A systematic review on outcome reporting in randomised controlled trials on surgical interventions for female stress urinary incontinence: a call to develop a core outcome set.

BACKGROUND: Several meta-analyses have identified methodological limitations in female stress urinary incontinence (SUI) trials, precluding the synthesis of primary studies and high-quality evidence. OBJECTIVES: Evaluation of outcome measure selection and outcome reporting in randomised controlled trials (RCTs) on surgery for SUI. SEARCH STRATEGY: Systematic review of RCTs identified from bibliographical databases, including Medline, Cochrane, and EMBASE. SELECTION CRITERIA: Randomised controlled trials evaluating the efficacy and safety of surgical interventions for the management of female SUI. DATA COLLECTION AND ANALYSIS: Two researchers independently assessed the included studies and documented outcomes. MAIN RESULTS: Overall, 108 studies were identified that included 422 reported outcomes and 119 outcome measures. The three most common outcomes were cure rates (87 studies), quality of life (85 studies), and overactive bladder (78 studies). The median methodological quality rating was 3 (range 0-3) and the outcome reporting quality rating was 3 (range 0-5). Multinomial logistic regression analysis revealed that the methodological quality and use of validated questionnaire were significant predictors of the quality of outcome reporting (ß = 0.538, P < 0.001; ß = 0.218, P = 0.011, respectively). CONCLUSIONS: Outcome reporting in SUI trials is highly variable. Until a core outcome set is developed and implemented, we propose an interim use of three commonly reported outcomes in each domain (treatment success rate - complete cure, partial improvement, or failure of response; urodynamic evaluation outcomes - overactive bladder (OAB), voiding dysfunction, and urodynamic stress incontinence; patient-reported outcomes - quality of life, sexual dysfunction, and patient satisfaction) with the use of validated questionnaires for patient-reported outcomes and subjective success rates. Complications should be also explicitly and comprehensively reported using validated outcome measures. TWEETABLE ABSTRACT: There is significant variation in outcome reporting in SUI trials. Our systematic review findings aim to form the basis for the development of a core outcome set.

In vitro activity of antimicrobial-impregnated catheters against biofilms formed by KPC-producing Klebsiella pneumoniae.

AIM: To evaluate the activity and effectiveness of impregnated central venous catheters (CVC) against Klebsiella pneumoniae biofilms. METHODS AND RESULTS: The antimicrobial activity and durability of impregnated-CVCs were evaluated over time and the size of zones of inhibition (ZI) was measured. Biofilm formation was observed by quantitative culture and also by scanning electron microscopy. The catheters impregnated with chlorhexidine/silver sulfadiazine (CHX/SS) reduced bacteria counts by 0·3 log and were most effective (P < 0·01) against Klebsiella pneumoniae biofilms N-acetylcysteine/levofloxacin (NAC/LEV) catheters. It was observed that the catheter impregnated with NAC/LEV had initially the largest average ZI size being statistically significant (P < 0·01). The NAC/LEV combination remained active until day 30, whereas the combination of CHX/SS was completely inactivated from day 15 on. CONCLUSIONS: The NAC/LEV combination showed greater durability on the catheters, but it was the CHX/SS combination that had the greater initial efficacy in bacterial inhibition. It was also observed that NAC/LEV-impregnated catheters do not prevent the emergence of resistant subpopulations inside the inhibition halos during antimicrobial susceptibility tests. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results highlighted that the in vitro efficacy of antimicrobial-impregnated CVCs is limited by time and that their colonization occurred earlier than expected. Our data also demonstrated that NAC/LEV remained active until day 30 of evaluation and CHX/SS combination was completely inactivated from day 15 on. Our findings suggested that implantable devices should be carefully used by medical community.

A phase 2a randomized controlled study to evaluate the pharmacokinetic, safety, tolerability and clinical effect of topically applied Umeclidinium in subjects with primary axillary hyperhidrosis.

