Considerations for the design and conduct of human gut microbiota intervention studies relating to foods.

With the growing appreciation for the influence of the intestinal microbiota on human health, there is increasing motivation to design and refine interventions to promote favorable shifts in the microbiota and their interactions with the host. Technological advances have improved our understanding and ability to measure this indigenous population and the impact of such interventions. However, the rapid growth and evolution of the field, as well as the diversity of methods used, parameters measured and populations studied, make it difficult to interpret the significance of the findings and translate their outcomes to the wider population. This can prevent comparisons across studies and hinder the drawing of appropriate conclusions. This review outlines considerations to facilitate the design, implementation and interpretation of human gut microbiota intervention studies relating to foods based upon our current understanding of the intestinal microbiota, its functionality and interactions with the human host. This includes parameters associated with study design, eligibility criteria, statistical considerations, characterization of products and the measurement of compliance. Methodologies and markers to assess compositional and functional changes in the microbiota, following interventions are discussed in addition to approaches to assess changes in microbiota-host interactions and host responses. Last, EU legislative aspects in relation to foods and health claims are presented. While it is appreciated that the field of gastrointestinal microbiology is rapidly evolving, such guidance will assist in the design and interpretation of human gut microbiota interventional studies relating to foods.

The Patient's Second Hip Fracture - One in Ten.

Introduction The number of fragility hip fractures (>60 years) are estimated to triple/quadruple by 2050. It is estimated that the prevalence of patient's contralateral hip fractures (HF2s) will increase also. Methods Single hospital, Retrospective review, 2013-2017, Radiograph review, n = 822. Results Management of patient's 2nd hip fractures accounted for 10.5% of all hip fracture surgeries. ~50% occurred within 3 years of the 1st hip fracture. There was no statistically significant difference in discharge destination, length-of-stay or mortality between the HF1 and HF2 cohorts. Discussion Patients with HF2s comprised a significant and stable proportion of all hip fractures treated. We advocate for the provision of a Fracture Liaison Service in each of the 16 hip fracture operating hospitals in Ireland to optimise the secondary prevention of hip fractures.

Hydrogen peroxide alters sternohyoid muscle function.

Upper airway (UA) dilator muscles are critical for the maintenance of airway patency. Injury or fatigue to this group of muscles, as observed in patients with obstructive sleep apnoea (OSA) and animal models of OSA, may leave the UA susceptible to collapse. Although the mechanisms underlying respiratory muscle dysfunction are not completely understood, there is strong evidence suggesting a link between increased production of reactive oxygen species and altered muscle function. The aim of this study was to examine the effects of H2O2 on rat sternohyoid muscle function in vitro. Sternohyoid contractile and endurance properties were examined at 35 °C under control or hypoxic conditions. Studies were conducted in the presence of varying concentrations of H2O2 (0, 0.01, 0.1 and 1 mM). Muscle function was also examined in the presence of antioxidants [desferoxamine (DFX), catalase] and the reducing agent dithiothreitol (DTT). H2O2 decreased muscle endurance in a concentration-dependent manner. This was partially reversed by catalase, DFX and DTT. Our results suggest that oxidants may contribute to UA respiratory muscle dysfunction with implications for the control of UA patency in vivo.

Predictors of mortality in adults with cystic fibrosis.

Assessment of prognostic indicators in patients with cystic fibrosis (CF) is important. The study's aim was to assess the relative contribution of gender, genetics and microbiology on survival in adults with CF. Adult patients were studied from 1995 to 2005 and data collected included FEV(1) (%predicted), body mass index (BMI), genetics, and microbiology. Data was available on 183 patients in 1995. Forty-five patients died in the subsequent 10 years. Patients who died during the study had lower mean (SD) FEV(1) %predicted in 1995 when compared to those remaining alive, 41.5 (15.2)% versus 69.8 (23.2)% predicted, respectively, P<0.001 and they had lower mean (SD) BMI in 1995, 19.2 (3.3) kg/m(2) in comparison to those remaining alive, 20.7 (3.4) kg/m(2), P=0.008. The proportion of patients infected with Pseudomonas aeruginosa and Burkholderia cepacia complex was higher in the group who died during the study compared to those remaining alive, odds ratio 20.9 P<0.0001 and 7.1 P<0.0001, respectively. The presence of the Delta F508 homozygous mutation did not alter survival, P=0.3. Patients infected with either P.aeruginosa or B.cepacia complex had reduced survival compared to those without infection, P=0.01 and P<0.0001, respectively. FEV(1)% (P<0.0001), infection with P.aeruginosa (P=0.005) or B.cepacia complex (P=0.03) were the only significant predictors of mortality. This study demonstrates adults who died were more likely to have worse lung function and be infected with either P.aeruginosa or B.cepacia complex. FEV(1)% and infection with P.aeruginosa or B.cepacia complex were the most significant predictors of survival in adults with CF.

