Baseline extent of damage predicts spinal radiographic progression in Korean patients with ankylosing spondylitis treated with golimumab.
For patients with ankylosing spondylitis (AS), golimumab has consistent efficacy in controlling disease activity over 5 years but its benefit in preventing radiographic progression was less clear at 4 years. To predict radiographic progression, we analyzed the baseline characteristics of AS patients in a Korean population. Sixty-eight Korean patients with AS participated in the phase 3, multicenter, randomized, placebo-controlled, double-blind trial (GO-RAISE) which has previously been described. Baseline modified stoke AS spine score (mSASSS) and change in mSASSS from baseline (ΔmSASSS) until week 208 were analyzed in the Korean patients enrolled in the GO-RAISE study. Although Korean patients had lower baseline mSASSS compared to non-Korean patients and received active management, radiographic progression was not prevented. Korean patients who did not undergo radiographic progression of spinal lesions of AS were younger and had shorter symptomatic duration, lower Bath AS functional and metrology indices, better chest expansion, and lower baseline mSASSS. The baseline mSASSS and ΔmSASSS were positively correlated in Korean AS patients (p < 0.001). Radiographic progression was more prevalent (80.0%) when baseline mSASSS > 10 and less common (13.0%) with baseline mSASSS = 0. In Korean AS patients, radiographic progression of the spine after 4 years was predicted effectively by the initial severity of the spinal lesion(s) in patients treated with golimumab.


A prospective evaluation of the Landing Error Scoring System (LESS) as a screening tool for anterior cruciate ligament injury risk.
Anterior cruciate ligament (ACL) injuries are immediately disabling, costly, take a significant amount of time to rehabilitate, and are associated with an increased risk of developing posttraumatic osteoarthritis of the knee. Specific multiplanar movement patterns of the lower extremity, such as those associated with the drop vertical jump (DVJ) test, have been shown to be associated with an increased risk of suffering noncontact ACL injuries. The Landing Error Scoring System (LESS) has been developed as a tool that can be applied to identify individuals who display at-risk movement patterns during the DVJ. An increase in LESS score is associated with an increased risk of noncontact ACL injury. Case-control study; Level of evidence, 3. Over a 3-year interval, 5047 high school and college participants performed preseason DVJ tests that were recorded using commercial video cameras. All participants were followed for ACL injury during their sports season, and video data from injured participants and matched controls were then assessed with the LESS. Conditional logistic regression analysis was used to examine the association between LESS score and ACL injury risk in all participants as well as subgroups of female, male, high school, and college participants. There was no relationship between the risk of suffering ACL injury and LESS score whether measured as a continuous or a categorical variable. This was the case for all participants combined (odds ratio, 1.04 per unit increase in LESS score; 95% confidence interval, 0.80-1.35) as well as within each subgroup (odds ratio range, 0.99-1.14). The LESS did not predict ACL injury in our cohort of high school and college athletes.


Tuberculosis control and directly observed therapy from the public health/human rights perspective.
Directly observed therapy, short course (DOTS) is the current international strategy for controlling tuberculosis. Decisions have been taken internationally about the increasing tuberculosis epidemic-what to do and why to do it. But do we know bow the DOTS strategy can be implemented most appropriately and what changes need to be made to ensure that it is effective? This paper uses the Public Health/Human Rights framework to discuss TB control from a human rights rather than the biomedical perspective. The aim is to introduce different approaches to the current DOTS strategy in order to find more effective and appropriate ways to treat and care for people with tuberculosis. The paper argues that key dimensions of social, economic and physical access to TB services need to be assessed and accounted for in programme design. This will require that TB control adopt a wider interdisciplinary and multisectoral perspective to complement the current biomedical orientation.


Microfluidics and microbial engineering.
The combination of microbial engineering and microfluidics is synergistic in nature. For example, microfluidics is benefiting from the outcome of microbial engineering and many reported point-of-care microfluidic devices employ engineered microbes as functional parts for the microsystems. In addition, microbial engineering is facilitated by various microfluidic techniques, due to their inherent strength in high-throughput screening and miniaturization. In this review article, we firstly examine the applications of engineered microbes for toxicity detection, biosensing, and motion generation in microfluidic platforms. Secondly, we look into how microfluidic technologies facilitate the upstream and downstream processes of microbial engineering, including DNA recombination, transformation, target microbe selection, mutant characterization, and microbial function analysis. Thirdly, we highlight an emerging concept in microbial engineering, namely, microbial consortium engineering, where the behavior of a multicultural microbial community rather than that of a single cell/species is delineated. Integrating the disciplines of microfluidics and microbial engineering opens up many new opportunities, for example in diagnostics, engineering of microbial motors, development of portable devices for genetics, high throughput characterization of genetic mutants, isolation and identification of rare/unculturable microbial species, single-cell analysis with high spatio-temporal resolution, and exploration of natural microbial communities.


Variability of pulmonary function in alpha-1-antitrypsin deficiency: clinical correlates.
To determine the range of pulmonary function variability in alpha-1-antitrypsin-deficient persons and to identify epidemiologic factors and pulmonary symptoms and conditions associated with this variability. Case series ascertained through investigation of extant obstructive lung disease (index cases, 22 subjects) or by other means (non-index cases, 30 subjects). Referral-based pulmonary division at a tertiary care medical center. Fifty-two alpha-1-antitrypsin-deficient persons of type Pi Z ascertained by: extant chronic obstructive pulmonary disease (22 cases), family studies (20 cases), liver disease (4 cases), population screening (4 cases), and other pulmonary problems (2 cases). Pulmonary function tests and a version of the American Thoracic Society 1978 standard respiratory epidemiology questionnaire were used. Persons of type Pi Z who were not specifically ascertained with chronic obstructive pulmonary disease had values of forced expiratory volume in 1 second over 65% of predicted in 20 out of 30 cases and frequently had normal lung function. Univariate and multivariate analyses of possible causes of lung disease showed that the following factors were significant (P less than 0.05): pulmonary symptoms (effects associated with chronic obstructive pulmonary disease), including dyspnea and chronic cough; age and pack-years of smoking (epidemiologic correlates); and other pulmonary conditions (potential causes or effects) including asthma, pneumonia, and episodes of increased cough and phlegm. Finally, we found a striking excess of questionnaire-reported parental emphysema in families of type Pi Z persons with chronic obstructive pulmonary disease compared with families of type Pi Z persons without disease. Many persons with alpha-1-antitrypsin deficiency do not have clinically significant lung function impairment: the perceived natural history of antitrypsin deficiency has been distorted by ascertainment bias. In addition to cigarette smoking, it appears that asthma, lower respiratory infections, and possibly some familial factors contribute to a severe clinical course. Follow-up of our cohort with widely varying lung function will provide insights into the natural history of the emphysema associated with alpha-1-antitrypsin deficiency.


Application of field methods to assess isometamidium resistance of trypanosomes in cattle in western Ethiopia.
This study assessed the degree of isometamidium resistance of trypanosomes infecting cattle in the upper Didessa valley of western Ethiopia. An initial prevalence study was conducted to identify sites with a high risk of trypanosmosis in cattle. The trypanosome prevalence varied widely, with two sites, Kone (21.3%) and Village 1 settlement (15%) having a relatively high prevalence based on the phase-contrast buffy-coat technique (BCT). In the highest risk area, the Kone settlement, an isometamidium block treatment study was conducted from April to June 2001. A total of 300 cattle were included in this study, 100 from each of three villages (Cheleleki, Kolu and Burka). At day minus 14 of the study, all 300 cattle were treated with diminazene aceturate at 7 mg/kg body weight. Subsequently, these cattle were ear-tagged and randomly assigned into two groups, 50 controls and 50 for isometamidium treatment in each village. Fourteen days later (day 0), the 50 treatment cattle were given isometamidium chloride at 1 mg/kg body weight. Both groups of cattle were then examined for trypanosome parasites using BCT every 14 days until 84 days. The two indices used in assessing isometamidium resistance, namely the proportion of infections during an 8-week follow-up period and the ratio of mean hazards in an isometamidium treated versus untreated group, provided consistent results across the three villages. In Burka village, both indices demonstrated the presence of isometamidium resistance trypanosome infections while in Cheleleki and Kolu villages, both indices did not indicate significant levels of resistance. There were significant differences between the Kaplan-Meier survival estimates of the control and treatment groups in Cheleleki (P < 0.01) and Kolu (P < 0.05) but not in Burka (P > 0.05).


Comparative cytophysiology of striated muscle with special reference to the role of the endoplasmic reticulum.
1. The structure and distribution of the components of striated muscle cells vary with the species and with the specialization of muscle fiber function. 2. There appear to be two, easily distinguishable, general categories of striated muscle structure. A. High frequency muscle (represented by flight muscle of higher insects and hummingbird, and cicada tympanal muscle) is characterized by widely spaced, non-branching fibrils of large diameter and short period, little endoplasmic reticulum, and large quantities of large mitochondria (low fibril-sarcoplasm ratio). This structure is correlated with heavy tracheolization or vascularization, high oxidative activity, and dark color as compared with other muscles of the same species. B. Low frequency muscle is characterized, in general, by high fibril-sarcoplasm ratio, relatively long period, few mitochondria increasing with activity and decreasing with absolute power of the fiber. Oxidative capacity and color are proportional to the quantity of mitochondria. These fibers are further differentiated into (a) fibrillar arrangement of contractile material which permits a regular pattern of interfibrillar and segmental reticulum, and (b) afibrillar arrangement of contractile material leading to an unsystematic distribution of reticulum. 3. The endoplasmic reticulum appears as a complex coordination system in the muscle fiber. Peripherally, it links the Z and M lines of the fibrils to the sarcolemma and between the fibrils it links the cross-bands, forming the Grundmembran of earlier authors. By longitudinal linkage, it connects with the sarcolemma at the muscle extremity to form a digital arrangement into which the tendon fibrils are spliced. The extent of its development and its position have a definite relationship to the degree and site of fiber shortening. At present the reticulum is the only structure that one can consider to be an internal conducting system. It may distribute the excitation transversely from fibril to fibril, and lengthwise saltatorially to the symmetry centers of the sarcomeres. 4. The nucleus is the mediating element between the cytoplasmic phases within and without the tubular system of the endoplasmic reticulum. A possible mechanism which correlates nucleus, adenylic acid system, ion exchange, and reticulum with the initiation of contraction is postulated.


