The ortho-plastic management of Gustilo grade IIIB fractures of the tibia in children: a systematic review of the literature.
The challenges of managing Gustilo IIIB tibial fractures in children are unique. A multi-disciplinary, evidence based approach is needed. We aimed to evaluate the evidence for the ortho-plastic management of Gustilo grade IIIB open tibial shaft fractures in children based on a review of all published data in order to rationalise the orthopaedic and plastic surgical approach to these complex injuries. A systematic review of the literature was performed. Gustilo grade IIIB tibial shaft fractures in pre-adolescent and adolescent children were identified and evaluated with regard to both the skeletal and soft tissue management, and the outcome. Of 54 children with grade IIIB tibial fractures, a mean union time of 31 weeks included 33 weeks for 42 adolescents and 23 weeks for 12 pre-adolescents. Faster union time in pre-adolescents tended towards significance. Delayed union occurred in 22%, nonunion in 13%, mostly in adolescents. Two of 45 covered by vascularised flaps and 3 of 9 treated without flaps developed deep infection (p=0.028). There was no correlation between method of skeletal fixation and union time. Gustilo IIIB tibial shaft fractures in pre-adolescents tended towards faster healing with fewer complications, irrespective of the method of skeletal fixation. In adolescents, healing times were similar to adults. Soft tissue closure without flaps was associated with deep infection in one-third of patients, requiring debridement and flap cover. Adequate debridement and flap cover is suggested in all cases, irrespective of age.


The diagnostic yield and safety of ultrasound-assisted transthoracic fine-needle aspiration of drowned lung.
Proximal lung tumors, though not discernable by means of transthoracic ultrasound (US), may cause varying degrees of pulmonary collapse and postobstructive pneumonitis which may give rise to a 'drowned lung' appearance on chest computed tomography (CT) and US. The diagnostic yield for malignancy of US-assisted transthoracic fine-needle aspiration (FNA) of these areas of drowned lung is unknown. We aimed to explore the feasibility of US-assisted FNA in this setting by prospectively investigating its diagnostic yield and safety. We enrolled 31 patients (aged 59.4 ± 9.7 years, 17 males) with central tumors and secondary drowned lung on CT scan. A respiratory physician performed transthoracic US to identify the target drowned lung tissue. Three US-assisted superficial FNA passes (≤20 mm from the pleura) were followed by 3 deeper FNA passes (>20 mm) aimed in the direction of a visible or approximated central mass. Rapid on-site evaluation of specimens was used. Superficial FNA was diagnostic in 11 patients (35.5%), whereas deeper FNA was diagnostic in 23 patients (74.2%, p = 0.002). Deeper FNA confirmed malignancy in all cases with diagnostic superficial FNA. We observed no pneumothoraces or major hemorrhage. All patients were ultimately diagnosed with malignancy (bronchogenic carcinoma, n = 30; lymphoma, n = 1). US-assisted FNA of drowned lung has an acceptable diagnostic yield and is safe.


Development of monocytes, macrophages, and dendritic cells.
Monocytes and macrophages are critical effectors and regulators of inflammation and the innate immune response, the immediate arm of the immune system. Dendritic cells initiate and regulate the highly pathogen-specific adaptive immune responses and are central to the development of immunologic memory and tolerance. Recent in vivo experimental approaches in the mouse have unveiled new aspects of the developmental and lineage relationships among these cell populations. Despite this, the origin and differentiation cues for many tissue macrophages, monocytes, and dendritic cell subsets in mice, and the corresponding cell populations in humans, remain to be elucidated.


Injuries caused by aquatic animals in Brazil: an analysis of the data present in the information system for notifiable diseases.
We present a review of injuries in humans caused by aquatic animals in Brazil using the Information System for Notifiable Diseases [ Sistema de Informação de Agravos de Notificação (SINAN)] database. A descriptive and retrospective epidemiological study was conducted from 2007 to 2013. A total of 4,118 accidents were recorded. Of these accidents, 88.7% (3,651) were caused by venomous species, and 11.3% (467) were caused by poisonous, traumatic or unidentified aquatic animals. Most of the events were injuries by stingrays (69%) and jellyfish (13.1%). The North region was responsible for the majority of reports (66.2%), with a significant emphasis on accidents caused by freshwater stingrays (92.2% or 2,317 cases). In the South region, the region with the second highest number of records (15.7%), jellyfish caused the majority of accidents (83.7% or 452 cases). The Northeastern region, with 12.5% of the records, was notable because almost all accidents were caused by toadfish (95.6% or 174 cases). Although a comparison of different databases has not been performed, the data presented in this study, compared to local and regional surveys, raises the hypothesis of underreporting of accidents. As the SINAN is the official system for the notification of accidents by venomous animals in Brazil, it is imperative that its operation be reviewed and improved, given that effective measures to prevent accidents by venomous animals depend on a reliable database and the ability to accurately report the true conditions.


Heteronormativity and sexual partnering among bisexual Latino men.
Our analyses address the question of how bisexual Latino men organize their sexual partnerships. Heteronormativity can be understood as the set of social norms and normative structures that guide sexual partnering among men and women. We provide descriptive statistics to describe bisexual Latino men's sexual partnerships. Logistic and linear regression modeling were used to explore bivariate and multivariate relationships. Of our total sample (N = 142), 41.6 % had unprotected vaginal intercourse 2 months prior to the interview; 21.8 % had unprotected anal intercourse with female partners; 37.5 % had unprotected insertive anal intercourse with male partners; and 22.5 % had unprotected receptive anal intercourse with male partners. In our multivariate model, machismo was directly associated with meeting female partners through formal spaces (workplace, school, and/or church), but inversely associated with meeting male partners in formal spaces. Machismo was positively associated with meeting male sex partners through social networks (i.e., friendship and kinship networks). The more comfortable men were with homosexuality the less likely they were to meet men online and the more likely they were to meet men through social networks of friends and kinship. Interventions to reduce sexually transmitted diseases that target bisexual behavior as an epidemiological "bridge" of transmission from homosexual to heterosexual networks might very well benefit from a more complex understanding of how Latino bisexuality is patterned. Thus, this exploratory analysis might lead to a rethinking of how to address risk and vulnerability among Latino bisexual men and their sexual networks.


Novel angiotensin I-converting enzyme inhibitory peptides derived from an edible mushroom, Pleurotus cystidiosus O.K. Miller identified by LC-MS/MS.
Angiotensin I-converting enzyme (ACE) inhibitors have been reported to reduce mortality in patients with hypertension. Compared to chemosynthetic drugs, ACE inhibitors derived from natural sources such as food proteins are believed to be safer for consumption and to have fewer adverse effects. Some edible mushrooms have been reported to significantly reduce blood pressure after oral administration. In addition, mushrooms are known to be rich in protein content. This makes them a potential source of ACE inhibitory peptides. Hence, the objective of the current study was to isolate and characterise ACE inhibitory peptides from an edible mushroom, Pleurotus cystidiosus. ACE inhibitory proteins were isolated from P. cystidiosus based on the bioassay guided purification steps, i.e. ammonium sulphate precipitation, reverse phase high performance liquid chromatography and size exclusion chromatography. Active fraction was then analysed by LC-MS/MS and potential ACE inhibitory peptides identified were chemically synthesized. Effect of in vitro gastrointestinal digestions on the ACE inhibitory activity of the peptides and their inhibition patterns were evaluated. Two potential ACE inhibitory peptides, AHEPVK and GPSMR were identified from P. cystidiosus with molecular masses of 679.53 and 546.36 Da, respectively. Both peptides exhibited potentially high ACE inhibitory activity with IC50 values of 62.8 and 277.5 μM, respectively. SEC chromatograms and BIOPEP analysis of these peptides revealed that the peptide sequence of the hexapeptide, AHEPVK, was stable throughout gastrointestinal digestion. The pentapeptide, GPSMR, was hydrolysed after digestion and it was predicted to release a dipeptide ACE inhibitor, GP, from its precursor. The Lineweaver-Burk plot of AHEPVK showed that this potent and stable ACE inhibitor has a competitive inhibitory effect against ACE. The present study indicated that the peptides from P. cystidiosus could be potential ACE inhibitors. Although these peptides had lower ACE inhibitory activity compared to commercial antihypertensive drugs, they are derived from mushroom which could be easily obtained and should have no side effects. Further in vivo studies can be carried out to reveal the clear mechanism of ACE inhibition by these peptides.


February precipitation in the wintering grounds of the lesser whitethroat, Sylvia curruca: is it a cue for migration onset?
Numerous studies report shifts in bird migration phenology, presumably owing to global warming. However, most studies focus on migration patterns in the Northern Hemisphere. In this study, we investigated associations between weather conditions in African wintering grounds of the lesser whitethroat, Sylvia curruca, and spring arrival time in Eilat, Israel. Using multivariate regression models, we analysed a 30-year dataset in order to examine correlations between median springtime arrival and 46 climate variables of the wintering quarters. The model obtained exhibited a highly statistical fit, involving mean precipitation in February and March with negative effects and number of wet days during November-February. February precipitation levels were also the major factor associated with the interquartile range of arrival time. Interestingly and contrary to published results, annual or seasonal precipitation showed no correlation with spring arrival time, nor did temperature. Moreover, winter in this region falls into dry season with negligible rainfall quantities. Hence, it is unlikely that precipitation effect on habitat productivity is a driving force of migration, as suggested by other studies. Instead, we propose that precipitation in February acts as a cue for the birds, indicating the approach of spring and migration time.


Differentiation of regulatory Foxp3+ T cells in the thymic cortex.
Regulatory Foxp3(+) T cells (T(R)) are indispensable for preventing autoimmune pathology in multiple organs and tissues. During thymic differentiation T cell receptor (TCR)-ligand interactions within a certain increased affinity range, in conjunction with gammac-containing cytokine receptor signals, induce Foxp3 expression and thereby commit developing thymocytes to the T(R) lineage. The contribution of distinct MHC class II-expressing accessory cell types to the differentiation process of Foxp3(+) thymocytes remains controversial, because a unique role in this process has been ascribed to either thymic dendritic cells (tDC) or to medullary thymic epithelial cells (mTEC). Furthermore, it was suggested that the thymic medulla, where the bulk of the negative selection of self-reactive thymocytes takes place, provides a specialized microenvironment supporting T(R) differentiation. Here, we report that the cortex, as defined by cortical thymic epithelial cells (cTEC), is sufficient for supporting T(R) differentiation. MHC class II expression restricted to both cTEC and mTEC or to cTEC alone did not significantly affect the numbers of Foxp3(+) thymocytes. Furthermore, genetic or pharmacologic blockade of thymocyte migration resulted in a prominent accumulation of Foxp3(+) thymocytes in the cortex, demonstrating that secondary signals required for Foxp3 up-regulation exist in the cortex. Our results suggest that mTEC or tDC do not serve as a cell type singularly responsible for T(R) differentiation and that neither the cortex nor the medulla exclusively provides an environment suitable for Foxp3 induction. Instead, multiple accessory cell types probably contribute to the thymic generation of regulatory Foxp3(+) T cells.