Clostridium difficile carriage in adult cystic fibrosis (CF); implications for patients with CF and the potential for transmission of nosocomial infection.

Clostridium difficile is an anaerobic Gram-positive, spore-forming, toxin-producing bacillus transmitted among humans through the faecal-oral route. Despite increasing carriage rates and the presence of C. difficile toxin in stool, patients with CF rarely appear to develop typical manifestations of C. difficile infection (CDI). In this study, we examined the carriage, toxin production, ribotype distribution and antibiotic susceptibility of C. difficile in a cohort of 60 adult patients with CF who were pre-lung transplant. C. difficile was detected in 50% (30/60) of patients with CF by culturing for the bacteria. C. difficile toxin was detected in 63% (19/30) of C. difficile-positive stool samples. All toxin-positive stool samples contained toxigenic C. difficile strains harbouring toxin genes, tcdA and tcdB. Despite the presence of C. difficile and its toxin in patient stool, no acute gastrointestinal symptoms were reported. Ribotyping of C. difficile strains revealed 16 distinct ribotypes (RT), 11 of which are known to be disease-causing including the hyper-virulent RT078. Additionally, strains RT002, RT014, and RT015, which are common in non-CF nosocomial infection were described. All strains were susceptible to vancomycin, metronidazole, fusidic acid and rifampicin. No correlation was observed between carriage of C. difficile or any characteristics of isolated strains and any recorded clinical parameters or treatment received. We demonstrate a high prevalence of hypervirulent, toxigenic strains of C. difficile in asymptomatic patients with CF. This highlights the potential role of asymptomatic patients with CF in nosocomial transmission of C. difficile.

A pilot study demonstrating the altered gut microbiota functionality in stable adults with Cystic Fibrosis.

Cystic Fibrosis (CF) and its treatment result in an altered gut microbiota composition compared to non-CF controls. However, the impact of this on gut microbiota functionality has not been extensively characterised. Our aim was to conduct a proof-of-principle study to investigate if measurable changes in gut microbiota functionality occur in adult CF patients compared to controls. Metagenomic DNA was extracted from faecal samples from six CF patients and six non-CF controls and shotgun metagenomic sequencing was performed on the MiSeq platform. Metabolomic analysis using gas chromatography-mass spectrometry was conducted on faecal water. The gut microbiota of the CF group was significantly different compared to the non-CF controls, with significantly increased Firmicutes and decreased Bacteroidetes. Functionality was altered, with higher pathway abundances and gene families involved in lipid (e.g. PWY 6284 unsaturated fatty acid biosynthesis (p = 0.016)) and xenobiotic metabolism (e.g. PWY-5430 meta-cleavage pathway of aromatic compounds (p = 0.004)) in CF patients compared to the controls. Significant differences in metabolites occurred between the two groups. This proof-of-principle study demonstrates that measurable changes in gut microbiota functionality occur in CF patients compared to controls. Larger studies are thus needed to interrogate this further.

Combining presentation high-sensitivity cardiac troponin I and glucose measurements to rule-out an acute myocardial infarction in patients presenting to emergency department with chest pain.

