Automated classification of parenchymal patterns in mammograms.
A method for automated determination of parenchymal patterns in mammograms has been developed that is insensitive to changes in the mammographic imaging technique. The method was designed to study the relation between breast cancer risk and changes of mammographic density. It includes a new method for automatic segmentation of the pectoral muscle in oblique mammograms, based on application of the Hough transform. The technique developed for classification of parenchymal patterns is based on a distance transform that subdivides the breast tissue area into regions in which distance to the skin line is approximately equal. Features are calculated from grey level histograms computed in these regions. In this way, dependency on varying tissue thickness in the peripheral zone of the breast is minimized. Additional features represent differences between tissue projected in pectoral and breast area. Robustness and classification performance were studied on a test set of 615 digitized mammograms, applying a kNN classifier and leave-one-out for training. Using four density categories in 67% of the cases an exact agreement was obtained with a subjective classification made by a radiologist. The number of cases for which classifications of the radiologist and the program differed by more that one category was only 2%. For more recent mammograms, recorded after 1991, an exact agreement of 80% was obtained.


Direct emergence of the dorsospinal artery from the aorta supplying the anterior spinal artery: report of two cases.
We report two cases of an isolated dorsospinal artery that emerged directly from the aorta to supply the anterior spinal artery, which in one case had another blood supply on the opposite side, in a lower position. It is important to identify this anatomic variant, otherwise spinal angiography might be incomplete, especially when the isolated dorsospinal artery supplies the anterior spinal artery.


Clinical outcome in patients with paroxysmal or persistent atrial fibrillation receiving bepridil.
It is unknown whether bepridil improves cardiovascular events in atrial fibrillation (AF) patients, so this study evaluated the clinical outcome in paroxysmal or persistent AF patients receiving bepridil. We conducted a cohort study of 284 consecutive patients who received bepridil for AF (25% female, 5913 years) with a median follow-up period of 17 months (4-157 months). A total of 135 (48%) patients had structural heart disease, and 231 patients (81%) had previously received class I or class III antiarrhythmic drugs. The cumulative rates for cardiovascular events were 2.4%, 8.1%, and 10.1% at 1, 3, and 5 years, respectively. The cumulative rates for a composite of mortality, cerebral infarction, systemic embolism, major bleeding and heart failure were 9.7%, 18.2%, and 29.6% at 1, 3, and 5 years, respectively. The probability of progression to permanent AF was 23.5% at 5 years. Sudden death occurred in a patient with a prior myocardial infarction who was taking 200mg daily, and torsade de pointes (Tdp) occurred in two patients without structural heart disease taking 200mg daily. Excessive corrected QT interval prolongation (>0.50s) was observed when plasma concentrations were higher than 800 ng/ml. Bepridil might not improve the clinical outcome in refractory AF patients. Bepridil-related adverse events, including QT prolongation and Tdp, occurred in a dose- and concentration-dependent manner.


AI2Seg: A Method and Tool for AI-based Annotation Inspection of Biomedical Instance Segmentation Datasets.
In biomedical engineering, deep neural networks are commonly used for the diagnosis and assessment of diseases through the interpretation of medical images. The effectiveness of these networks relies heavily on the availability of annotated datasets for training. However, obtaining noise-free and consistent annotations from experts, such as pathologists, radiologists, and biologists, remains a significant challenge. One common task in clinical practice and biological imaging applications is instance segmentation. Though, there is currently a lack of methods and open-source tools for the automated inspection of biomedical instance segmentation datasets concerning noisy annotations. To address this issue, we propose a novel deep learning-based approach for inspecting noisy annotations and provide an accompanying software implementation, AI2Seg, to facilitate its use by domain experts. The performance of the proposed algorithm is demonstrated on the medical MoNuSeg dataset and the biological LIVECell dataset.


Deuterium Depleted Water Inhibits the Proliferation of Human MCF7 Breast Cancer Cell Lines by Inducing Cell Cycle Arrest.
Recent studies have shown that the depletion of naturally occurring deuterium can result in tumor regression. The aim of the present study was to show the growth inhibitory effects of DDW discretely and in combination with 5-FU on MCF-7 breast cancer cells and to determine possible mechanisms underlying these changes. MCF7 cells were grown in RPMI medium with decreasing deuterium concentrations of DDW individually, 5-FU alone and both for 24, 48, and 72 h. Cell viability was determined with the MTT assay. The cell cycle and antioxidant enzymes status were measured using flow cytometry and quantitative luminescence methods, respectively. Our results showed that treatment with DDW especially in 30-100 ppm concentrations imposed the highest cell growth inhibitory effect. The cell cycle analysis revealed that DDW caused the cell cycle arrest in the G1/S transition, reduced the number of the cells in the S phase and significantly increased the population of cells in the G1 phase in MCF-7 cells. The activities of superoxide dismutase (SOD) and catalase (CAT) enzymes also increased in the same low concentrations of DDW. In conclusion, DDW can open new strategic approach in breast cancer therapy. Highlights DDW cause lethality in cancer cells. DDW augmented 5-FU inhibitory effects on breast cancer cell lines. Cell inhibitory results lead to the discovery of synergic effects of DDW-drug combinations Synergistic anti cancer effects of DDW with 5-FU is enhanced by decreasing deuterium content of the DDW. DDW exerts effects on the cell cycle, changes in cell configuration and induces antioxidant enzymes in vitro. DDW can be considered as an adjuvant to conventional anticancer agents in future trials.


On the Path to Thermo-Stable Collagen: Culturing the Versatile Sponge Chondrosia reniformis.
The collagen proteins family is sought-after in the pharmaceuticals, cosmetics, and food industries for various biotechnological applications. The most abundant sources of collagen are pigs and cows, but due to religious restrictions and possible disease transmission, they became less attractive. An alternative source can be found in marine invertebrates, specifically in sponges. Alas, two problems arise: (1). Growing sponges is complicated. (2). Sponge collagen has low heat tolerance, which can impose a problem for human biotechnological usage. To fill these gaps, we studied the collagen-abundant sponge Chondrosia reniformis. Two culture experiments were conducted: (1). A sea-based system examined the difference in growth rates of C. reniformis from different habitats, growing under natural seasonal conditions; (2). A land-based controlled system, which assessed the growth-rate of C. reniformis at different temperatures. The results reveal that C. reniformis from shallow habitats are growing larger and faster than individuals from colder, deeper habitats, and that the optimal temperature for C. reniformis growth is 25 °C. The results demonstrate that C. reniformis is highly fit for culture and can produce thermally stable collagen. Further research is needed to determine the best conditions for C. reniformis culture for collagen extract and other exciting materials for bioprospecting.


Feasibility of localized immunosuppression: 1. Exploratory studies with glucocorticoids in a biohybrid device designed for cell transplantation.
Emerging biotechnologies, such as the use of biohybrid devices for cellular therapies, are showing increasing therapeutic promise for the treatment of various diseases, including type 1 diabetes mellitus. The functionality of such devices could be greatly enhanced if successful localized immunosuppression regimens could be established, since they would eliminate the many otherwise unavoidable side effects of currently used systemic immunosuppressive therapies. The existence of local immune privilege at some specialized tissues, such as the eye, CNS, or pregnant uterus, supports the feasibility of localized immunomodulation, and such an approach is particularly well-suited for cell transplant therapies where all transplanted tissue is localized within a device. Following the success of syngeneic transplantation in a subcutaneous prevascularized device as a bioartificial pancreas in a rodent model, we now report the first results of exploratory in vivo islet allograft studies in rats using locally delivered glucocorticoids (dexamethasone phosphate and the soft steroid loteprednol etabonate). Following in vitro assessments, in silico drug distribution models were used to establish tentative therapeutic dose ranges. Sustained local delivery was achieved via implantable osmotic mini-pumps through a central sprinkler, as well as with a sustained-delivery formulation for loteprednol etabonate using poly(D,L-lactic) acid (PLA) microspheres. Doses delivered locally were approximately hundred-fold smaller than those typically used in systemic treatments. While several solubility, stability, and implantation problems still remain to be addressed, both compounds showed promise in their ability to prolong graft survival after tapering of systemic immunosuppression, compared to control groups.


Therapeutic pancreatic endoscopy.
Therapeutic endoscopy in pancreatitis has included biliary or pancreatic sphincterotomy, or both, for microlithiasis and sphincter dysfunction. Alternatively, endoscopic treatment has been used to treat ductal disruption, including external or internal fistulas (high amylase effusions, pancreatic ascites, pancreaticoenteric communications, or pseudocysts). Finally, endoscopy has been used to treat obstructive disease, including both stones and strictures. This article reviews the state of the art in this field over the past year. Unfortunately, this is an area in which the appropriate approaches are still poorly defined, and prospective studies randomly assigning patients to endoscopic treatment in comparison with medical or surgical therapy remain few and far between.


Heparin requirements for full anticoagulation are higher for patients on dabigatran than for those on warfarin - a model-based study.
Dabigatran (D) is increasingly used for chronic anticoagulation in place of warfarin (W). These patients may present for catheter-based procedures requiring full anticoagulation with heparin. This study compares the heparin sensitivity of patients previously on dabigatran, on warfarin, or on no chronic anticoagulant during ablation of atrial fibrillation. In a retrospective study of patients treated with D, W, or neither drug (N) undergoing atrial ablation, the timing of heparin doses and resulting activated clotting times were collected. First, the initial activated clotting time response to the first heparin bolus was compared. Then, a non-linear mixed effects modelling (NONMEM) analysis was performed, fitting a pharmacokinetic and -dynamic model to the entire anticoagulation course of each patient. Resulting model coefficients were used to compare the different patient groups. Data for 66 patients on dabigatran, 95 patients on warfarin, and 27 patients on no anticoagulation were retrieved. The last dose of dabigatran or warfarin had occurred 27 hours and 15 hours before the procedure. Groups D and N both responded significantly less (P<0.05) to the initial heparin bolus than Group W (approximately 50%). Likewise, the model coefficients resulting from the fit to each group reflected a significantly lower heparin sensitivity in groups D and N compared to W. Clearances of the heparin effect in the model did not differ significantly among groups. Patients on warfarin with an average INR of 1.5 or higher are more sensitive to heparin than patients not previously anticoagulated or patients who discontinued dabigatran 27 hours earlier (approximately two half-lives) warfarin.