BACKGROUND: Hyperhidrosis is a common medical condition which can have a significant impact on quality of life. Umeclidinium (UMEC) is a long-acting muscarinic antagonist (LAMA) developed as a dermal formulation. OBJECTIVES: This 2-week, double-blind, randomized, vehicle-controlled study evaluated systemic exposure, safety and tolerability of topically administered UMEC in subjects with primary axillary hyperhidrosis. Clinical effect was a secondary objective, measured by gravimetry and the hyperhidrosis disease severity scale (HDSS). Vehicle was included to evaluate safety. METHODS: Twenty-three subjects were randomized to either 1.85% UMEC (N = 18) or vehicle (N = 5) once daily. RESULTS: Measurable plasma concentrations were observed in 78% of subjects after the treatment. Nine subjects (50%) on UMEC and two subjects (40%) on vehicle reported AEs, most commonly application site reactions. At Day 15, seven subjects (41%) in UMEC and two subjects (40%) in vehicle had at least a 50% reduction in sweat production. Eight subjects (47%) in UMEC and one subject (20%) in vehicle had at least a two-point reduction in HDSS. No comparisons of treatment arms were planned prospectively. CONCLUSIONS: The measurable exposure, acceptable safety and preliminary clinical activity observed in this proof-of-concept study suggest the potential clinical utility of topical UMEC in subjects with axillary hyperhidrosis.

Marker of coxsackievirus-B4 infection in saliva of patients with type 1 diabetes.

BACKGROUND: Coxsackieviruses B (CV-B) are enteroviruses that have been reported to play a role in the pathogenesis of type 1 diabetes. Enteroviral RNA was detected in the gut mucosa of patients. The mucosal immunity is an interconnected network; therefore, the response to enteroviruses possibly present in the gastrointestinal mucosa can be reflected by specific antibodies in the saliva. In the present study, the anti-CV-B neutralizing activity of saliva samples from patients with type 1 diabetes was investigated. METHODS: Saliva samples were collected from patients and controls of 3 countries, and plasma was obtained from some of them. The anti-CV-B activity of clinical samples was determined by neutralization of the cytopathic effect induced by challenging viruses in vitro and expressed as titre value. RESULTS: Overall prevalence and levels of anti-CV-B4 activity of saliva were higher in patients (n = 181) than in controls (n = 135; P = .0002; titre values ≥ 16: odds ratio = 4.22 95% CI: 1.90-9.38 P = .0002). It has been shown that IgA1 played a role in this activity. There was no correlation between the saliva and the plasma anti-CV-B4 neutralizing activity. The neutralizing activity of saliva against CV-B1, CV-B2, CV-B3, and CV-B5 existed rarely, if at all. Increased levels of anti-CV-B4 activity were observed all along a 4 year follow-up period in patients but not in matched controls (P = .01). CONCLUSION: There is an anti-CV-B4 activity in saliva of patients with type 1 diabetes that may be a useful marker to study the role of CV-B in the pathogenesis of the disease.

In-MOFs based on amide functionalised flexible linkers.

Two new amide functionalised metal-organic frameworks, In(OH)CSA and In(OH)PDG, were synthesized using two flexible linkers, N-(4-carboxyphenyl)succinamic acid (CSA) and N,N'-(1,4-phenylenedicarbonyl)diglycine (PDG), respectively. Both structures consist of corner-sharing {InO4(OH)2} octahedra in the form of trans indium hydroxide chains, which are interconnected by the dicarboxylate linkers to form stacked 2-dimensional layers. The different symmetries and configurations of the flexible and rigid features on the linkers results in different supramolecular interactions dominating between linkers, resulting in different shaped pores and functional group orientation. In(OH)CSA lacks hydrogen bonding between linkers, which results in close packing between the layers and very small solvent accessible pores running perpendicular to the plane of the layers. In(OH)PDG exhibits strong intra- and interlayer hydrogen bonding, which prevents the layers from close packing and results in larger cylindrical pores running parallel to the indium hydroxide chains, producing a total accessible volume of 25% of the unit cell volume.