Comparison of Methods for Assessing Resistance to Meloidogyne arenaria in Peanut.
Use of resistant cultivars is a desirable approach to manage the peanut root-knot nematode (Meloidogyne arenaria). To incorporate resistance into commercially acceptable cultivars requires reliable, efficient screening methods. To optimize the resistance screening protocol, a series of greenhouse tests were done using seven genotypes with three levels of resistance to M. arenaria. The three resistance levels could be separated based on gall indices as early as two weeks after inoculation (WAI) using 8,000 eggs of M. arenaria per plant, while four or more weeks were needed when 1,000-6,000 eggs/plant were used. High inoculum densities (over 8,000 eggs/plant) were needed to separate the three resistance levels based on eggs per gram of root within eight WAI. A gall index based on percentage of galled roots could separate the three resistance levels at lower inoculum levels and earlier harvest dates than other assessment methods. The use of eggs vs. second-stage juveniles (J2) as inoculum provided similar results; however, it took three to five more days to collect J2 than to collect eggs from roots. Plant age affected gall index and nematode reproduction on peanut, especially on the susceptible genotypes AT201 and D098. The genotypes were separated into their correct resistance classes when inoculated 10 to 30 days after planting, but were not separated correctly when inoculated on day 40.


STATE INSTITUTION «NATIONAL RESEARCH CENTER FOR RADIATION MEDICINE OF THE NATIONAL ACADEMY OF MEDICAL SCIENCES OF UKRAINE» - RESEARCH ACTIVITIES AND SCIENTIFIC ADVANCE IN 2019.
Research activities and scientific advance achieved in 2019 at the State Institution «National Research Center forRadiation Medicine of the National Academy of Medical Sciences of Ukraine» (NRCRM) concerning medical problemsof the Chornobyl disaster, radiation medicine, radiobiology, radiation hygiene and epidemiology in collaborationwith the WHO network of medical preparedness and assistance in radiation accidents are outlined in the annualreport. The report presents the results of fundamental and applied research works of the study of radiation effectsand health effects of the Chornobyl accident.The report also shows the results of scientific-organizational and health care work, staff training.The Scientific Council meeting of NAMS approved the NRCRM Annual Report.


Intrabacterial proton-dependent CagA transport system in Helicobacter pylori.
Helicobacter pylori CagA modifies the signalling of host cells and causes gastric diseases. Although CagA is injected into gastric epithelial cells through the type IV secretion machinery, it remains unclear how CagA is transported towards the machinery in the bacterial cytoplasm. In this study, it was determined that the proton-dependent intracytoplasmic transport system correlates with the priming of CagA secretion from H. pylori. The cytotoxicity of neutral-pH- and acidic-pH-treated H. pylori was examined in the AGS cell line. The amount of phosphorylated CagA in AGS cells incubated with acidic-pH- and neutral-pH-treated H. pylori was determined by enzyme immunoassay and Western blot. The production of CagA and adherence of the treated bacteria were examined by enzyme immunoassay and light microscopy, respectively. To clarify how CagA is transported towards the inner membrane of the treated bacteria, the localization of CagA was analysed by immunoelectron microscopy. The proportion of hummingbird cells in the AGS cell line rapidly increased following the inoculation of acidic-pH-treated H. pylori but increased more slowly with neutral-pH-treated H. pylori, and the phenomenon correlated with the amount of phosphorylated CagA in AGS cells. CagA was densely localized near the inner membrane in the acidic-pH-treated bacterial cytoplasm, but this localization was not observed in the neutral-pH-treated bacterial cytoplasm, suggesting that CagA shifts from the centre to the peripheral portion of the cytoplasm as a result of an extracellular decrease in pH. This phenomenon depended on the presence of UreI, a proton-dependent urea channel, but not on the presence of urea. The pH treatments did not enhance CagA production or the adherence of the bacterium to AGS cells. The authors propose that H. pylori possesses a proton-dependent intracytoplasmic transport system that probably accelerates priming for CagA injection.


Traditional and non-traditional strategies to optimize catheter function: go with more flow.
In the United States, over 340,000 patients have end-stage renal disease treated by hemodialysis (HD) and are dependent on a reliable vascular access. In over 80% of patients initiating HD, this access is the central venous catheter (CVC). Although the CVC has many advantages that make it desirable for dialysis initiation-ease of insertion, unnecessary maturation time, and availability for immediate use-it is not without significant disadvantages. The substantial morbidity and mortality associated with CVC use has been well documented in the literature. Initiating and maintaining HD patients using a CVC is suboptimal from the perspective of both patient care and associated long-term costs. Yet, in the United States, the most common HD access-related event is replacement of any vascular access type with a CVC. Although in recent years greater effort has be made to reduce CVC use, some patients are unable to have a functioning arteriovenous fistula or graft created due to exhaustion of vessels from previous permanent accesses or limiting comorbidities. In patients dependent on long-term CVC use, the primary problems are due to malfunction ('poor flows') or infection. Catheter malfunction leads to inadequate dialysis, the need for costly and inconvenient intervention, and reduced quality of life. This review will focus on the etiology, prevention, and management of CVC-related malfunction.


Heparan sulfate proteoglycans mediate interstitial flow mechanotransduction regulating MMP-13 expression and cell motility via FAK-ERK in 3D collagen.
Interstitial flow directly affects cells that reside in tissues and regulates tissue physiology and pathology by modulating important cellular processes including proliferation, differentiation, and migration. However, the structures that cells utilize to sense interstitial flow in a 3-dimensional (3D) environment have not yet been elucidated. Previously, we have shown that interstitial flow upregulates matrix metalloproteinase (MMP) expression in rat vascular smooth muscle cells (SMCs) and fibroblasts/myofibroblasts via activation of an ERK1/2-c-Jun pathway, which in turn promotes cell migration in collagen. Herein, we focused on uncovering the flow-induced mechanotransduction mechanism in 3D. Cleavage of rat vascular SMC surface glycocalyx heparan sulfate (HS) chains from proteoglycan (PG) core proteins by heparinase or disruption of HS biosynthesis by silencing N-deacetylase/N-sulfotransferase 1 (NDST1) suppressed interstitial flow-induced ERK1/2 activation, interstitial collagenase (MMP-13) expression, and SMC motility in 3D collagen. Inhibition or knockdown of focal adhesion kinase (FAK) also attenuated or blocked flow-induced ERK1/2 activation, MMP-13 expression, and cell motility. Interstitial flow induced FAK phosphorylation at Tyr925, and this activation was blocked when heparan sulfate proteoglycans (HSPGs) were disrupted. These data suggest that HSPGs mediate interstitial flow-induced mechanotransduction through FAK-ERK. In addition, we show that integrins are crucial for mechanotransduction through HSPGs as they mediate cell spreading and maintain cytoskeletal rigidity. We propose a conceptual mechanotransduction model wherein cell surface glycocalyx HSPGs, in the presence of integrin-mediated cell-matrix adhesions and cytoskeleton organization, sense interstitial flow and activate the FAK-ERK signaling axis, leading to upregulation of MMP expression and cell motility in 3D. This is the first study to describe a flow-induced mechanotransduction mechanism via HSPG-mediated FAK activation in 3D. This study will be of interest in understanding the flow-related mechanobiology in vascular lesion formation, tissue morphogenesis, cancer cell metastasis, and stem cell differentiation in 3D, and also has implications in tissue engineering.


Genetics and cardiovascular disease: Design and development of a DNA biobank.
Coronary artery disease (CAD) is a complex disease with environmental and genetic determinants. Many other cardiovascular (CV) conditions also have a genetic basis. A positive family history of CV disease in first-degree relatives is a strong independent risk factor for CAD as well as several other cardiac disorders. This genetic susceptibility to CV diseases will be understood more clearly when combined with genomics, proteomics and genotyping.The Department of Cardiology at Gold Coast Hospital (Queensland, Australia) with the Faculty of Health, Science and Medicine at Bond University (Queensland, Australia) established the Gold Coast Cardiovascular DNA bank in 2006. The dataset on each individual volunteer includes coronary angiograms, clinical information (including a coronary risk factor profile), biochemical (including cardiac biomarkers) and hematological parameters, and electrocardiograms and echocardiograms. The establishment of the DNA biobank was associated with several key challenges, both technical and logistic.Given the comprehensive nature of the information gathered, the present study has the added potential of identifying genes associated with nonischemic cardiomyopathies, valvular heart disease, congenital heart diseases and other cardiomyopathies. Pooling data from results obtained here with multiple existing DNA biobanks and registries will help in finding answers to the genetic conundrum in CV diseases. The present DNA biobank will serve as a resource well into the future as the technology and science of medical genetics evolve.The most frequent pathology encountered in the biobank is CAD. The significance of the familial occurrence of CAD has been the focus of research for at least 50 years, with a positive family history of CV disease emerging as an independent predictor of risk in the development of CAD. By applying the knowledge learned from studies on CV genetics together with the data from the DNA biobank, scientists may be able to effectively prevent and treat CV diseases in the future.