Rhinovirus infections in the upper airway.
The majority of cold and flulike illnesses are caused by human rhinoviruses (HRVs). Improved detection of HRV has shown that HRVs are also associated with more serious illness, such as exacerbation of asthma, wheezing illnesses in children, chronic obstructive pulmonary disease, cardiopulmonary disease, and fatal pneumonia in immune-compromised patients. HRV is a major cause of acute viral respiratory tract infections in hospitalized children and is among the leading causes of childhood mortality worldwide. Detection of the HRV genome by reverse transcriptase-polymerase chain reaction and genomic sequencing has brought to light a new clade, HRV-C, to the already recognized HRV-A and HRV-B clades. The clinical complications related to all rhinovirus infections include acute otitis media, acute sinusitis, and acute bronchitis. The enormous public health implications from those diseases far overshadow those of the common cold. This article provides an overview of the pathogenesis of rhinovirus infection in the upper airways. Most research has been done in young healthy adults with self-limiting experimental and natural rhinovirus infections, and this may set the stage for understanding rhinovirus infections in the ear, sinus, and lower airways.


Increase in acute malnutrition in children following the 2014-2015 Ebola outbreak in rural Sierra Leone.
Setting: All health facility and community malnutrition screening programmes in Tonkolili, a rural Ebola-affected district in Sierra Leone. Objectives: Before the Ebola disease outbreak, Sierra Leone had set a goal to reduce the prevalence of severe acute malnutrition (SAM) in children aged <5 years to <0.2%. We compared the number of children screened, diagnosed and treated for malnutrition before, during and after the outbreak (2013-2016). Design: This was a retrospective cross-sectional study. Results: Health facility screening declined from 16 805 children per month pre-outbreak to 13 510 during the outbreak (P = 0.02), and returned to pre-outbreak levels after the outbreak. Community-based screening remained stable during the outbreak, and increased by 30% post-outbreak (P < 0.001). The proportion diagnosed with moderate acute malnutrition using mid-upper arm circumference increased from respectively 3.6% and 5.1% pre-outbreak in the community and health facilities to 8.2% and 7.9% post-outbreak (P < 0.001, P = 0.003). The proportion of children diagnosed with SAM using a weight-for-age ratio at health facilities increased from 1.5% pre-outbreak to 3.5% post-outbreak (P = 0.003). On average, for every four children diagnosed with SAM per month, one child completed SAM treatment. Conclusion: After a decline in screening during the Ebola outbreak, diagnoses of acute malnutrition increased post-outbreak. Nutrition programmes need to be strengthened to pre-empt such effects in the event of future Ebola outbreaks.


[Diagnostic significance of pathologic synkinesis for detection of pyramidal pathology].
Five types of pathological synkinesis (++blepharo-ocular, ++blepharo-facial, ++bucco-manual, ++digito-digital on the hands, ++pedo-digital) are described. They are of definite importance for revealing pyramidal pathology including its early stages as well as for objective evaluation and observation of the time-course of changes in the illness.


Flavonoids as Human Intestinal α-Glucosidase Inhibitors.
Certain flavonoids can influence glucose metabolism by inhibiting enzymes involved in carbohydrate digestion and suppressing intestinal glucose absorption. In this study, four structurally-related flavonols (quercetin, kaempferol, quercetagetin and galangin) were evaluated individually for their ability to inhibit human α-glucosidases (sucrase, maltase and isomaltase), and were compared with the antidiabetic drug acarbose and the flavan-3-ol(-)-epigallocatechin-3-gallate (EGCG). Cell-free extracts from human intestinal Caco-2/TC7 cells were used as the enzyme source and products were quantified chromatographically with high accuracy, precision and sensitivity. Acarbose inhibited sucrase, maltase and isomaltase with IC50 values of 1.65, 13.9 and 39.1 µM, respectively. A similar inhibition pattern, but with comparatively higher values, was observed with EGCG. Of the flavonols, quercetagetin was the strongest inhibitor of α-glucosidases, with inhibition constants approaching those of acarbose, followed by galangin and kaempferol, while the weakest were quercetin and EGCG. The varied inhibitory effects of flavonols against human α-glucosidases depend on their structures, the enzyme source and substrates employed. The flavonols were more effective than EGCG, but less so than acarbose, and so may be useful in regulating sugar digestion and postprandial glycaemia without the side effects associated with acarbose treatment.


Multidisciplinary care for people with Parkinson's disease: the new kids on the block!
Parkinson's disease (PD) is a chronic multisystem disorder that causes a wide variety of motor and non-motor symptoms. Over time, the progressive nature of the disease increases the risk of complications such as falls and loss of independence, having a profound impact on quality of life. The complexity and heterogeneity of symptoms therefore warrant a holistic, multidisciplinary approach. Specific healthcare professionals, e.g. the movement disorders neurologist and the PD nurse specialist, are considered essential members of this multidisciplinary team. However, with our increasing knowledge about different aspects of the disease, other disciplines are also being recognized as important contributors to the healthcare team. Areas covered: The authors describe a selection of these relatively newly-recognized disciplines, including the specialist in vascular medicine, gastroenterologist, pulmonologist, neuro-ophthalmologist, urologist, geriatrician/elderly care physician, palliative care specialist and the dentist. Furthermore, they share the view of a person with PD on how patients and caregivers should be involved in the multidisciplinary team. Finally, they have included a perspective on the new role of the movement disorder neurologist, with care delivery via 'tele-neurology'. Expert commentary: Increased awareness about the potential role of these 'new' professionals will further improve disease management and quality of life of PD patients.


Conduction studies of the medial cutaneous nerve of the forearm.
The forearm medial cutaneous nerve is a pure sensory branch from the medial cord of the brachial plexus. Its fibers are derived from the eighth cervical and first thoracic nerves. A simple, easily reproducible antidromic technique for studying conduction of the forearm medial cutaneous nerve is described with the knowledge (obtained from cadaver dissections) of its exact topography. Sixty nerves were studied in 30 able-bodied adults, 15 women and 15 men. Surface stimulation was done over the medial aspect of the middle of the arm at a level where the nerve pierces the deep fascia. Surface recordings were made 18cm distally over the course of its volar branch. Action potentials were obtained in 100% of the subjects without electronic averaging. Mean values obtained were as follows: latency to onset 2.73 +/- 0.17 msec; latency to peak 3.31 +/- 0.19 msec; conduction velocity of the fastest fibers 65.9 +/- 4.3 m/s; amplitude 15.4 +/- 4.1 microvolts, and the mean difference in latency between the right and left nerves in the same subject was 0.1 msec. Conduction studies of this nerve should be useful in electrodiagnostic evaluation of peripheral neuropathy (particularly in below amputees), local neuropathic conditions, and entrapment syndromes involving the medial cord of the brachial plexus.


Efficacy of the lactate dehydrogenase (LDH)/lymphocyte ratio (LLR) to reduce the need for X-ray in pregnant patients with COVID-19.
Pregnancy carries a significant risk for coronavirus disease-2019 (COVID-19) due to natural immunosuppression. A previous study from our center has shown that the lactate dehydrogenase (LDH)/lymphocyte ratio (LLR) can be used in the early diagnosis of COVID-19 and predicting mortality. Based on this, we aimed to determine the effect of LLR on early detection of critical pregnant women and mortality in COVID-19. The data of 145 patients who were admitted to our hospital between March and December 2020; diagnosed with COVID-19 and hospitalized, were retrospectively analyzed. The median gestation period was 31 weeks (range: 5-41), 30.3% (n: 44) gave birth and 68.3% (n: 99) were pregnant. Median LLR was 0.13 (range: 0.04-0.70). The rate of cough (47% vs. 22.8%; p=0.003) was found to be high in patients with LLR>0.13. The patients were divided into subgroups. The proportion of patients without active complaints was higher in the Q1, followed by the Q4. The proportion of patients with an initial complaint of cough increased as LLR from Q1 to Q4, the distribution of other complaints did not differ between the quartiles. The higher rate of cough in the group with high LLR indicates that it may be an important indicator of lung involvement during pregnancy. The highest rate of non-treatment follow-up in the lowest LLR group proved that the LLR value at the time of diagnosis can be used as an important clinical marker in pregnant women.


Evaluating Chondroitin Sulfate and Dermatan Sulfate Expression in Glial Scar to Determine Appropriate Intervention Time in Rats.
The proteoglycans of the extracellular matrix increases in the glial scar during spinal cord injury and significantly affects the inhibition of axonal regeneration. The results of injury therapies are limited due to the lack of identifying a timely therapeutic intervention. The present study aimed to investigate the glial scar Chondroitin Sulfate (CS) and Dermatan Sulfate (DS) levels at different post-injury times to determine the appropriate time for therapeutic intervention. By this experimental study, 72 Wistar rats were randomly divided into 12 groups, as follows: control, sham, injured animals at 1, 2, 4, and 8 days, as well as 2, 4, 8, 12, 16, and 20 weeks post-injury. The animals in the injured groups were contused in the T10 segment of the spinal cord. The motor function of animals was assessed using the Basso, Beattie, and Bresnahan (BBB) test. Besides, the histological assessment was performed using Luxol Fast Blue and Bielshovisky Staining. The CS and DS levels of lesions were measured using the Enzyme-Linked Immunosorbent Assay (ELISA) method. The motor function assessment indicated a relative recovery over time. Histological results confirmed some regeneration in the injury site at 20 weeks post-injury. The ELISA results demonstrated a much higher level of DS than that of CS in the glial scar. Considering high levels of DS, compared to CS in the glial scar and its reduction from second weeks after SCI onwards, the second week after SCI seems to be the best time for therapeutic interventions in terms of scar permeability.


[Dysbacteriosis--infectious processes of mixed etiology].
At present dysbacteriosis or altered normal and sometimes very individual proportion of microorganisms in various macroorganism microbiotopes is considered as an infectious disease since it results from pathogenic action of microbes, is characterized by a set of features relating to pyoinflammatory processes and even can be contagious.


The timing and sequence of appearance of the distal branches of the human embryonic vestibular ganglion.
Vestibular ganglion was investigated in human embryos during developmental stages l3-23 (34-56 postovulatory days). Branches of the vestibular ganglion forming ampullar nerves were observed in ernbryos at stage I5. During the last embryonic stages (stage 22 and 23) the utriculo-ampullar, posterior ampullar, and saccular nerves were clearly distinguished.


