Genome sequence based, comparative analysis of the fluorescent amplified fragment length polymorphisms (FAFLP) of tubercle bacilli from seals provides molecular evidence for a new species within the Mycobacterium tuberculosis complex.

Tuberculosis in seals is caused by a member of the Mycobacterium tuberculosis complex referred to as the 'seal bacillus'. Fluorescent amplified-fragment length polymorphism (FAFLP) analysis was applied to isolates from four Australian and six Argentinean seals and compared with FAFLP pattern for standard strains belonging to the M. tuberculosis complex. The FAFLP profiles derived from EcoRI/MseI restricted fragments of blind coded DNA samples differentiated the seal bacillus from other members of the M. tuberculosis complex. According to the phylogenetic analysis performed using FAFLP data, seal bacilli appear to have diverged significantly from other members of the M. tuberculosis complex. We describe the suitability of a panel of 19 highly polymorphic markers for rapid identification and comparative genomic analyses of the seal bacillus strains. It is likely that these bacilli got separated from the M. tuberculosis lineage as a result of different insertion deletion events occurring on a genome wide scale. Our analysis reveals that the seal bacillus and M. bovis are genetically related and therefore, might have originated from a common ancestor. Our data additionally support the hypothesis that seal bacillus occupies a unique taxonomic position within the M. tuberculosis complex.     

Genome and transcriptome scale portrait of sigma factors in Mycobacterium avium subsp. paratuberculosis.

Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of Johne's disease (JD), a chronic gastroenteritis of ruminants and other animals, including primates. Many evidences suggested association of MAP to Crohn's disease, a chronic granulomatous gastrointestinal disease of humans with strong similarities with JD. The present study attempts to evaluate global gene regulation in MAP, which has not been addressed previously, despite the availability of MAP genome sequence. For this purpose, we investigated: (i) the presence of sigma factors and their relationship to sigma factors of other mycobacteria (M. avium subsp.avium, M. tuberculosis, M. bovis, M. leprae and M. smegmatis), and (ii) their expression during different growth conditions and in vitro infection of intestinal epithelial Caco2 cells. MAP genome contains 19 putative sigma factor, but only 12 belong to gene families common to other mycobacteria. Gene expression was evaluated with Real-Time PCR during growth in 7H9 medium and mycobactin J, in 7H9 medium plus mycobactin J and lisozyme, and during infection of Caco2 cells: very different expression patterns were observed and, on the whole, only 7 sigma factors were found to be expressed. sigJ was upregulated during the infection of Caco2 cells. Even if only few sigma factors were expressed in the three conditions tested, the overall high numbers of MAP sigma factors suggests a noteworthy flexibility of this pathogen. Thus, this first report on expression of MAP sigma factors opens the way to an extensive characterization of global gene regulation, as a key to understand strategies of survival and mechanisms of infections used by this organism.     

Analysis of genomic downsizing on the basis of region-of-difference polymorphism profiling of Mycobacterium tuberculosis patient isolates reveals geographic partitioning

Mycobacterium tuberculosis, the etiological agent of tuberculosis, has lost many coding and noncoding regions in its genome during the course of evolution. We performed region-of-difference (RD) analysis using PCR-based genotyping of 131 M. tuberculosis clinical isolates obtained from four different countries, namely, India, Peru, Libya, and Angola. Our studies revealed that RD patterns are often distinct for strains circulating in specific geographical regions and can be used to trace the descent and spread of an isolate from its original reservoir. We describe our findings, which show that no single isolate from the four countries (n = 131) had all the 15 RDs either deleted or retained. Tuberculosis-specific deletion 1 (TbD1) was found to be conserved in 23% of the Indian isolates, indicating their possible ancient origin. RD9 was the most conserved region, RD11 was predominantly deleted, and RD6 was the most variable among the isolates in our collection irrespective of their geographic region. In contrast to earlier reports, our results demonstrate that the deletion of RD1 does not correlate with a decrease in the virulence potential of M. tuberculosis, as Indian isolates (n = 30) examined by us were from diseased individuals and yet had lost the RD1 region. Our results further illustrated that the intactness of the RD5 region may be associated with increased virulence of the organism. This study highlights that the RDs in M. tuberculosis genomes are geographically distributed and specific and may possibly be associated with virulence spectrum.     

