The human gastrointestinal tract (GIT) represents one of the most densely populated microbial ecosystems studied to date. Although this microbial consortium has been recognized to have a crucial ...impact on human health, its precise composition is still subject to intense investigation. Among the GIT microbiota, bifidobacteria represent an important commensal group, being among the first microbial colonizers of the gut. However, the prevalence and diversity of members of the genus Bifidobacterium in the infant intestinal microbiota has not yet been fully characterized, while some inconsistencies exist in literature regarding the abundance of this genus.
In the current report, we assessed the complexity of the infant intestinal bifidobacterial population by analysis of pyrosequencing data of PCR amplicons derived from two hypervariable regions of the 16 S rRNA gene. Eleven faecal samples were collected from healthy infants of different geographical origins (Italy, Spain or Ireland), feeding type (breast milk or formula) and mode of delivery (vaginal or caesarean delivery), while in four cases, faecal samples of corresponding mothers were also analyzed.
In contrast to several previously published culture-independent studies, our analysis revealed a predominance of bifidobacteria in the infant gut as well as a profile of co-occurrence of bifidobacterial species in the infant's intestine.
•HT qPCR detected all 73 targeted ARGs, while metagenomics detected 491 ARGs.•HT qPCR was more sensitive to low abundance genes, detecting all target ARGs.•Both methods enabled the spatiotemporal ...separation of hospital and WWTP resistomes.•Metagenomics provided contextual data making it more suitable for risk assessment.•HT qPCR permitted more sensitive quantification of clinically relevant AMR genes.
Wastewater serves as an important reservoir of antimicrobial resistance (AMR), and its surveillance can provide insights into population-level trends in AMR to inform public health policy. This study compared two common high-throughput screening approaches, namely (i) high-throughput quantitative PCR (HT qPCR), targeting 73 antimicrobial resistance genes, and (ii) metagenomic sequencing. Weekly composite samples of wastewater influent were taken from 47 wastewater treatment plants (WWTPs) across Wales, as part of a national AMR surveillance programme, alongside 4 weeks of daily wastewater effluent samples from a large municipal hospital. Metagenomic analysis provided more comprehensive resistome coverage, detecting 545 genes compared to the targeted 73 genes by HT qPCR. It further provided contextual information critical to risk assessment (i.e. potential bacterial hosts). In contrast, HT qPCR exhibited higher sensitivity, quantifying all targeted genes including those of clinical relevance present at low abundance. When limited to the HT qPCR target genes, both methods were able to reflect the spatiotemporal dynamics of the complete metagenomic resistome, distinguishing that of the hospital and the WWTPs. Both approaches revealed correlations between resistome compositional shifts and environmental variables like ammonium wastewater concentration, though differed in their interpretation of some potential influencing factors. Overall, metagenomics provides more comprehensive resistome profiling, while qPCR permits sensitive quantification of genes significant to clinical resistance. We highlight the importance of selecting appropriate methodologies aligned to surveillance aims to guide the development of effective wastewater-based AMR monitoring programmes.
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Gut microbiome manipulation with the aim of altering the respiratory immune response is a novel approach to protect the host against respiratory infections. Recent clinical trials of probiotics show ...promise in this regard in healthy adults and children. However, few data exist regarding the potential role of probiotics in preventing respiratory infections in overweight/obese and older people, despite these groups having markedly elevated risks of viral upper respiratory tract infections (URTI) in particular.
We recently performed a randomised double-blind, placebo-controlled trial to investigate the impact of probiotics on free-living overweight/obese people (220 participants, BMI 25-34.9kg/m2, aged 30-65 years), with primary outcomes focused upon any probiotic effect upon weight, BMI and other anthropometric measures (PROMAGEN trial, trial number: ISRCTN12562026). Participants were randomised 1:1 to either daily Lab4P probiotic (50 billion colony-forming units, containing three strains of Lactobacilli and two strains of Bifidobacteria) or placebo for six months. We analyzed self-reported daily symptom diaries recorded contemporaneously by study participants during the trial, focusing on five key symptomatic indicators of URTI, namely: cough, sore throat, headache, muscle ache and wheeze (‘URTI symptoms’). Poisson regression analysis and log-rank test were used to statistically investigate the impact of probiotics, BMI and age upon rates of URTI symptoms.