Donepezil for dementia due to Alzheimer's disease.
Alzheimer's disease is the most common cause of dementia in older people. One approach to symptomatic treatment of Alzheimer's disease is to enhance cholinergic neurotransmission in the brain by blocking the action of the enzyme responsible for the breakdown of the neurotransmitter acetylcholine. This can be done by a group of drugs known as cholinesterase inhibitors. Donepezil is a cholinesterase inhibitor.This review is an updated version of a review first published in 1998. To assess the clinical efficacy and safety of donepezil in people with mild, moderate or severe dementia due to Alzheimer's disease; to compare the efficacy and safety of different doses of donepezil; and to assess the effect of donepezil on healthcare resource use and costs. We searched Cochrane Dementia and Cognitive Improvement's Specialized Register, MEDLINE, Embase, PsycINFO and a number of other sources on 20 May 2017 to ensure that the search was as comprehensive and up-to-date as possible. In addition, we contacted members of the Donepezil Study Group and Eisai Inc. We included all double-blind, randomised controlled trials in which treatment with donepezil was administered to people with mild, moderate or severe dementia due to Alzheimer's disease for 12 weeks or more and its effects compared with those of placebo in a parallel group of patients, or where two different doses of donepezil were compared. One reviewer (JSB) extracted data on cognitive function, activities of daily living, behavioural symptoms, global clinical state, quality of life, adverse events, deaths and healthcare resource costs. Where appropriate and possible, we estimated pooled treatment effects. We used GRADE methods to assess the quality of the evidence for each outcome. Thirty studies involving 8257 participants met the inclusion criteria of the review, of which 28 studies reported results in sufficient detail for the meta-analyses. Most studies were of six months' duration or less. Only one small trial lasted 52 weeks. The studies tested mainly donepezil capsules at a dose of 5 mg/day or 10 mg/day. Two studies tested a slow-release oral formulation that delivered 23 mg/day. Participants in 21 studies had mild to moderate disease, in five studies moderate to severe, and in four severe disease. Seventeen studies were industry funded or sponsored, four studies were funded independently of industry and for nine studies there was no information on source of funding.Our main analysis compared the safety and efficacy of donepezil 10 mg/day with placebo at 24 to 26 weeks of treatment. Thirteen studies contributed data from 3396 participants to this analysis. Eleven of these studies were multicentre studies. Seven studies recruited patients with mild to moderate Alzheimer's disease, two with moderate to severe, and four with severe Alzheimer's disease, with a mean age of about 75 years. Almost all evidence was of moderate quality, downgraded due to study limitations.After 26 weeks of treatment, donepezil compared with placebo was associated with better outcomes for cognitive function measured with the Alzheimer's Disease Assessment Scale-Cognitive (ADAS-Cog, range 0 to 70) (mean difference (MD) -2.67, 95% confidence interval (CI) -3.31 to -2.02, 1130 participants, 5 studies), the Mini-Mental State Examination (MMSE) score (MD 1.05, 95% CI 0.73 to 1.37, 1757 participants, 7 studies) and the Severe Impairment Battery (SIB, range 0 to 100) (MD 5.92, 95% CI 4.53 to 7.31, 1348 participants, 5 studies). Donepezil was also associated with better function measured with the Alzheimer's Disease Cooperative Study activities of daily living score for severe Alzheimer's disease (ADCS-ADL-sev) (MD 1.03, 95% CI 0.21 to 1.85, 733 participants, 3 studies). A higher proportion of participants treated with donepezil experienced improvement on the clinician-rated global impression of change scale (odds ratio (OR) 1.92, 95% CI 1.54 to 2.39, 1674 participants, 6 studies). There was no difference between donepezil and placebo for behavioural symptoms measured by the Neuropsychiatric Inventory (NPI) (MD -1.62, 95% CI -3.43 to 0.19, 1035 participants, 4 studies) or by the Behavioural Pathology in Alzheimer's Disease (BEHAVE-AD) scale (MD 0.4, 95% CI -1.28 to 2.08, 194 participants, 1 study). There was also no difference between donepezil and placebo for Quality of Life (QoL) (MD -2.79, 95% CI -8.15 to 2.56, 815 participants, 2 studies).Participants receiving donepezil were more likely to withdraw from the studies before the end of treatment (24% versus 20%, OR 1.25, 95% CI 1.05 to 1.50, 2846 participants, 12 studies) or to experience an adverse event during the studies (72% vs 65%, OR 1.59, 95% 1.31 to 1.95, 2500 participants, 10 studies).There was no evidence of a difference between donepezil and placebo for patient total healthcare resource utilisation.Three studies compared donepezil 10 mg/day to donepezil 5 mg/day over 26 weeks. The 5 mg dose was associated with slightly worse cognitive function on the ADAS-Cog, but not on the MMSE or SIB, with slightly better QoL and with fewer adverse events and withdrawals from treatment. Two studies compared donepezil 10 mg/day to donepezil 23 mg/day. There were no differences on efficacy outcomes, but fewer participants on 10 mg/day experienced adverse events or withdrew from treatment. There is moderate-quality evidence that people with mild, moderate or severe dementia due to Alzheimer's disease treated for periods of 12 or 24 weeks with donepezil experience small benefits in cognitive function, activities of daily living and clinician-rated global clinical state. There is some evidence that use of donepezil is neither more nor less expensive compared with placebo when assessing total healthcare resource costs. Benefits on 23 mg/day were no greater than on 10 mg/day, and benefits on the 10 mg/day dose were marginally larger than on the 5 mg/day dose, but the rates of withdrawal and of adverse events before end of treatment were higher the higher the dose.


Genomic expression libraries for the identification of cross-reactive orthopoxvirus antigens.
Increasing numbers of human cowpox virus infections that are being observed and that particularly affect young non-vaccinated persons have renewed interest in this zoonotic disease. Usually causing a self-limiting local infection, human cowpox can in fact be fatal for immunocompromised individuals. Conventional smallpox vaccination presumably protects an individual from infections with other Orthopoxviruses, including cowpox virus. However, available live vaccines are causing severe adverse reactions especially in individuals with impaired immunity. Because of a decrease in protective immunity against Orthopoxviruses and a coincident increase in the proportion of immunodeficient individuals in today's population, safer vaccines need to be developed. Recombinant subunit vaccines containing cross-reactive antigens are promising candidates, which avoid the application of infectious virus. However, subunit vaccines should contain carefully selected antigens to confer a solid cross-protection against different Orthopoxvirus species. Little is known about the cross-reactivity of antibodies elicited to cowpox virus proteins. Here, we first identified 21 immunogenic proteins of cowpox and vaccinia virus by serological screenings of genomic Orthopoxvirus expression libraries. Screenings were performed using sera from vaccinated humans and animals as well as clinical sera from patients and animals with a naturally acquired cowpox virus infection. We further analyzed the cross-reactivity of the identified immunogenic proteins. Out of 21 identified proteins 16 were found to be cross-reactive between cowpox and vaccinia virus. The presented findings provide important indications for the design of new-generation recombinant subunit vaccines.


A randomized comparison of TR band and radistop hemostatic compression devices after transradial coronary intervention.
The transradial route for coronary intervention has proven to be safe, effective, and widely applicable in different clinical situations. Several compressive hemostatic devices have been introduced that have shown to be safe and are effective in achieving hemostasis. Seven hundred ninety patients were randomly assigned to receive either TR band or Radistop hemostatic compression devices after transradial coronary procedure. The outcome measures were patient tolerance of the device, local vascular complications, and the time taken to achieve hemostasis. The mean age was 62.88 years, and 74.2% of the patients were men. Patient age, height, weight, wrist circumference, body mass index, male sex, hypertension, diabetes, hypercholesterolemia, and smoking incidences were similar in both groups. There were significantly more patients reporting no discomfort in the TR band group compared to the Radistop group (77% vs. 61%; P = 0.0001). Patients in the Radistop group reported significantly more pain across all categories of severity and three patients in the Radistop group were crossed over to TR band because of severe discomfort. Oozing and ecchymosis were seen in about 16% of the patients. Local small hematoma and large hematoma were seen in 5.4% and 2.2% patients respectively, and similar in both groups. Radial artery occlusion at the time of discharge was seen in 9.2% of the patients though only 6.8% showed persistent occlusion at the time of follow-up. The time taken to achieve hemostasis was significantly longer in the TR Band group (5.32 ± 2.29 vs. 4.83 ± 2.23 hr; P = 0.004). There was significantly higher incidence of radial artery occlusion in patients with smaller wrist circumference, the patients who experienced radial artery spasm during the procedure, and patients with no heparin administration during the procedure. We have shown in a randomized comparison of Radistop and TR band that both devices are safe and effective as hemostatic compression devices following transradial procedures. However, more patients felt discomfort with the Radistop device and the time taken to achieve hemostasis was longer with TR band. © 2010 Wiley-Liss, Inc.


Effects of Vedolizumab in Patients With Primary Sclerosing Cholangitis and Inflammatory Bowel Diseases.
Gut-homing lymphocytes that express the integrin α4β7 and CCR9 might contribute to development of primary sclerosing cholangitis (PSC). Vedolizumab, which blocks the integrin α4β7, is used to treat patients with inflammatory bowel diseases (IBD), but there are few data on its efficacy in patients with PSC. We investigated the effects of vedolizumab in a large international cohort of patients with PSC and IBD. We collected data from European and North American centers participating in the International PSC Study Group from patients with PSC and IBD who received at least 3 doses of vedolizumab (n = 102; median vedolizumab treatment duration, 412 days). Demographic and clinical data were collected from baseline and during the follow-up period (until liver transplantation, death, or 56 days after the final vedolizumab infusion). We analyzed overall changes in biochemical features of liver and proportions of patients with reductions in serum levels of alkaline phosphatase (ALP) of 20% or more, from baseline through last follow-up evaluation. Other endpoints included response of IBD to treatment (improved, unchanged, or worsened, judged by the treating clinician, as well as endoscopic score) and liver-related outcomes. In the entire cohort, the median serum level of ALP increased from 1.54-fold the upper limit of normal at baseline to 1.64-fold the upper limit of normal at the last follow-up examination (P = .018); serum levels of transaminases and bilirubin also increased by a small amount between baseline and the last follow-up examination. Serum levels of ALP decreased by 20% or more in 21 patients (20.6%); only the presence of cirrhosis (odds ratio, 4.48; P = .019) was independently associated with this outcome. Of patients with available endoscopic data, 56.8% had a response of IBD to treatment. Liver-related events occurred in 21 patients (20.6%), including bacterial cholangitis, cirrhosis decompensation, or transplantation. In an analysis of patients with PSC and IBD in an international study group, we found no evidence for a biochemical response to vedolizumab, although serum level of ALP decreased by 20% or more in a subset of patients. Vedolizumab appears to be well tolerated and the overall response of IBD was the same as expected for patients without PSC.