Maintenance of hardware after early postoperative infection following fracture internal fixation.
The development of a deep wound infection in the presence of hardware after open reduction and internal fixation presents a clinical dilemma, and there is scant literature to aid in decision-making. The purpose of the present study was to determine the prevalence of osseous union with maintenance of hardware after the development of postoperative infection within six weeks after internal fixation of a fracture. The present study included 121 patients from three level-I trauma centers, retrospectively identified from billing and trauma registries, in whom 123 postoperative wound infections with positive intraoperative cultures had developed within six weeks after internal fixation of acute fractures. The incidence of fracture union without hardware removal was calculated, and the parameters that predicted success or failure were evaluated. Eighty-six patients (eighty-seven fractures; 71%) had fracture union with operative débridement, retention of hardware, and culture-specific antibiotic treatment and suppression. Predictors of treatment failure were open fracture (p = 0.03) and the presence of an intramedullary nail (p = 0.01). Several variables were not significant but trended toward an association with failure, including smoking, infection with Pseudomonas species, and involvement of the femur, tibia, ankle, or foot. Deep infection after internal fixation of a fracture can be treated successfully with operative débridement, antibiotic suppression, and retention of hardware until fracture union occurs. These results may be improved by patient selection based on certain risk factors and the specific bacteria and implants involved.


In which regions is breast-feeding safer from the impact of toxic elements from the environment?
Owing to its unique nutritional and immunological characteristics, breast milk is the most important food source for infants. But, children are at greater risk for exposure to environmental toxicants from breast milk. The aim of this study was to evaluate the influence of environmental pollution on essential and toxic element contents of breast milk and determine the risky locations in our population. This study was conducted on women who were breastfeeding (n=90). Milk samples were collected at three locations in Marmara region, Turkey: highly industrialized region highly affected by pollution, urbanized region moderately and rural area that is affected little. Breast milk samples (5 mL) were collected at approximately one month postpartum (mature milk). The concentrations of cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), iron (Fe), manganese (Mn), nickel (Ni), lead (Pb) and zinc (Zn) in milk samples were compared to the milk samples coming from different locations.Lead, cadmium, nickel, chromium, iron and manganese levels in the breast milk are highest and engrossing especially in rural areas compared to the other regions but cobalt, copper, zinc levels are highest in highly industrial areas. The levels of essential and toxic elements in breast milk can vary in different regions. The levels presented in our study are above some countries' data albeit not at toxic levels. Because of global effects, environmental pollution is not the problem for industrializing regions only. Rural area also may not be safe for breastfeed babies.


Object specificity and personal relevance in long-term visual remembering.
The personal relevance of an object is multi-faceted, each facet being capable of contributing to the effects on object memory attributed to personal relevance. An object's status as an individual object (object specificity), rather than just a category of object, is one such facet and its impact on the long-term visual remembering of everyday objects is assessed in two experiments. Images and drawings were produced under generic (e.g., "Please draw a bed") and personal exemplar (e.g., "Please draw your bed") instructions, and participants indicated the degree to which the image on which their drawing was based was of a specific object or a generic object. Object specificity induced a sense of time and place for a remembered object, the most recent encounter with the object being most salient. Other aspects of personal relevance collectively facilitated the retrieval of an object's category-irrelevant features (thereby increasing the vividness of the object image), the other objects with which it was seen, and a more general episodic sense of place. Against a broader theoretical perspective, it is proposed that visual episodic memory and visual knowledge are primary sources of information for specific personally relevant objects and generic objects, respectively.


Analysis of Massive Online Medical Consultation Service Data to Understand Physicians' Economic Return: Observational Data Mining Study.
Online health care consultation has become increasingly popular and is considered a potential solution to health care resource shortages and inefficient resource distribution. However, many online medical consultation platforms are struggling to attract and retain patients who are willing to pay, and health care providers on the platform have the additional challenge of standing out in a crowd of physicians who can provide comparable services. This study used machine learning (ML) approaches to mine massive service data to (1) identify the important features that are associated with patient payment, as opposed to free trial-only appointments; (2) explore the relative importance of these features; and (3) understand how these features interact, linearly or nonlinearly, in relation to payment. The dataset is from the largest China-based online medical consultation platform, which covers 1,582,564 consultation records between patient-physician pairs from 2009 to 2018. ML techniques (ie, hyperparameter tuning, model training, and validation) were applied with four classifiers-logistic regression, decision tree (DT), random forest, and gradient boost-to identify the most important features and their relative importance for predicting paid vs free-only appointments. After applying the ML feature selection procedures, we identified 11 key features on the platform, which are potentially useful to predict payment. For the binary ML classification task (paid vs free services), the 11 features as a whole system achieved very good prediction performance across all four classifiers. DT analysis further identified five distinct subgroups of patients delineated by five top-ranked features: previous offline connection, total dialog, physician response rate, patient privacy concern, and social return. These subgroups interact with the physician differently, resulting in different payment outcomes. The results show that, compared with features related to physician reputation, service-related features, such as service delivery quality (eg, consultation dialog intensity and physician response rate), patient source (eg, online vs offline returning patients), and patient involvement (eg, provide social returns and reveal previous treatment), appear to contribute more to the patient's payment decision. Promoting multiple timely responses in patient-provider interactions is essential to encourage payment.


Reconstruction of the Acetabulum in Developmental Dysplasia of the Hip in total hip replacement.
Developmental dysplasia of the hip (DDH) or congenital hip dysplasia (CDH) is the most prevalent developmental childhood hip disorder. It includes a wide spectrum of hip abnormalities ranging from dysplasia to subluxation and complete dislocation of the hip joint. The natural history of neglected DDH in adults is highly variable. The mean age of onset of symptoms is 34.5 years for dysplastic DDH, 32.5 years for low dislocation, 31.2 years for high dislocation with a false acetabulum, and 46.4 years for high dislocation without a false acetabulum. Thorough understanding of the bony and soft tissue deformities induced by dysplasia is crucial for the success of total hip arthroplasty. It is important to evaluate the existing acetabular deformity three-dimensionally, and customize the correction in accordance with the quantity and location of ace tabular deficiencies. Acetabular reconstruction in patients with DDH is challenging. Interpretation of published data is difficult and should be done with caution because most series include patients with different types of hip disease. In general, the complication rate associated with THA is higher in patients with hip dysplasia than it is in patients with osteoarthritis. Overall, clinical and functional outcomes following THA in patients hip dysplasia (DDH) differ from those treated for primary hip osteoarthritis, possibly due to the lower age and level of activity. Although function scores decline with age, the scores for pain and range of motion presented with a statistically significant improvement in the long-term.


[Arthroscopic Treatment of Chronic Retrocalcaneal Bursitis - Endoscopic Calcaneoplasty].
INTRODUCTION The authors evaluate the results of endoscopic treatment and analyse the causes of persisting difficulties in retrocalcaneal bursitis unresponsive to conservative treatment. Stated as one of the causes is the possible chronic irritation of minor/partial tears of anterior parts of the distal portion of the Achilles tendon, which have no chance to heal due to continued overload and impingement syndrome of the superior prominence of the calcaneal tubercle. MATERIAL AND METHODS The authors evaluate the group of 24 patients (10 women and 14 men) diagnosed with retrocalcaneal bursitis, who reported more than 12 months of difficulties and in whom the conservative therapy options have already been exhausted. The decision to perform a revision arthroscopy was made by the authors based on chronic long-lasting difficulties, clinical examination, calcaneus radiograph, ultrasound examination and negative response to the performed conservative treatment. The MRI scan was obtained in 4 patients. In 11 patients during arthroscopy the Achilles tendon was intact, in 13 patients (54.2%) minor tears were detected on the anterior aspect of the distal Achilles tendon, not diagnosed preoperatively (by ultrasound examination). In all the patients, resection of the prominence of the calcaneal tubercle was performed, whereas in patients with affected Achilles tendon also the loose tendon fragments were resected. RESULTS A total of 23 patients reported an improvement of their condition, the pain subsided at 21-43 days. Altogether 21 patients returned to activities carried out before the onset of pain. The AOFAS score values measured preoperatively in patients with an intact tendon were 59.5 ± 15.0, in patients with an injured tendon it was 45.57 ± 9.6, while 6 months after the surgery the values were 95.7 ± 6.2, or 88.71 ± 7.8 respectively. In 1 female patient, with one prior surgery, diagnosed with a partial tear of the Achilles tendon, the effect of the surgery was unsatisfactory. After six months the patient underwent an open revision surgery and the Achilles tendon reinsertion. No intraoperative or postoperative complications were observed. DISCUSSION The endoscopic technique facilitates a more visible bone resection, with better control of resection, which is less invasive than the open technique. The source of irritation can be removed, which is not always only the prominence of the calcaneal tubercle, but another cause of persisting problems can also be a microtrauma affecting the anterior portion of the Achilles tendon, with a reactive hyperaemia in bursal tissue. Compared to literature and also based on the results of the authors of this retrospective study, the endoscopic calcaneoplasty is less invasive than the open surgery. It is always necessary to properly diagnose the causes of difficulties and to timely respond to persisting symptoms in patients, not responding satisfactorily to conservative therapy. CONCLUSIONS The findings presented by the authors provide a new perspective on the causes of chronic problems such as the "posterior heel pain" and tend to give preference to the active endoscopic approach in patients with persisting problems, not responding to conservative treatment, predisposed based on the radiological examination and with a positive finding on ultrasound or MRI scan. Key words:retrocalcaneal bursitis, endoscopic calcaneoplasty, arthroscopy.


Evolutionary dynamics of complex networks of HIV drug-resistant strains: the case of San Francisco.
Over the past two decades, HIV resistance to antiretroviral drugs (ARVs) has risen to high levels in the wealthier countries of the world, which are able to afford widespread treatment. We have gained insights into the evolution and transmission dynamics of ARV resistance by designing a biologically complex multistrain network model. With this model, we traced the evolutionary history of ARV resistance in San Francisco and predict its future dynamics. By using classification and regression trees, we identified the key immunologic, virologic, and treatment factors that increase ARV resistance. Our modeling shows that 60% of the currently circulating ARV-resistant strains in San Francisco are capable of causing self-sustaining epidemics, because each individual infected with one of these strains can cause, on average, more than one new resistant infection. It is possible that a new wave of ARV-resistant strains that pose a substantial threat to global public health is emerging.