As previously reported, at the study's end, significant between group decreases were identified in body weight (1.3kg, p<0.0001) and BMI (0.045kg/m2, p<0.0001), both favouring the probiotic arm. Participants taking probiotic experienced a 27% lower overall incidence of URTI symptoms vs those taking placebo (Incidence Rate Ratio (IRR): 0.73, 95% confidence intervals (CI): 0.63, 0.84, p<0.0001; Figure 1). Poisson regression identified significant interactions for symptom incidence with age (p=0.0071) and BMI (p=0.0393), with the greater reduction in URTI symptoms observed in participants taking probiotics who were >45 years old compared to younger participants (IRR: 0.60, p<0.0001 vs IRR: 0.90, p=0.3336), and obese participants compared to overweight (IRR: 0.57, p<0.0001 vs IRR: 0.78, p=0.0059; Figure 1). After only 1-2 weeks supplementation, there was a clear indication of a divergence between groups in the time taken to record the first URTI symptoms, favouring the probiotic, in two subgroups (i.e. those ≥ 45 years old and the obese; Figure 2).
Probiotic use is associated with a more marked reduction in URTI symptoms in overweight/obese people than in their counterparts. Such apparent benefits in preventing URTIs are most notable in obese and/or older participants, and occur rapidly after commencing probiotics.
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Mutations in the epidermal growth factor receptor gene (EGFR) represent one of the most frequent “actionable” alterations in non–small cell lung cancer (NSCLC). Typified by high response rates to ...targeted therapies, EGFR tyrosine kinase inhibitors (TKIs) are now established first-line treatment options and have transformed the treatment paradigm for NSCLC. With the recent breakthrough designation and approval of the third-generation EGFR TKI osimertinib, available systemic and local treatment options have expanded, requiring new clinical algorithms that take into account individual patient molecular and clinical profiles. In this International Association for the Study of Lung Cancer commissioned consensus statement, key pathologic, diagnostic, and therapeutic considerations, such as optimal choice of EGFR TKI and management of brain metastasis, are discussed. In addition, recommendations are made for clinical guidelines and research priorities, such as the role of repeat biopsies and use of circulating free DNA for molecular studies. With the rapid pace of progress in treating EGFR-mutant NSCLC, this statement provides a state-of-the-art review of the contemporary issues in managing this unique subgroup of patients.
Laboratory-based studies on neuromuscular control after concussion and epidemiological studies suggest that concussion may increase the risk of subsequent musculoskeletal injury.
The purpose of this ...study was to determine if athletes have an increased risk of lower extremity musculoskeletal injury after return to play from a concussion.
Injury data were collected from 2006 to 2013 for men's football and for women's basketball, soccer and lacrosse at a National Collegiate Athletic Association Division I university. Ninety cases of in-season concussion in 73 athletes (52 male, 21 female) with return to play at least 30 days prior to the end of the season were identified. A period of up to 90 days of in-season competition following return to play was reviewed for time-loss injury. The same period was studied in up to two control athletes who had no concussion within the prior year and were matched for sport, starting status and position.
Lower extremity musculoskeletal injuries occurred at a higher rate in the concussed athletes (45/90 or 50 %) than in the non-concussed athletes (30/148 or 20 %; P < 0.01). The odds of sustaining a musculoskeletal injury were 3.39 times higher in the concussed athletes (95 % confidence interval 1.90-6.05; P < 0.01). Overall, the number of days lost because of injury was similar between concussed and non-concussed athletes (median 9 versus 15; P = 0.41).
The results of this study demonstrate a relationship between concussion and an increased risk of lower extremity musculoskeletal injury after return to play, and may have implications for current medical practice standards regarding evaluation and management of concussion injuries.
The increased detection of small-sized peripheral non-small-cell lung cancer (NSCLC) has renewed interest in sublobar resection in lieu of lobectomy.
We conducted a multicenter, noninferiority, phase ...3 trial in which patients with NSCLC clinically staged as T1aN0 (tumor size, ≤2 cm) were randomly assigned to undergo sublobar resection or lobar resection after intraoperative confirmation of node-negative disease. The primary end point was disease-free survival, defined as the time between randomization and disease recurrence or death from any cause. Secondary end points were overall survival, locoregional and systemic recurrence, and pulmonary functions.