Modelling soil erosion response to sustainable landscape management scenarios in the Mo River Basin (Togo, West Africa).
The rural landscapes in Central Togo are experiencing severe land degradation, including soil erosion. However, spatially distributed information has scarcely been produced to identify the effects of landscape pattern dynamics on ecosystem services, especially the soil erosion control. In addition, relevant information for sustainable land and soil conservation is still lacking at watershed level. On this basis, using the LAndscape Management and Planning Tool for the Mo River basin (LAMPT_Mo), we (1) modelled soil erosion patterns in relation with land use/cover change (LUCC), land protection regime, and landforms, and (2) examined the efficiency of landscape redesign options on soil erosion amounts at basin scale. We found that Simulated historical net soil loss (NSL) for the Mo basin were approximately 26, 23, 27, and 44t/ha/yr, for 1972, 1987, 2000, and 2014, respectively. These simulated NSLs were higher than the tolerable soil loss limits for the Tropics. Steep slopes (≥15°), poorly covered lands (croplands and savannas), and riversides (distances ≤100m) are critical areas of sediment sources. The local appraisal of soil loss was in line with the simulated outputs even though quantification was not accounted for when dealing with rural illiterate people. Furthermore, results showed that the examined management measures, such as controlling the identified erosion hotspots through land protective measures, could help reduce the NSL up to 70%, to values closer to the tolerable limits for the Tropics. The model implementation in the basin showed insights for identifying erosion hotspots and targeting soil conservation planning and landscape restoration measures.


Determinants of gait speed in COPD.
Measures of physical function, daily physical activity, and exercise capacity have been proposed for the care of patients with COPD but are not used routinely in daily office care. Gait speed is a powerful and simple measure of physical function in elderly patients and seems to be a promising measure for the daily care of patients with COPD. The objective of this study was to comprehensively evaluate the determinants and factors influencing gait speed in COPD, particularly the association of gait speed with objectively measured physical activity and the most used exercise capacity field test in cardiopulmonary disease: the 6-min walk test (6MWT). One hundred thirty patients with stable COPD performed two different 4-m gait speed protocols (usual and maximal pace). We modeled gait speed using demographics, lung function, dyspnea, quality of life, physical activity monitoring, exercise capacity, mood, cognitive function, and health-care use. Gait speed was independently associated with 6MWT but not with daily physical activity. The correlation between gait speed and 6MWT was high regardless of protocol used (r = 0.77-0.80). Both 6MWT and gait speed shared similar constructs. Gait speed had an excellent ability to predict poor (≤ 350 m) or very poor (≤ 200 m) 6MWT distances (areas under the curve, 0.87 and 0.98, respectively). Gait speed was not independently associated with quality of life, mood, or cognitive function. Gait speed is more indicative of exercise capacity (6MWT) than daily physical activity in COPD. Despite its simplicity, gait speed has outstanding screening properties for detecting poor and very poor 6MWT performance, making it a useful and informative tool for the clinical care of patients with COPD.


[Primary adrenal carcinoma].
A series of 17 patients with primary adrenal cortical carcinoma is presented. Of 14 patients without evidence of disseminated disease prior to radical surgery, 6 developed distant metastases during follow-up. The mean interval free of disease in these patients was 12 months (range 3-40) and the median survival 15 months. Of the six patients in whom adjuvant external beam radiotherapy was delivered, no evidence of local recurrence was observed. Of the patients treated without external beam radiotherapy, 3 local failures were observed. The projected 5 years survival was 35%, which is similar to that reported in the literature. To date, there is no evidence that chemotherapy nor radiotherapy could improve the survival, however, radiotherapy might increase the local control in selected cases.


Biomarkers in Lung Cancer Screening: Achievements, Promises, and Challenges.
The present review is an update of the research and development efforts regarding the use of molecular biomarkers in the lung cancer screening setting. The two main unmet clinical needs, namely, the refinement of risk to improve the selection of individuals undergoing screening and the characterization of undetermined nodules found during the computed tomography-based screening process are the object of the biomarkers described in the present review. We first propose some principles to optimize lung cancer biomarker discovery projects. Then, we summarize the discovery and developmental status of currently promising molecular candidates, such as autoantibodies, complement fragments, microRNAs, circulating tumor DNA, DNA methylation, blood protein profiling, or RNA airway or nasal signatures. We also mention other emerging biomarkers or new technologies to follow, such as exhaled breath biomarkers, metabolomics, sputum cell imaging, genetic predisposition studies, and the integration of next-generation sequencing into study of circulating DNA. We also underline the importance of integrating different molecular technologies together with imaging, radiomics, and artificial intelligence. We list a number of completed, ongoing, or planned trials to show the clinical utility of molecular biomarkers. Finally, we comment on future research challenges in the field of biomarkers in the context of lung cancer screening and propose a design of a trial to test the clinical utility of one or several candidate biomarkers.


Is current practice around late termination of pregnancy eugenic and discriminatory? Maternal interests and abortion.
The attitudes of Australian practitioners working in clinical genetics and obstetrical ultrasound were surveyed on whether termination of pregnancy (TOP) should be available for conditions ranging from mild to severe fetal abnormality and for non-medical reasons. These were compared for terminations at 13 weeks and 24 weeks. It was found that some practitioners would not facilitate TOP at 24 weeks even for lethal or major abnormalities, fewer practitioners support TOP at 24 weeks compared with 13 weeks for any condition, and the difference in attitudes to TOP between 13 weeks and 24 weeks is most marked for pregnancies which are normal or involve a mild disorder. It is argued that a fetal abnormality criterion for late TOP is inconsistently applied, discriminatory and eugenic. Four possible moral justifications for current practice are examined, each of which would require significant changes to current practice. I argue in favour of a maternal interests criterion for any TOP.


Development of colorimetric microtiter plate assay for assessment of antimicrobials against Acanthamoeba.
We have developed and optimized a 96-well microtiter plate assay, based on the reduction of alamarBlue, to assess the efficacies of much needed new antimicrobials against Acanthamoeba species. This assay has been optimized for determination of drug efficacy against two potentially pathogenic species, Acanthamoeba castellanii and Acanthamoeba polyphaga, and has been validated by comparison of their relative susceptibilities to chlorhexidine, a drug widely used to treat Acanthamoeba keratitis. The results demonstrate that the assay is comparable to a manual counting assay and that A. polyphaga is more resistant to chlorhexidine than A. castellanii. Thus, by use of the manual counting assay, 3.125 microM chlorohexidine was almost completely effective against A. castellanii, whereas this concentration was less than 20% effective against A. polyphaga. Similar results were obtained by the alamarBlue assay. The new assay was used to determine the relative susceptibilities of A. castellanii and A. polyphaga to the alkylphosphocholines (APCs) hexadecylphosphocholine (hexadecyl-PC; miltefosine) and octadecylphosphocholine (octadecyl-PC) as well as an alkylgycerolphosphocholine, edelfosine. Both APCs studied were equally effective against A. castellanii, but octadecyl-PC was less effective than hexadecyl-PC against A. polyphaga. Both APCs were more effective than edelfosine against both Acanthamoeba species. A. polyphaga was found to be significantly less susceptible to each of the phosphocholine analogues. The newly described assay offers a number of advantages over those described previously. It is less labor-intensive than previously described assays and is sensitive and rapid, and the results can be read in a nonsubjective manner. As it is based on a standard 96-well, microtiter plate, it is amenable to automation and high throughput.


Population heterogeneity in Plasmodium vivax relapse risk.
A key characteristic of Plasmodium vivax parasites is their ability to adopt a latent liver-stage form called hypnozoites, able to cause relapse of infection months or years after a primary infection. Relapses of infection through hypnozoite activation are a major contributor to blood-stage infections in P vivax endemic regions and are thought to be influenced by factors such as febrile infections which may cause temporary changes in hypnozoite activation leading to 'temporal heterogeneity' in reactivation risk. In addition, immunity and variation in exposure to infection may be longer-term characteristics of individuals that lead to 'population heterogeneity' in hypnozoite activation. We analyze data on risk of P vivax in two previously published data sets from Papua New Guinea and the Thailand-Myanmar border region. Modeling different mechanisms of reactivation risk, we find strong evidence for population heterogeneity, with 30% of patients having almost 70% of all P vivax infections. Model fitting and data analysis indicates that individual variation in relapse risk is a primary source of heterogeneity of P vivax risk of recurrences. Trial Registration: ClinicalTrials.gov NCT01640574, NCT01074905, NCT02143934.


Pharmacology of epidermal growth factor inhibitors.
Research into the molecular bases of malignant diseases has yielded the development of many novel agents with potential antitumor activity. Evidence for a causative role for the epidermal growth factor receptor (EGFR), which is now regarded as an excellent target for cancer chemotherapy in human cancer, leads to the development of EGFR inhibitors. Two classes of anti-EGFR agents are currently in clinical use: monoclonal antibodies directed at the extracellular domain of the receptor, and the low-molecular-weight receptor tyrosine kinase inhibitors acting intracellularly by competing with adenosine triphosphate for binding to the tyrosine kinase portion of the EGFR. The effect on the receptor interferes with key biological functions including cell cycle arrest, potentiation of apoptosis, inhibition of angiogenesis and cell invasion and metastasis. Cetuximab, a monoclonal antibody, and the receptor tyrosine kinase inhibitors gefitinib and erlotinib are currently approved for the treatment of patients with cancer. New agents with clinical activity are entering the clinic, and new combinatorial approaches are being explored with the aim of improving the potency and pharmacokinetics of EGFR inhibition, to increase the synergistic activity in combination with chemotherapy and overcome resistance to the EGFR inhibitors.


Targeted capture and sequencing of gene-sized DNA molecules.
Targeted capture provides an efficient and sensitive means for sequencing specific genomic regions in a high-throughput manner. To date, this method has mostly been used to capture exons from the genome (the exome) using short insert libraries and short-read sequencing technology, enabling the identification of genetic variants or new members of large gene families. Sequencing larger molecules results in the capture of whole genes, including intronic and intergenic sequences that are typically more polymorphic and allow the resolution of the gene structure of homologous genes, which are often clustered together on the chromosome. Here, we describe an improved method for the capture and single-molecule sequencing of DNA molecules as large as 7 kb by means of size selection and optimized PCR conditions. Our approach can be used to capture, sequence, and distinguish between similar members of the NB-LRR gene family-key genes in plant immune systems.