In Vitro and In Vivo Effects of Jia-Wei-Xiao-Yao-San in Human Breast Cancer MCF-7 Cells Treated With Tamoxifen.
There is epidemiological evidence that Jia-Wei-Xiao-Yao-San (JWXYS) is the most common Chinese medicine decoction coprescribed with tamoxifen (Tam) when breast cancer is treated by hormonal therapy. However, whether there is interaction between JWXYS and Tam remains to be clarified. The aim of this study was to investigate the in vitro and in vivo effects of JWXYS on human breast cancer MCF-7 cells treated with Tam. In vitro cultured MCF-7 cells were cotreated with JWXYS and Tam. This was followed by MTT ([4,5-cimethylthiazol-2-yl]- 2,5-diphenyl tetrazolium bromide) assays and cell cycle analysis to assess cell proliferation; Western blot analysis was used to analyze the expression of various proteins involved in growth-related signal pathways. In addition, immunohistochemistry was used to detect autophagy among the cancer cells. In vivo analysis used female athymic nude mice implanted with MCF-7 cells; these mice were randomly assigned to 6 groups. All mice were killed humanely after 21 days of treatment; body weight, tumor volume, and tumor weight were then measured. JWXYS was not cytotoxic to MCF-7 cells, based on the fact that there were no statistically significant changes between the JWXYS + Tam groups and the Tam-alone group in cell numbers, cell cycle progression, and cell proliferation signals, the latter including the expression levels of AKT, ERK, P38, p27(Kip1), and light chain (LC3)-I, II. Furthermore, using the MCF-7 xenograft mouse model, there were no significant changes between the JWXYS (1.3-3.9 gm/kg) + Tam groups and the Tam-alone group in terms of tumor weight and the protein expression levels of AKT, ERK, P38, and p27 (Kip1). However, there was a significant decrease in LC3-II protein expression with the low-dose JWXYS + Tam group but not with the middle- or high-dose JWXYS + Tam groups compared with the Tam-alone group. Based on in vitro studies and in vivo functional studies, there is no obvious interaction between JWXYS and Tam. However, the presence of interference at the molecular level in relation to LC3-II expression provides important information and may affect treatment strategies when physicians have patients with estrogen receptor-α(+) or progesterone receptor(+) breast cancers.


Suberoylanilide hydroxamic acid (SAHA; vorinostat) causes bone loss by inhibiting immature osteoblasts.
Histone deacetylase (Hdac) inhibitors are used clinically to treat cancer and epilepsy. Although Hdac inhibition accelerates osteoblast maturation and suppresses osteoclast maturation in vitro, the effects of Hdac inhibitors on the skeleton are not understood. The purpose of this study was to determine how the pan-Hdac inhibitor, suberoylanilide hydroxamic acid (SAHA; a.k.a. vorinostat or Zolinza(TM)) affects bone mass and remodeling in vivo. Male C57BL/6J mice received daily SAHA (100mg/kg) or vehicle injections for 3 to 4weeks. SAHA decreased trabecular bone volume fraction and trabecular number in the distal femur. Cortical bone at the femoral midshaft was not affected. SAHA reduced serum levels of P1NP, a bone formation marker, and also suppressed tibial mRNA levels of type I collagen, osteocalcin and osteopontin, but did not alter Runx2 or osterix transcripts. SAHA decreased histological measures of osteoblast number but interestingly increased indices of osteoblast activity including mineral apposition rate and bone formation rate. Neither serum (TRAcP 5b) nor histological markers of bone resorption were affected by SAHA. P1NP levels returned to baseline in animals which were allowed to recover for 4weeks after 4weeks of daily SAHA injections, but bone density remained low. In vitro, SAHA suppressed osteogenic colony formation, decreased osteoblastic gene expression, induced cell cycle arrest, and caused DNA damage in bone marrow-derived adherent cells. Collectively, these data demonstrate that bone loss following treatment with SAHA is primarily due to a reduction in osteoblast number. Moreover, these decreases in osteoblast number can be attributed to the deleterious effects of SAHA on immature osteoblasts, even while mature osteoblasts are resistant to the harmful effects and demonstrate increased activity in vivo, indicating that the response of osteoblasts to SAHA is dependent upon their differentiation state. These studies suggest that clinical use of SAHA and other Hdac inhibitors to treat cancer, epilepsy or other conditions may potentially compromise skeletal structure and function.


Validation of quantitative Ronchi test through numerical propagation.
We validate the quantitative analysis of Ronchigrams for wavefront sensing through detailed numerical simulations. Analysis of the experimental Ronchigrams provides the wavefront aberrations, the F/# of the beam and the distance of the Ronchi ruling from the paraxial focus. These retrieved parameters are used to numerically simulate the Ronchigrams with excellent agreement. This favorable comparison validates the accuracy of the wavefront recovery and provides a tool to examine the accuracy and robustness of this wavefront measurement technique.


Tityustoxin-K alpha, a structurally novel and highly potent K+ channel peptide toxin, interacts with the alpha-dendrotoxin binding site on the cloned Kv1.2 K+ channel.
The interaction between two nonhomologous K+ channel toxins, Tityus serrulatus (scorpion) toxin tityustoxin-K alpha (TsTX-K alpha) and Dendroaspis angusticeps (snake) toxin dendrotoxin (alpha-DTX), was investigated on K+ currents in B82 fibroblast cells transformed to express the Kv1.2 K+ channel. As demonstrated previously, alpha-DTX was a potent blocker of the K+ current (Kd, 2.8 nM). Recombinant TsTX-K alpha produced a similar block of the current but was 1 order of magnitude more potent (Kd, 0.21 nM). TsTX-K alpha did not affect the kinetic properties of the current or its voltage dependence of activation. Experiments with excised and cell-attached patch recordings demonstrated that TsTX-K alpha blocks the K+ channel by binding to an extracellular site. In the presence of TsTX-K alpha the blocking potency of alpha-DTX was reduced, whereas the potency of 4-aminopyridine, which also blocks the channel, was unaffected. alpha-DTX caused a rightward shift in the scaled concentration-response curve for TsTX-K alpha, the magnitude of which was reasonably well predicted by a model in which there is a competitive interaction between the two peptide toxins. We conclude that TsTX-K alpha and alpha-DTX block the Kv1.2 K+ channel by binding to the same or closely related sites.


Fine mapping the MHC region identified rs4997052 as a new variant associated with nonobstructive azoospermia in Han Chinese males.
To investigate the association between genetic variants in the major histocompatibility complex (MHC) region and nonobstructive azoospermia (NOA) susceptibility. MHC region fine-mapping analysis based on previous NOA genome-wide association study (GWAS) data. Medical university. Nine hundred and eighty-one men with NOA and 1,657 normal fertile male controls. None. The MHC region imputation assessed with SNP2HLA software, taking the specific Han-MHC database as a reference panel; statistical significance of the MHC variants calculated using logistic regression models; functional annotation based on online public databases; and phenotypic variances explained by specific groups of genetic variants estimated using the fixed effects model from individual associations. Two independent risk loci, rs7194 (odds ratio [OR] 1.37) at MHC class II molecules and rs4997052 (OR 1.30) at MHC class I molecules, were identified. Functional annotation showed rs7194 may tag the effect of multiple amino acid residues and the expression of HLA-DQB1 and HLA-DRB1; while rs4997052 showed the effect of amino acid changes of HLA-B at position 116 as well as the expression of HLA-B and CCHCR1, which coexpressed with genes enriched in pathways of spermatogenesis and male gamete generation. The novel variant rs4997052 identified in our study can explain another approximately 0.66% of the phenotypic variances of NOA. We fine-mapped the MHC region and identified two loci that independently drove NOA susceptibility. These results provide a deeper understanding of the association mechanisms of MHC and NOA risk.


Inviting consumers to downsize fast-food portions significantly reduces calorie consumption.
Policies that mandate calorie labeling in fast-food and chain restaurants have had little or no observable impact on calorie consumption to date. In three field experiments, we tested an alternative approach: activating consumers' self-control by having servers ask customers if they wanted to downsize portions of three starchy side dishes at a Chinese fast-food restaurant. We consistently found that 14-33 percent of customers accepted the downsizing offer, and they did so whether or not they were given a nominal twenty-five-cent discount. Overall, those who accepted smaller portions did not compensate by ordering more calories in their entrées, and the total calories served to them were, on average, reduced by more than 200. We also found that accepting the downsizing offer did not change the amount of uneaten food left at the end of the meal, so the calorie savings during purchasing translated into calorie savings during consumption. Labeling the calorie content of food during one of the experiments had no measurable impact on ordering behavior. If anything, the downsizing offer was less effective in changing customers' ordering patterns with the calorie labeling present. These findings highlight the potential importance of portion-control interventions that specifically activate consumers' self-control.


Incorporating basic needs to reconcile poverty and ecosystem services.
Conservation managers frequently face the challenge of protecting and sustaining biodiversity without producing detrimental outcomes for (often poor) human populations that depend on ecosystem services for their well-being. However, mutually beneficial solutions are often elusive and can mask trade-offs and negative outcomes for people. To deal with such trade-offs, ecological and social thresholds need to be identified to determine the acceptable solution space for conservation. Although human well-being as a concept has recently gained prominence, conservationists still lack tools to evaluate how their actions affect it in a given context. We applied the theory of human needs to conservation by building on an extensive historical application of need approaches in international development. In an innovative participatory method that included focus groups and household surveys, we evaluated how human needs are met based on locally relevant thresholds. We then established connections between human needs and ecosystem services through key-informant focus groups. We applied our method in coastal East Africa to identify households that would not be able to meet their basic needs and to uncover the role of ecosystem services in meeting these. This enabled us to identify how benefits derived from the environment were contributing to meeting basic needs and to consider potential repercussions that could arise through changes to ecosystem service provision. We suggest our approach can help conservationists and planners balance poverty alleviation and biodiversity protection and ensure conservation measures do not, at the very least, cause serious harm to individuals. We further argue it can be used as a basis for monitoring the impacts of conservation on multidimensional poverty.


Structure of force networks in tapped particulate systems of disks and pentagons. I. Clusters and loops.
The force network of a granular assembly, defined by the contact network and the corresponding contact forces, carries valuable information about the state of the packing. Simple analysis of these networks based on the distribution of force strengths is rather insensitive to the changes in preparation protocols or to the types of particles. In this and the companion paper [Kondic et al., Phys. Rev. E 93, 062903 (2016)10.1103/PhysRevE.93.062903], we consider two-dimensional simulations of tapped systems built from frictional disks and pentagons, and study the structure of the force networks of granular packings by considering network's topology as force thresholds are varied. We show that the number of clusters and loops observed in the force networks as a function of the force threshold are markedly different for disks and pentagons if the tangential contact forces are considered, whereas they are surprisingly similar for the network defined by the normal forces. In particular, the results indicate that, overall, the force network is more heterogeneous for disks than for pentagons. Such differences in network properties are expected to lead to different macroscale response of the considered systems, despite the fact that averaged measures (such as force probability density function) do not show any obvious differences. Additionally, we show that the states obtained by tapping with different intensities that display similar packing fraction are difficult to distinguish based on simple topological invariants.