Ultrastructural immunocytochemical localization of the N-methyl-D-aspartate receptor and tyrosine hydroxylase in the shell of the rat nucleus accumbens.
The N-methyl-D-aspartate (NMDA)-type glutamate receptors in the shell region of the nucleus accumbens (ACB) have been implicated in the modulation of dopamine release and in amphetamine-induced neurotoxicity. We used electron microscopic immunocyto-chemistry to determine the anatomical sites for NMDA-mediated effects of glutamate and for their potential interactions with dopaminergic afferents identified by the presence of tyrosine hydroxylase (TH) in this region of the rat brain. Immunogold and immunoperoxidase methods were used to localize antisera against the R1 subunit of the NMDA receptor (NMDAR1) alone or combined with TH. In single labeling experiments, approximately half of the NMDAR1-like immunoreactivity (NMDAR1-LI) was localized to extrasynaptic plasma membranes of neuronal processes, many (92 out of 215) of which were dendrites, and only 33 out of 215 were unmyelinated axons or terminals. Surprisingly, the neuronal labeling of NMDAR1 was almost equaled by that seen in astrocytic processes (88 out of 215). Dual labeling for TH and NMDAR1 was rarely observed and was only seen in axons. However, in favorable planes of section, NMDAR1 was noted along intervaricose segments of axons in which TH was more readily seen in the varicosity. This differential intra-axonal distribution suggests an underestimation of dual labeling in single coronal sections through unmyelinated axons and terminals. The TH-immunoreactive terminals were more often seen apposed to NMDA-immunoreactive astrocytic processes and dendrites. These results provide the first ultrastructural evidence for presynaptic modulation of dopamine release by NMDA receptors in the shell of the nucleus accumbens. They also indicate that NMDA receptors modulate postsynaptic neurons receiving input from the dopaminergic afferents and suggest a previously unsuspected functional association involving glial NMDA receptors and dopaminergic afferents in this brain region.


Dual function of Slit2 in repulsion and enhanced migration of trunk, but not vagal, neural crest cells.
Neural crest precursors to the autonomic nervous system form different derivatives depending upon their axial level of origin; for example, vagal, but not trunk, neural crest cells form the enteric ganglia of the gut. Here, we show that Slit2 is expressed at the entrance of the gut, which is selectively invaded by vagal, but not trunk, neural crest. Accordingly, only trunk neural crest cells express Robo receptors. In vivo and in vitro experiments demonstrate that trunk, not vagal, crest cells avoid cells or cell membranes expressing Slit2, thereby contributing to the differential ability of neural crest populations to invade and innervate the gut. Conversely, exposure to soluble Slit2 significantly increases the distance traversed by trunk neural crest cells. These results suggest that Slit2 can act bifunctionally, both repulsing and stimulating the motility of trunk neural crest cells.


The serum- and glucocorticoid-induced kinase is a physiological mediator of aldosterone action.
Aldosterone plays a major role in regulating sodium and potassium flux in epithelial tissues such as kidney and colon. Recent evidence suggests that serum- and glucocorticoid-regulated kinase (SGK) is induced by aldosterone and acts as a key mediator of aldosterone action in epithelial tissues. Induction of SGK messenger RNA (mRNA) has previously been shown within 30 min of addition of supraphysiological doses of aldosterone to Xenopus A6 cells and within 4 h in rat kidney in vivo. In this study we determined the time course of SGK induction, at doses of aldosterone in the physiological range, in rat kidney and colon, using Northern and Western blot analyses and in situ hybridization and determined concurrent changes in urinary sodium and potassium excretion by Kagawa bioassay. On Northern blot analysis, SGK mRNA levels were significantly elevated in both kidney and colon 60 min after the injection of aldosterone. SGK protein in late distal colon was significantly elevated 2 and 4 h after aldosterone treatment. In situ hybridization showed SGK mRNA to be induced in renal collecting ducts and distal tubular elements in both cortex and medulla by doses of aldosterone of 0.1 microg/100 g BW or more within 30 min of steroid treatment. Significant changes in urinary composition were similarly seen with an aldosterone dose of 0.1 microg/100 g BW from 90 min after aldosterone injection. The early onset of SGK induction in kidney and colon and the correlation with urinary changes in terms of both time course and dose response suggest that SGK plays an important role in mediating the effects of aldosterone on sodium homeostasis in vivo.


Antibodies to rat soluble IL-6 receptor stimulate B9 hybridoma cell proliferation.
Interleukin-6 mediates its pleiotropic effects by interacting with its membrane bound receptor (gp80) or the soluble counterpart gp54, resulting in activation of a complex that includes the transducer protein gp130. We have generated a polyclonal antibody against the rat soluble IL-6 receptor (anti-rat sIL-6R) in rabbits. By Western blot analysis we show that purified anti-rat sIL-6R IgG antibody reacts specifically with recombinant rat sIL-6R generated from E. coli, baculovirus or adenovirus expression systems. Anti-rat sIL-6R inhibited IL-6-induced acute phase protein synthesis in rat (H35) but not human (HepG2) hepatoma cells, and did not affect stimulation of those cells by Oncostatin-M. Conversely, on the mouse hybridoma B9 cell line, IgG anti-rat sIL-6R showed a dose-dependent stimulation of proliferation. Fab fragments of this antibody did not stimulate, but abrogated IL-6-mediated hepatoma cell stimulation and B9 cell proliferation. Gel shift analysis of STAT nuclear factors showed activation of STAT DNA binding in nuclei of B9 cells treated with IgG anti-rat sIL-6R, whereas in H35, NIH-3T3 and M1 cells, only IL-6 could trigger a similar STAT activation. Our data suggest that mechanisms of IL-6 receptor activation and signalling in mouse B9 hybridoma cells show subtle but important differences from other IL-6-responsive cells.


How effective has the Clean Water Act been at reducing pollutant mass emissions to the Southern California Bight over the past 35 years?
The Clean Water Act (CWA) has regulated discharges of contaminants since 1972. However, evaluations of the CWA's effectiveness at improving regional water quality are lacking, primarily because integration of monitoring data from multiple dischargers to assess cumulative effects is not required. A rare opportunity exists to assess CWA effectiveness by integrating mass emissions data from all major sources of contaminants to the Southern California Bight from 1971 to 2000. While the coastal population grew by 56% and total effluent volume increased 31% since 1971, mass emissions of nearly all constituents decreased since passage of the CWA, most by greater than 65%. Publicly owned treatment works were the dominant point source of many contaminants, but also accounted for the greatest reductions in pollutant discharge since 1971. As point source treatment has improved, the relative contribution of non-point sources, such as storm water runoff has increased. Despite the increased importance of storm water discharges, regional monitoring and data compilation of this source is lacking, making it difficult to accurately assess trends in non-point source discharge.


Giardiasis and diarrhea in dogs: Does the microbiome matter?
Giardia duodenalis (Gd) causes intestinal parasitosis. The involvement of the intestinal microbiome in determining the infection's clinical phenotype is unknown. Investigate the fecal microbiome features in dogs with giardiasis. Cross-sectional study, including fecal samples of kenneled dogs with Gd diagnosed by fecal Giardia antigen dot ELISA. The fecal microbial compositional characteristics and dysbiosis index (DI) were compared between diarrheic and nondiarrheic dogs. Fecal samples of 38 Gd-infected dogs (diarrheic, 21; nondiarrheic, 17) were included. No differences were found in Faith's phylogenic diversity and beta diversity (weighted UniFrac distances) and in specific taxa abundances at the phylum, genus, and species levels, as well as in alpha and beta diversities between diarrheic and nondiarrheic dogs, and also when divided by sex or age. Among diarrheic dogs, alpha diversity was higher in males than in females (pairwise Kruskal-Wallis, q = 0.01). Among males, fecal abundances of the genus Clostridium (W = 19) and Clostridium spiroforme species (W = 33) were higher in diarrheic compared to nondiarrheic dogs. In diarrheic dog fecal samples, Proteobacteria were more prevalent (W = 1), whereas Verrucomicrobia were less prevalent in dogs <1 year of age than in older dogs. The fecal sample DI of 19 diarrheic and 19 nondiarrheic dogs was similar (median, -0.2; range, -4.3 to 4.5 and median, -1.0; range, -4.3 to 5.8, respectively). The fecal microbial composition of symptomatic and asymptomatic dogs with giardiasis is similar. Based on fecal DI, giardiasis is not characterized by prominent dysbiosis. Other host and parasite characteristics might determine the severity of giardiasis in dogs.


Clinical factors associated with work disability in epilepsy: A cross-sectional study at a tertiary referral hospital.
The impact of epilepsy on work disability remains unclear. The aim of this study was to determine the percentage of patients with epilepsy who are unemployed or on temporary or permanent disability leave and to analyze associated clinical factors. We performed an observational cross-sectional study of consecutively recruited patients with epilepsy seen at a specialized epilepsy unit or admitted to the epilepsy monitoring unit of a tertiary referral hospital. We analyzed the percentage of patients who were actively employed, unemployed, and on temporary or permanent disability leave. The groups were compared for sociodemographic data (age, sex, marital status, and type of work), clinical data (type of epilepsy, disease duration, monthly seizure frequency, and presence of anxiety or depression), treatment-related factors, and quality of life. We included 742 patients (53% male, mean age 44.3 ± 13.7 years old): 40.5% were employed, 29.2% were unemployed, 19% were on temporary disability leave, and 11.1% had a permanent work disability. Depressive symptoms and poorer quality of life were associated with unemployment (OR 2.3, p = 0.02 and OR 1.8, p = 0.01), temporary disability leave (OR 1.4, p = 0.05 and OR 1.7, p = 0.02), and permanent work disability (OR 1.9, p = 0.01 and OR 2.2, p = 0.01). Low-skilled work was also predictive of unemployment (OR 1.9, p = 0.04), temporary disability leave (OR 2.8, p = 0.03), and permanent work disability (OR 1.7, p = 0.04). A higher monthly seizure frequency was associated with permanent work disability (OR 2.01, p = 0.02). Less than 50% of patients with epilepsy in our setting are working. Factors associated with unemployment and work disability are a higher frequency of seizures, low-skilled work, depressive symptoms, and poor quality of life.