BACKGROUND AND AIMS: The use of high sensitivity troponin (hs-Tn) may enable early rule out of acute myocardial infarction (AMI) for patients presenting to the emergency department (ED) with chest pain. This study evaluated two approaches to the early rule out of AMI; a combination of a presentation hs-Tn <4ng/L and normal glucose at presentation (dual testing) and a presentation hs-Tn troponin below the limit of detection (LoD). METHODS: We utilised prospectively collected data on adult patients presenting with suspected ACS in two EDs in Australia and New Zealand. Blood samples were taken on presentation and tested for glucose and high sensitivity troponin I. The primary endpoint was index AMI and the secondary endpoint was 30-day acute coronary syndrome (ACS). Sensitivity, specificity, positive and negative predictive values were used to assess the diagnostic accuracy of the dual testing and LoD approaches. RESULTS: Of the 1412 participants, 182 (12.9%) had index AMI. The LoD and the dual testing approach were 100% sensitive for index AMI. The specificity of the dual testing approach (25.2%) was slightly higher than that of the LoD (20.4%). Sensitivity for ACS was similar for the two approaches (96.5% for dual testing and 98.1% for the LoD). CONCLUSIONS: The dual testing and LoD approach identified all patients with index AMI and could be used to reduce the proportion of patients requiring lengthy assessment and inpatient admission. Further investigation is still required to rule out unstable angina pectoris in patients identified as low risk.

Assessment of sodium and potassium intakes.

The sodium, potassium and creatinine contents of three non-consecutive 24-h urine samples collected by 34 selected adult individuals (10 m; 24 f) living in Cork City were determined. The pooled mean 24-h excretion of sodium and potassium in collections adjudged to be complete were 152 mmol and 78 mmol, respectively. There was no significant difference between group average weekday and weekend-day excretion of Na or K, for either males or females. This suggests that weekend 24-h urinary collections, which most subjects find more convenient, are suitable for studies of sodium and potassium intakes of groups. The ratios of intra- to inter-individual variation for 24-h urinary sodium were 1.4 and 2.1 for males and females, respectively. The corresponding ratios for 24-h urinary potassium were 6.6 for males and 4.9 for females. These ratios indicated that there were large individual day-to-day variations in urinary sodium and potassium excretion in this group. It was estimated that a sample size of 35-60 individuals would be required to estimate group mean sodium and potassium intakes by means of single 24-h urine collections.

Influence of dietary sodium intake on urinary calcium excretion in selected Irish individuals.

The relationship between dietary sodium intake (as assessed by the 24-h urinary excretion of sodium) and urinary calcium excretion was examined in a selected group of free-living Irish individuals (n = 94), under conditions in which the completeness of urine collections was assessed. Results indicated that there was a significant positive correlation between 24-h urinary sodium and calcium excretion for males (r = 0.56, P less than 0.001) and females (r = 0.35, P less than 0.01). In addition, linear regression analysis indicated that mean 24-h urinary calcium excretion increased by 39 mg and 26 mg for a 1 g increment in sodium intake for males and females, respectively. The relationship between dietary sodium intake and urinary calcium excretion was also examined in 12 healthy young adults under controlled dietary conditions. Urinary sodium and calcium excretion values over 24 h were positively correlated for males (r = 0.44, P less than 0.05) and females (r = 0.80, P less than 0.001). However, the relationship was only statistically significant for 5 of the 12 subjects when the data were analysed on an individual basis. Results of this study suggest that salt intake may be an important determinant of urinary calcium loss in certain individuals and, as such, may be a risk factor in the development of osteoporosis. However, further studies are required to establish whether low salt intakes have a beneficial effect on calcium balance in free-living individuals.

Selenium status of a group of Scottish adults.

OBJECTIVE: To examine dietary selenium intake and indices of selenium status (plasma and red blood cell selenium and glutathione peroxidase activities) in apparently healthy Scottish individuals. DESIGN AND SUBJECTS: One hundred subjects, aged between 40 and 60 y, completed a seven day weighed food intake and provided blood samples for selenium status analysis. SETTING: Inverurie, Aberdeenshire, Scotland. RESULTS: Average reported selenium intake was low (43 micrograms/d). A significant number of subjects had reported intakes below the RNI. Low levels of plasma selenium were also found but no subject had values below 40 micrograms/1. Red blood cell selenium was within the reference range established for a healthy UK population. Smoking status had no consistent effect on selenium status. CONCLUSIONS: The results of the present study suggest that selenium status of certain Scottish individuals may be compromised and that further studies are warranted. SPONSORSHIP: BASF, Germany; The Tobacco Products Research Trust, UK; Scottish Office Agriculture Environment and Fisheries Department.