From June 2007 through March 2017, a total of 697 patients were assigned to undergo sublobar resection (340 patients) or lobar resection (357 patients). After a median follow-up of 7 years, sublobar resection was noninferior to lobar resection for disease-free survival (hazard ratio for disease recurrence or death, 1.01; 90% confidence interval CI, 0.83 to 1.24). In addition, overall survival after sublobar resection was similar to that after lobar resection (hazard ratio for death, 0.95; 95% CI, 0.72 to 1.26). The 5-year disease-free survival was 63.6% (95% CI, 57.9 to 68.8) after sublobar resection and 64.1% (95% CI, 58.5 to 69.0) after lobar resection. The 5-year overall survival was 80.3% (95% CI, 75.5 to 84.3) after sublobar resection and 78.9% (95% CI, 74.1 to 82.9) after lobar resection. No substantial difference was seen between the two groups in the incidence of locoregional or distant recurrence. At 6 months postoperatively, a between-group difference of 2 percentage points was measured in the median percentage of predicted forced expiratory volume in 1 second, favoring the sublobar-resection group.
In patients with peripheral NSCLC with a tumor size of 2 cm or less and pathologically confirmed node-negative disease in the hilar and mediastinal lymph nodes, sublobar resection was not inferior to lobectomy with respect to disease-free survival. Overall survival was similar with the two procedures. (Funded by the National Cancer Institute and others; CALGB 140503 ClinicalTrials.gov number, NCT00499330.).
Purpose Active surveillance represents a strategy to address the overtreatment of prostate cancer, yet uncertainty regarding individual patient outcomes remains a concern. We evaluated outcomes in a ...prospective multicenter study of active surveillance. Materials and Methods We studied 905 men in the prospective Canary PASS enrolled between 2008 and 2013. We collected clinical data at study entry and at prespecified intervals, and determined associations with adverse reclassification, defined as increased Gleason grade or greater cancer volume on followup biopsy. We also evaluated the relationships of clinical parameters with pathology findings in participants who underwent surgery after a period of active surveillance. Results At a median followup of 28 months 24% of participants experienced adverse reclassification, of whom 53% underwent treatment while 31% continued on active surveillance. Overall 19% of participants received treatment, 68% with adverse reclassification, while 32% opted for treatment without disease reclassification. In multivariate Cox proportional hazards modeling the percent of biopsy cores with cancer, body mass index and prostate specific antigen density were associated with adverse reclassification (p=0.01, 0.04, 0.04, respectively). Of 103 participants subsequently treated with radical prostatectomy 34% had adverse pathology, defined as primary pattern 4-5 or nonorgan confined disease, including 2 with positive lymph nodes, with no significant relationship between risk category at diagnosis and findings at surgery (p=0.76). Conclusions Most men remain on active surveillance at 5 years without adverse reclassification or adverse pathology at surgery. However, clinical factors had only a modest association with disease reclassification, supporting the need for approaches that improve the prediction of this outcome.
Malignant pleural effusions (MPE) complicate malignancies and portend worse outcomes. MPE is comprised of various components, including immune cells, cancer cells, and cell-free DNA/RNA. There have ...been investigations into using these components to diagnose and prognosticate MPE. We hypothesize that the microbiome of MPE is unique and may be associated with diagnosis and prognosis. We compared the microbiota of MPE against microbiota of pleural effusions from non-malignant and paramalignant states. We collected a total of 165 pleural fluid samples from 165 subjects; Benign (n = 16), Paramalignant (n = 21), MPE-Lung (n = 57), MPE-Other (n = 22), and Mesothelioma (n = 49). We performed high throughput 16S rRNA gene sequencing on pleural fluid samples and controls. We showed that there are compositional differences among pleural effusions related to non-malignant, paramalignant, and malignant disease. Furthermore, we showed differential enrichment of bacterial taxa within MPE depending on the site of primary malignancy. Pleural fluid of MPE-Lung and Mesothelioma were associated with enrichment with oral and gut bacteria that are commonly thought to be commensals, including Rickettsiella, Ruminococcus, Enterococcus, and Lactobacillales. Mortality in MPE-Lung is associated with enrichment in Methylobacterium, Blattabacterium, and Deinococcus. These observations lay the groundwork for future studies that explore host-microbiome interactions and their influence on carcinogenesis.
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