Chylous ascites following abdominal aortic surgery.
Three patients, two women, one man (mean age 74 years), who had abdominal aortic aneurysms (2) or aortobifemoral surgery (1), developed chylous ascites postoperatively. They were studied to determine their clinical course and develop a plan for management of this complication. In each patient, the ascites was not manifest until abdominal swelling developed two weeks after operation, and the problem was confirmed by the finding of milky fluid on paracentesis. A low serum albumin (mean 2.6 gm) was also characteristic. The ascites was not altered by parenteral nutrition or reduction of dietary fat and ingestion of medium chain triglycerides. In one patient (man, age 93) the ascites resolved spontaneously two months after abdominal aortic aneurysm surgery. Another (woman, age 70) was cured following operative ligation of a lymphatic fistula identified at operation five weeks after abdominal aortic aneurysm repair. In the third (woman, age 60), the ascites resolved immediately following placement of a peritoneal venous shunt six weeks after an aortobifemoral bypass. Chylous ascites is rare after aortic surgery and manifests itself about two weeks after operation, at times after discharge from hospital. It has an indolent course, but may resolve spontaneously up to two months after operation. Its course appears not to be foreshortened by diet, including omission of fat, but can be successfully treated surgically with a shunt or fistula ligation. If done early a protracted hospital course may be avoided.


Compliance with Smoke-Free Policy and Challenges in Implementation: Evidence from Bengkulu, Indonesia.
Smoking is among the top contributors to deaths and disability-adjusted life years in Indonesia, particularly among males. In 2012, a presidential decree encouraged provinces and districts to implement a smoke-free policy (SFP). This study aims to evaluate compliance and explore the challenges in the implementation. Through a mixed-methods design, we used quantitative methods to examine the compliance with six criteria including signage, no active smoking, no selling, no advertisement, no smoke, and no ashtray at SFP facilities in Bengkulu city. We observed SFP compliance at 105 facilities, including health/educational facilities, places of worship, workplaces, and indoor/outdoor public facilities. We also used a qualitative method to explore challenges in the implementation through interviews with the government and legislators. The compliance rate to all six criteria was 38% overall, ranging from 17% at outdoor public facilities to 67% at health facilities. We found no spatial patterning, as shown by non-significant differences in compliance rates between SFP facilities inside and outside of 1-kilometer around the provincial and city health offices. Implementation challenges included lack of sensitization, lack of coordination, and limited budget. The compliance was relatively low due to several challenges, which could serve as a tobacco control policy lesson in a lower-middle-income country.


Efficient singlet fission discovered in a disordered acene film.
Singlet exciton fission is a process that occurs in select organic semiconductors and entails the splitting of a singlet excited state into two lower triplet excitons located on adjacent chromophores. Research examining this phenomenon has recently seen a renaissance due to the potential to exploit singlet fission within the context of organic photovoltaics to prepare devices with the ability to circumvent the Shockley-Queisser limit. To date, high singlet fission yields have only been reported for crystalline or polycrystalline materials, suggesting that molecular disorder inhibits singlet fission. Here, we report the results of ultrafast transient absorption and time-resolved emission experiments performed on 5,12-diphenyl tetracene (DPT). Unlike tetracene, which tends to form polycrystalline films when vapor deposited, DPT's pendant phenyl groups frustrate crystal growth, yielding amorphous films. Despite the high level of disorder in these films, we find that DPT exhibits a surprisingly high singlet fission yield, with 1.22 triplets being created per excited singlet. This triplet production occurs over two principal time scales, with ~50% of the triplets appearing within 1 ps after photoexcitation followed by a slower phase of triplet growth over a few hundred picoseconds. To fit these kinetics, we have developed a model that assumes that due to molecular disorder, only a subset of DPT dimer pairs adopt configurations that promote fission. Singlet excitons directly excited at these sites can undergo fission rapidly, while singlet excitons created elsewhere in the film must diffuse to these sites to fission.


Manipulation of central blood volume and implications for respiratory control function.
The respiratory operating point (ventilatory or arterial PCO2 response) is determined by the intersection point between the controller and plant subsystem elements within the respiratory control system. However, to what extent changes in central blood volume (CBV) influence these two elements and the corresponding implications for the respiratory operating point remain unclear. To examine this, 17 apparently healthy male participants were exposed to water immersion (WI) or lower body negative pressure (LBNP) challenges to manipulate CBV and determine the corresponding changes. The respiratory controller was characterized by determining the linear relationship between end-tidal PCO2 (PetCO2 ) and minute ventilation (Ve) [Ve = S × (PetCO2 - B)], whereas the plant was determined by the hyperbolic relationship between Ve and PetCO2 (PetCO2 = A/Ve + C). Changes in Ve at the operating point were not observed under either WI or LBNP conditions despite altered PetCO2 (P < 0.01), indicating a moving respiratory operating point. An increase (WI) and a decrease (LBNP) in CBV were shown to reset the controller element (PetCO2 intercept B) rightward and leftward, respectively (P < 0.05), without any change in S, whereas the plant curve remained unaltered at the operating point. Collectively, these findings indicate that modification of the controller element rather than the plant element is the major factor that contributes toward an alteration of the respiratory operating point during CBV shifts.


Helping and Cooperation at 14 Months of Age.
Two experiments investigated the proclivity of 14-month-old infants (a) to altruistically help others toward individual goals, and (b) to cooperate toward a shared goal. The infants helped another person by handing over objects the other person was unsuccessfully reaching for, but did not help reliably in situations involving more complex goals. When a programmed adult partner interrupted a joint cooperative activity at specific moments, infants sometimes tried to reengage the adult, perhaps indicating that they understood the interdependency of actions toward a shared goal. However, as compared to 18- and 24-month-olds, their skills in behaviorally coordinating their actions with a social partner remained rudimentary. Results are integrated into a model of cooperative activities as they develop over the 2nd year of life.


Guide: a desktop application for analysing gene expression data.
Multiplecompeting bioinformatics tools exist for next-generation sequencing data analysis. Many of these tools are available as R/Bioconductor modules, and it can be challenging for the bench biologist without any programming background to quickly analyse genomics data. Here, we present an application that is designed to be simple to use, while leveraging the power of R as the analysis engine behind the scenes. Genome Informatics Data Explorer (Guide) is a desktop application designed for the bench biologist to analyse RNA-seq and microarray gene expression data. It requires a text file of summarised read counts or expression values as input data, and performs differential expression analyses at both the gene and pathway level. It uses well-established R/Bioconductor packages such as limma for its analyses, without requiring the user to have specific knowledge of the underlying R functions. Results are presented in figures or interactive tables which integrate useful data from multiple sources such as gene annotation and orthologue data. Advanced options include the ability to edit R commands to customise the analysis pipeline. Guide is a desktop application designed to query gene expression data in a user-friendly way while automatically communicating with R. Its customisation options make it possible to use different bioinformatics tools available through R/Bioconductor for its analyses, while keeping the core usage simple. Guide is written in the cross-platform framework of Qt, and is freely available for use from http://guide.wehi.edu.au.


Reliability and concurrent validity of the Infant Motor Profile.
The Infant Motor Profile (IMP) is a qualitative assessment of motor behaviour in infancy. It consists of five domains: movement variation, variability, fluency, symmetry, and performance. The aim of this study was to assess interobserver reliability and concurrent validity of the IMP with the Alberta Infant Motor Scale (AIMS) and an age-specific neurological examination. Fifty-nine preterm infants (25 females, 34 males; median gestational age 29.7wks, median birthweight 1285g) and 146 term infants (74 females, 72 males; median gestational age 40.1wks, birthweight 3500g) were included. Assessments were performed at corrected ages of 4, 6, 10, 12, and 18 months and consisted of the IMP, AIMS, and an age-specific neurological examination. Interobserver reliability was investigated on a sample of 25 video recordings. Non-parametric statistics were used to analyse the data. Interobserver reliability was high (intraclass correlation coefficient 0.95). At all ages, AIMS scores correlated weakly to fairly with total IMP scores (Spearman's ρ 0.36-0.55), but moderately to strongly with scores on the performance domain of the IMP (Spearman's ρ 0.47-0.84). A clear relation was found between total IMP score and outcome of the neurological examination (Kruskal-Wallis p<0.001 at all ages). Interobserver reliability of the IMP is good. Concurrent validity with the AIMS is best for the IMP performance domain. Concurrent validity with age-specific neurological examination is very good.


Emergency management of pediatric convulsive status epilepticus: a multicenter study of 542 patients.
To perform a multicenter study examining the presentations and emergency management of children with convulsive status epilepticus (CSE) to sites within the Paediatric Research in Emergency Departments International Collaborative. Retrospective review of children presenting to emergency departments (EDs) with convulsive seizures of at least 10 minutes' duration. Eight sites within the Paediatric Research in Emergency Departments International Collaborative network in Australia and New Zealand participated. Patients were identified through a search of ED electronic records for the period January 2000 to December 2004. Data were obtained from 542 eligible episodes of CSE. Demographics and seizure history were similar across all sites. One third of children with CSE presented with their first seizure. A preexisting diagnosis that predisposed to seizures was present in 59%. Median duration of seizures before hospitalization was 45 minutes, and median duration of treatment in ED before termination was 30 minutes. Prehospital duration did not seem to influence the timing of key ED interventions such as the administration of second-line anticonvulsants or progression to rapid sequence induction (RSI) of anesthesia and intubation. Convulsive status epilepticus was terminated after first-line treatment in 42%, second-line treatment in 35%, and RSI in 22%. One third of the patients had persistent seizure activity beyond 40 minutes of ED treatment. Marked variation in the use of RSI for refractory seizures was observed between sites. Convulsive status epilepticus is an important neurological emergency, with many children experiencing prolonged seizures in both the prehospital and hospital phases. Persistent seizure activity beyond 40 minutes contrasts with current published guidelines. There is a need to adopt a widely accepted approach to the management of children who fail to respond to standard anticonvulsant therapy.


[Distorsion or rupture of ankle ligaments?].
In the Netherlands, almost 500,000 patients visit a physician each year because of an ankle inversion trauma. Most people are seen by a general practitioner, but a considerable number go directly to an emergency department. Recently a nation-wide accepted strategy for diagnosis and treatment of these lesions was formulated. This strategy includes the use of the Ottawa ankle rules, a second physical examination after four to five days if there is severe swelling of the ankle and functional treatment with inelastic tape bandage or brace application in case of an established rupture of the lateral band complex. At present, however, the efficacy of a second physical examination in primary health care has not been analysed and the economic consequences of the new strategy are unknown. From a scientific point of view none of the types of conservative treatment is preferable to another. Further scientific studies are needed.