Detectability of the degree of freeze damage in meat depends on analytic-tool selection.
Novel freezing solutions are constantly being developed to reduce quality loss in meat production chains. However, there is limited focus on identifying the sensitive analytical tools needed to directly validate product changes that result from potential improvements in freezing technology. To benchmark analytical tools relevant to meat research and production, we froze pork samples using traditional (-25 °C, -35 °C) and cryogenic freezing (-196 °C). Three classes of analyses were tested for their capacity to separate different freeze treatments: thaw loss testing, bioelectrical spectroscopy (nuclear magnetic resonance, microwave, bioimpedance) and low-temperature microscopy (cryo-SEM). A general effect of freeze treatment was detected with all bioelectrical methods. Yet, only cryo-SEM resolved quality differences between all freeze treatments, not only between cryogenic and traditional freezing. The detection sensitivity with cryo-SEM may be explained by testing meat directly in the frozen state without prior defrosting. We discuss advantages, shortcomings and cost factors in using analytical tools for quality monitoring in the meat sector.


Design of nanowire-induced nanocavities in grooved 1D and 2D SiN photonic crystals for the ultra-violet and visible ranges.
Nanowire-induced SiN photonic crystal (PhC) nanocavities specifically designed for the ultra-violet and visible range are investigated by three-dimensional finite-difference time-domain calculations. As opposed to their silicon PhC counterpart, we find that the formation of nanowire-induced two-dimensional (2D) SiN PhC nanocavities is more challenging because of the low refractive index of SiN. We thus discuss optimization strategies to circumvent such difficulties and we investigate the influence of critical design parameters such as PhC geometry, as well as nanowire geometry and position. We also propose a novel nanowire-induced cavity design based on one-dimensional (1D) nanobeam PhCs. We finally report on nanowire-induced nanocavity designs in 1D (resp. 2D) PhCs presenting quality factors as high as Qc = 5.1 x 104 (resp. Qc = 2.5 x 104 with a mode volume Vm=1.8(λ/nrNW)3 (resp. Vm=5.1(λ/nrNW)3), which show good prospects for light-matter interaction in the near-ultraviolet and visible ranges.


The beta/alpha peak height ratio of ATP. A measure of free [Mg2+] using 31P NMR.
From 31P NMR measurements made in vitro at 38 degrees C, I = 0.25, pH 5. 75-8.5, and calculated free [Mg2+] from 0 to 5 mM, we show that, within the physiological range of cytosolic free [Mg2+] from 0.25 to 1.5 mM, the chemical shift difference between the alpha- and beta-ATP resonances, deltaalphabeta, changes by only 0.6 ppm. Consequently, we developed new formalisms from known acid and Mg2+ dissociation constants by which the observed chemical shift of Pi, deltaPi, and the peak height ratio of the beta- and alpha-ATP resonances, hbeta/alpha, could be related to free [Mg2+] by simultaneous solution of: [equation: see text] We found that hbeta/alpha changed 2.5-fold as free [Mg2+] varied from 0.25 to 1.5 mM, providing a more sensitive and accurate measure of free cytosolic [Mg2+]. In working rat heart perfused with glucose, free [Mg2+] was 1.0 +/- 0.1 from hbeta/alpha and 1.2 +/- 0.03 from measured [citrate]/[isocitrate] but 0.51 +/- 0.1 from deltaalphabeta. Addition of ketone bodies to the perfusate decreased free [Mg2+] estimated from hbeta/alpha to 0.61 +/- 0.02 and 0.74 +/- 0.11 by [citrate]/[isocitrate] but the estimate from deltaalphabeta was unchanged at 0.46 +/- 0.04 mM. Such differences in estimated free [Mg2+] alter the apparent Keq of the creatine kinase reaction and hence the estimated cytosolic free [SigmaADP].


Multiculturalism as a dimension of school climate: the impact on the academic achievement of Asian American and Hispanic youth.
Multiculturalism constitutes an important element of school climate, but the relation between perceived multiculturalism and academic achievement has not been widely studied. This study examined the influence of students' perceptions of school support for multiculturalism on academic achievement among 280 Asian American and Hispanic youth, including ethnic identity and ethnocultural empathy as potential mediators. Results of structural equation modeling revealed that perceived multiculturalism was significantly positively related to ethnocultural empathy for Asian Americans and Hispanics, and that ethnocultural empathy, in turn, was predictive of academic achievement for Hispanics only. Results of bootstrapping to test for mediation effects revealed ethnocultural empathy to be a salient mediator for Hispanic youth. Although ethnic identity did not mediate the link between multiculturalism and academic achievement, ethnic identity was significantly predictive of achievement for Hispanics. On the whole, these findings suggest that fostering a school climate supportive of multiculturalism may improve empathy toward ethnic out-groups. Furthermore, schools that promote compassion and tolerance for diverse ethnic groups may achieve better academic outcomes among Hispanic youth.


State-of-the-art techniques in operative dentistry: contemporary teaching of posterior composites in UK and Irish dental schools.
Advances of composite systems and their application have revolutionised the management of posterior teeth affected by caries, facilitating a minimally invasive approach. Previous surveys have indicated that the teaching of posterior composites within dental schools was developing, albeit not keeping pace with clinical evidence and the development of increasingly predictable techniques and materials. Concurrently, surveys of dental practice indicate that dental amalgam still predominates as the 'material of choice' for the restoration of posterior teeth within UK general dental practice. In light of such considerations, the aim of this study was to investigate current teaching of posterior composites in Irish and UK dental schools. An online questionnaire which sought information in relation to the current teaching of posterior composites was developed and distributed to the 17 established Irish and UK dental schools with undergraduate teaching programmes in late 2009. Completed responses were received from all 17 schools (response rate = 100%). All 17 schools taught the placement of occlusal and two-surface occlusoproximal composites in premolar and permanent molar teeth. Two schools did not teach placement of three-surface occlusoproximal composites in either premolars or molars. In their preclinical courses, ten schools taught posterior composites before teaching dental amalgams. Fifty-five percent of posterior restorations placed by dental students were of composite (range = 10-90%) and 44% amalgam (range = 10-90%), indicating an increase of 180% in the numbers of posterior composites placed over the past five years. Diversity was noted in the teaching of clinical techniques and students at different schools are trained with different composites and bonding systems. Some cause for concern was noted in the teaching of certain techniques that were not in keeping with existing best evidence, such as the teaching of transparent matrix bands and light-transmitting wedges for occluso-proximal composites (eight schools) and the teaching of bevels on the cavosurface enamel margins of both the occlusal and proximal box margins (three schools). The teaching of posterior composites in the Irish and UK dental schools has substantially increased over the last five years. Dental students in these schools often gain more experience in the placement of posterior composites than amalgam. However, practice trends indicate that a majority of GDPs continue to place amalgam in preference to composite, thereby suggesting a source of tension as current dental students emerge into the dental workforce over the coming years. There is, as a consequence, a challenge to the dental profession and its funding agencies in the UK to encourage more of a shift towards the minimally interventive use of composite systems in the restoration of posterior teeth, in particular among established practitioners.


Genetic and phenotypic spectrum associated with IFIH1 gain-of-function.
IFIH1 gain-of-function has been reported as a cause of a type I interferonopathy encompassing a spectrum of autoinflammatory phenotypes including Aicardi-Goutières syndrome and Singleton Merten syndrome. Ascertaining patients through a European and North American collaboration, we set out to describe the molecular, clinical and interferon status of a cohort of individuals with pathogenic heterozygous mutations in IFIH1. We identified 74 individuals from 51 families segregating a total of 27 likely pathogenic mutations in IFIH1. Ten adult individuals, 13.5% of all mutation carriers, were clinically asymptomatic (with seven of these aged over 50 years). All mutations were associated with enhanced type I interferon signaling, including six variants (22%) which were predicted as benign according to multiple in silico pathogenicity programs. The identified mutations cluster close to the ATP binding region of the protein. These data confirm variable expression and nonpenetrance as important characteristics of the IFIH1 genotype, a consistent association with enhanced type I interferon signaling, and a common mutational mechanism involving increased RNA binding affinity or decreased efficiency of ATP hydrolysis and filament disassembly rate.


Evaluation of the monitor cursor-line method for measuring pulmonary artery and central venous pressures.
To determine if the monitor cursor-line feature on bedside monitors is accurate for measuring central venous and pulmonary artery pressures in cardiac surgery patients. Central venous and pulmonary artery pressures were measured via 3 methods (end-expiratory graphic recording, monitor cursor-line display, and monitor digital display) in a convenience sample of postoperative cardiac surgery patients. Pressures were measured twice during both mechanical ventilation and spontaneous breathing. Analysis of variance was used to determine differences between measurement methods and the percentage of monitor pressures that differed by 4 mm Hg or more from the measurement obtained from the graphic recording. Significance level was set at P less than .05. Twenty-five patients were studied during mechanical ventilation (50 measurements) and 21 patients during spontaneous breathing (42 measurements). Measurements obtained via the 3 methods did not differ significantly for either type of pressure (P > .05). Graphically recorded pressures and measurements obtained via the monitor cursor-line or digital display methods differed by 4 mm Hg or more in 4% and 6% of measurements, respectively, during mechanical ventilation and 4% and 11%, respectively, during spontaneous breathing. The monitor cursor-line method for measuring central venous and pulmonary artery pressures may be a reasonable alternative to the end-expiratory graphic recording method in hemodynamically stable, postoperative cardiac surgery patients. Use of the digital display on the bedside monitor may result in larger discrepancies from the graphically recorded pressures than when the cursor-line method is used, particularly in spontaneously breathing patients.