[Evaluation of the management of soft tissue sarcomas in Franche-Comté since the establishment of a multidisciplinary meeting at University Hospital. About 47 cases]. / Évaluation de la prise en charge des sarcomes des tissus mous en Franche-Comté depuis l'instauration d'une réunion de concertation pluridisciplinaire au CHRU. À propos de 47 cas.

GOALS OF STUDY: A multidisciplinary meeting (RCP) dedicated to the treatment of sarcoma was established in Franche-Comte in 2010. The goals of the study are: (a) To evaluate the treatment of sarcomas by confrontation with the existing literature; (b) To evaluate the influence of the multidisciplinary meeting on the management of sarcomas by hospitals at the regional level. MATERIALS AND METHODS: This is a retrospective single center study from 2010 to 2015 on patients with sarcoma and peripheral soft tissue drawn from a Netsarc database (National Network of sarcomas) and communicating cancer record. A database Cleanweb especially dedicated is created. RESULTS: Forty-seven patients were included: ten sarcomas at the upper member 26 to the lower limbs, 11 on the trunk. Forty patients were operated on: ten out of the university hospital, 28 at the university hospital and two in a coordinating center. Ninety percent of patients treated at the university hospital were in accordance with the recommandations. None of the patients operated out of the university hospital benefited from medical care in accordance to the recommendations. There is an increase in the number of files sent by the hospitals out of the university hospital discussed in multidisciplinary meeting, before treatment. CONCLUSION: The creation of a dedicated multidisciplinary meeting sarcoma improves the medical management of these tumors and decreases inappropriate medical managements thanks to a better education of the regional physicians.

Nanoscale Structure of the Oil-Water Interface.

X-ray reflectivity (XR) and atomistic molecular dynamics (MD) simulations, carried out to determine the structure of the oil-water interface, provide new insight into the simplest liquid-liquid interface. For several oils (hexane, dodecane, and hexadecane) the XR shows very good agreement with a monotonic interface-normal electron density profile (EDP) broadened only by capillary waves. Similar agreement is also found for an EDP including a sub-Å thick electron depletion layer separating the oil and the water. The XR and MD derived depletions are much smaller than reported for the interface between solid-supported hydrophobic monolayers and water.

Susceptibility of Maize to Stalk Rot Caused by Fusarium graminearum Deoxynivalenol and Zearalenone Mutants.

Fusarium graminearum is a destructive pathogen of cereals that can cause stalk rot in maize. Stalk rot results in yield losses due to impaired grain filling, premature senescence, and lodging, which limits production and harvesting of ears. In addition, mycotoxins can make infected tissues unfit for silage. Our objectives were to evaluate the natural variation in stalk rot resistance among maize inbreds, to establish whether deoxynivalenol (DON)- and zearalenone (ZEA)-deficient strains are pathogenic on a panel of diverse inbreds, and to quantify the accumulation of DON in infected stalk tissue. Wild-type F. graminearum and mycotoxin mutants (DON and ZEA) were used to separately inoculate stalks of 9-week-old plants of 20 inbreds in the greenhouse. Plants were evaluated for lesion area at the inoculation point at 0, 2, 14, and 28 days postinoculation and tissues around lesions were sampled to determine the DON content. Regardless of their ability to produce DON or ZEA, all tested F. graminearum strains caused stalk rot; however, significant differences in disease levels were detected. Among the tested inbreds, Mp717 was resistant to all three F. graminearum strains while Mp317 and HP301 were only partially resistant. Accumulation of DON was significantly lower in infected stalks of the resistant and partially resistant inbreds than the susceptible inbreds. Analysis of the 20 inbreds using data from 17 simple-sequence repeats revealed population structure among the individuals; however, there was no association between genetic clustering and stalk rot resistance. These findings are an additional step toward breeding maize inbreds suitable for planting in fields infested with F. graminearum.

[Large surgical resection of a shoulder sarcoma's and secure one stage reconstruction with lattissimi dorsi and serratus anterior flap. Updating about a case]. / Exérèse large d'un sarcome de l'épaule et reconstruction en un temps par double lambeau de grand dorsal et de grand dentelé pédiculé. Discussion à propos d'un cas.

