Intragenomic and intraspecific heterogeneity of the 16S rRNA gene in seven bacterial species from the respiratory tract of cystic fibrosis patients assessed by PCR-Temporal Temperature Gel Electrophoresis

16S rRNA gene-based cultivation-independent methods are increasingly used to study the diversity of microbiota during health and disease. One bias of these methods is the variability of 16S rRNA gene that may exist among strains of a same species (intraspecific heterogeneity) or between rrs copies in a genome (intragenomic heterogeneity). We evaluated the level of intraspecific and intragenomic 16S rDNA variability in seven species frequently encountered in respiratory tract samples in cystic fibrosis (CF). A total of 179 strains were subjected to V3 region 16S rDNA PCR-TTGE. Using this easy-to-perform and rapid method, different levels of V3 region rrs heterogeneity were demonstrated. No intraspecific and intragenomic rrs heterogeneity was demonstrated for Moraxella catarrhalis (n=16), Pseudomonas aeruginosa (n=31) and Streptococcus pneumoniae (n=14) showing a single PCR-TTGE band characteristic of the species. Low level of intraspecific heterogeneity was observed for Staphylococcus aureus (n=30), Stenotrophomonas maltophilia (n=29) and Achromobacter xylosoxidans (n=28), and 17%, 38% and 96% of these strains showed intragenomic heterogeneity (two to four different rrs copies), respectively. Haemophilus influenzae (n=31) displayed the higher level of intraspecific variability with 23 different PCR-TTGE patterns and 61% of the strains showed intragenomic rrs heterogeneity (two to four different rrs copies). Although only one hypervariable region of the 16S rRNA gene was explored, intraspecific and intragenomic rrs heterogeneity was frequently observed in this study and should be taken into consideration for a better interpretation of 16S rRNA gene-based diversity profiles in denaturing gels and to avoid any overestimation of the respiratory microbiota diversity in CF.     

Perception of health care quality by patients with chronic conditions

BackgroundPatient opinion is becoming ever more important when considering healthcare quality and the reforms required to improve healthcare quality. The main aim of this study was to explore factors determining perceived healthcare quality among patients with chronic diseases.MethodsData are drawn from the survey carried out in 2008 by the Commonwealth Fund, in partnership with the French Superior Health Authority (Haute Autorité de santé). The prospective telephone survey targeted adults in eight countries who had serious health problems (chronic or severe disease, declared poor state of health, hospital admission or major surgery). Of the 1202 French respondents, 851 had at least one diagnosed chronic disease. A multinomial logistic model was used to identify the relationship between perceived healthcare quality and patients’ recent experience with the healthcare system.ResultsPeople with chronic disease in general perceived that healthcare quality was excellent (45%) or good (44%). Only 11% of respondents judged it to be average or poor. There was a hint of “could do better”, for example when considering podology and ophthalmology follow-up in diabetes or the management of multiple medications. The explanatory model revealed a positive correlation between excellent perceived healthcare quality and a strong doctor–patient relationship, taking into account both the length of this relationship and the ability of the doctor to involve the patient at all stages of decision-making concerning therapeutic management. There was no major link between the perceived quality of care and objective care quality, the quality of procedures, the cost of care to the patient or how frequently patients access the healthcare system.ConclusionThe quality of the relationship between the patient and his/her doctor is a determining factor in the patient's judgement of the quality of healthcare he/she receives.     

A differing pattern of association between dietary fish and allergen-specific subgroups of atopy

BACKGROUND: We examined the role of fish intake in the development of atopic disease with particular reference to the possibility of differential effects on allergen-specific subgroups of sensitization. METHODS: The exposure of interest was parental report of fish intake by children aged 8 years at the 1997 Childhood Allergy and Respiratory Health Study (n = 499). The outcomes of interest were subgroups of atopy: house dust mite (HDM)-pure sensitization [a positive skin-prick test (SPT) > or = 2 mm to Der p or Der f only], ryegrass-pure sensitization (a positive SPT > or = 2 mm to ryegrass only); asthma and hay fever by allergen-specific sensitization. RESULTS: A significant association between fish intake and ryegrass-pure [adjusted odds ratio (AOR) 0.37 (0.15-0.90)] but not HDM-pure sensitization [AOR 0.87 (0.36-2.13)] was found. Fish consumption significantly decreased the risk for ryegrass-pure sensitization in comparison with HDM-pure sensitization [AOR 0.20 (0.05-0.79)]. CONCLUSIONS: We have demonstrated a differential effect of fish intake for sensitization to different aeroallergens. This may be due to the different timing of allergen exposure during early life. Further investigation of the causes of atopic disease should take into account allergen-specific subgroups.     

Inflammatory complications of CGRP monoclonal antibodies: a case series

BackgroundCalcitonin gene-related peptide (CGRP) is expressed throughout the body and is a known mediator of migraine, exerting this biological effect through activation of trigeminovascular, meningeal and associated neuronal pathways located in close proximity to the central nervous system. Monoclonal antibodies (mAb) targeting the CGRP pathway are an effective new preventive treatment for migraine, with a generally favourable adverse event profile. Pre-clinical evidence supports an anti-inflammatory/immunoregulatory role for CGRP in other organ systems, and therefore inhibition of the normal action of this peptide may promote a pro-inflammatory response.CasesWe present a case series of eight patients with new or significantly worsened inflammatory pathology in close temporal association with the commencement of CGRP mAb therapy.ConclusionThis case series provides novel insights on the potential molecular mechanisms and side-effects of CGRP antagonism in migraine and supports clinical vigilance in patient care going forward.