Gene expression changes induced by unilateral ureteral obstruction in mice.
Loss of renal function is often the impetus for operative intervention in renal obstruction cases. Obstructive nephropathy is characterized by discrete morphological and physiological changes, including tubular dilatation, apoptosis and atrophy as well as interstitial cellular infiltration and progressive interstitial fibrosis. We hypothesized that gene expression alterations correlate with obstructive nephropathy and could serve as biomarkers for early intervention. C57BL/6 mice were subjected to unilateral ureteral obstruction or sham surgery at postnatal day 21. Kidneys were harvested 1, 2, 5 and 9 days postoperatively. RNA was extracted from kidneys and comprehensive gene expression profiling was performed with microarrays. IPA® pathway analysis software was used to analyze the biological function and gene networks of gene expression data. Microarray analysis revealed more than 1,800 transcripts that were up-regulated or down-regulated during days 1 through 9 after obstruction, including many previously reported transcripts (FOS, CD44, CLU, SPP1 and EGF). Pathway analysis showed significant enrichment of transcripts in cell activation/differentiation, immune/inflammatory responses, cell cycle, metabolic process and transport. Network analysis using IPA showed that transcriptional regulatory pathways involving CEBPB and HNF4A are involved in obstructive nephropathy. This data set provides a foundation for development of biomarkers for obstructive nephropathy.


TRPV4 antagonists ameliorate ventriculomegaly in a rat model of hydrocephalus.
Hydrocephalus is a serious condition that impacts patients of all ages. The standards of care are surgical options to divert, or inhibit production of, cerebrospinal fluid; to date, there are no effective pharmaceutical treatments, to our knowledge. The causes vary widely, but one commonality of this condition is aberrations in salt and fluid balance. We have used a genetic model of hydrocephalus to show that ventriculomegaly can be alleviated by inhibition of the transient receptor potential vanilloid 4, a channel that is activated by changes in osmotic balance, temperature, pressure and inflammatory mediators. The TRPV4 antagonists do not appear to have adverse effects on the overall health of the WT or hydrocephalic animals.


SU-E-I-25: Performance Evaluation of a Proposed CMOS-Based X-Ray Detector Using Linear Cascade Model Analysis.
The need for high-resolution, dynamic x-ray imaging capability for neurovascular applications has put an ever increasing demand on x-ray detector technology. Present state-of-the-art detectors such as flat panels have limited resolution and noise performance. A linear cascade model analysis was used to estimate the theoretical performance for a proposed CMOS-based detector. The proposed CMOS-based detector was assumed to have a 300-micron thick HL type CsI phosphor, 35-micron pixels, a variable gain light image intensifier (LU), and 400 electron readout noise. The proposed detector has a CMOS sensor coupled to an LII which views the output of the CsI phosphor. For the analysis the whole imaging chain was divided into individual stages characterized by one of the basic processes (stochastic/deterministic blurring, binomial selection, quantum gain, additive noise). Standard linear cascade modeling was used for the propagation of signal and noise through the stages and an RQA5 spectrum was assumed. The gain, blurring or transmission of different stages was either measured or taken from manufacturer's specifications. The theoretically calculated MTF and DQE for the proposed detector were compared with a high-resolution, high-sensitive Micro-Angio Fluoroscope (MAF), predecessor of the proposed detector. Signal and noise for each of the 19 stages in the complete imaging chain were calculated and showed improved performance. For example, at 5 cycles/mm the MTF and DQE were 0.08 and 0.28, respectively, for the CMOS detector compared to 0.05 and 0.07 for the MAF detector. The proposed detector will have improved MTF and DQE and slimmer physical dimension due to the elimination of the large fiber-optic taper used in the MAF. Once operational, the proposed CMOS detector will serve as a further improvement over standard flat panel detectors compared to the MAF which is already receiving a very positive reception by neuro-vascular interventionalists. (Support:NIH-Grant R01EB002873) NIH Grants R01- EB008425, R01-EB002873 and an equipment grant from Toshiba Medical Systems Corp.


Demethylation of the TSDR is a marker of squamous cell carcinoma in transplant recipients.
Malignancy is an important cause of death in transplant recipients. Cutaneous squamous cell carcinoma (cSCC) causes significant morbidity and mortality as 30% of transplant recipients will develop cSCC within 10 years of transplantation. Previously we have shown that high numbers of regulatory T cells (Tregs) are associated with the development of cSCC in kidney transplant recipients (KTRs). Demethylation analysis of the Treg-specific demethylated region (TSDR) provides a more accurate association with cSCC risk after transplantation. Age, gender and duration of immunosuppression matched KTRs with (n=32) and without (n=27) cSCC, were re-analyzed for putative clinical and immunological markers of cancer risk. The proportion of FOXP3+ CD4+ cells was higher in the population with a previous SCC. Major T cell subsets remained stable over time; although B cell, CD8 and CD4 subpopulations demonstrated age-related changes. TSDR methylation analysis allowed clarification of Treg numbers, enhancing the association of high Treg levels in KTRs with cSCC compared to the cSCC-free cohort. These data validate and expand on previous findings in long-term KTRs, and show that immune markers remain stable over time. TSDR demethylation analysis provides a more accurate biomarker of cancer posttransplantation.


Biomolecular network motif counting and discovery by color coding.
Protein-protein interaction (PPI) networks of many organisms share global topological features such as degree distribution, k-hop reachability, betweenness and closeness. Yet, some of these networks can differ significantly from the others in terms of local structures: e.g. the number of specific network motifs can vary significantly among PPI networks. Counting the number of network motifs provides a major challenge to compare biomolecular networks. Recently developed algorithms have been able to count the number of induced occurrences of subgraphs with k < or = 7 vertices. Yet no practical algorithm exists for counting non-induced occurrences, or counting subgraphs with k > or = 8 vertices. Counting non-induced occurrences of network motifs is not only challenging but also quite desirable as available PPI networks include several false interactions and miss many others. In this article, we show how to apply the 'color coding' technique for counting non-induced occurrences of subgraph topologies in the form of trees and bounded treewidth subgraphs. Our algorithm can count all occurrences of motif G' with k vertices in a network G with n vertices in time polynomial with n, provided k = O(log n). We use our algorithm to obtain 'treelet' distributions for k < or = 10 of available PPI networks of unicellular organisms (Saccharomyces cerevisiae Escherichia coli and Helicobacter Pyloris), which are all quite similar, and a multicellular organism (Caenorhabditis elegans) which is significantly different. Furthermore, the treelet distribution of the unicellular organisms are similar to that obtained by the 'duplication model' but are quite different from that of the 'preferential attachment model'. The treelet distribution is robust w.r.t. sparsification with bait/edge coverage of 70% but differences can be observed when bait/edge coverage drops to 50%.


Markers for primary care: missed opportunities to immunize and screen for lead and tuberculosis by private physicians serving large numbers of inner-city Medicaid-eligible children.
This study examines coverage levels for immunization, missed opportunities to immunize, and extent of lead and tuberculosis screening in inner-city storefront physician offices and then relates child, visit, and physician characteristics to missed opportunities. With the use of a nested sampling strategy, 232 charts were selected for review in 31 physicians' offices. Charts selected were for children 0 to 35 months of age who had three or more visits in more than 3 months. Physicians were selected from those in specific low-income New York inner-city neighborhoods who submitted large volumes of Medicaid billing claims. Variables examined were missed opportunities to immunize, immunization coverage levels, lead, and tuberculosis screening. The outcome measure was missed opportunities to immunize. Only 26% of the children were up to date for their age for diphtheria, tetanus, pertussis (DTP), oral polio vaccine (OPV), and measles, mumps, rubella (MMR) compared with a city-wide coverage level of 49%. Children who were not up to date for immunization coverage were more likely not to be up to date for lead (RR = 1.24, CI 0.96 to 1.60) or tuberculosis (RR = 1.54, CI 1.14 to 2.08) screening. Physicians miss opportunities to immunize in 84% of the eligible visits. Opportunities to immunize are missed more frequently at sick care or follow-up visits (95% and 91% missed opportunities) than at well care visits (41% missed opportunities). The quality of pediatric primary care given by these inner-city storefront physicians is suboptimal. Sick and follow-up visits predominate; well care visits are infrequent. If care is to be improved, Medicaid reimbursement policies, which make delivery of well care unprofitable, will need to be changed. In addition, monitoring the quality of care will need to be more aggressive. In the near future children who receive Medicaid in New York will be in managed care. If reimbursement and monitoring policies that provide incentives for delivering pediatric primary care are to be in place, it will be the managed care plans that implement this.


Colostrum and milk can transmit jaagsiekte retrovirus to lambs.
Ovine pulmonary adenocarcinoma (OPA) is a contagious disease caused by jaagsiekte sheep retrovirus (JSRV). In the three studies performed, we have obtained data of the importance of colostrum/milk (C/M) in the transmission of JSRV. In the first study, a group of sheep from a flock with a long history of OPA, samples from colostrum and peripheral blood leucocytes (PBLs) were collected. Two specific PCRs (U3-LTR and env of the JSRV) were carried out. Using U3PCR 8/34 sheep were positive in colostrum whereas with envPCR 7/34 were positive. From these animals only one was positive with U3PCR in the PBLs. Evidence of the transmission of JSRV infection by C/M was obtained in two more separate studies. In the second study, PBLs from five lambs from JSRV+ ewes and two from JSRV-ewes were tested by the U3PCR. They were fed C/M by their mothers during 3 months and slaughtered 7 months after birth. Three out of five lambs from the JSRV+ sheep become PBL positive at 3-4 months old and the other two were also positive at 4-6 months of age. One lamb of the JSRV-sheep became also PBL positive at an age of 3 months. In the third study, a group of lambs from JSRV negative mothers were fed with C/M from JSRV+ sheep and housed in separate unit. For comparison, another group of the same origin and maintained in another different unit, were fed with C/M containing a JSRV virus preparation. All lambs were blood sampled monthly and JSRV infection was detected as early as 15 days and several times onwards in both groups. Control groups fed with C/M from JSRV free flock and JSRV blood test negative sheep were always negative. Together these results indicate that suckling is an important natural transmission route for JSRV.


Development and evaluation of in situ nasal gel formulations of loratadine.
The objective of the present work was to formulate and evaluate mucoadhesive in situ nasal gels of loratadine. This drug delivery system may overcome the first-pass metabolism and subsequently improve the bioavailability of the drug. A total of 16 formulations of in situ nasal gels were prepared using different polymeric ratios of hydroxypropyl methylcellulose (HPMC K-100) and xanthan gum. All formulations had a clear appearance in the sol form, with gelling temperature of the nasal gels ranging between 33.1 ± 0.43 and 34.8 ± 0.82 °C. The gelling time of all the formulations varied from 4.0 ± 0.21 to 11.3 ± 0.22 s; the drug content was >95%. The pH of the formulations ranged between 5.6 ± 0.004 and 6.0 ± 0.003, i.e. no mucosal irritation is expected as the pH was in the acceptable range. Mucoadhesive strength was adequate (3010.89 ± 1.21-6678.89 ± 0.45 dyne/cm(2)) to provide prolonged adhesion. In vitro drug release studies showed that the prepared formulations could release the drug for up to 10 h with all of them following Higuchi kinetics. The accelerated stability studies indicated that the gels were stable over the six months test period. The DSC and XRD analysis revealed that there was no drug-polymer interaction. From these findings it can be concluded that in situ nasal gels may be potential drug delivery systems for loratadine to overcome first-pass metabolism and thereby to improve the bioavailability.