Back pain is also improved by lumbar disc herniation surgery

Background and purpose — Indication for lumbar disc herniation (LDH) surgery is usually to relieve sciatica. We evaluated whether back pain also decreases after LDH surgery.Patients and methods — In the Swedish register for spinal surgery (SweSpine) we identified 14,097 patients aged 20–64 years, with pre- and postoperative data, who in 2000–2016 had LDH surgery. We calculated 1-year improvement on numeric rating scale (rating 0–10) in back pain (N_back) and leg pain (N_leg) and by negative binomial regression relative risk (RR) for gaining improvement exceeding minimum clinically important difference (MCID).Results — N_leg was preoperatively (mean [SD]) 6.7 (2.5) and N_back was 4.7 (2.9) (p < 0.001). Surgery reduced N_leg by mean 4.5 (95% CI 4.5–4.6) and N_back by 2.2 (CI 2.1–2.2). Mean reduction in N_leg) was 67% and in N_back 47% (p < 0.001). Among patients with preoperative pain ≥ MCID (that is, patients with significant baseline pain and with a theoretical possibility to improve above MCID), the proportion who reached improvement ≥ MCID was 79% in N_leg and 60% in N_back. RR for gaining improvement ≥ MCID in smokers compared with non-smokers was for N_leg 0.9 (CI 0.8–0.9) and ­N_back 0.9 (CI 0.8–0.9), and in patients with preoperative duration of back pain 0–3 months compared with > 24 months for N_leg 1.3 (CI 1.2–1.5) and for N_back 1.4 (CI 1.2–1.5).Interpretation — LDH surgery improves leg pain more than back pain; nevertheless, 60% of the patients with significant back pain improved ≥ MCID. Smoking and long duration of pain is associated with inferior recovery in both N_leg and N_back.

The most common indication for lumbar disc herniation (LDH) surgery is persistent sciatica that does not respond to nonoperative treatment (Blamoutier 2013). However, most patients who undergo LDH surgery also suffer from back pain (Hakkinen et al. 2003, Stromqvist et al. 2017), on a national level reported in 93% of patients having LDH surgery (Stromqvist et al. 2017). Decades ago, Mixter (1937) therefore argued that LDH extirpation should be accompanied by fusion to minimize postoperative back pain. Recent studies have opposed this view, showing that LDH surgery is not followed by increased back pain when only removing the hernia (Pearson et al. 2008, Owens et al. 2018), and in many cases even improvement of back pain seems sustainable over time.Most studies that evaluate the outcome of LDH surgery focus on the relief from sciatica and improvement in patient-reported outcome measures (PROMs) (Weber 1983, Atlas et al. 2005, Peul et al. 2007, Weinstein et al. 2008, Lurie et al. 2014). A few studies have focused on back pain or included back pain in the evaluation (Kotilainen et al. 1993, Hakkinen et al. 2003, Toyone et al. 2004, Atlas et al. 2005, Pearson et al. 2008, Owens et al. 2018). While some of these infer that back pain is improved by the LDH surgery (Hakkinen et al. 2003, Toyone et al. 2004, Pearson et al. 2008, Owens et al. 2018) others report inconclusive results (Kotilainen et al. 1993, Atlas et al. 2005). There is a lack of consensus on the expected level of back pain reduction with LDH surgery.It would also be of clinical interest to identify preoperative factors that are associated with favorable reduction of back pain following LDH surgery such as age, sex, smoking, preoperative health, and duration of pain (Nygaard et al. 2000, Jansson et al. 2005, Stromqvist et al. 2016, Wilson et al. 2016, Hareni et al. 2019).We (i) evaluated whether back pain is reduced after LDH surgery and if so, to what extent compared with the reduction in leg pain and (ii) what proportion of patients gain improvement in back and leg pain exceeding minimum clinically important difference (MCID). The secondary aim was to identify factors associated with improvement in back pain exceeding MCID.      