INTRODUCTION: The soft tissue sarcomas (STM) are tumors developed at the expense of connective tissue. They are rare and have severe prognosis. The principles of management are recalled through an extended case of shoulder sarcoma. CLINICAL CASE: A 48-year-old patient has a sarcoma of the right pectoralis major muscle confirmed by biopsy. After multidisciplinary meeting, a wide surgical excision exposing the subclavian vessels and brachial plexus is performed with double cover flap pedicled latissimus dorsi and serratus and thin skin graft. RESULT: Healing process is acquired at 3 months. Adjuvant chemotherapy is established (adriamycin, ifosfamide) along with radiotherapy (54Gy). CONCLUSION: Sarcomas treatment has to be realized after considering multidisciplinary meeting (RCP) in dedicated structures. Surgery is the main treatment, it should ideally be R0, that is to say, integral with healthy tissue margin around the tumor (or healthy anatomical barrier). Optimal surgery performs a resection "without seen tumor". The diagnosis has to be made with a biopsy before the surgical treatment. Healing is quickly obtained due to adjuvant treatments.

Flow cytometric analysis: Interdependence of healthy and infected udder quarters.

An important question about intramammary infections that is still debated in the literature is the independence or interdependence of the quarters of dairy cows. The present study sought to explore milk neutrophil function and the milk lymphocyte profile of uninfected quarters from uninfected and infected (one infected quarter per cow) udders to evaluate interdependence of the quarters. Thus, 32 (8 cows) and 18 (6 cows) uninfected quarters from uninfected and infected udders were used, respectively. Using flow cytometry, we evaluated the percentage of milk neutrophils and their expression of adhesion molecules L-selectin (CD62L), ß2-integrin (CD11b), and an endothelial-selectin ligand (CD44); levels of intracellular reactive oxygen species (ROS); phagocytosis of Staphylococcus aureus by milk neutrophils; and neutrophil viability. Furthermore, we assessed the percentage of B-cell (CD21(+)) and T-lymphocyte subsets (CD3(+)/CD4(+)/CD8(-), CD3(+)/CD8(+)/CD4(-), CD3(+)/CD4(+)/CD25(-), CD3(+)/CD4(+)/CD25(+), and CD3(+)/CD4(-)/CD25(-)) using flow cytometry with monoclonal antibodies. The infected quarter did not affect somatic cell count or the percentage of neutrophils in the neighboring uninfected quarters. Furthermore, the infected quarter did not influence neutrophil viability, intracellular reactive oxygen species production, or phagocytosis of S. aureus by milk neutrophils. Conversely, the expression of adhesion molecules CD11b, CD62L, and CD44 by milk neutrophils differed between uninfected quarters from infected versus uninfected udders. The lymphocyte subsets did not differ between groups, except for a higher percentage of B cells in uninfected quarters from infected udders than in those from uninfected udders. Thus, our study strongly supports the hypothesis of interdependence of quarters based on the influence of infection on both the percentage of B cells and the expression of adhesion molecules by milk neutrophils in the neighboring uninfected quarters.

The efficiency of wire nets in enhancing the biosecurity of poultry in Brazil.

The aim of this study was to evaluate the efficiency of the use of wire nets of various mesh sizes to enhance biosecurity in the poultry industry in Brazil by preventing other bird species from entering chicken houses. The Brazilian poultry industry is technologically advanced and employs updated technology. The current Brazilian guidelines recommend the use of 25.40 mm mesh. However, scientific evidence of the efficiency of the nets recommended by these guidelines is lacking. In this study, a bird biometric methodology was developed to evaluate bird species. The methodology was based on the body dimensions of the animal, and it employed a new statistical design to analyse the data. Three groups of bird species were designated according to their importance. The value of this criterion (the importance of the species) was estimated by assessing the ability of birds to pass through the net. The paradigm was used to study 23 wild avian species that are naturally present in Brazil. The best results were observed for nets with a mesh size < or = 19.11 mm. This mesh size was able to efficiently restrain all of the species studied. However, in the same test, the net with 25.40 mm mesh could not restrain 11 bird species, one of which was Passer domesticus, which is found worldwide. On the basis of these results, the use of 19.11 mm mesh should be strongly recommended in order to achieve biosecurity of poultry houses.