Epicardial adipose stem cell sheets results in greater post-infarction survival than intramyocardial injections.
Intramyocardial injections of cells can damage tissue and enhance dissociation-induced cell death. We assessed whether epicardial delivery of cell sheets could overcome these issues in a rat model of chronic myocardial infarction. Eighty-two rats that had undergone coronary ligation and simultaneous harvest of fat tissue to yield the adipose-derived stromal cell (ADSC) fraction were randomized 1 month after infarction to receive injections of either control medium (n= 24) or 10 × 10(6) autologous ADSC (n= 37) or the epicardial deposit, onto the infarcted area, of a trilayered ADSC sheet (10 × 10(6), n= 21) prepared by culturing cells on temperature-sensitive dishes. Some treated rats received green fluorescent protein labelled ADSC. Survival, function, and cell engraftment were blindly assessed after 2 months. Prior to implantation, cell sheets and suspended cells were assessed for the expression of extracellular matrix constituents and molecules involved in angiogenesis and cardiac remodelling. The survival rate of rats receiving the cell sheets was significantly higher than after cell injections (73 vs. 41%, P = 0.01). This correlated with the absence of left ventricular (LV) remodelling in the cell sheet group, as end-diastolic volume only increased by 2.8% compared with baseline [95% confidence interval (CI): -18.7%; +30.0%, P = 0.81] vs. increases of 25.9% (-0.4%; +59.2%, P = 0.05) and 51.2% (+18.6%; +92.8, P = 0.001) in the cell and medium injection groups, respectively. Sheets also resulted in a greater cell engraftment possibly related to the greater expression of extracellular matrix constituents. The better preservation of LV geometry afforded by ADSC sheets is associated with increased survival and engraftment, which supports the concept of an epicardial delivery of cell-seeded biomaterials.


Translationally controlled tumour protein TCTP is induced early in human colorectal tumours and contributes to the resistance of HCT116 colon cancer cells to 5-FU and oxaliplatin.
Translationally controlled tumour protein TCTP is an anti-apoptotic protein frequently overexpressed in cancers, where high levels are often associated with poor patient outcome. TCTP may be involved in protecting cancer cells against the cytotoxic action of anti-cancer drugs. Here we study the early increase of TCTP levels in human colorectal cancer (CRC) and the regulation of TCTP expression in HCT116 colon cancer cells, in response to treatment with the anti-cancer drugs 5-FU and oxaliplatin. Using immunohistochemistry, we assessed TCTP levels in surgical samples from adenomas and adenocarcinomas of the colon, compared to normal colon tissue. We also studied the regulation of TCTP in HCT116 colon cancer cells in response to 5-FU and oxaliplatin by western blotting. TCTP mRNA levels were assessed by RT-qPCR. We used mTOR kinase inhibitors to demonstrate mTOR-dependent translational regulation of TCTP under these conditions. Employing the Real-Time Cell Analysis (RTCA) System and the MTS assay, we investigated the effect of TCTP-knockdown on the sensitivity of HCT116 cells to the anti-cancer drugs 5-FU and oxaliplatin. 1. TCTP levels are significantly increased in colon adenomas and adenocarcinomas, compared to normal colon tissue. 2. TCTP protein levels are about 4-fold upregulated in HCT116 colon cancer cells, in response to 5-FU and oxaliplatin treatment, whereas TCTP mRNA levels are down regulated. 3. mTOR kinase inhibitors prevented the up-regulation of TCTP protein, indicating that TCTP is translationally regulated through the mTOR complex 1 signalling pathway under these conditions. 4. Using two cellular assay systems, we demonstrated that TCTP-knockdown sensitises HCT116 cells to the cytotoxicity caused by 5-FU and oxaliplatin. Our results demonstrate that TCTP levels increase significantly in the early stages of CRC development. In colon cancer cells, expression of this protein is largely upregulated during treatment with the DNA-damaging anti-cancer drugs 5-FU and oxaliplatin, as part of the cellular stress response. TCTP may thus contribute to the development of anti-cancer drug resistance. These findings indicate that TCTP might be suitable as a biomarker and that combinatorial treatment using 5-FU/oxaliplatin, together with mTOR kinase inhibitors, could be a route to preventing the development of resistance to these drugs.


Improved Self-Acceptance, Quality of Life, and Stress Level from Participation in a Worksite Yoga Foundations Program: A Pilot Study.
Yoga is increasing in popularity in the United States and across the globe. However, most yoga programs are provided outside the worksite; although many companies offer worksite wellness programs, at present there is limited documentation regarding the potential benefits of participating in a worksite yoga program. Therefore, the purpose of this project was to examine the potential effect of a worksite yoga program on self-acceptance, quality of life, and perceived stress. A prospective cohort pilot study that examined a structured worksite yoga program was designed and tailored to individuals new to yoga. The 8-week Yoga Foundations program was conducted at an academic medical center's worksite wellness center with 86 subjects. Outcome measures were the 36-item Self-Acceptance Scale; a six-item quality-of-life measure that assesses overall, social, mental, physical, emotional, and spiritual well-being; and the ten-item Perceived Stress Scale. Participants demonstrated significant improvement in their overall self-acceptance ( p < 0.001), quality of life ( p < 0.001), and perceived stress ( p < 0.001) levels. They also highly rated the yoga instructors and the weekly format of the program. Participation in a Yoga Foundations program was associated with improvements in self-acceptance, quality of life and stress levels in worksite wellness center members. Future studies should use randomized designs and examine other wellness domains to learn more about the potential benefits of worksite yoga programs.


Specific induction of apoptosis by 1,8-cineole in two human leukemia cell lines, but not a in human stomach cancer cell line.
We have investigated the effects of 1,8-cineole [the main component of essential oil prepared from bay-leaves Laurus nobilis L.)] on DNA of human leukemia cell lines, Molt 4B, HL-60 and stomach cancer KATO III cells. Specific induction of apoptosis by 1,8-cineole was observed in human leukemia Molt 4B and HL-60 cells, but not in human stomach cancer KATO III cells. Morphological changes showing apoptotic bodies were observed in the human leukemia HL-60 cells treated with 1,8-cineole. The fragmentations of DNA by cineole to oligonucleosomal-sized fragments that is a characteristic of apoptosis were concentration- and time-dependent in Molt 4B and HL-60 cells, but not in KATO III cells. The present study shows that the suppression growth by 1,8-cineole in the leukemia cell lines results from the induction of apoptosis by this compound.


Genetic differences in human circadian clock genes among worldwide populations.
The daily biological clock regulates the timing of sleep and physiological processes that are of fundamental importance to human health, performance, and well-being. Environmental parameters of relevance to biological clocks include (1) daily fluctuations in light intensity and temperature, and (2) seasonal changes in photoperiod (day length) and temperature; these parameters vary dramatically as a function of latitude and locale. In wide-ranging species other than humans, natural selection has genetically optimized adaptiveness along latitudinal clines. Is there evidence for selection of clock gene alleles along latitudinal/photoperiod clines in humans? A number of polymorphisms in the human clock genes Per2, Per3, Clock, and AANAT have been reported as alleles that could be subject to selection. In addition, this investigation discovered several novel polymorphisms in the human Arntl and Arntl2 genes that may have functional impact upon the expression of these clock transcriptional factors. The frequency distribution of these clock gene polymorphisms is reported for diverse populations of African Americans, European Americans, Ghanaians, Han Chinese, and Papua New Guineans (including 5 subpopulations within Papua New Guinea). There are significant differences in the frequency distribution of clock gene alleles among these populations. Population genetic analyses indicate that these differences are likely to arise from genetic drift rather than from natural selection.


Autologous Bone Marrow Mononuclear Cells Exert Broad Effects on Short- and Long-Term Biological and Functional Outcomes in Rodents with Intracerebral Hemorrhage.
Autologous bone marrow-derived mononuclear cells (MNCs) are a potential therapy for ischemic stroke. However, the effect of MNCs in intracerebral hemorrhage (ICH) has not been fully studied. In this study, we investigated the effects of autologous MNCs in experimental ICH. ICH was induced by infusion of autologous blood into the left striatum in young and aged male Long Evans rats. Twenty-four hours after ICH, rats were randomized to receive an intravenous administration of autologous MNCs (1 × 10(7) cells/kg) or saline. We examined brain water content, various markers related to the integrity of the neurovascular unit and inflammation, neurological deficit, neuroregeneration, and brain atrophy. We found that MNC-treated young rats showed a reduction in the neurotrophil infiltration, the number of inducible nitric oxide synthase-positive cells, and the expression of inflammatory-related signalings such as the high-mobility group protein box-1, S100 calcium binding protein B, matrix metalloproteinase-9, and aquaporin 4. Ultimately, MNCs reduced brain edema in the perihematomal area compared with saline-treated animals at 3 days after ICH. Moreover, MNCs increased vessel density and migration of doublecortin-positive cells, improved motor functional recovery, spatial learning, and memory impairment, and reduced brain atrophy compared with saline-treated animals at 28 days after ICH. We also found that MNCs reduced brain edema and brain atrophy and improved spatial learning and memory in aged rats after ICH. We conclude that autologous MNCs can be safely harvested and intravenously reinfused in rodent ICH and may improve long-term structural and functional recovery after ICH. The results of this study may be applicable when considering future clinical trials testing MNCs for ICH.


The role of demography and markets in determining deforestation rates near Ranomafana National Park, Madagascar.
The highland forests of Madagascar are home to some of the world's most unique and diverse flora and fauna and to some of its poorest people. This juxtaposition of poverty and biodiversity is continually reinforced by rapid population growth, which results in increasing pressure on the remaining forest habitat in the highland region, and the biodiversity therein. Here we derive a mathematical expression for the subsistence of households to assess the role of markets and household demography on deforestation near Ranomafana National Park. In villages closest to urban rice markets, households were likely to clear less land than our model predicted, presumably because they were purchasing food at market. This effect was offset by the large number of migrant households who cleared significantly more land between 1989-2003 than did residents throughout the region. Deforestation by migrant households typically occurred after a mean time lag of 9 years. Analyses suggest that while local conservation efforts in Madagascar have been successful at reducing the footprint of individual households, large-scale conservation must rely on policies that can reduce the establishment of new households in remaining forested areas.