Genomic and Functional Portrait of a Highly Virulent,CTX-M-15-Producing H30-Rx Subclone of Escherichia coli Sequence Type 131

Escherichia coli sequence type 131 (ST131) is a pandemic clone associated with multidrug-resistant, extraintestinal infections, attributable to the presence of the CTX-M-15 extended-spectrum β-lactamase gene and mutations entailing fluoroquinolone resistance. Studies on subclones within E. coli ST131 are critically required for targeting and implementation of successful control efforts. Our study comprehensively analyzed the genomic and functional attributes of the H30-Rx subclonal strains NA097 and NA114, belonging to the ST131 lineage. We carried out whole-genome sequencing, comparative analysis, phenotypic virulence assays, and profiling of the antibacterial responses of THP1 cells infected with these subclones. Phylogenomic analysis suggested that the strains were clonal in nature and confined entirely to a single clade. Comparative genomic analysis revealed that the virulence and resistance repertoires were comparable among the H30-Rx ST131 strains except for the commensal ST131 strain SE15. Similarly, seven phage-specific regions were found to be strongly associated with the H30-Rx strains but were largely absent in the genome of SE15. Phenotypic analysis confirmed the virulence and resistance similarities between the two strains. However, NA097 was found to be more robust than NA114 in terms of virulence gene carriage (dra operon), invasion ability (P < 0.05), and antimicrobial resistance (streptomycin resistance). RT² gene expression profiling revealed generic upregulation of key proinflammatory responses in THP1 cells, irrespective of ST131 lineage status. In conclusion, our study provides comprehensive, genome-inferred insights into the biology and immunological properties of ST131 strains and suggests clonal diversification of genomic and phenotypic features within the H30-Rx subclone of E. coli ST131.     

A review of ibuprofen and acetaminophen use in febrile children and the occurrence of asthma-related symptoms

BACKGROUND: Although many studies have investigated the safety and tolerability of ibuprofen or acetaminophen (paracetamol) use in children, few have specifically examined the association of ibuprofen or acetaminophen and the occurrence of asthma in pediatric populations. OBJECTIVES: The primary objective of this literature review was to ascertain whether ibuprofen use exacerbates the symptoms of asthma or asthma-related adverse events in febrile children, and how it compares with acetaminophen use. The secondary objective was to develop an algorithm that allows for the consideration of ibuprofen treatment in children by health care professionals. METHODS: Twelve electronic databases (MEDLINE, EMBASE, Cochrane Database, DARE, British Nursing Index, CBIB, Derwent Drug File, International Pharmaceutical Abstracts, Pharm-Line, CINAHL, PASCAL, SCZZ-SciSearch) were searched from their year of inception to June 2007, to identify English-language articles pertaining to ibuprofen or acetaminophen use in the asthmatic pediatric population. The following search terms were used: asthma, child, pediatric, pediatrics, ibuprofen, Nurofen, Brufen, Motrin, Advil, propionic acid, paracetamol, and acetaminophen. RESULTS: Of 472 articles retrieved, 3 were relevant for the development of the algorithm. Two were subanalyses of a major randomized controlled trial (RCT), the Boston University Fever Study. Therefore, some overlap should be noted. The third article was another RCT. Other studies and review articles identified were used for the discussion. Findings from the literature analysis indicated that the use of ibuprofen in the pediatric population does not exacerbate asthma morbidity. Two of the studies demonstrated that ibuprofen was associated with a lower risk for asthma morbidity in febrile children with or without asthma compared with acetaminophen. In one study, ibuprofen use was associated with a lower relative risk for hospitalization (0.63) and outpatient visits (0.56) for asthma compared with acetaminophen. In the second study, acetaminophen use was associated with the exacerbation of wheezing in febrile children. This observation was corroborated by the findings of other studies that revealed an increased risk for asthma, wheezing, and other atopic outcomes with acetaminophen use. CONCLUSIONS: The evidence reviewed in this article suggests a low risk for asthma-related morbidity associated with ibuprofen use in children and a possible protective and therapeutic effect compared with acetaminophen. The findings also suggest that acetaminophen use in children is associated with an increased risk for wheezing. The pediatric algorithm developed might serve as a guide for health care professionals in assessing suitability for ibuprofen use in children.     