General Practitioners' Perception of Risk for Travelers Visiting Friends and Relatives.
General practitioners (GPs) are an important source of pre-travel health advice for travelers; however, only a few studies have investigated primary healthcare provider-related barriers to the provision of pre-travel health advice, particularly to travelers visiting friends and relatives (VFR). We aimed to investigate Australian GPs' knowledge, attitudes, and practices with regard to VFR travelers. A postal survey was sent to randomly sampled GPs in Sydney, Australia, in 2012. The questionnaire investigated GPs' perception of risk and barriers to the provision of advice to VFR travelers. Of 563 GPs, 431 (76.6%) spoke a language other than English (LOTE) with 361 (64.1%) consulting in a LOTE. Overall, 222 (39.4%) GPs considered VFR travelers to be at higher risk than holiday travelers, with GPs consulting in English only [adjusted odds ratio (aOR) 1.65, 95% confidence interval (CI) 1.11-2.44, p = 0.01] and GPs considering long-term migrants as VFR travelers (aOR 1.86 95% CI 1.07-3.23, p = 0.03) remaining significant on multivariate analysis. Multilingual GPs are a valuable resource to reducing language and cultural barriers to healthcare. Targeted education of this subgroup of GPs may assist in promoting pre-travel health assessments for VFR travelers. Awareness of the need for opportunistic targeting of migrants for pre-travel consultation through routine identification of future travel is needed.


Resequencing PNMT in European hypertensive and normotensive individuals: no common susceptibilily variants for hypertension and purifying selection on intron 1.
Human linkage and animal QTL studies have indicated the contribution of genes on Chr17 into blood pressure regulation. One candidate gene is PNMT, coding for phenylethanolamine-N-methyltransferase, catalyzing the synthesis of epinephrine from norepinephrine. Fine-scale variation of PNMT was screened by resequencing hypertensive (n = 50) and normotensive (n = 50) individuals from two European populations (Estonians and Czechs). The resulting polymorphism data were analyzed by statistical genetics methods using Genepop 3.4, PHASE 2.1 and DnaSP 4.0 software programs. In silico prediction of transcription factor binding sites for intron 1 was performed with MatInspector 2.2 software. PNMT was characterized by minimum variation and excess of rare SNPs in both normo- and hypertensive individuals. None of the SNPs showed significant differences in allelic frequencies among population samples, as well as between screened hypertensives and normotensives. In the joint case-control analysis of the Estonian and the Czech samples, hypertension patients had a significant excess of heterozygotes for two promoter region polymorphisms (SNP-184; SNP-390). The identified variation pattern of PNMT reflects the effect of purifying selection consistent with an important role of PNMT-synthesized epinephrine in the regulation of cardiovascular and metabolic functions, and as a CNS neurotransmitter. A striking feature is the lack of intronic variation. In silico analysis of PNMT intron 1 confirmed the presence of a human-specific putative Glucocorticoid Responsive Element (GRE), inserted by Alu-mediated transfer. Further analysis of intron 1 supported the possible existence of a full Glucocorticoid Responsive Unit (GRU) predicted to consist of multiple gene regulatory elements known to cooperate with GRE in driving transcription. The role of these elements in regulating PNMT expression patterns and thus determining the dynamics of the synthesis of epinephrine is still to be studied. We suggest that the differences in PNMT expression between normotensives and hypertensives are not determined by the polymorphisms in this gene, but rather by the interplay of gene expression regulators, which may vary among individuals. Understanding the determinants of PNMT expression may assist in developing PNMT inhibitors as potential novel therapeutics.


Crossover from band-like to thermally activated charge transport in organic transistors due to strain-induced traps.
The temperature dependence of the charge-carrier mobility provides essential insight into the charge transport mechanisms in organic semiconductors. Such knowledge imparts critical understanding of the electrical properties of these materials, leading to better design of high-performance materials for consumer applications. Here, we present experimental results that suggest that the inhomogeneous strain induced in organic semiconductor layers by the mismatch between the coefficients of thermal expansion (CTE) of the consecutive device layers of field-effect transistors generates trapping states that localize charge carriers. We observe a universal scaling between the activation energy of the transistors and the interfacial thermal expansion mismatch, in which band-like transport is observed for similar CTEs, and activated transport otherwise. Our results provide evidence that a high-quality semiconductor layer is necessary, but not sufficient, to obtain efficient charge-carrier transport in devices, and underline the importance of holistic device design to achieve the intrinsic performance limits of a given organic semiconductor. We go on to show that insertion of an ultrathin CTE buffer layer mitigates this problem and can help achieve band-like transport on a wide range of substrate platforms.


Genome-wide identification of conserved intronic non-coding sequences using a Bayesian segmentation approach.
Computational identification of non-coding RNAs (ncRNAs) is a challenging problem. We describe a genome-wide analysis using Bayesian segmentation to identify intronic elements highly conserved between three evolutionarily distant vertebrate species: human, mouse and zebrafish. We investigate the extent to which these elements include ncRNAs (or conserved domains of ncRNAs) and regulatory sequences. We identified 655 deeply conserved intronic sequences in a genome-wide analysis. We also performed a pathway-focussed analysis on genes involved in muscle development, detecting 27 intronic elements, of which 22 were not detected in the genome-wide analysis. At least 87% of the genome-wide and 70% of the pathway-focussed elements have existing annotations indicative of conserved RNA secondary structure. The expression of 26 of the pathway-focused elements was examined using RT-PCR, providing confirmation that they include expressed ncRNAs. Consistent with previous studies, these elements are significantly over-represented in the introns of transcription factors. This study demonstrates a novel, highly effective, Bayesian approach to identifying conserved non-coding sequences. Our results complement previous findings that these sequences are enriched in transcription factors. However, in contrast to previous studies which suggest the majority of conserved sequences are regulatory factor binding sites, the majority of conserved sequences identified using our approach contain evidence of conserved RNA secondary structures, and our laboratory results suggest most are expressed. Functional roles at DNA and RNA levels are not mutually exclusive, and many of our elements possess evidence of both. Moreover, ncRNAs play roles in transcriptional and post-transcriptional regulation, and this may contribute to the over-representation of these elements in introns of transcription factors. We attribute the higher sensitivity of the pathway-focussed analysis compared to the genome-wide analysis to improved alignment quality, suggesting that enhanced genomic alignments may reveal many more conserved intronic sequences.


The topology of the triacylglycerol synthesizing enzyme Lro1 indicates that neutral lipids can be produced within the luminal compartment of the endoplasmatic reticulum: Implications for the biogenesis of lipid droplets.
Eukaryotes store metabolic energy in form of neutral lipids, which are deposited within a dedicated organelle, termed lipid droplet (LD). While neutral lipids are synthesized by ER localized integral membrane proteins, the fate of these lipids after their synthesis and the mechanism resulting in their accumulation in LDs are not well understood. We have recently shown that LDs are functionally connected to the ER membrane allowing for a bidirectional and energy-independent transport of integral membrane proteins and possibly lipids between the two compartments during lipogenesis or lipolysis. To further characterize the nature of this connection, we investigated the topology of triacylglycerol (TAG) formation. Here we show that the active site residues of the TAG biosynthetic enzyme in yeast, Lro1, a homolog of the lecithin cholesterol acyltransferase (LCAT)-related proteins, are located within the ER luminal domain of the enzyme, suggesting that TAG formed by Lro1 is initially present in the ER luminal leaflets of the ER membrane. The topology of TAG formed by Lro1 thus contrasts that of the second TAG biosynthetic enzyme, Dga1, which has a cytosolic acyl-CoA binding domain and thus is likely to catalyze TAG formation in the cytosolic leaflet of the ER membrane. Since TAG formed by either Dga1 or Lro1 can be efficiently packed into LDs we conclude that neutral lipids from both the cytosolic as well as the luminal leaflets of the ER membrane can be concentrated and packed into LDs.


Histocompatibility gene mutation rates in the mouse: a 25-year review.
Mutation rates of H2 and non-H2 histocompatibility genes in the mouse are examined over a 25-year period. Detected by skin graft rejections, the mutations were screened in inbred and hybrid mice from a continuously maintained and monitored colony and from a regularly supplied set of mice provided from the National Cancer Institute for monitoring of genetic integrity. Twenty-five H2 mutations were recovered, involving the K, D, L, and Ab loci, as well as over 80 mutations of non-H2 histocompatibility genes. Aside from a single allele at a single locus (H2-Kb), the spontaneous mutation rate of H2 class I genes appears to be equivalent to that found estimated for non-H2 histocompatibility genes, and comparable to rates reported for a variety of mouse genes. This is in contrast with previous suggestions that H2 genes mutate at orders of magnitude greater than do "average" mammalian genes. The discrepancy is attributed to the H2-Kb gene which accounts for over half of all reported H2 mutations and which mutates spontaneously at a rate of 1-2 x 10(-4) per gene per generation. Furthermore, over half of the spontaneous H2-Kb mutations result in a single mutant phenotype (the "bg" group) which involve similar changes at amino acid residues 116 and 121. Thus, the high spontaneous mutation rate for H2-Kb appears to be the exception among major histocompatibility genes, rather than the rule.


Evaluation of Cellular Responses for the Diagnosis of Allergic Bronchopulmonary Mycosis: A Preliminary Study in Cystic Fibrosis Patients.
Background: Allergic bronchopulmonary mycosis (ABPM) is an underestimated allergic disease due to fungi. Most reported cases are caused by Aspergillus fumigatus (Af) and are referred to as allergic bronchopulmonary aspergillosis (ABPA). The main risk factor of ABPA is a history of lung disease, such as cystic fibrosis, asthma, or chronic obstructive pulmonary disease. The main diagnostic criteria for ABPA rely on the evaluation of humoral IgE and IgG responses to Af extracts, although these cannot discriminate Af sensitization and ABPA. Moreover, fungi other than Af have been incriminated. Flow cytometric evaluation of functional responses of basophils and lymphocytes in the context of allergic diseases is gaining momentum. Objectives: We hypothesized that the detection of functional responses through basophil and lymphocyte activation tests might be useful for ABPM diagnosis. We present here the results of a pilot study comparing the performance of these cellular assays vs. usual diagnostic criteria in a cystic fibrosis (CF) cohort. Methods:Ex vivo basophil activation test (BAT) is a diagnostic tool highlighting an immediate hypersensitivity mechanism against an allergen, e.g., through CD63 upregulation as an indirect measure of degranulation. Lymphocyte stimulation test (LST) relies on the upregulation of activation markers, such as CD69, after incubation with allergen(s), to explain delayed hypersensitivity. These assays were performed with Af, Penicillium, and Alternaria extracts in 29 adult CF patients. Results: BAT responses of ABPA patients were higher than those of sensitized or control CF patients. The highest LST result was for a woman who developed ABPA 3 months after the tests, despite the absence of specific IgG and IgE to Af at the time of the initial investigation. Conclusion: We conclude that basophil and lymphocyte activation tests could enhance the diagnosis of allergic mycosis, compared to usual humoral markers. Further studies with larger cohorts and addressing both mold extracts and mold relevant molecules are needed in order to confirm and extend the application of this personalized medicine approach.