DENND2B activates Rab13 at the leading edge of migrating cells and promotes metastatic behavior.
The small guanosine triphosphatase Rab13 functions in exocytic vesicle trafficking in epithelial cells. Alterations in Rab13 activity have been observed in human cancers, yet the mechanism of Rab13 activation and its role in cancer progression remain unclear. In this paper, we identify the DENN domain protein DENND2B as the guanine nucleotide exchange factor for Rab13 and develop a novel Förster resonance energy transfer-based Rab biosensor to reveal activation of Rab13 by DENND2B at the leading edge of migrating cells. DENND2B interacts with the Rab13 effector MICAL-L2 at the cell periphery, and this interaction is required for the dynamic remodeling of the cell's leading edge. Disruption of Rab13-mediated trafficking dramatically limits the invasive behavior of epithelial cells in vitro and the growth and migration of highly invasive cancer cells in vivo. Thus, blocking Rab13 activation by DENND2B may provide a novel target to limit the spread of epithelial cancers.


Identification of major components of larviposition pheromone from larvae of tsetse fliesGlossina morsitans morsitans Westwood andGlossina morsitans centralis Machado.
The presence of pheromones produced by larvae ofGlossina morsitans morsitans andG. m. centralis, respectively, which attract gravid females and result in aggregation of pupae, is confirmed. Behavioral experiments indicated that females preferred to larviposit over moist sand conditioned by previously allowing larvae to pupariate in it. Similar results were obtained with filter papers contaminated with the prepupariation excretions of larvae and with volatiles collected from larvae prior to pupariation.n-Pentadecane andn-dodecane were identified as the dominant electrophysiologically active components of the larviposition pheromones ofG. m. morsitans andG. m. centralis, respectively, by GC-EAD and GC-MS analysis of the trapped larval volatiles. Both identified compounds were shown to significantly attract gravid females to larviposition sites in laboratory behavioral assays.


[Report of four clinical cases of filariasis in Alto Nanay, Loreto].
This study describes the clinical, parasitological and laboratory findings of four patients who lived in the district of Alto Nanay, Maynas, Loreto, Peru and were infected with Mansonella ozzardi microfilariae. Clinical examinations by ophthalmologists, cardiologists and nephrologists were performed. In case 1, the presence of 2 subcutaneous lumps was the most important finding, one at dorsal level and the other in the lower third of the left leg; in case 2, there was a sensation of tingling or coldness in the legs; in case 3, an associated chronic hepatitis B infection was found, and in case 4, a large lump was detected in the left lumbar region. All 4 patients were infected with Mansonella ozzardi and had eosinophilia, 3 of them in percentages of 20%. The lump found in case 4 was due to a herniation of bowel content. Further clinical studies and an evaluation of the actual pathogenic effect of microfilariae are to be performed. A study of the genetic diversity of filariae in the Peruvian Amazon would also be important.


Dehydroepiandrosterone suppresses the elevated hepatic glucose-6-phosphatase and fructose-1,6-bisphosphatase activities in C57BL/Ksj-db/db mice: comparison with troglitazone.
The effect of dehydroepiandrosterone (DHEA) on the hepatic and muscle glucose metabolizing enzymes and on blood glucose were investigated in insulin-resistant diabetic C57BL/KsJ-db/db mice and their heterozygote littermates (db/+m). The results were compared with those after troglitazone administration under the same conditions. Despite hyperinsulinemia, hepatic glucose-6-phosphatase (G6Pase) and fructose-1,6-bisphosphatase (FBPase) activities are higher in db/db than in db/+m mice. Dietary administration of DHEA and that of troglitazone for 15 days to respective groups of five mice each significantly decreased blood glucose in db/db mice and hepatic G6Pase and FBPase activities in both db/db and db/+m mice. Hepatic G6Pase and FBPase activities showed a linear relationship with blood glucose in all groups of mice, suggesting that the activities of G6Pase and FBPase are closely related to blood glucose levels. Because androstenedione, a DHEA metabolite, barely affected either of these enzyme activities or blood glucose in db/db mice, the actions of DHEA, which are similar to those of troglitazone, are presumed to be caused by DHEA itself. DHEA is considered to be a modulating agent for the activities of hepatic gluconeogenic enzymes in db/db mice.


Molecular docking and 3D-QSAR studies on beta-phenylalanine derivatives as dipeptidyl peptidase IV inhibitors.
Three-dimensional quantitative structure-activity relationship (3D-QSAR) and molecular docking studies were carried out to explore the binding of 73 inhibitors to dipeptidyl peptidase IV (DPP-IV), and to construct highly predictive 3D-QSAR models using comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA). The negative logarithm of IC(50) (pIC(50)) was used as the biological activity in the 3D-QSAR study. The CoMFA model was developed by steric and electrostatic field methods, and leave-one-out cross-validated partial least squares analysis yielded a cross-validated value (r(2)(cv)) of 0.759. Three CoMSIA models developed by different combinations of steric, electrostatic, hydrophobic and hydrogen-bond fields yielded significant r(2)(cv) values of 0.750, 0.708 and 0.694, respectively. The CoMFA and CoMSIA models were validated by a structurally diversified test set of 18 compounds. All of the test compounds were predicted accurately using these models. The mean and standard deviation of prediction errors were within 0.33 and 0.26 for all models. Analysis of CoMFA and CoMSIA contour maps helped identify the structural requirements of inhibitors, with implications for the design of the next generation of DPP-IV inhibitors for the treatment of type 2 diabetes.


A Phase II Exploratory Study of PXD-101 (Belinostat) Followed by Zevalin in Patients with Relapsed Aggressive High-Risk Lymphoma.
Aggressive lymphomas (aNHL) including diffuse large B-cell lymphoma (DLBCL) have poor outcomes in relapsed refractory patients. Prior studies have demonstrated that loss of major histocompatibility complex class II (MHCII) expression in DLBCL is associated with poor survival. The objective of this single-arm phase II study was to evaluate if PXD-101 would increase MHCII expression, synergize with Zevalin, and improve clinical outcomes. This was a single-center open-label phase II trial (NCT01686165) geared toward heavily pretreated patients with CD20-positive aNHL. The primary endpoint was overall response rate (ORR) in aNHL patients treated with 2 cycles of PXD-101 followed by restaging CT and 1 cycle of Zevalin. Five patients were enrolled, and all were heavily pretreated. Therapy was well tolerated, with nausea and vomiting being the most frequent adverse events. All patients progressed after receiving therapy; the study did not achieve the required ORR to proceed to the next stage. The pleotropic effects of histone deacetylase inhibition and lack of clinical biomarkers have precluded a priori identification of responding patients. Thus, while we report a negative trial of PXD-101 in combination with Zevalin, this study highlights the importance of a clinically feasible biomarker.


Resurfacing and reconstruction of the glans penis.
To describe the techniques and results of surgical reconstruction of glans penis lesions. Seventeen patients (mean age: 53.2 yr) were treated by resurfacing or reconstruction of the glans penis for benign, premalignant and malignant penile lesions. The aetiology of the lesions was one Zoon's balanitis, four lichen sclerosus, one carcinoma in situ, five squamous cell carcinomas, and six squamous cell carcinomas associated with lichen sclerosus. Five cases were treated by glans skinning and resurfacing; five cases by glans amputation and reconstruction of the neoglans, and seven cases by partial penile amputation and reconstruction of the neoglans. Glans resurfacing and reconstruction were performed with the use of a skin graft harvested from the thigh. The mean follow-up was 32 mo. All patients were free of local premalignant/malignant recurrence. Patients who underwent glans resurfacing reported glandular sensory restoration and complete sexual ability. Patients who underwent glansectomy or partial penectomy with neoglans reconstruction maintained sexual function and activity, although sensitivity was reduced as a consequence of glans/penile amputation. In selected cases of benign, premalignant or malignant penile lesions, glans resurfacing or reconstruction can ensure a normal appearing and functional penis, without jeopardizing cancer control.


Members of the Hsp70 Family Recognize Distinct Types of Sequences to Execute ER Quality Control.
Protein maturation in the endoplasmic reticulum is controlled by multiple chaperones, but how they recognize and determine the fate of their clients remains unclear. We developed an in vivo peptide library covering substrates of the ER Hsp70 system: BiP, Grp170, and three of BiP's DnaJ-family co-factors (ERdj3, ERdj4, and ERdj5). In vivo binding studies revealed that sites for pro-folding chaperones BiP and ERdj3 were frequent and dispersed throughout the clients, whereas Grp170, ERdj4, and ERdj5 specifically recognized a distinct type of rarer sequence with a high predicted aggregation potential. Mutational analyses provided insights into sequence recognition characteristics for these pro-degradation chaperones, which could be readily introduced or disrupted, allowing the consequences for client fates to be determined. Our data reveal unanticipated diversity in recognition sequences for chaperones; establish a sequence-encoded interplay between protein folding, aggregation, and degradation; and highlight the ability of clients to co-evolve with chaperones, ensuring quality control.


Advanced Uses of Pulse Oximetry for Monitoring Mechanically Ventilated Patients.
Pulse oximetry is an undisputable standard of care in clinical monitoring. It combines a spectrometer to detect hypoxemia with a plethysmograph for the diagnosis, monitoring, and follow-up of cardiovascular diseases. These pulse oximetry capabilities are extremely useful for assessing the respiratory and circulatory status and for monitoring of mechanically ventilated patients. On the one hand, the key spectrography-derived function of pulse oximetry is to evaluate a patient's gas exchange that results from a particular ventilatory treatment by continuously and noninvasively measuring arterial hemoglobin saturation (SpO2). This information helps to maintain patients above the hypoxemic levels, leading to appropriate ventilator settings and inspired oxygen fractions. However, whenever higher than normal oxygen fractions are used, SpO2 can mask existing oxygenation defects in ventilated patients. This limitation, resulting from the S shape of the oxyhemoglobin saturation curve, can be overcome by reducing the oxygen fraction delivered to the patient in a controlled and stepwise manner. This results in a SpO2/FIO2 diagram, which allows a rough characterization of a patient's gas exchange, shunt, and the amount of lung area with a low ventilation/perfusion ratio without the need of blood sampling. On the other hand, the photoplethysmography-derived oximeter function has barely been exploited for the purpose of monitoring hemodynamics in mechanically ventilated patients. The analysis of the photoplethysmography contour provides useful real-time and noninvasive information about the interaction of heart and lungs during positive pressure ventilation. These hemodynamic monitoring capabilities are related to both the assessment of preload dependency-mainly by analyzing the breath-by-breath variation of the photoplethysmographic signals-and the analysis of arterial impedance, which examines the changes in the plethysmographic amplitude, contour, and derived indexes. In this article, we present and describe these extended monitoring capabilities and propose a more holistic monitoring concept that takes advantage of these advanced uses of pulse oximetry in the monitoring of ventilated patients. Today's monitors need to be improved if such novel functionalities were to be offered for clinical use. Future developments and clinical evaluations are needed to establish the true potential of these advanced monitoring uses of pulse oximetry.