Variation in serum ionized calcium on cardiopulmonary resuscitation.
Changes in serum ionized calcium (Ca(++)) levels during cardiopulmonary resuscitation (CPR) and before and after CaCl(2) administration have been examined and investigated in 30 patients with cardiopulmonary arrest on arrival (dead on arrival patients) when a significant negative correlation was found to exist between the transportation time and aCa(++), as the aCa(++) level decreased with an increase in the transportation time. Upon arrival, the pH had fallen due to acidosis so that aCa(++) and cCa(++) levels were virtually normal. After admission, the pH rose as a result of CPR, resulting in a significant drop in both Ca(++) levels, so that in most cases resuscitation was not possible. Those successfully resuscitated took over 60 min to return to normal Ca(++) levels. Administration of approximately 6.6 mg/kg of CaCl(2) led to significant increases in aCa(++) and cCa(++) to essentially normal levels, even with some patients recording extremely elevated Ca levels, even with some patients recording extremely elevated Ca levels. However, the success rate of resuscitation was not found to show any significant difference according to whether CaCl(2) had or had not been administrated.Thus, it is felt necessary to re-examine the use of calcium chloride on CPR.


Lower cerebral vasoreactivity as a predictor of gait speed decline in type 2 diabetes mellitus.
Gait speed is an indicator of overall functional health and is correlated with survival in older adults. We prospectively evaluated the long-term association between cerebral vasoreactivity and gait speed during normal walking (NW) and dual-task walking (DTW) in older adults with and without type 2 diabetes mellitus (T2DM). 40 participants (aged 67.3 ± 8.8 years, 20 with T2DM) completed a 2-year prospective study consisting of MRI, blood sampling, and gait assessments. The whole brain vasoreactivity was quantified using continuous arterial spin labeling MRI. Gait speed during DTW was assessed by subtracting serial sevens. Dual-task cost was calculated as the percent change in gait speed from NW to DTW. In the entire cohort, higher glycemic profiles were associated with a slower gait speed. In the diabetic group, lower vasoreactivity was associated with a slower gait speed during NW ([Formula: see text] = 0.30, p = 0.019) and DTW ([Formula: see text] = 0.35, p = 0.01) and a higher dual-task cost ([Formula: see text] = 0.69, p = 0.009) at 2-year follow-up. The participants with T2DM and lower cerebral vasoreactivity had a greater decrease in gait speed during NW and DTW after the 2-year follow-up ([Formula: see text] = 0.17, p = 0.04 and [Formula: see text] = 0.28, p = 0.03, respectively). Longer diabetes duration was associated with a higher dual-task cost ([Formula: see text] = 0.19, p = 0.04) and a greater decrease in gait speed during NW ([Formula: see text] = 0.17, p = 0.02). These findings indicate that in older adults with type 2 diabetes, gait performance is highly dependent on the integrity of cerebrovascular regulation.


Identifying zoonotic origin of SARS-CoV-2 by modeling the binding affinity between Spike receptor-binding domain and host ACE2.
Despite considerable research progress on SARS-CoV-2, the direct zoonotic origin (intermediate host) of the virus remains ambiguous. The most definitive approach to identify the intermediate host would be the detection of SARS-CoV-2-like coronaviruses in wild animals. However, due to the high number of animal species, it is not feasible to screen all the species in the laboratory. Given that the recognition of the binding ACE2 proteins is the first step for the coronaviruses to invade host cells, we proposed a computational pipeline to identify potential intermediate hosts of SARS-CoV-2 by modeling the binding affinity between the Spike receptor-binding domain (RBD) and host ACE2. Using this pipeline, we systematically examined 285 ACE2 variants from mammals, birds, fish, reptiles, and amphibians, and found that the binding energies calculated on the modeled Spike-RBD/ACE2 complex structures correlate closely with the effectiveness of animal infections as determined by multiple experimental datasets. Built on the optimized binding affinity cutoff, we suggested a set of 96 mammals, including 48 experimentally investigated ones, which are permissive to SARS-CoV-2, with candidates from primates, rodents, and carnivores at the highest risk of infection. Overall, this work not only suggested a limited range of potential intermediate SARS-CoV-2 hosts for further experimental investigation; but more importantly, it proposed a new structure-based approach to general zoonotic origin and susceptibility analyses that are critical for human infectious disease control and wildlife protection.


Ingested inorganic arsenic and prevalence of diabetes mellitus.
To examine the association between ingested inorganic arsenic and prevalence of diabetes mellitus, in 1988, the authors studied 891 adults residing in villages in southern Taiwan where arseniasis is hyperendemic. The status of diabetes mellitus was determined by an oral glucose tolerance test and a history of diabetes regularly treated with sulfonylurea or insulin. The cumulative arsenic exposure in parts per million-years was calculated from the detailed history of residential addresses and duration of drinking artesian well water obtained through standardized interviews based on a structured questionnaire and the arsenic concentration in well water. The body mass index was derived from body height and weight measured according to a standard protocol, while the physical activity at work was also obtained by questionnaire interviews. Residents in villages where the chronic arseniasis was hyperendemic had a twofold increase in age- and sex-adjusted prevalence of diabetes mellitus compared with residents in Taipei City and the Taiwan area. There was a dose-response relation between cumulative arsenic exposure and prevalence of diabetes mellitus. The relation remained significant after adjustment for age, sex, body mass index, and activity level at work by a multiple logistic regression analysis giving a multivariate-adjusted odds ratio of 6.61 and 10.05, respectively, for those who had a cumulative arsenic exposure of 0.1-15.0 and greater than 15.0 ppm-year compared with those who were unexposed. These results suggest the chronic arsenic exposure may induce diabetes mellitus in humans.


The annual cycles of phytoplankton biomass.
Terrestrial plants are powerful climate sentinels because their annual cycles of growth, reproduction and senescence are finely tuned to the annual climate cycle having a period of one year. Consistency in the seasonal phasing of terrestrial plant activity provides a relatively low-noise background from which phenological shifts can be detected and attributed to climate change. Here, we ask whether phytoplankton biomass also fluctuates over a consistent annual cycle in lake, estuarine-coastal and ocean ecosystems and whether there is a characteristic phenology of phytoplankton as a consistent phase and amplitude of variability. We compiled 125 time series of phytoplankton biomass (chlorophyll a concentration) from temperate and subtropical zones and used wavelet analysis to extract their dominant periods of variability and the recurrence strength at those periods. Fewer than half (48%) of the series had a dominant 12-month period of variability, commonly expressed as the canonical spring-bloom pattern. About 20 per cent had a dominant six-month period of variability, commonly expressed as the spring and autumn or winter and summer blooms of temperate lakes and oceans. These annual patterns varied in recurrence strength across sites, and did not persist over the full series duration at some sites. About a third of the series had no component of variability at either the six- or 12-month period, reflecting a series of irregular pulses of biomass. These findings show that there is high variability of annual phytoplankton cycles across ecosystems, and that climate-driven annual cycles can be obscured by other drivers of population variability, including human disturbance, aperiodic weather events and strong trophic coupling between phytoplankton and their consumers. Regulation of phytoplankton biomass by multiple processes operating at multiple time scales adds complexity to the challenge of detecting climate-driven trends in aquatic ecosystems where the noise to signal ratio is high.


Fusarium Wilt of Tomato Caused by Fusarium oxysporum f. sp. lycopersici Race 3 in Baja California Sur, Mexico.
Vascular wilting diseases have become one of the most serious diseases of tomato (Lycopersicon esculentum) throughout the Baja California Peninsula. Since the winter of 2004, a disease with symptoms characteristic of those caused by a Fusarium species has been observed in commercial fields near La Paz and Todos Santos in the state of Baja California Sur (BCS). Symptoms include typical one-sided wilting and dark brown vascular discoloration. Upper stem tissues and wilted seedlings were disinfested by immersion in a 1.0% aqueous solution of sodium hypochlorite for 2 min, rinsed in sterile water, and placed on Komada's medium (pH 6.8) at 22 ± 3°C. After 72 h, hyphal growth was recovered and subcultured on carnation leaf agar and potato dextrose agar and incubated at 25°C in 12-h light/dark cycles. Identification was based on colony morphology, conidial characteristics, and molecular techniques. White cottony mycelium, reddish coloration of the medium, ovoid two-celled macroconidia, and large macroconidia, all characteristic of F. oxysporum, were observed (2). Polymerase chain reaction and restriction fragment length polymorphisms with restriction enzymes EcoRI, RsaI, and HaeIII were used to characterize 24 isolates (sampled during January 2005) from La Paz (Fol-LaP) as formae speciales lycopersici and assigned to vegetative compatibility group 0030 (1). Confirmation of pathogenicity and race determination for the Fol-LaP isolates were as described previously (3). Mexican isolates of races 2 and 3 (one each) were included as positive controls. Conidial suspensions of 7 × 105 CFU/ml were used to inoculate differential tomato cvs. Bonny Best (Millington Co., universally susceptible), Tequila F1 (Vilmorin, race 1 resistant), Rio Grande (Harris Moran, race 1 and 2 resistant), and Sebring (Rogers, race 1, 2, and 3 resistant). Plants at the first true-leaf stage were inoculated by dipping their roots in the conidial suspension. Inoculated seedlings were transplanted into pots containing a sterile 5:1:1 mixture of sand/vermiculite/soil (v/v/v) and maintained in the greenhouse at 25 to 28°C under natural daylight. An equal number of plants of each cultivar dipped in water were used as controls. The experimental design was a completely randomized type with six replications (pots) containing four seedlings per pot. The test was done twice. The most susceptible plants inoculated by root-dipping developed typical symptoms of wilt, slight vein clearing on outer leaflets, stunting, dark brown vascular discoloration, and death. F. oxysporum was recovered from all symptomatic plants, whereas noninoculated tomato seedlings showed no symptoms. According to differential infection and symptomatology observed on infected cultivars, 62.5% of the isolated strains were identified as race 2, 25% as race 3, and 12.5% as an undetermined race isolated from Sebring. The presence of race 3 in BCS has important epidemiological implications since it has been reported on tomato in Sinaloa (4). The potential spread of the pathogen on introduced transplants represents a risk to tomato crops on the peninsula. To our knowledge, this is the first report of F. oxysporum f. sp. lycopersici race 3 in the state of BCS, Mexico. References: (1) G. Cai et al. Phytopathology 93:1014, 2003. (2) P. E. Nelson et al. Fusarium species. Pennsylvania State University Press, University Park, 1983. (3) B. A. Summerell et al. Plant Dis. 87:117, 2003. (4) J. G. Valenzuela-Ureta et al. Plant Dis. 80:105, 1996.