Placental site trophoblastic tumours and the concept of fertility preservation.

The standard management of placental site trophoblastic tumours (PSTTs) is a radical hysterectomy with pelvic lymph node sampling. We present five cases to demonstrate a modified Strassman procedure (MSP), which is an alternative fertility-sparing technique. Each had a presumed solitary uterine PSTT. Following surgery, one patient remained in remission with her fertility intact. The other four underwent a completion hysterectomy because of incomplete excision of the disease. No residual disease was later found in two of these four uteri. This treatment should only be offered after extensive counselling. We intend to investigate the use of intraoperative frozen section analysis with cold-knife dissection in future.

Intravesical treatment of painful bladder syndrome: a systematic review and meta-analysis.

INTRODUCTION AND HYPOTHESIS: The objective of the study was to assess the effectiveness of intravesical treatment for painful bladder syndrome (PBS). METHODS: A systematic review was performed until December 31, 2010. The selection criteria included only randomized controlled trials of PBS patients who received intravesical treatment. The primary outcomes measures were clinical and urodynamic parameters. Relative risk and mean differences were used for binary and continuous outcomes respectively, with confidence interval of 95%. RESULTS: The search strategy identified 770; however, only 28 eligible trials met methodological requirements for complete analysis. Altogether, the review included four treatment modalities: resiniferatoxin, Bacillus Calmette-Guérin (BCG), oxybutynin, and alkalinized lidocaine. Meta-analysis of BCG therapy showed improvement in symptoms according to the Wisconsin Interstitial Cystitis Symptom Inventory, but no difference in 24-h urinary frequency. CONCLUSIONS: Meta-analysis showed an improvement exclusively of the symptoms as measured by the Wisconsin Interstitial Cystitis Inventory, but not in 24-h urinary frequency, with BCG therapy. Further randomized clinical trials, including trials of more recent drugs, are required for evaluation of intravesical therapies for PBS.

AUGS-IUGA Joint Clinical Consensus Statement on Enhanced Recovery After Urogynecologic Surgery: Developed by the Joint Writing Group of the International Urogynecological Association and the American Urogynecologic Society. Individual writing group members are noted in the Acknowledgements section.

INTRODUCTION AND HYPOTHESIS: Enhanced recovery after surgery (ERAS) evidence-based protocols for perioperative care can lead to improvements in clinical outcomes and cost savings. This article aims to present consensus recommendations for the optimal perioperative management of patients undergoing urogynecological surgery. METHODS: A review of meta-analyses, randomized clinical trials, large nonrandomized studies, and review articles was conducted via PubMed and other databases for ERAS and urogynecological surgery. ERAS protocol components were established, and then quality of the evidence was both graded and used to form consensus recommendations for each topic. These recommendations were developed and endorsed by the writing group, which is comprised of the American Urogynecologic Society and the International Urogynecological Association members. RESULTS: All recommendations on ERAS protocol items are based on best available evidence. The level of evidence for each item is presented accordingly. The components of ERAS with a high level of evidence to support their use include fasting for 6 h and taking clear fluids up to 2 h preoperatively, euvolemia, normothermia, surgical site preparation, antibiotic and antithrombotic prophylaxis, strong antiemetics and dexamethasone to reduce postoperative nausea and vomiting, multimodal analgesia and restrictive use of opiates, use of chewing gum to reduce ileus, removal of catheter as soon as feasible after surgery and avoiding systematic use of drains/vaginal packs. CONCLUSIONS: The evidence base and recommendations for a urogynecology-relevant ERAS perioperative care pathway are presented in this consensus review. There are several elements of ERAS with strong evidence of benefit in urogynecological surgery.