Preoperative autologous blood donation in adult bone marrow donors: reappraisal of a single-centre experience.
To avoid risk for allogeneic transfusions in healthy bone marrow (BM) donors, 1-2 preoperative autologous blood donations (PAD) are usually collected before the BM harvest. We analysed the haematological parameters in BM donors before and after the harvest, to assess the efficacy of this practice in limiting the postharvest anaemia. Overall, 102 consecutive donors underwent BM harvest preceded by one (26 cases) or two PAD (76 cases), which were infused during BM collection. We analysed the parameters related to donors, PAD timing and BM graft characteristics. PAD induced a significant decrease in Hb (from 14·6 g/dl, IQ range 13·3-15·5 to12·9 g/dl, IQ range 11·8-13·9; P < 0·0001) in all donors, with a median Hb loss at day -1 of 10·9% (IQ range 6·8-14·2). The PAD-related Hb decrease was independent of sex or number of PAD, and was inversely related to the time elapsed from first or last PAD. In comparison with values recorded at day-1, BM harvest produced an additional Hb decrease, accounting for a median Hb loss of 18·9% (IQ range 14·9-24·4). Overall, in comparison with pre-PAD values, Hb levels at day +1 were reduced of 28·9% (IQ range 23·6-32·2), independently if donors had 1 or 2 PAD reinfused. In conclusion, these data show that two PAD do not carry any advantage over one PAD. An eventual benefit of PAD can be achieved only if an adequate interval between PAD and BM harvest elapses. Prospective randomized studies could be worth to establish if any role for PAD does exist in BM donors.


Reduced diversity of faecal microbiota in Crohn's disease revealed by a metagenomic approach.
A role for the intestinal microbial community (microbiota) in the onset and chronicity of Crohn's disease (CD) is strongly suspected. However, investigation of such a complex ecosystem is difficult, even with culture independent molecular approaches. We used, for the first time, a comprehensive metagenomic approach to investigate the full range of intestinal microbial diversity. We used a fosmid vector to construct two libraries of genomic DNA isolated directly from faecal samples of six healthy donors and six patients with CD. Bacterial diversity was analysed by screening the two DNA libraries, each composed of 25,000 clones, for the 16S rRNA gene by DNA hybridisation. Among 1190 selected clones, we identified 125 non-redundant ribotypes mainly represented by the phyla Bacteroidetes and Firmicutes. Among the Firmicutes, 43 distinct ribotypes were identified in the healthy microbiota, compared with only 13 in CD (p<0.025). Fluorescent in situ hybridisation directly targeting 16S rRNA in faecal samples analysed individually (n=12) confirmed the significant reduction in the proportion of bacteria belonging to this phylum in CD patients (p<0.02). The metagenomic approach allowed us to detect a reduced complexity of the bacterial phylum Firmicutes as a signature of the faecal microbiota in patients with CD. It also indicated the presence of new bacterial species.


Designing and pilot testing a novel high-definition transcranial burst electrostimulation device for neurorehabilitation.
Objective.Non-invasive brain stimulation has been promoted to facilitate neuromodulation in treating neurological diseases. Recently, high-definition (HD) transcranial electrical stimulation and a novel electrical waveform combining a direct current (DC) and theta burst stimulation (TBS)-like protocol were proposed and demonstrated high potential to enhance neuroplastic effects in a more-efficient manner. In this study, we designed a novel HD transcranial burst electrostimulation device and to preliminarily examined its therapeutic potential in neurorehabilitation.Approach.A prototype of the transcranial burst electrostimulation device was developed, which can flexibly output a waveform that combined a DC and TBS-like protocol and can equally distribute the current into 4 × 1 HD electrical stimulation by automatic impedance adjustments. The safety and accuracy of the device were then validated in a series ofin vitroexperiments. Finally, a pilot clinical trial was conducted to assess its clinical safety and therapeutic potential on upper-extremity rehabilitation in six patients with chronic stroke, where patients received either active or sham HD transcranial burst electrostimulation combined with occupational therapy three times per week for four weeks.Main results.The prototype was tested, and it was found to comply with all safety requirements. The output parameters were accurate and met the clinical study needs. The pilot clinical study demonstrated that the active HD transcranial burst electrostimulation group had greater improvement in voluntary motor function and coordination of the upper extremity than the sham control group. Additionally, no severe adverse events were noted, but slight skin redness under the stimulus electrode immediately after stimulation was seen.Conclusions.The results demonstrated the feasibility of incorporating the HD electrical DC and TBS-like protocol in our device; and the novel neuromodulatory device produced positive neurorehabilitation outcomes in a safe fashion, which could be the basis for the future clinical implementation for treating neurological diseases.Trial registration:ClinicalTrials.gov Identifier: NCT04278105. Registered on 20 February 2020.


Characterization of GH-1 mutations in children with isolated growth hormone deficiency in the Turkish population.
Isolated growth hormone deficiency (IGHD) is defined as a medical condition associated with growth failure due to insufficient production of growth hormone (GH) or lack of growth hormone action. It occurs with an incidence of between 1/4,000 and 1/10,000 live births. Most cases are sporadic and idiopathic but 5-30% of growth hormone deficiency (GHD) has genetic etiology. Mutations in the GH encoding gene (GH-1) have been detected in patients with IGHD. The purpose of this study was to characterize mutations of the GH-1 gene in children with IGHD in the Turkish population. We found four missense mutations (E33G, N47D, T-24A and A13S), one nonsense mutation (W-7X), one insertion and two deletions in nine patients out of seventy-five patients with IGHD. The missense mutation A13S, GAAA insertion at intron 1 (+178A), and the deletions of +83C in intron 1 and deltaF166 in exon 5 are novel mutations.


Functional Outcome of Intravenous Thrombolysis in Patients With Lacunar Infarcts in the WAKE-UP Trial.
The rationale for intravenous thrombolysis in patients with lacunar infarcts is debated, since it is hypothesized that the microvascular occlusion underlying lacunar infarcts might not be susceptible to pharmacological reperfusion treatment. To study the efficacy and safety of intravenous thrombolysis among patients with lacunar infarcts. This exploratory secondary post hoc analysis of the WAKE-UP trial included patients who were screened and enrolled between September 2012 and June 2017 (with final follow-up in September 2017). The WAKE-UP trial was a multicenter, double-blind, placebo-controlled randomized clinical trial to study the efficacy and safety of intravenous thrombolysis with alteplase in patients with an acute stroke of unknown onset time, guided by magnetic resonance imaging. All 503 patients randomized in the WAKE-UP trial were reviewed for lacunar infarcts. Diagnosis of lacunar infarcts was based on magnetic resonance imaging and made by consensus of 2 independent investigators blinded to clinical information. The primary efficacy variable was favorable outcome defined by a score of 0 to 1 on the modified Rankin Scale at 90 days after stroke, adjusted for age and severity of symptoms. Of the 503 patients randomized in the WAKE-UP trial, 108 patients (including 74 men [68.5%]) had imaging-defined lacunar infarcts, whereas 395 patients (including 251 men [63.5%]) had nonlacunar infarcts. Patients with lacunar infarcts were younger than patients with nonlacunar infarcts (mean age [SD], 63 [12] years vs 66 [12] years; P = .003). Of patients with lacunar infarcts, 55 (50.9%) were assigned to treatment with alteplase and 53 (49.1%) to receive placebo. Treatment with alteplase was associated with higher odds of favorable outcome, with no heterogeneity of treatment outcome between lacunar and nonlacunar stroke subtypes. In patients with lacunar strokes, a favorable outcome was observed in 31 of 53 patients (59%) in the alteplase group compared with 24 of 52 patients (46%) in the placebo group (adjusted odds ratio [aOR], 1.67 [95% CI, 0.77-3.64]). There was 1 death and 1 symptomatic intracranial hemorrhage according to Safe Implementation of Thrombolysis in Stroke-Monitoring Study criteria in the alteplase group, while no death and no symptomatic intracranial hemorrhage occurred in the placebo group. The distribution of the modified Rankin Scale scores 90 days after stroke also showed a nonsignificant shift toward better outcomes in patients with lacunar infarcts treated with alteplase, with an adjusted common odds ratio of 1.94 (95% CI, 0.95-3.93). While the WAKE-UP trial was not powered to demonstrate the efficacy of treatment in subgroups of patients, the results indicate that the association of intravenous alteplase with functional outcome does not differ in patients with imaging-defined lacunar infarcts compared with those experiencing other stroke subtypes.


[New therapeutic options in chronic hepatitis C virus infection].
Chronic hepatitis C virus (HCV) infection is the major etiology and the reason of chronic liver disease, liver cirrhosis, hepatic decompensation, hepatocellular cancer and liver transplantation. Less than half of patients with HCV-related chronic hepatitis achieve sustained viral clearance with current pegylated interferon and ribavirin (P+R) combination therapy. Due to the insufficient treatment success, an extended search for new, direct acting anti-HCV agents (DAAs) is ongoing, already leading to submissions of applications for marketing authorization of the protease-inhibitors boceprevir and telaprevir. Both are effective only in triple combinations with P+R. Studies demonstrate a 50% success rate advantage for triple therapies above current standards. In addition, treatment duration can be shortened, and half of the patients who failed previous therapy with P+R can be cured with triple therapies. A major concern with new DAAs is rapid development of DAA-resistant viral mutants, a reason as well as a consequence of insufficient triple therapy. Clinical studies with boceprevir and telaprevir are reviewed in this paper.