Laser nanosurgery of cerebellar axons in vivo.
Only a few neuronal populations in the central nervous system (CNS) of adult mammals show local regrowth upon dissection of their axon. In order to understand the mechanism that promotes neuronal regeneration, an in-depth analysis of the neuronal types that can remodel after injury is needed. Several studies showed that damaged climbing fibers are capable of regrowing also in adult animals. The investigation of the time-lapse dynamics of degeneration and regeneration of these axons within their complex environment can be performed by time-lapse two-photon fluorescence (TPF) imaging in vivo. This technique is here combined with laser surgery, which proved to be a highly selective tool to disrupt fluorescent structures in the intact mouse cortex. This protocol describes how to perform TPF time-lapse imaging and laser nanosurgery of single axonal branches in the cerebellum in vivo. Olivocerebellar neurons are labeled by anterograde tracing with a dextran-conjugated dye and then monitored by TPF imaging through a cranial window. The terminal portion of their axons are then dissected by irradiation with a Ti:Sapphire laser at high power. The degeneration and potential regrowth of the damaged neuron are monitored by TPF in vivo imaging during the days following the injury.


Changes in physical activity in leisure time and the risk of myocardial infarction, ischemic heart disease, and all-cause mortality.
Physical activity is associated to a lower risk of mortality from all-causes and from coronary heart disease. The long-term effects of changes in physical activity on coronary heart disease are, however, less known. We examined the association between changes in leisure time physical activity and the risk of myocardial infarction (MI), ischemic heart disease (IHD), and all-cause mortality as well as changes in blood pressure in 4,487 men and 5,956 women in the Copenhagen City Heart Study. Physical activity was measured in 1976-1978 and 1981-1983 and participants were followed in nation-wide registers until 2009. Men who decreased physical activity by at least two levels and women who decreased by one level had a higher risk of MI relatively to an unchanged physical activity level (hazard ratio [HR] = 1.74, 95% confidence interval [95% CI]: 1.17-2.60 and HR = 1.30, 95% CI: 1.03-1.65). Similar associations were found for IHD although only significant in women. In all-cause mortality, men who increased physical activity had a lower risk and both men and women who reduced physical activity had a higher risk compared to an unchanged physical activity level. No association between changes in physical activity and blood pressure was observed. Findings from this prospective study suggest that changes in physical activity affect the risk of MI, IHD and all-cause mortality. A decrease in physical activity was associated to a higher risk of coronary heart disease.


Intramuscular ziprasidone compared with intramuscular haloperidol in the treatment of acute psychosis. Ziprasidone I.M. Study Group.
This 7-day, randomized, open-label, multicenter, international study compared the efficacy and tolerability of intramuscular (i.m.) ziprasidone with haloperidol i.m. and the transition from i.m. to oral treatment in hospitalized patients with acute psychotic agitation (related to DSM-III-R diagnoses). Patients received up to 3 days of flexible-dose ziprasidone i.m. (N = 90) or haloperidol i.m. (N = 42) followed by oral treatment to day 7. After an initial ziprasidone i.m. dose of 10 mg, subsequent i.m. doses of 5 to 20 mg could be given every 4 to 6 hours (maximum daily dose = 80 mg) if needed, followed by oral ziprasidone, 80-200 mg/day. Haloperidol i.m. doses of 2.5 to 10 mg were given on entry, followed by 2.5 to 10 mg i.m. every 4 to 6 hours (maximum daily dose = 40 mg) if needed, then by oral haloperidol, 10-80 mg/day. The mean reductions in Brief Psychiatric Rating Scale (BPRS) total, BPRS agitation items, and Clinical Global Impressions-Severity scale scores were statistically significantly greater (p < .05, p < .01, and p < .01, respectively) after ziprasidone i.m. treatment compared with haloperidol i.m. treatment. Further reductions in these scores also occurred in both groups following transition to oral treatment. Ziprasidone was associated with a lower incidence of movement disorders and a reduced requirement for anticholinergic medication during both i.m. and oral treatment compared with haloperidol. Movement disorder scale scores improved with ziprasidone i.m. and oral treatment, but deteriorated with haloperidol. Other adverse events were rare with both treatments. Ziprasidone i.m. was significantly more effective in reducing the symptoms of acute psychosis and was better tolerated than haloperidol i.m., particularly in movement disorders. The transition from ziprasidone i.m. to oral ziprasidone was effective and well tolerated.


Computational analysis of spoken language in acute psychosis and mania.
This study aimed to (1) determine the feasibility of collecting behavioral data from participants hospitalized with acute psychosis and (2) begin to evaluate the clinical information that can be computationally derived from such data. Behavioral data was collected across 99 sessions from 38 participants recruited from an inpatient psychiatric unit. Each session started with a semi-structured interview modeled on a typical "clinical rounds" encounter and included administration of the Positive and Negative Syndrome Scale (PANSS). We quantified aspects of participants' verbal behavior during the interview using lexical, coherence, and disfluency features. We then used two complementary approaches to explore our second objective. The first approach used predictive models to estimate participants' PANSS scores from their language features. Our second approach used inferential models to quantify the relationships between individual language features and symptom measures. Our predictive models showed promise but lacked sufficient data to achieve clinically useful accuracy. Our inferential models identified statistically significant relationships between numerous language features and symptom domains. Our interview recording procedures were well-tolerated and produced adequate data for transcription and analysis. The results of our inferential modeling suggest that automatic measurements of expressive language contain signals highly relevant to the assessment of psychosis. These findings establish the potential of measuring language during a clinical interview in a naturalistic setting and generate specific hypotheses that can be tested in future studies. This, in turn, will lead to more accurate modeling and better understanding of the relationships between expressive language and psychosis.


Lithium reduces tau phosphorylation: effects in living cells and in neurons at therapeutic concentrations.
The mechanism of action of lithium remains to be determined satisfactorily. Recent studies suggested a possible role in inhibiting glycogen synthase kinase-3 (GSK-3), previously shown to phosphorylate the protein tau. Tau is expressed mainly in neurons, where it functions to stabilize microtubules in a phosphorylation-dependent manner. Neurons and transfected non-neuronal cells were treated with lithium and the phosphorylation of tau at multiple epitopes examined by western blotting and by immunocytochemistry. Using green fluorescent protein as a tag we examined the effects of lithium on phosphorylated tau in living cells. Lithium reversibly reduced tau phosphorylation at therapeutic concentrations, and even at high concentrations did not alter neuronal morphology. Green fluorescent protein tagged-tau when phosphorylated by GSK-3 was diffusely distributed; treatment with lithium resulted in association with microtubules and then bundle formation. Removing lithium allowed observation of the dissolution of bundles and gradual dissociation of tau from microtubules in living cells. Lithium may have multiple effects in brain, but at least one action is demonstrated to be a relative inhibition of GSK-3-induced tau phosphorylation. These results carry implications for future studies of the actions of mood-stabilizing drugs and indeed of the molecular mechanisms of affective disorders.


LSD Flashbacks - The Appearance of New Visual Imagery Not Experienced During Initial Intoxication: Two Case Reports.
A side effect associated with the use of synthetic hallucinogens such as lysergic acid diethylamide-(LSD) is the partial or total recurrence of perceptual disturbances which previously appeared during intoxication, despite absence of recent use. These are commonly referred to as "flashbacks" or Hallucinogen Persisting Perception Disorder (HPPD). Here we present two cases of patients with a prior history of LSD use who turned to psychiatric consultation following brief episodes of HPPD. Surprisingly, in both cases new visual imagery appeared during episodes of flashbacks which was not experienced during primary LSD use. Both subjects reported the ability to discern between LSD-associated visual disturbances and new visual imagery. This phenomenon did not cause functional impairment and in both cases caused gradual concern due to its persistence. Both patients refused medical treatment and continued psychiatric follow-up. At one year follow-up both patients reported almost complete spontaneous remission. To the best of our knowledge these are the first reported cases of LSD-related benign flashbacks in which new imagery is experienced. Reasons for this reversible and apparently harmless side effect are proposed. Conclusions from case reports should be taken with caution.


A predictive multiscale model for simulating flow-induced platelet activation: Correlating in silico results with in vitro results.
Flow-induced platelet activation prompts complex filopodial formation. Continuum methods fail to capture such molecular-scale mechanisms. A multiscale numerical model was developed to simulate this activation process, where a Dissipative Particle Dynamics (DPD) model of viscous blood flow is interfaced with a Coarse Grained Molecular Dynamics (CGMD) platelet model. Embedded in DPD blood flow, the macroscopic dynamic stresses are interactively transferred to the CGMD model, inducing intra-platelet associated events. The platelets activate by a biomechanical transductive linkage chain and dynamically change their shape in response. The models are fully coupled via a hybrid-potential interface and multiple time-stepping (MTS) schemes for handling the disparity between the spatiotemporal scales. Cumulative hemodynamic stresses that may lead to platelet activation are mapped on the surface membrane and simultaneously transmitted to the cytoplasm and cytoskeleton. Upon activation, the flowing platelets lose their quiescent discoid shape and evolve by forming filopodia. The model predictions were validated by a set of in vitro experiments, Platelets were exposed to various combinations of shear stresses and durations in our programmable hemodynamic shearing device (HSD). Their shape change was measured at multiple time points using scanning electron microscopy (SEM). The CGMD model parameters were fine-tuned by interrogating a parameter space established in these experiments. Segmentation of the SEM imaging streams was conducted by a deep machine learning system. This model can be further employed to simulate shear mediated platelet activation thrombosis initiation and to study the effects of modulating platelet properties to enhance their shear resistance via mechanotransduction pathways.