Epidemiology and clinical impact of Pseudomonas aeruginosa infection in cystic fibrosis using AP-PCR fingerprinting.

Arbitrarily primed PCR (AP-PCR) was utilized to genetically fingerprint 252 Pseudomonas aeruginosa strains isolated from the sputa of 50 cystic fibrosis (CF) patients attending the Cork CF clinic over a period of 3 years. Ten distinct P. aeruginosa strains were identified and the distribution, temporal trends and clinical impact of colonization with these individual P. aeruginosa clones was studied. A number of random isolates from each AP-PCR group were analysed using pulsed field gel electrophoresis (PFGE) in order to confirm the discriminatory power of the AP-PCR technique. The majority of patients were colonized with a single strain over the time period of the study, but it was also possible to harbour two or more strains transiently or simultaneously. Four main strains were relatively evenly distributed throughout the CF population, and it was noted that patients from the same family or attending the same school tended to harbour the same P. aeruginosa clone. Disease severity was significantly associated with the age of the patient (P < 0.001), clearly indicating an increase in severity with increase in age. The general clinical status of the CF patients was not significantly associated with the P. aeruginosa variant isolated from their sputa. Lung status was defined by FEV1 measurement and chest X-ray score (CXR). The non parametric Kruskal-Wallis significance test of FEV1, CXR and age by colonizing P. aeruginosa clone indicated that FEV1 (P = 0.017), but not CXR (P = 0.19) or age (P = 0.842), differed significantly across the clones of P. aeruginosa isolated. Patients harbouring P. aeruginosa strains B, F or G clearly had lower FEV1 scores while those harbouring clones A, C, D or H generally had higher FEV1 scores. Thus, the sub-species variant of P. aeruginosa colonizing CF patients may be associated with the severity of progressive lung disease.

Home intravenous therapy using a silastic long line catheter in cystic fibrosis patients.

Young patients with cystic fibrosis are now accustomed to having regular periods of intravenous therapy to help in the management of their recurring respiratory infections particularly due to pseudomonas organisms. Most patients will eventually experience difficulties in accepting recurring intravenous cannulations and the life span of the conventional intravenous cannula is frequently brief. We have compared percutaneous long line silastic catheters with conventional intravenous cannulas as intravenous access in children with cystic fibrosis being treated for pulmonary exacerbations. We conclude that silastic long lines are superior to conventional cannulas in terms of patient tolerance, reduction in the number of hospital admissions, reduction in the number of repeat venepunctures/cannulations and reduction in local complications. The high degree of patient acceptability has been impressive and has increased with the enthusiasm of children with cystic fibrosis for more frequent intravenous therapy.

Diabetes mellitus in childhood cystic fibrosis.

Since 1984, five patients in the cystic fibrosis (CF) clinic at Cork Regional Hospital have developed diabetes mellitus (DM) and were treated with Insulin. None had received systemic corticosteroids but two had high calorie naso-gastric feeding regimes. Two died from lung disease. A fifteen year old boy developed bilateral cataracts. In nine other paediatric CF clinics in the Republic of Ireland (total: 420 patients), three patients have DM, two receiving Insulin. Abnormal glucose tolerance is becoming more common in CF as patients survive longer. The possible role of corticosteroid treatment and intensive carbohydrate feeding regimes in development of glucose intolerance must be considered. DM in CF differs from the usual childhood DM. Regular screening and early Insulin supplementation may be beneficial.

Immediate direct peripheral vasoconstriction in response to hyperinsulinemia and metformin in the anesthetized pig.