Reprint of: Highthroughtput analysis of behavior for drug discovery.
Drug testing with traditional behavioral assays constitutes a major bottleneck in the development of novel therapies. PsychoGenics developed three comprehensive highthroughtput systems, SmartCube(®), NeuroCube(®) and PhenoCube(®) systems, to increase the efficiency of the drug screening and phenotyping in rodents. These three systems capture different domains of behavior, namely, cognitive, motor, circadian, social, anxiety-like, gait and others, using custom-built computer vision software and machine learning algorithms for analysis. This review exemplifies the use of the three systems and explains how they can advance drug screening with their applications to phenotyping of disease models, drug screening, selection of lead candidates, behavior-driven lead optimization, and drug repurposing.


Strategies for skeletal muscle targeting in drug discovery.
The targeting of drugs to skeletal muscle is an emerging area of research. Driven by the need for new therapies to treat a range of muscle-associated diseases, these strategies aim to provide improved drug exposure at the site of action in skeletal muscle with reduced concentration in other tissues where unwanted side effects could occur. By interacting with muscle-specific cell surface recognition elements, both tissue localization and selective uptake into skeletal muscle cells can be achieved. The design of molecules that are substrates for muscle uptake transporters can provide concentration in m uscle tissue. For example, drug conjugates with carnitine can provide improved muscle uptake via OCTN2 transport. Binding to muscle surface recognition elements followed by endocytosis can allow even large molecules such as antibodies to enter muscle cells. Monoclonal antibody 3E10 demonstrated selective uptake into skeletal muscle in vivo. Hybrid adeno-associated viral vectors have recently shown promise for high skeletal muscle selectivity in gene transfer applications. Delivery technology methods, including electroporation of DNA plasmids, have also been investigated for selective muscle uptake. This review discusses challenges and opportunities for skeletal muscle targeting, highlighting specific examples and areas in need of additional research.


Greek Physiotherapists' Perspectives on Rehabilitation Following Total Knee Replacement: a Descriptive Survey.
In Greece, as in many countries, there is a scarcity of evidence in the type of physiotherapy services offered for the rehabilitation of total knee replacement (TKR). Despite the number of TKRs annually performed in Greece (over 10,000), there are no available clinical guidelines as to the content of best physiotherapy practice. The aim of this nationwide survey undertaken by physiotherapists treating TKR patients post-operatively was to record standard practice and services available in Greece. Design: cross-country survey Ten per cent of all registered physiotherapists working in public/private sectors were recruited. The developed survey comprised of questions regarding therapists' profile, protocols implemented at different stages of rehabilitation and the aims and modalities used. A 58.7% response rate was achieved, where 36% (47/132) of respondents were treating patients in the inpatient phase and 64% (85/132) after hospital discharge. Patients in Greece are discharged with a home-based exercise program (56.7%) and, to a lesser extent, are referred to rehabilitation centres (13.3%). Strengthening, range of movement and functionality seemed to be the primary goals especially in the inpatient phase, whereas in the outpatient phase, apart from the larger differences identified, functionality and balance training were more frequently reported. No significant variations in practice were found during inpatient rehabilitation, whilst there seemed to be diversity across outpatient physiotherapy programs. The current survey suggests that patient's general health and psychological and behavioural issues are the criteria by which physiotherapists select the volume of implemented exercise and progression. However, no specific guidelines were followed. Copyright © 2016 John Wiley & Sons, Ltd.


Wide-Area Composite Load Parameter Identification Based on Multi-Residual Deep Neural Network.
Accurate and practical load modeling plays a critical role in the power system studies including stability, control, and protection. Recently, wide-area measurement systems (WAMSs) are utilized to model the static and dynamic behavior of the load consumption pattern in real-time, simultaneously. In this article, a WAMS-based load modeling method is established based on a multi-residual deep learning structure. To do so, a comprehensive and efficient load model founded on combination of impedance-current-power and induction motor (IM) is constructed at the first step. Then, a deep learning-based framework is developed to understand the time-varying and complex behavior of the composite load model (CLM). To do so, a residual convolutional neural network (ResCNN) is developed to capture the spatial features of the load at different location of the large-scale power system. Then, gated recurrent unit (GRU) is used to fully understand the temporal features from highly variant time-domain signals. It is essential to provide a balance between fast and slow variant parameters. Thus, the designed structure is implemented in a parallel manner to fulfill the balance and moreover, weighted fusion method is used to estimate the parameters, as well. Consequently, an error-based loss function is reformulated to improve the training process as well as robustness in the noisy conditions. The numerical experiments on IEEE 68-bus and Iranian 95-bus systems verify the effectiveness and robustness of the proposed load modeling approach. Furthermore, a comparative study with some relevant methods demonstrates the superiority of the proposed structure. The obtained results in the worst-case scenario show error lower than 0.055% considering noisy condition and at least 50% improvement comparing the several state-of-art methods.


Quantitative trait loci mapping and the genetic basis of heterosis in maize and rice.
Despite its importance to agriculture, the genetic basis of heterosis is still not well understood. The main competing hypotheses include dominance, overdominance, and epistasis. NC design III is an experimental design that has been used for estimating the average degree of dominance of quantitative trait loci (QTL) and also for studying heterosis. In this study, we first develop a multiple-interval mapping (MIM) model for design III that provides a platform to estimate the number, genomic positions, augmented additive and dominance effects, and epistatic interactions of QTL. The model can be used for parents with any generation of selfing. We apply the method to two data sets, one for maize and one for rice. Our results show that heterosis in maize is mainly due to dominant gene action, although overdominance of individual QTL could not completely be ruled out due to the mapping resolution and limitations of NC design III. For rice, the estimated QTL dominant effects could not explain the observed heterosis. There is evidence that additive x additive epistatic effects of QTL could be the main cause for the heterosis in rice. The difference in the genetic basis of heterosis seems to be related to open or self pollination of the two species. The MIM model for NC design III is implemented in Windows QTL Cartographer, a freely distributed software.


Effectiveness of ezetimibe alone or in combination with twice a week Atorvastatin (10 mg) for statin intolerant high-risk patients.
This study was undertaken to investigate the effect of ezetimibe (10 mg/day) alone or in combination with atorvastatin (10 mg twice a week) on hypercholesterolemia in 56 high-risk patients intolerant to daily statin use. Ezetimibe monotherapy was well tolerated (2 withdrawals) and induced a mean reduction in low-density lipoprotein (LDL) cholesterol of 20% (p <0.05) at the third month. However, of the 54 patients still taking ezetimibe, only 5 (9%) were at their LDL cholesterol targets. Atorvastatin 10 mg twice a week was then added to ezetimibe and was well tolerated (3 withdrawals). This combination reduced LDL cholesterol (in a treatment-based analysis) by 37% compared with baseline (p <0.001), with 43 (84%) patients reaching their LDL cholesterol goals. When patients (n = 34, 25 men) with baseline serum creatinine values in the upper 2 tertiles were analyzed separately, there was a significant (p = 0.041) decrease in serum creatinine levels after 6 months of treatment. In conclusion, the combination of ezetimibe plus atorvastatin 10 mg twice a week might be a therapeutic option for high-risk patients intolerant to daily statin monotherapy.


Novel Genetically Encoded Bright Positive Calcium Indicator NCaMP7 Based on the mNeonGreen Fluorescent Protein.
Green fluorescent genetically encoded calcium indicators (GECIs) are the most popular tool for visualization of calcium dynamics in vivo. However, most of them are based on the EGFP protein and have similar molecular brightnesses. The NTnC indicator, which is composed of the mNeonGreen fluorescent protein with the insertion of troponin C, has higher brightness as compared to EGFP-based GECIs, but shows a limited inverted response with an ΔF/F of 1. By insertion of a calmodulin/M13-peptide pair into the mNeonGreen protein, we developed a green GECI called NCaMP7. In vitro, NCaMP7 showed positive response with an ΔF/F of 27 and high affinity (Kd of 125 nM) to calcium ions. NCaMP7 demonstrated a 1.7-fold higher brightness and similar calcium-association/dissociation dynamics compared to the standard GCaMP6s GECI in vitro. According to fluorescence recovery after photobleaching (FRAP) experiments, the NCaMP7 design partially prevented interactions of NCaMP7 with the intracellular environment. The NCaMP7 crystal structure was obtained at 1.75 Å resolution to uncover the molecular basis of its calcium ions sensitivity. The NCaMP7 indicator retained a high and fast response when expressed in cultured HeLa and neuronal cells. Finally, we successfully utilized the NCaMP7 indicator for in vivo visualization of grating-evoked and place-dependent neuronal activity in the visual cortex and the hippocampus of mice using a two-photon microscope and an NVista miniscope, respectively.


Hindsight bias, outcome knowledge and adaptive learning.
The ubiquitous nature of hindsight bias is a cause for concern for those engaged in investigations and retrospective analysis of medical error. Hindsight does not equal foresight. Investigations that are anchored to outcome knowledge run the risk of not capturing the complexities and uncertainties facing sharp end personnel and why their actions made sense at the time. Important lessons go unlearned if the exercise is simply to back track someone else's decision landmarks. Outcome knowledge can also bias our thinking on the quality of the processes that led to the outcome. This paper examines the influence of outcome knowledge in relation to reconstructive memory and legal testimony, ways for reducing the impact of outcome knowledge, and an adaptive learning framework that places hindsight bias in a broader context of rapid updating of knowledge.