A Single Meal Containing Raw, Crushed Garlic Influences Expression of Immunity- and Cancer-Related Genes in Whole Blood of Humans.
Preclinical and epidemiologic studies suggest that garlic intake is inversely associated with the progression of cancer and cardiovascular disease. We designed a study to probe the mechanisms of garlic action in humans. We conducted a randomized crossover feeding trial in which 17 volunteers consumed a garlic-containing meal (100 g white bread, 15 g butter, and 5 g raw, crushed garlic) or a garlic-free control meal (100 g white bread and 15 g butter) after 10 d of consuming a controlled, garlic-free diet. Blood was collected before and 3 h after test meal consumption for gene expression analysis in whole blood. Illumina BeadArray was used to screen for genes of interest, followed by real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) on selected genes. To augment human study findings, Mono Mac 6 cells were treated with a purified garlic extract (0.5 μL/mL), and mRNA was measured by qRT-PCR at 0, 3, 6, and 24 h. The following 7 genes were found to be upregulated by garlic intake: aryl hydrocarbon receptor (AHR), aryl hydrocarbon receptor nuclear translocator (ARNT), hypoxia-inducible factor 1α (HIF1A), proto-oncogene c-Jun (JUN), nuclear factor of activated T cells (NFAT) activating protein with immunoreceptor tyrosine-based activation motif 1 (NFAM1), oncostatin M (OSM), and V-rel avian reticuloendotheliosis viral oncogene homolog (REL). Fold-increases in mRNA transcripts ranged from 1.6 (HIF1A) to 3.0 (NFAM1) (P < 0.05). The mRNA levels of 5 of the 7 genes that were upregulated in the human trial were also upregulated in cell culture at 3 and 6 h: AHR, HIF1A, JUN, OSM, and REL. Fold-increases in mRNA transcripts in cell culture ranged from 1.7 (HIF1A) to 12.1 (JUN) (P < 0.01). OSM protein was measured by ELISA and was significantly higher than the control at 3, 6, and 24 h (24 h: 19.5 ± 1.4 and 74.8 ± 1.4 pg/mL for control and garlic, respectively). OSM is a pleiotropic cytokine that inhibits several tumor cell lines in culture. These data indicate that the bioactivity of garlic is multifaceted and includes activation of genes related to immunity, apoptosis, and xenobiotic metabolism in humans and Mono Mac 6 cells. This trial is registered at clinicaltrials.gov as NCT01293591.


Orthosiphon stamineus Proteins Alleviate Pentylenetetrazol-Induced Seizures in Zebrafish.
The anticonvulsive potential of proteins extracted from Orthosiphon stamineus leaves (OSLP) has never been elucidated in zebrafish (Danio rerio). This study thus aims to elucidate the anticonvulsive potential of OSLP in pentylenetetrazol (PTZ)-induced seizure model. Physical changes (seizure score and seizure onset time, behavior, locomotor) and neurotransmitter analysis were elucidated to assess the pharmacological activity. The protective mechanism of OSLP on brain was also studied using mass spectrometry-based label-free proteomic quantification (LFQ) and bioinformatics. OSLP was found to be safe up to 800 µg/kg and pre-treatment with OSLP (800 µg/kg, i.p., 30 min) decreased the frequency of convulsive activities (lower seizure score and prolonged seizure onset time), improved locomotor behaviors (reduced erratic swimming movements and bottom-dwelling habit), and lowered the excitatory neurotransmitter (glutamate). Pre-treatment with OSLP increased protein Complexin 2 (Cplx 2) expression in the zebrafish brain. Cplx2 is an important regulator in the trans-SNARE complex which is required during the vesicle priming phase in the calcium-dependent synaptic vesicle exocytosis. Findings in this study collectively suggests that OSLP could be regulating the release of neurotransmitters via calcium-dependent synaptic vesicle exocytosis mediated by the "Synaptic Vesicle Cycle" pathway. OSLP's anticonvulsive actions could be acting differently from diazepam (DZP) and with that, it might not produce the similar cognitive insults such as DZP.


Photonic bandgap Bragg fiber sensors for bending/displacement detection.
We demonstrate an amplitude-based bending/displacement sensor that uses a plastic photonic bandgap Bragg fiber with one end coated with a silver layer. The reflection intensity of the Bragg fiber is characterized in response to different displacements (or bending curvatures). We note that the Bragg reflector of the fiber acts as an efficient mode stripper for the wavelengths near the edge of the fiber bandgap, which makes the sensor extremely sensitive to bending or displacements at these wavelengths. Besides, by comparison of the Bragg fiber sensor to a sensor based on a standard multimode fiber with similar outer diameter and length, we find that the Bragg fiber sensor is more sensitive to bending due to the presence of a mode stripper in the form of a multilayer reflector. Experimental results show that the minimum detection limit of the Bragg fiber sensor can be as small as 3 μm for displacement sensing.


Orlistat, sibutramine, or combination therapy: which performs better on waist circumference in relation with body mass index in obese patients?
The aim of this study was to evaluate decrease in waist circumference in obese patients receiving different anti-obesity treatments. The study was designed as a short-term (12 weeks), open-label, and randomized trial. Eighty six patients (70 females, 81.4%; mean age 41.09+/-8.73 years, mean BMI 36.1+/-4.3 kg/m2) were randomized to four different therapy groups. The primary outcome parameters were waist circumference and body mass index (BMI). The therapy groups were a) diet+sibutramine 1 x 10 mg/d (n=22), b) diet+orlistat 3 x 120 mg/d (n=25), c) combination of diet+sibutramine+orlistat (n=20) and d) diet (n=19). Combination therapy was more effective than diet and orlistat mono-therapy (p<0.0001 for all), but not significantly superior to sibutramine mono-therapy (p=0.072) in decreasing BMI. Sibutramine mono-therapy was significantly more effective in inducing BMI decrease compared with orlistat mono-therapy (p=0.039). The association between change in BMI and change in waist circumference was strongest in the orlistat mono-therapy group (P interaction=0.003). This means that patients taking orlistat experienced more decrease in waist circumference (3.4 cm, R2=0.29) per unit decrease in BMI compared to patients under combination therapy (2.6 cm, R2=0.25, P interaction = 0.015) and patients taking sibutramine (1.8 cm, R2=0.19, P interaction=0.026). In the diet therapy group decline in waist circumference was independent of BMI (1.9 cm, R2=0.02, P interaction=0.076). Although combination therapy and sibutramine mono-therapy were more effective in decreasing BMI, reduction in waist circumference and BMI was most significantly associated with the orlistat mono-therapy group. This may hint at the possibility of orlistat inducing weight loss mainly in the abdominal area targeted to reduce cardiovascular risk.


Phosphorus binding sites in proteins: structural preorganization and coordination.
Phosphorus is a ubiquitous element of the cell, which is found throughout numerous key molecules related to cell structure, energy and information storage and transfer, and a diverse array of other cellular functions. In this work, we adopt an approach often used for characterizing metal binding and selectivity of metalloproteins in terms of interactions in a first shell (direct residue interactions with the metal) and a second shell (residue interactions with first shell residues) and use it to characterize binding of phosphorus compounds. Similar analyses of binding have previously been limited to individual structures that bind to phosphate groups; here, we investigate a total of 8307 structures obtained from the RCSB Protein Data Bank (PDB). An analysis of the binding site amino acid propensities reveals very characteristic first shell residue distributions, which are found to be influenced by the characteristics of the phosphorus compound and by the presence of cobound cations. The second shell, which supports the coordinating residues in the first shell, is found to consist mainly of protein backbone groups. Our results show how the second shell residue distribution is dictated mainly by the first shell of the binding site, especially by cobound cations and that the main function of the second shell is to stabilize the first shell residues.


Intraoperative near-infrared imaging can identify pulmonary nodules.
Over 80,000 people undergo pulmonary resection for a lung nodule in the United States each year. Small nodules are frequently missed or difficult to find despite preoperative imaging. We hypothesized that near-infrared (NIR) imaging technology could be used to identify and locate lung nodules during surgery. We enrolled 18 patients who were diagnosed with a pulmonary nodule that required resection. All patients had a fine-cut 1-mm computed tomography scan preoperatively. The patients were given systemic 5 mg/kg indocyanine green and then underwent an open thoracotomy 24 hours later. The NIR imaging was used to identify the primary nodule and search for additional nodules that were not found by visual inspection or manual palpation of the ipsilateral lung. Manual palpation and visual inspection identified all 18 primary pulmonary nodules and no additional lesions. Intraoperative NIR imaging detected 16 out of the 18 primary nodules. The NIR imaging also identified 5 additional subcentimeter nodules; 3 metastatic adenocarcinomas and 2 metastatic sarcomas. This technology could identify nodules as small as 0.2 cm and as deep as 1.3 cm from the pleural surface. This approach discovered 3 nodules that were in different lobes than the primary tumor. Nodule fluorescence was independent of size, metabolic activity, histology, tumor grade and vascularity. This is the first-in-human demonstration of identifying pulmonary nodules during thoracic surgery with NIR imaging without a priori knowledge of their location or existence. The NIR imaging can detect pulmonary nodules during lung resections that are poorly visualized on computed tomography and difficult to discriminate on finger palpation.


Parallel comparison of three methodologies for measuring functional C1-inhibitor in Hereditary angioedema patients.
Hereditary angioedema (HAE) types I and II are characterized by functional C1 inhibitor (fC1-INH) deficiency which results in bradykinin overproduction. Sensitive, specific and robust methods to quantitate fC1-INH in human samples are required for diagnosing HAE and/or to measure pharmacodynamic activity of C1-INH drugs in clinical studies. To date, three methods have been reported in literature to measure fC1-INH: conventional chromogenic assay measuring residual C1-esterase activity, and immunoassays based on functional binding to either activated complement C1s or Factor XIIa/kallikrein. We used three qualified/validated fit-for purpose methods to quantitate fC1-INH in human plasma and to conduct a parallel comparison for diagnostic purposes and as a read-out for pharmacodynamic activity. Sensitivity and specificity were determined from the Receiver Operator Characteristics (ROC) curve analysis of the three fC1-INH methods through testing of fifty healthy control vs. HAE plasma samples. fC1-INH profile of fifteen HAE subjects, who underwent different treatment regimen in a cross-over Shire C1-INH clinical study, was analyzed in these three methods in parallel. A correlation analysis performed between these methods using data generated from clinical samples showed that profiles obtained from different fC1-INH methods matched for individual HAE subjects. Our findings suggest that functional binding immunoassay methods serve as reliable alternates for conventional chromogenic method to quantitate fC1-INH in human plasma samples with a better dynamic range of detection and ease of use. Of the two immunoassays used in this study, FXIIa-binding method gave better sensitivity, specificity, and correlation to the chromogenic method as a diagnostic method to distinguish HAE samples from healthy controls.