Elevated levels of insulin have been reported to induce both an arterial vasodilation mediated by nitric oxide (NO), and vasoconstriction mediated by endothelin and reactive oxygen radicals. Metformin, used to control blood glucose levels in type 2 diabetes, has also been shown to cause NO-mediated dilation of conduit arteries. It is possible that these contradictory vascular effects are due to a non-direct action on arteries. Therefore, the direct effect of high levels of insulin and metformin infusion on resistance artery diameter was evaluated. Experiments were carried out on the anesthetized pig; blood flow and pressure were measured in the iliac artery. An adjustable snare was applied to the iliac above the pressure and flow measurement site to induce step decreases (3-4 occlusions at 5 min intervals were performed for each infusion) in blood flow, and hence iliac pressure, and the conductance (deltaflow / deltapressure) calculated. Saline, insulin (20 and 40 mUSP/l/min), and metformin (1 microg/ml/min) were infused separately downstream of the adjustable snare and their effect on arterial conductance assessed. Insulin at both infusion rates and metformin caused a significant reduction in peripheral vascular conductance. In conclusion, hyperinsulinemia and metformin infusion constrict resistance arterial vessels in vivo.

Inhaled versus nebulised tobramycin: a real world comparison in adult cystic fibrosis (CF).

BACKGROUND: There are no published data on real-life clinical experience comparing inhaled antibiotic therapy via new rapid delivery systems with nebulised antibiotic therapy in CF. This real world study compares safety, effectiveness and tolerability using tobramycin inhaled powder (TIP) versus tobramycin inhaled solution (TIS). METHODS: Adult patients with CF commencing TIP (n=78) completed a questionnaire assessing safety, efficacy, tolerability, patient-satisfaction and self-reported adherence to TIS at baseline and during 12 months of TIP therapy. FEV1% predicted and exacerbation rate were recorded at each visit. RESULTS: There was a significant improvement in adherence scores, with a significant decrease in the number of intravenous antibiotic courses received during 12 months of TIP compared with the preceding 12 months using TIS. 94% of patients who had previously used TIS preferred TIP therapy over TIS. CONCLUSIONS: Inhaled powder tobramycin in CF is associated with improved adherence, tolerability and decreased exacerbation rates compared to nebulised treatment in real-life practice.

Responses of iliac conduit artery and hindlimb resistance vessels to luminal hyperfructosemia in the anaesthetized pig.

AIMS: High fructose levels are found in diabetes mellitus, associated with high corn syrup diets, and have been claimed to cause hypertension. As the direct effects on conduit and resistance arteries have not been previously reported, we measured these in vivo in the anaesthetized pig with instrumented iliac arteries. METHODS: Experiments were performed on the iliac artery preparation in the anaesthetized pig: blood flow, diameter and pressure were measured in the iliac. RESULTS: The change in diameter of an occluded iliac artery segment filled with hyperfructosemic (15 µm) blood was 89.5 ± 22.1 µm (mean ± SE), contrasted with 7.7 ± 13.06 µm control (P = 0.005, paired t-test, n = 6). There was no significant difference when compared with blood containing both hyperfructosemic blood and the nitric oxide synthesis inhibitor, N(G)-nitro-l-arginine methyl ester (250 µg mL(-1)). Step changes in pressure and flow were achieved by progressive arterial stenosis during control saline and 15 µm min(-1) fructose downstream intra-arterial infusions. Linear regression of the step changes in blood pressure versus the instantaneous step changes in blood flow showed a statistically significant decrease in slope of the conductance (P < 0.001, analysis of covariance), indicating an increase in instantaneous peripheral vascular resistance. Peripheral autoregulation and conduit artery shear-stress-mediated dilatation were not significantly altered. CONCLUSION: An elevated level of fructose caused dilatation of a conduit artery but constriction of resistance vessels. The latter effect could account, if maintained long-term, for the hypertension claimed to be due to hyperfuctosemia.

Accuracy of whole-body low-dose multidetector CT (WBLDCT) versus skeletal survey in the detection of myelomatous lesions, and correlation of disease distribution with whole-body MRI (WBMRI).