Functional MRI detection of activation in the primary gustatory cortices in humans.
Magnetoencephalography (MEG) has recently revealed that the transitions between the parietal operculum (Pop) and the insula (area G) and the ventral end of the central sulcus (cs) were activated with the shortest latency by instrumental gustatory stimulation, which suggests that the location of the primary gustatory area is in these two regions. However, studies using other noninvasive brain-imaging methods such as positron-emission tomography or functional magnetic resonance imaging (fMRI) with manual application of tastants into the mouth have been unable to confirm this. The present study examined cortical activation by repetitive stimulation of the tongue tip with 1 M NaCl with a computer-controlled stimulator and used fMRI to detect it. In individual brains, activations were detected with multiple comparisons (false discovery rate) across the whole brain corrected (threshold at P < 0.05) at both area G and frontal operculum (Fop) in 8 of 11 subjects and at the rolandic operculum (Rop) in 7 subjects. Activations were also found at the ventral end of the cs (n = 3). Group analysis with random-effect models (multiple comparison using familywise error in regions of interest, P < 0.02) revealed activation at area G in both hemispheres and in the Fop, Rop, and ventral end of the cs on the left side. The present study revealed no activation on the gyrus of the external cerebral surface except for the Rop. Taking MEG findings into consideration, the present findings strongly indicate that the primary gustatory area is present at both the transition between the Pop and insula and the Rop including the gray matter within a ventral part of the cs.


An update regarding the treatment of nonallergic rhinitis.
The review presents some information on known options for treatment of nonallergic rhinitis (NAR) with introduction to new therapies. The merits and limitations of recent advancements in pharmacotherapy of this common problem are briefly discussed as well. Intranasal corticosteroids are first-line therapy for NAR. Fluticasone propionate and beclomethasone remain the only topical corticosteroids approved for NAR. The use of azelastine - another first-line option - has also been found to be effective even though NAR is a nonallergic entity by definition. Combination of fluticasone propionate and azelastine is a promising option in achieving better symptom reduction. Coadministration of intranasal corticosteroid and topical decongestants is an attractive topic that requires additional safety studies before recommending treatment. Although promising, no scientifically valid recommendation can be made for treatment of NAR with capsaicin. Surgical options in patients with refractory NAR are limited. New studies demonstrate a lack of correlation between objective outcome of radiofrequency ablation of the inferior turbinate and subjective patient symptoms. The heterogeneity in clinical presentation makes NAR treatment a daily challenge for otolaryngologists. The diversity of clinical studies with use of unique outcome measures limit systematic reviews which may be instrumental in providing strong recommendations.


The MLO1 powdery mildew susceptibility gene in Lathyrus species: The power of high-density linkage maps in comparative mapping and synteny analysis.
Powdery mildews are major diseases for a range of crops. The loss of function of specific Mildew Locus O (MLO) genes has long been associated with pre-haustorial plant resistance to powdery mildew and has proven to be durable in several species. Erysiphe pisi is the major causal agent of powdery mildew in pea (Pisum sativum L.) and in the closely related Lathyrus sativus L. and Lathyrus cicera L. PsMLO1 has been extensively studied in pea. However, no MLO gene family members have been isolated and characterized in Lathyrus species so far. In this study, MLO1 genes were isolated and characterized in L. sativus and L. cicera genotypes with varied levels of partial resistance against powdery mildew. Phylogenetic analyses confirmed that Lathyrus MLO1 belongs to Clade V, like all dicot MLO proteins associated with powdery mildew susceptibility. A L. sativus recombinant inbred line population (RIL) was genotyped by sequencing to develop a high-density L. sativus genetic linkage map. DNA sequence polymorphisms between the analyzed genotypes allowed the location of MLO1 in the newly developed L. sativus RIL genetic linkage map. Subsequent comparative mapping between L. sativus and L. cicera genetic maps and P. sativum, Lens culinaris Medik., and Medicago truncatula Gaertn. reference genomes revealed important aspects of the conservation of the MLO1 locus position and of the overall chromosomal rearrangements occurring during legume evolution, with relevance to legume disease resistance breeding programs.


Cementless and cemented total knee arthroplasty in patients younger than fifty five years. Which is better?
The purpose of this prospective, randomised study was to evaluate long-term clinical results, radiographic findings, complications and revision and survivorship rates in patients <55 years at a minimum of 16 years after undergoing bilateral, sequential, simultaneous, cemented and cementless total knee arthroplasties (TKAs) in the same patients. Bilateral, sequential, simultaneous TKAs were performed in 80 patients (160 knees). There were 63 women and 17 men with a mean age of 54.3 years (range 49-55), who received a cementless prosthesis in one knee and a cemented prosthesis in the other. The mean follow-up was 16.6 years (range 16-17). At final review, the mean Knee Society (KS) knee scores (95.8 versus 96.9), Western Ontario and McMaster Universities (WOMAC) osteoarthritis (OA) index (25.4 versus 25.9), range of motion (ROM) (125° versus 128°), patient satisfaction (8.1 versus 8.3) and radiological results were similar in both groups. Femoral component survival rate was 100% in both groups at 17 years; at 17 years, the cemented tibial component survival rate was 100% and the cementless tibial component 98.7%. No osteolysis was identified in either group. Long-term results of both cementless and cemented TKAs were encouraging in patients with OA who were <55 years. However, we found no evidence to prove the superiority of cementless over cemented TKAs.


7 tesla FMRI reveals systematic functional organization for binocular disparity in dorsal visual cortex.
The binocular disparity between the views of the world registered by the left and right eyes provides a powerful signal about the depth structure of the environment. Despite increasing knowledge of the cortical areas that process disparity from animal models, comparatively little is known about the local architecture of stereoscopic processing in the human brain. Here, we take advantage of the high spatial specificity and image contrast offered by 7 tesla fMRI to test for systematic organization of disparity representations in the human brain. Participants viewed random dot stereogram stimuli depicting different depth positions while we recorded fMRI responses from dorsomedial visual cortex. We repeated measurements across three separate imaging sessions. Using a series of computational modeling approaches, we report three main advances in understanding disparity organization in the human brain. First, we show that disparity preferences are clustered and that this organization persists across imaging sessions, particularly in area V3A. Second, we observe differences between the local distribution of voxel responses in early and dorsomedial visual areas, suggesting different cortical organization. Third, using modeling of voxel responses, we show that higher dorsal areas (V3A, V3B/KO) have properties that are characteristic of human depth judgments: a simple model that uses tuning parameters estimated from fMRI data captures known variations in human psychophysical performance. Together, these findings indicate that human dorsal visual cortex contains selective cortical structures for disparity that may support the neural computations that underlie depth perception.


Oral mucositis in pediatric patients undergoing hematopoietic stem cell transplantation: clinical outcomes in a context of specialized oral care using low-level laser therapy.
OM is a painful inflammatory condition of the oral mucosa, derived from the toxic effects of chemotherapy and radiotherapy. High OM severity is frequently present in HSCT pediatric patients, who exhibit multiple painful ulcers that limit their mastication and swallowing, leading to poor nutritional status. Few studies have demonstrated OM clinical outcomes in young patients undergoing HSCT. Feasibility of oral care and LLLT on OM prophylaxis and treatment is also poorly discussed. The aim of this study was to describe a specialized oral care protocol that included LLLT for pediatric patients undergoing transplantation and to demonstrate the clinical outcomes after OM prevention and treatment. Data from OM-related morbidity were collected from 51 HSCT pediatric patients treated daily with LLLT, followed by standard oral care protocols. All the patients, even infants and young children, accepted the daily oral care and LLLT well. The majority (80.0%) only exhibited erythema in the oral mucosa, and the maximum OM degree was WHO II. Patients who had undergone autologous and HLA-haploidentical transplants showed OM with the lowest severity. The frequency of total body irradiation and methotrexate prescriptions was higher in adolescents when compared with infants (p = 0.044), and adolescents also exhibited OM more severely than infants and young children. We found that good clinical outcomes were obtained using this therapy, mainly in regard to the control of OM severity and pain reduction in the oral cavity. Specialized oral care, including LLLT, is feasible and affordable for HSCT pediatric patients, although some adaptation in the patient's oral hygiene routine must be adopted with help from parents/companions and clinical staff.


[Heterotopic pregnancy following ovulation induction by clomiphene citrate and prednisone. Report of a case].
Heterotopic pregnancy (HP) is a very rare pathologic event. The incidence of ectopic pregnancy has increased as the consequence of assisted reproduction and the wider use of ovulation induction agents. The diagnosis of HP is frequently done not as earlier as it should be, and it has serious repercussions. The concept of screening, to find relevant rise factors and the correct diagnosis of HP should be pursued as early as possible. In this report we describe an unusual case of a female patient with therapeutic failure to clomiphene citrate. She was treated with two cycles of combined clomiphene citrate and prednisone, and ovulation induction assent. The after she developed HP. Overall, we conclude that the possible association observed could be an accident. The land of investigation is open, to fill cover. The holes on understanding that we still have.


Microsatellite markers and multiple paternity in the garter snake thamnophis sirtalis
Data from four microsatellite loci developed for the common garter snake, Thamnophis sirtalis, show that multiple paternity is common in a natural population on Beaver Island, Michigan. Six of eight litters tested, and all litters with five or more neonates, were multiply sired. At least triple paternity was documented in the largest litter examined (n = 13 neonates). Inheritance patterns and genotype frequencies in the wild population indicate the presence of null allele(s) at one of the microsatellite loci. Garter snakes are widely used in quantitative genetics research, and paternity testing is essential in studies that rely on sibling analysis.


