying a wind instrument was not associated with a lower risk of OSA.     

Asystole during laryngoscopy of a patient with pleural and pericardial effusions: a case report

A 53-year-old woman presented to the operating room for surgical correction of pericardial and pleural effusions. Her history included stage IV breast cancer, well-controlled hypertension, and diverticulitis. Although her baseline blood pressure, heart rate, and respirations were normal, she was short of breath with diminished breath sounds on the left side of the lungs and required oxygen, 2 L/min via nasal cannula. The nurse anesthesia student, under the direction of the Certified Registered Nurse Anesthetist (CRNA) and anesthesiologist, induced general anesthesia with etomidate, fentanyl, lidocaine, and succinylcholine. During placement of a double-lumen endotracheal tube, the patient became asystolic. The nurse anesthesia student immediately withdrew the laryngoscope, and the patient returned to normal sinus rhythm. A second attempt at laryngoscopy produced asystole as well. Again, the laryngoscope was withdrawn, and the patient returned to normal sinus rhythm. After resuming ventilation with 100% oxygen and administering atropine, 0.4 mg, the next intubation was successful, producing no untoward effects. Reintubation at the end of the case with a single lumen endotracheal tube was uneventful. The patient was transported to the intensive care unit and mechanically ventilated overnight. The next morning, she was extubated with no further anesthetic complications.     

Impact of rapid molecular screening for meticillin-resistant Staphylococcus aureus in surgical wards

Background:

This study aimed to establish the feasibility and cost‐effectiveness of rapid molecular screening for hospital‐acquired meticillin‐resistant Staphylococcus aureus (MRSA) in surgical patients within a teaching hospital.

Methods:

In 2006, nasal swabs were obtained before surgery from all patients undergoing elective and emergency procedures, and screened for MRSA using a rapid molecular technique. MRSA‐positive patients were started on suppression therapy of mupirocin nasal ointment (2 per cent) and undiluted chlorhexidine gluconate bodywash.

Results:

A total of 18 810 samples were processed, of which 850 (4·5 per cent) were MRSA positive. In comparison to the annual mean for the preceding 6 years, MRSA bacteraemia fell by 38·5 per cent (P < 0·001), and MRSA wound isolates fell by 12·7 per cent (P = 0·031). The reduction in MRSA bacteraemia and wound infection was equivalent to a saving of 3·78 beds per year (£276 220), compared with the annual mean for the preceding 6 years. The cost of screening was £302 500, making a net loss of £26 280. Compared with 2005, however, there was a net saving of £545 486.

Conclusion:

Rapid MRSA screening of all surgical admissions resulted in a significant reduction in staphylococcal bacteraemia during the screening period, although a causal link cannot be established. Copyright © 2007 British Journal of Surgery Society Ltd. Published by John Wiley & Sons, Ltd.     

Screening for childhood anxiety symptoms in primary care: integrating child and parent reports

OBJECTIVE: Parents' concerns typically determine the focus of a primary care visit. This study examined which information is lost if child reports are excluded from screening for anxiety. It also explores the use in primary care of the Screen for Child Anxiety Related Emotional Disorders (SCARED) and the Pediatric Symptom Checklist (PSC-17). METHOD: Two hundred thirty-six children (8-12 years 11 months) and their parents completed SCARED and PSC-17 before a primary care visit occurring during discrete periods between June 1999 and March 2001. RESULTS: Child reports yielded higher SCARED scores than parent reports (mean=18.12, SD=12.14 versus 14.43, SD=10.34, p <.001). Somatic/panic and separation anxiety accounted for 73.8% of the excess score from children's reports. The level of parent-reported symptoms did not vary with demographics. Female gender and younger age predicted greater excess reporting by children. Parent and child scores were moderately to highly correlated (R=0.55 total score; 0.40-0.58 subfactors). CONCLUSIONS: There are discrete anxiety domains in which children's reports are likely to yield more information than that of parents. This phenomenon is almost entirely attributable to variations in the level of symptoms reported by children. Studies are needed to design brief screening procedures that integrate parent and child reports and carry age- and gender-adjusted thresholds.     

Concurrent and discriminant validity of Spanish language instruments for measuring functional health status

BACKGROUND: Questionnaires translated into languages other than English are often not validated to the same extent as the English versions. This study examined the concurrent and discriminant validity of selected domains related to physical function from Spanish language versions of the Child Health Questionnaire (CHQ), Pediatric Outcomes Data Collection Instrument (PODCI), and Pediatric Evaluation and Disability Inventory (PEDI). METHODS: Concurrent validity was examined in 93 children with cerebral palsy by correlating questionnaire domain scores with Gross Motor Function Measure and Gillette Functional Assessment Questionnaire walking scale scores. Discriminant validity with respect to Gross Motor Function Classification System (GMFCS) level was examined using analysis of variance and nonparametric discriminant analysis. RESULTS: Concurrent validity was demonstrated for 3 domains from the PEDI (Mobility functional skills, tau = 0.62; Mobility caregiver assistance, tau = 0.46-0.55; and Self-care functional skills, tau = 0.30-0.36), 3 domains from the PODCI (Sports and physical function, tau = 0.48-0.51; Transfer and basic mobility, tau = 0.48-0.51; and Upper extremity physical function, tau = 0.28), and 1 domain from the CHQ (Physical function, tau = 0.31-0.36). Discriminant validity was demonstrated for the same domains based on significant decreases in domain scores with increasing GMFCS level. Discriminant validity was highest for the PODCI, which correctly classified 98% (91/93) of subjects into the correct GMFCS level when all 3 domains were considered. CONCLUSIONS: For the first time, concurrent validity and discriminant validity have been demonstrated for the physical function domains of Spanish language versions of the PODCI, PEDI, and CHQ questionnaires. PODCI and PEDI had the highest concurrent validity, and PODCI had the best discriminant ability. CLINICAL RELEVANCE: It is important to examine the validity of instruments when they have been translated from English into other languages. This importance will only increase as the population of non-English-speaking patients expands.