AIM: The aim of the study is to assess the feasibility of whole-body low-dose computed tomography (WBLDCT) in the diagnosis and staging of multiple myeloma and compare to skeletal survey (SS), using bone marrow biopsy and whole-body magnetic resonance imaging (WBMRI; where available) as gold standard. MATERIALS AND METHODS: Patients referred over an 18-month period for investigation of suspected multiple myeloma or restaging of myeloma were randomized to undergo one of two WBLDCT protocols using high kVp, low mAs technique (140 kVp, 14 mAs; or 140 kVp, 25 mAs). Recent WBMRI scans were reviewed in 23 cases. Each imaging modality was assessed by two radiologists in consensus and scored from 0-3 (0 = normal, 1 = 1-4 lesions, 2 = 5-20 lesions, 3 >or= 20 lesions/diffuse disease) in ten anatomical areas. Overall stage of disease, image quality score, and the degree of confidence of diagnosis were recorded. Diagnostic accuracy of skeletal survey and WBLDCT were determined using a gold standard of bone marrow biopsy and distribution of disease was compared to WBMRI. RESULTS: Thirty-nine patients were evaluated. WBLDCT identified more osteolytic lesions than skeletal survey with a greater degree of diagnostic confidence and led to restaging in 18 instances (16 upstaged, two downstaged). In those with recent WBMRI, distribution of disease on WBLDCT showed superior correlation with WBMRI when compared with SS. Overall reader impression of stage on WBLDCT showed significant correlation with WBMRI (kappa = 0.454, p < 0.05). WBLDCT provided complementary information to WBMRI in nine patients with normal marrow signal following treatment response, but which were shown to have diffuse residual cortical abnormalities on CT. CONCLUSION: WBLDCT at effective doses lower than previously reported, is superior to SS at detecting osteolytic lesions and at determining overall stage of multiple myeloma, and provides complementary information to WBMRI.

Radiation protection to the eye and thyroid during diagnostic cerebral angiography: a phantom study.

We measured radiation doses to the eye and thyroid during diagnostic cerebral angiography to assess the effectiveness of bismuth and lead shields at dose reduction. Phantom head angiographic studies were performed with bismuth (study 1) and lead shields (study 2). In study 1 (12 phantoms), thermoluminescent dosimeters (TLD) were placed over the eyes and thyroid in three groups: (i) no shields (four phantoms); (ii) anterior bismuth shields (four phantoms) and (iii) anterior and posterior bismuth shields (four phantoms). In a second study (eight phantoms), lead shields were placed over the thyroid only and TLD dose measurements obtained in two groups: (i) no shielding (four phantoms) and (ii) thyroid lead shielding (four phantoms). A standard 4-vessel cerebral angiogram was performed on each phantom. Study 1 (bismuth shields) showed higher doses to the eyes compared with thyroid (mean 13.03 vs 5.98 mSv, P < 0.001) and a higher eye dose on the X-ray tube side. Overall, the use of bismuth shielding did not significantly reduce dose to either eyes or thyroid in the measured TLD positions. In study 2, a significant thyroid dose reduction was found with the use of lead shields (47%, mean 2.46 vs 4.62 mSv, P < 0.001). Considerable doses to the eyes and thyroid highlight the need for increased awareness of patient protection. Eye shielding is impractical and interferes with diagnostic capability. Thyroid lead shielding yields significant protection to the thyroid, is not in the field of view and should be used routinely.

Staff radiation doses to the lower extremities in interventional radiology.

The purpose of this study was to investigate the radiation doses to the lower extremities in interventional radiology suites and evaluate the benefit of installation of protective lead shielding. After an alarmingly increased dose to the lower extremity in a preliminary study, nine interventional radiologists wore thermoluminescent dosimeters (TLDs) just above the ankle, over a 4-week period. Two different interventional suites were used with Siemens undercouch fluoroscopy systems. A range of procedures was carried out including angiography, embolization, venous access, drainages, and biopsies. A second identical 4-week study was then performed after the installation of a 0.25-mm lead curtain on the working side of each interventional table. Equivalent doses for all nine radiologists were calculated. One radiologist exceeded the monthly dose limit for a Category B worker (12.5 mSv) for both lower extremities before lead shield placement but not afterward. The averages of both lower extremities showed a statistically significant dose reduction of 64% (p < 0.004) after shield placement. The left lower extremity received a higher dose than the right, 6.49 vs. 4.57 mSv, an increase by a factor of 1.42. Interventional radiology is here to stay but the benefits of interventional radiology should never distract us from the important issue of radiation protection. All possible measures should be taken to optimize working conditions for staff. This study showed a significant lower limb extremity dose reduction with the use of a protective lead curtain. This curtain should be used routinely on all C-arm interventional radiologic equipment.