A qualitative study of stakeholder views and experiences of minor ailment services in the United Kingdom

BackgroundAn international strategy designed to promote access to primary care is the utilisation of community pharmacy to deliver structured minor ailment services (MASs). An understanding of key implementation features of MASs will support effective service delivery and implementation, promote MAS viability, sustainability and overall improvement.AimThe aim of this study is to explore the views and experiences of a range of stakeholders concerning the implementation of MASs in the United Kingdom.MethodsA qualitative approach was used to obtain data. Participants were recruited using purposeful and snowball sampling. Stakeholders from five different regions were included. Using the digital recordings of the interviews, thematic content analysis was undertaken.ResultsThirty-three participants agreed to be interviewed. Twenty-nine semi-structured interviews were conducted. Thematic content analysis yielded three major themes, including (1) benefits of MASs, (2) structural challenges associated with MAS design and (3) other implementation factors associated with MAS delivery. Stakeholders recognised the positive impact of the service to improve patient access and care, promote efficiencies, and promote the professional role of the pharmacist. Nevertheless barriers do exist to service delivery and implementation. Stakeholders identified the need to potentially increase the population groups served by MASs, increase the conditions treated and widen their formulary lists. Similarly, marketing strategies needed to be improved to enhance consumer awareness. Stakeholders presented mixed views about whether pharmacists needed to complete clinical training and the need to increase pharmacist's remuneration. In addition the level of healthcare collaboration needed to improve.ConclusionSeveral concepts emerged from the investigation to facilitate service delivery. Barriers to service implementation had a variable impact on implementation. Service delivery should function to meet all stakeholder needs and can be achieved through stakeholder collaboration. However, improved marketing to promote consumer awareness together with better collaborative processes can potentially improve MAS implementation.     

Expression analysis of microRNA-155 in type 2 diabetes in Kashmiri population

International Journal of Diabetes in Developing Countries - The present study aimed to evaluate the expression of microRNA-155 (miR-155) in type 2 diabetes mellitus (T2DM) and assess its...     

Mutational analysis of the RB1 gene in Indian patients with retinoblastoma

Twenty-one probands, twelve with bilateral and nine with unilateral retinoblastoma, were screened for mutations in the RB1 gene using genomic DNA from peripheral blood leukocytes as well as tumors. Amplification of individual exons and flanking regions of the RB1 gene were carried out, followed by direct sequencing of the amplified products. Sequences of affected individuals were compared with those of controls. Mutations were identified in seven patients, five with bilateral and two with unilateral retinoblastoma. Six out of seven mutations involved the formation of premature termination codons by means of single base substitutions (2), frameshifts due to splice-site mutations (2), or deletion and duplication (2). One missense mutation was identified. Of the remaining fourteen patients, seven with bilateral disease had no mutations in peripheral blood (7 cases) or tumors (3/7 cases). Analysis of the peripheral blood of seven patients with unilateral disease also showed no mutations. Mutations were detected in about one-third of the cases, suggesting that hemizygous deletions at the RB1 locus or mutations outside the coding regions of RB1 may be responsible for the disease in the remaining patients.