Comparative measurements of mineral elements in milk powders with laser-induced breakdown spectroscopy and inductively coupled plasma atomic emission spectroscopy.

Mineral elements contained in commercially available milk powders, including seven infant formulae and one adult milk, were analyzed with inductively coupled plasma atomic emission spectrometry (ICP-AES) and laser-induced breakdown spectroscopy (LIBS). The purpose of this work was, through a direct comparison of the analytical results, to provide an assessment of the performance of LIBS, and especially of the procedure of calibration-free LIBS (CF-LIBS), to deal with organic compounds such as milk powders. In our experiments, the matrix effect was clearly observed affecting the analytical results each time laser ablation was employed for sampling. Such effect was in addition directly observed by determining the physical parameters of the plasmas induced on the different samples. The CF-LIBS procedure was implemented to deduce the concentrations of Mg and K with Ca as the internal reference element. Quantitative analytical results with CF-LIBS were validated with ICP-AES measurements and nominal concentrations specified for commercial milks. The obtained good results with the CF-LIBS procedure demonstrate its capacity to take into account the difference in physical parameters of the plasma in the calculation of the concentrations of mineral elements, which allows a significant reduction of the matrix effect related to laser ablation. We finally discuss the way to optimize the implementation of the CF-LIBS procedure for the analysis of mineral elements in organic materials.

A matrix effect and accuracy evaluation for the determination of elements in milk powder LIBS and laser ablation/ICP-OES spectrometry.

Laser ablation coupled to inductively coupled plasma optical emission spectrometry (LA-ICP-OES) and laser-induced breakdown spectroscopy (LIBS) were investigated for the determination of Ca, Mg, Zn and Na in milk samples. The accuracy of both methods was evaluated by comparison of the concentration found using LA-ICP-OES and LIBS with classical wet digestion associated with ICP-OES determination. The results were not fully acceptable, with biases from less than 1% to more than 60%. Matrix effects were also investigated. The sample matrix can influence the temperature, electron number density (n (e)) and other excitation characteristics in the ICP. These ICP characteristics were studied and evaluated during ablation of eight milk samples. Differences in n (e) (from 8.9 to 13.8 × 10(14) cm(-3)) and rotational temperature (ranging from 3,400 to 4,400 K) occurred with no correlation with trueness. LIBS results obtained after classical external calibration procedure gave degraded accuracy, indicating a strong matrix effect. The LIBS measurements clearly showed that the major problem in LA-ICP was related to the ablation process and that LIBS spectroscopy is an excellent diagnostic tool for LA-ICP techniques.

Frequency of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Mycoplasma hominis and Ureaplasma species in cervical samples.

We investigated the relative frequencies of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Mycoplasma hominis and Ureaplasma sp. in cervical samples. PCR analyses were performed in ectocervical and endocervical samples from 224 patients attending public health services in Belo Horizonte and Contagem, Minas Gerais Brazil. A high prevalence of colonisation of the cervix (6.3% for C. trachomatis, 4.0% for N. gonorrhoeae, 0.9% for M. genitalium, 21.9% for M. hominis, 38.4% for Ureaplasma sp.) was demonstrated not only for pathogens classically associated to cervicitis (C. trachomatis and N. gonorrhoeae), but also for M. hominis and Ureaplasma sp. These findings may be useful to guide more adequate diagnosis to interrupt transmission and to avoid negative impacts on the female reproductive tract.

Is the addition of vaginal electrical stimulation to transcutaneous tibial nerve electrical stimulation more effective for overactive bladder treatment? A randomized controlled trial. / ¿Es más efectivo el tratamiento combinado de electroestimulación transcutánea del nervio tibial y electroestimulación vaginal que el tratamiento en monoterapia de electroestimulación del nervio tibial para la vejiga hiperactiva? Un ensayo controlado aleatorizado.