Yam genomics supports West Africa as a major cradle of crop domestication.
While there has been progress in our understanding of the origin and history of agriculture in sub-Saharan Africa, a unified perspective is still lacking on where and how major crops were domesticated in the region. Here, we investigated the domestication of African yam (Dioscorea rotundata), a key crop in early African agriculture. Using whole-genome resequencing and statistical models, we show that cultivated yam was domesticated from a forest species. We infer that the expansion of African yam agriculture started in the Niger River basin. This result, alongside with the origins of African rice and pearl millet, supports the hypothesis that the vicinity of the Niger River was a major cradle of African agriculture.


Physiological Response in a Specialist Paramedic During Helicopter Winch Rescue in Remote Wilderness and Extreme Heat.
Tasks performed by search and rescue (SAR) teams can be physically demanding. SAR organizations are faced with mounting challenges due to increased participation in recreation in remote locations and more frequent extreme weather. We sought to describe the physiological response and the methods for data collection during helicopter emergency medical service (HEMS) winch rescue from remote wilderness in extreme heat. A flight paramedic sustained 81% of maximum heart rate (VO₂ ~44.8 mL/kg/min) for ~10 minutes at a rate of perceived exertion of 19/20, and a relative heart rate of 77.5% in 37.1°C. Maximal acceptable work time for this task was calculated at 37.7 minutes. Our data collection methods were feasible, and the data captured demonstrated the level of physiological strain that may be encountered during HEMS SAR operations in austere environments and hot climate. It is essential that SAR teams that perform physically demanding tasks use a scientific approach to adapt and evolve. This is necessary to ensure personnel are appropriately selected, trained, and equipped to respond in an era of increasing demand and extreme environments.


Patches of different types for carotid patch angioplasty.
Carotid patches for carotid endarterectomy may be made from an autologous vein or synthetic material. To assess the safety and efficacy of different materials for carotid patch angioplasty. We searched the Cochrane Stroke Group trials register (last searched 3 August 2009), the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library Issue 3, 2009), MEDLINE (1966 to November 2008), EMBASE (1980 to November 2008) and Index to Scientific and Technical Proceedings (1980 to 2008). We handsearched relevant journals and conference proceedings, checked reference lists, and contacted experts in the field. Randomised and quasi-randomised trials comparing one type of carotid patch with another for carotid endarterectomy. Two review authors independently assessed eligibility, trial quality, and extracted data. We included 13 trials involving a total of 2083 operations; seven trials compared vein closure with PTFE closure, and six compared Dacron grafts with other synthetic materials. In most trials a patient could be randomised twice and have each carotid artery randomised to different treatment groups. There were no significant differences in the outcomes between vein patches and synthetic materials apart from pseudoaneurysms where there were fewer associated with synthetic patches than vein patches (odds ratio (OR) 0.09, 95% confidence interval (CI) 0.02 to 0.49). However, the numbers involved were small and the clinical significance of this finding is uncertain. Compared to other synthetic patches, Dacron was associated with a higher risk of: perioperative combined stroke and transient ischaemic attack (P = 0.03); restenosis at 30 days (P = 0.004); perioperative stroke (P = 0.07) and perioperative carotid thrombosis (P = 0.1). During follow-up for more than one year, there were also significantly more strokes (P = 0.03), stroke/death (P = 0.02) and arterial restenoses (P < 0.0001) with Dacron but the numbers of outcomes were small and the significance of this finding is uncertain. The number of outcome events is too small to allow reliable conclusions to be drawn and more trial data are required to establish whether any differences do exist. Nevertheless, there is some evidence that other synthetic (e.g. PTFE) patches may be superior to collagen impregnated Dacron grafts in terms of perioperative stroke rates and restenosis. Pseudoaneurysm formation may be more common after use of a vein patch compared with a synthetic patch.


Effects of emulsification, purity, and fluorination of silicone oil on human retinal pigment epithelial cells.
When silicone oil is used as a vitreous substitute, reproliferation of vitreoretinal membranes beneath the oil occurs frequently. Nevertheless, the effects of various properties of silicone oils such as purity, viscosity, fluorination, or emulsification on cellular proliferation have not been established. Human retinal pigment epithelial (RPE) cells were grown to confluence on filters, and then covered with silicone oil. The cellular monolayers were fed from below. At 72 hr and 14 days a proliferation index was determined by measuring 3H-thymidine incorporation into the cells. An assay for the enzyme gamma-glutamyl-transpeptidase (gamma GTP) was also done to assess cell polarization under some oils. A total of 14 different oils were studied. At 72 hr, emulsified oil was associated with significantly less proliferation than unemulsified oil, a difference that disappeared at 2 weeks. Neither fluorination nor viscosity had a significant effect on RPE proliferation. In addition, RPE proliferation indices were not significantly different from one another when purified oils were compared with most commercial-grade oils. However, a very contaminated oil was associated with a significantly higher proliferation index compared with severe purified or medical-grade oils.


A novel upstream enhancer of FOXP3, sensitive to methylation-induced silencing, exhibits dysregulated methylation in rheumatoid arthritis Treg cells.
Treg-cell function is compromised in rheumatoid arthritis (RA). As the master regulator of Treg cells, FOXP3 controls development and suppressive function. Stable Treg-cell FOXP3 expression is epigenetically regulated; constitutive expression requires a demethylated Treg-specific demethylated region. Here, we hypothesised that methylation of the FOXP3 locus is altered in Treg cells of established RA patients. Methylation analysis of key regulatory regions in the FOXP3 locus was performed on Treg cells from RA patients and healthy controls. The FOXP3 Treg-specific demethylated region and proximal promoter displayed comparable methylation profiles in RA and healthy-donor Treg cells. We identified a novel differentially methylated region (DMR) upstream of the FOXP3 promoter, with enhancer activity sensitive to methylation-induced silencing. In RA Treg cells we observed significantly reduced DMR methylation and lower DNA methyltransferase (DNMT1/3A) expression compared with healthy Treg cells. Furthermore, DMR methylation negatively correlated with FOXP3 mRNA expression, and Treg cells isolated from rheumatoid factor negative RA patients were found to express significantly higher levels of FOXP3 than Treg cells from RhF-positive patients, with an associated decrease in DMR methylation. In conclusion, the novel DMR is involved in the regulation of Treg-cell FOXP3 expression, but this regulation is lost post-transcriptionally in RA Treg cells.


Authentication of Primary Murine Cell Lines by a Microfluidics-Based Lab-On-Chip System.
The reliable authentication of cell lines is a prerequisite for the reproducibility and replicability of experiments. A common method of cell line authentication is the fragment length analysis (FLA) of short-tandem repeats (STR) by capillary electrophoresis. However, this technique is not always accessible and is often costly. Using a microfluidic electrophoresis system, we analyzed the quality and integrity of different murine cell lines by STR profiling. As a proof of concept, we isolated and immortalized hematopoietic progenitor cells (HPC) of various genotypes through retroviral transduction of the fusion of the estrogen receptor hormone-binding domain with the coding sequence of HoxB8. Cell lines were maintained in the HPC state with Flt3 ligand (FL) and estrogen treatment and could be characterized upon differentiation. In a validation cohort, we applied this technique on primary mutant Kras-driven pancreatic cancer cell lines, which again allowed for clear discrimination. In summary, our study provides evidence that FLA of STR-amplicons by microfluidic electrophoresis allows for stringent quality control and the tracking of cross-contaminations in both genetically stable HPC lines and cancer cell lines, making it a simple and cost-efficient alternative to traditional capillary electrophoresis.


Internal maxillary artery bypass for the treatment of complex middle cerebral artery aneurysms.
OBJECTIVEThe rapid innovation of the endovascular armamentarium results in a decreased number of indications for a classic surgical approach. However, a middle cerebral artery (MCA) aneurysm remains the best example of one for which results have favored microsurgery over endovascular intervention. In this study, the authors aimed to evaluate the experience and efficacy regarding surgical outcomes after applying internal maxillary artery (IMA) bypass for complex MCA aneurysms (CMCAAs).METHODSAll IMA bypasses performed between January 2010 and July 2018 in a single-center, single-surgeon practice were screened.RESULTSIn total, 12 patients (9 males, 3 females) with CMCAAs managed by high-flow IMA bypass were identified. The mean size of CMCAAs was 23.7 mm (range 10-37 mm), and the patients had a mean age of 31.7 years (range 14-56 years). The aneurysms were proximally occluded in 8 cases, completely trapped in 3 cases, and completely resected in 1 case. The radial artery was used as the graft vessel in all cases. At discharge, the graft patency rate was 83.3% (n = 10), and all aneurysms were completely eliminated (83.3%, n = 10) or greatly diminished (16.7%, n = 2) from the circulation. Postoperative ischemia was detected in 2 patients as a result of graft occlusion, and 1 patient presenting with subarachnoid hemorrhage achieved improved modified Rankin Scale scores compared to the preoperative status but retained some neurological deficits. Therefore, neurological assessment at discharge showed that 9 of the 12 patients experienced unremarkable outcomes. The mean interval time from bypass to angiographic and clinical follow-up was 28.7 months (range 2-74 months) and 53.1 months (range 19-82 months), respectively. Although 2 grafts remained occluded, all aneurysms were isolated from the circulation, and no patient had an unfavorable outcome.CONCLUSIONSThe satisfactory result in the present study demonstrated that IMA bypass is a promising method for the treatment of CMCAAs and should be maintained in the neurosurgical armamentarium. However, cases with intraoperative radical resection or inappropriate bypass recipient selection such as aneurysmal wall should be meticulously chosen with respect to the subtype of MCA aneurysm.