The intracoelomic route: a new approach for in utero human cord blood stem cell transplantation.
The intracoelomic route for in utero hematopoietic stem cell transplantation has been evaluated in pre-immune fetal sheep and the engraftment characteristics defined. Twelve ovine fetuses (gestational ages: 40-45 days) received intracoelomic transplants of human CD3-depleted (50 x 10(6) per lamb) or CD34-selected (1-2 x 10(5) per lamb) cord blood hematopoietic stem cells. Engraftment was evaluated from cell suspension of the liver, spleen, bone marrow and thymus by flow cytometry, cloning assays and polymerase chain reaction (PCR) analysis for human beta(2)-microglobulin gene. The engraftment of liver samples was also evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR), fluorescent in situ hybridization (FISH) and immunohistochemistry. Four fetuses (33%) aborted shortly after intracoelomic transplantation and were not evaluable for engraftment. Engraftment was detected in 4 fetuses obtained from cesarean delivery on day 70 after transplantation of CD3-depleted cord blood cells. The degree of engraftment in these 4 fetuses ranged from 6 to 22% in the different organs (as revealed by antigenic analysis of human CD45 with flow cytometry). Three fetuses obtained after cesarean section at 102 (No. 435184) and 105 (Nos 915293, 037568) days and 1 fetus delivered at term, which received CD34-selected cord blood cells, had human engraftment with 10, 32, 20 and 10% CD45+ cells in bone marrow, respectively. A further check of human chimerism was done at 1 year after birth of the fetus delivered at term and 7.6% of bone marrow chimerism was detected. In 6 out of 8 fetuses evaluable for human engraftment, chimerism was confirmed by PCR analysis for human beta(2)-microglobulin which also identified human cells in brain, spinal cord, heart, lung and skeletal muscle. On liver samples, FISH and RT-PCR confirmed the xenograft of human cells and the immunohistochemical analysis detected human markers of hematopoietic and hepatic lineage of differentiation. This preliminary study indicates that intracoelomic transplantation of human hematopoietic stem cells in fetal lambs is feasible and effective in terms of hematopoietic engraftment.


Perioperative and continence outcomes of robotic radical prostatectomy in elderly Indian men (≥70 years): A sub-group analysis.
Many healthy elderly Indian men seek surgical treatment for localized prostate cancer. Quite often, radical surgery is not offered to the patients over 70 years of age due to the perception of increased side-effects and complications. We have previously reported our results of robotic radical prostatectomy in a study comprising 150 Indian patients, where almost a quarter of patients were elderly. This subgroup analysis was therefore focused on evaluating perioperative and continence outcomes in elderly men (≥70 years) with localized prostate cancer. Between April 2010 and August 2013, 153 men had robot-assisted radical prostatectomy performed by two surgeons. Of the 150 men analyzed, 39 (26%) were aged ≥70 years. All patients underwent robotic prostatectomy using a 4 arm da Vinci surgical system. Pre-operative, intraoperative and post-operative parameters were studied. Check cystogram was performed in all patients prior to catheter removal. Complications were categorized using the Clavien-Dindo classification system. Continence was defined as use of "no pad" or security liner only. All data were recorded prospectively and analyzed using SPSS version 20. There were no significant intraoperative or perioperative complications in this group. Median blood loss during surgery was 150 mL. None of the patient required blood transfusion. There were two minor complications (5.1%) within the first 30 days of surgery: Minimal anastomotic site leak (one patient) requiring replacement and prolongation of Foley's drainage by 1 week and ileus (one patient). No patient had any cardiopulmonary or vascular complications in the post-operative period. The median duration of hospital stay was 3 days. The median duration of catheterization was 7 days. No patient had problem of bladder neck stenosis in the follow-up period. At 1 month, 3 months, 6 months and 1 year of follow-up, 66.7% (n = 26), 74.3% (n = 29), 87.9% (n = 34) and 94.8% (n = 37), respectively, were continent. Robotic surgery is safe and feasible in a select group of elderly patients. It has acceptable and minimal perioperative complications along with good continence outcome.


Volume navigation with contrast enhanced ultrasound and image fusion for percutaneous interventions: first results.
Assessing the feasibility and efficiency of interventions using ultrasound (US) volume navigation (V Nav) with real time needle tracking and image fusion with contrast enhanced (ce) CT, MRI or US. First an in vitro study on a liver phantom with CT data image fusion was performed, involving the puncture of a 10 mm lesion in a depth of 5 cm performed by 15 examiners with US guided freehand technique vs. V Nav for the purpose of time optimization. Then 23 patients underwent ultrasound-navigated biopsies or interventions using V Nav image fusion of live ultrasound with ceCT, ceMRI or CEUS, which were acquired before the intervention. A CEUS data set was acquired in all patients. Image fusion was established for CEUS and CT or CEUS and MRI using anatomical landmarks in the area of the targeted lesion. The definition of a virtual biopsy line with navigational axes targeting the lesion was achieved by the usage of sterile trocar with a magnetic sensor embedded in its distal tip employing a dedicated navigation software for real time needle tracking. The in vitro study showed significantly less time needed for the simulated interventions in all examiners when V Nav was used (p<0.05). In the study involving patients, in all 10 biopsies of suspect lesions of the liver a histological confirmation was achieved. We also used V Nav for a breast biopsy (intraductal carcinoma), for a biopsy of the abdominal wall (metastasis of ovarial carcinoma) and for radiofrequency ablations (4 ablations). In 8 cases of inflammatory abdominal lesions 9 percutaneous drainages were successfully inserted. Percutaneous biopsies and drainages, even of small lesions involving complex access pathways, can be accomplished with a high success rate by using 3D real time image fusion together with real time needle tracking.


Survival outcomes for patients with surgically induced end-stage renal disease.
While medically induced end-stage renal disease (m-ESRD) has been well-studied, outcomes in patients with surgically induced ESRD (s-ESRD) are unknown. We sought to quantitatively compare the non-oncological outcomes for s-ESRD and m-ESRD in a large, population-based cohort. Medicare patients >65 years old initiating hemodialysis were identified using the U.S. Renal Data System database (2000-2012). Metastatic cancer, prior transplant history, and nephrectomy for polycystic kidney disease were exclusion criteria. Patients were classified as having s-ESRD or m-ESRD based on hospital and physician claims for nephrectomy within a year preceding the onset of maintenance hemodialysis. Outcomes included non-cancer mortality (NCM), overall survival (OS), cardiovascular event (CVE), and renal transplantation. Time-to-event analyses were performed using Kaplan-Meier and cumulative incidence curves, and multivariable Cox and Fine-and-Grey regression models. The cohort included 312 612 patients, of whom 1648 (0.53%) had s-ESRD. Compared to m-ESRD patients, s-ESRD patients had a significantly lower five-year cumulative incidence of NCM (68% vs. 80%; p<0.001) and CVE (62% vs. 68%; p<0.001), with a correspondingly higher probability of OS (22% vs. 17%; p<0.001) and rate of renal transplantation (3.6% vs. 2.0%; p<0.001). On multivariable analyses, s-ESRD remained associated with lower risks of NCM (p<0.001) and CVE (p<0.001), improved OS (p<0.001), and higher chance of renal transplantation (p<0.001). While outcomes for s-ESRD appear more favorable than m-ESRD, s-ESRD is still associated with a substantial risk of NCM and CVE, and a low incidence of renal transplantation in Medicare patients >65 years old. These non-oncological outcomes are worth considering in patients potentially facing postoperative ESRD.


Characterization of a putative periplasmic transport system for octopine accumulation encoded by Agrobacterium tumefaciens Ti plasmid pTiA6.
Neoplastic crown gall tumors incited by Agrobacterium tumefaciens release novel amino acid or sugar derivatives known as opines, whose synthesis is directed by genes transferred to plant cells. Agrobacterium cells can transport and catabolize these compounds as sources of carbon and nitrogen. This article describes a region of the pTiA6 plasmid which is required for catabolism of the opine octopine and whose transcription is induced by octopine. This region of the plasmid contains four open reading frames, occQ, occM, occP, and occJ, which show homology to the family of so-called shock-sensitive permeases. TnphoA mutagenesis demonstrated that the OccJ and OccM proteins lie fully or partly in the periplasmic space. The OccJ protein was identified by electrophoresis and found to be fully localized in the periplasmic space. When these proteins were expressed in Escherichia coli, radiolabeled octopine became cell-associated.


Altered hippocampal kainate-receptor mRNA levels in temporal lobe epilepsy patients.
This study determined whether hippocampal kainate (KA) receptor mRNA levels were increased or decreased in temporal lobe epilepsy patients compared with nonseizure autopsies. Hippocampal sclerosis (HS; n = 17), nonsclerosis (non-HS; n = 11), and autopsy hippocampi (n = 9) were studied for KA1-2 and GluR5-7 mRNA levels using semiquantitative in situ hybridization techniques, along with neuron densities. Compared with autopsy hippocampi, HS and non-HS cases showed decreased GluR5 and GluR6 hybridization densities per CA2 and/or CA3 pyramid. Furthermore, HS patients demonstrated increased KA2 and GluR5 hybridization densities per granule cell compared with autopsy hippocampi. These findings indicate that chronic temporal lobe seizures were associated with differential changes in hippocampal KA1-2 and GluR5-7 hybridization densities that vary by subfield and pathology group. In temporal lobe epilepsy patients, these results support the hypothesis that pyramidal cell GluR5 and GluR6 mRNA levels are decreased as a consequence of seizures, and in HS patients granule cell KA2 and GluR5 mRNA levels are increased in association with aberrant fascia dentata mossy fiber sprouting and/or hippocampal neuronal loss.


Morphofunctional analysis of antigen uptake mechanisms following sublingual immunotherapy with beads in mice.
Recently, sublingual immunotherapy (SLIT) has been used as a safe and efficient method for the treatment of and immunization against asthma and various allergies. However, the routes of antigen/allergen (particulate antigen) uptake through the mucosa of the oral cavity remain incompletely understood, as do the roles of sex and age in the process. For this purpose, to elucidate the mechanism and efficacy of SLIT among different sexes and ages, microbeads were dripped into the sublingual region to mimic particulate antigen uptake by the sublingual mucosa. Twenty microliters of either phosphate buffered saline (PBS) or fluorescently labelled microbeads (latex and silica beads) were placed under the tongue of both male and female C57BL/6 mice at young (3 months) and old (6 months) ages. The lower jaw was examined 30 min after administration, and beads were detected with a fluorescence stereomicroscope. Morphological observations of the mucosa of the fluorescent areas were made with a scanning electron microscope (SEM) and an all-in-one light fluorescence microscope (LM). Fluorescence intensity was compared between both sexes and ages. Stereomicroscopic observation revealed fluorescent illuminations in three compartments of the sublingual mucosa: the sublingual caruncles (SC), the oral rostral mucosa (OR) and the buccal mucosa (BM). Interestingly, the fluorescence intensity tended to be higher among females than among males in the SC region in particular. However, there were no significant age-related differences. SEM and LM revealed beads in the lumina of both mandibular ducts and sublingual ducts (Sd). Additionally, the apical cytoplasm of some Sd cells contained silica beads. However, there was no specification in the OR mucosa or BM. This study reveals the major role Sd plays in local immunity via the antigen uptake mechanisms. Furthermore, our data suggest that the efficacy of SLIT in humans could be affected by sex.