INTRODUCTION AND OBJECTIVES: Overactive bladder (OAB) is a prevalent disorder that increases with age and impairs patients' quality of life. Guidelines recommend behavior modifications as the first-line treatment; however, physiotherapy has also been used with success, safety, and low cost. Transcutaneous tibial nerve electrical stimulation (TTNS) and vaginal electrical stimulation (VS) are being used in clinical physiotherapy practice. This study aimed to verify whether the addition of VS to TTNS is more beneficial than TTNS alone for women with OAB. PATIENTS AND METHODS: In all, 106 women aged >18 years diagnosed with OAB or mixed urinary incontinence with prevalent OAB symptoms were randomly divided into 2 groups: Group 1: TTNS (n = 52); Group 2: TTNS + VS (n = 54). The 3 day voiding diary, pelvic floor muscle strength (Ortiz Scale), King's Health Questionnaire, and Overactive Bladder Questionnaire were assessed before and after treatment. Urinary frequency was considered the primary outcome, and a reduction of ≥ 3 micturitions/day was considered clinically relevant. Mixed linear models were used to compare the 2groups. RESULTS: Initially, the groups were similar in age, body mass index, number of pregnancies, time of OAB onset, and prevalence of OAB symptoms. After treatment, a reduction in urinary frequency of 1.5 micturitions was observed in Group 2, which was not clinically relevant despite being statistically significant. CONCLUSIONS: The addition of VS to TTNS for the treatment of OAB was not more effective than TTNS as a single therapy.

Comparison of three methods of DNA extraction from peripheral blood mononuclear cells and lung fragments of equines.

We compared three different protocols for DNA extraction from horse peripheral blood mononuclear cells (PBMC) and lung fragments, determining average final DNA concentration, purity, percentage of PCR amplification using beta-actin, and cost. Thirty-four samples from PBMC, and 33 samples from lung fragments were submitted to DNA extraction by three different protocols. Protocol A consisted of a phenol-chloroform and isoamylic alcohol extraction, Protocol B used alkaline extraction with NaOH, and Protocol C used the DNAzol((R)) reagent kit. Protocol A was the best option for DNA extraction from lung fragments, producing high DNA concentrations, with high sensitivity in PCR amplification (100%), followed by Protocols C and B. On the other hand, for PBMC samples, Protocol B gave the highest sensitivity in PCR amplification (100%), followed by Protocols C and A. We conclude that Protocol A should be used for PCR diagnosis from lung fragment samples, while Protocol B should be used for PBMC.

Gait characteristics during inadvertent obstacle contacts in young, middle-aged and older adults.

BACKGROUND: When stepping over obstacles, analyses have focused on the successful trials to understand adaptive gait. However, examination of the inadvertent trips that occasionally occur in the laboratory can provide a rich source of information regarding the gait characteristics underlying trip-related falls. RESEARCH QUESTION: What gait variables during obstacle crossing are associated with inadvertent obstacle contacts, and are these variables different across the lifespan? METHODS: Three age groups included: young adults (20-35 years, N = 20), middle-aged adults (50-64 years, N = 15), and older adults (65-79 years, N = 19). A stationary, visible obstacle (26 cm tall) was placed in the middle of a walkway. Foot trajectories and head angles were compared between contact and non-contact trials. RESULTS: Twelve participants contacted the obstacle: seven young adults (3.5% of young adult trials), two middle-aged adults (1.3%), and three older adults (1.6%). Young and middle-aged adults contacted primarily with the trail limb, while older adults contacted primarily with the lead limb. Contacts occurred for different reasons: Most young adult contact trials had appropriate foot placement, but inadequate elevation; middle-aged and older adults demonstrated inappropriate foot placement before the obstacle, leading to foot contact during the swing phase. SIGNIFICANCE: Lower contact rates in the middle-aged and older adults indicates that the cautious strategies adopted during obstacle crossing are effective. Higher contact rates in young adults may indicate trial-and-error exploratory behavior. Inappropriate foot placement in the middle-aged and older adults may indicate impaired ability to gather obstacle position information during the approach phase.