Increased cortisol response to exogenous adrenocorticotropic hormone in chronically stressed pigs: influence of housing conditions.
In a longitudinal experiment, the influence of tethered housing (a condition of chronic stress) on the reactivity of the adrenal cortex to exogenous ACTH was investigated in gilts. To that end, the plasma cortisol response to synthetic ACTH (1-24; 10 micrograms/kg of BW; i.v. bolus injection via a permanent catheter) was determined before and after prolonged tethered housing. Two systems for tethered housing were used, one more restrictive than the other with regard to possibilities for visual and tactile contacts with conspecifics and visual control over the environment. The ACTH treatment induced a marked, transient plasma cortisol response in all gilts studied, irrespective of their housing conditions. Long-term tethered housing increased the ACTH-induced cortisol response. Possible effects of the experimental procedure or age-related effects could be excluded, because in control gilts, which were housed loose during the entire experimental period, the cortisol response to ACTH remained unaltered. The chronic stress-induced increase in the ACTH-induced cortisol response was considerably more pronounced and persistent in gilts that were deprived of possibilities for social contacts with conspecifics and visual control over the environment than in gilts with such possibilities. These data indicate that in tethered gilts adaptational changes occur at the level of the adrenal cortex that affect the ACTH-induced adrenocortical response. In addition, not only physical restraint but also restriction of social contact and visual control play an important role in the development of these changes.


The polarized carbon nanotube thin film LED.
We demonstrate a light emitting p-i-n diode made of a highly aligned film of separated (99%) semiconducting carbon nanotubes, self-assembled from solution. By using a split gate technique, we create p- and n-doped regions in the nanotube film that are separated by a micron-wide gap. We inject p- and n-type charge carriers into the device channel from opposite contacts and investigate the radiative recombination using optical micro-spectroscopy. We find that the threshold-less light generation efficiency in the intrinsic carbon nanotube film segment can be enhanced by increasing the potential drop across the junction, demonstrating the LED-principle in a carbon nanotube film for the first time. The device emits infrared light that is polarized along the long axes of the carbon nanotubes that form the aligned film.


Expression of BMP-4 and Smad1 in patients with Hirschsprung disease and its clinical significance.
Expression and clinical significance of bone morphogenetic protein (BMP)-4 and Smad1 in patients with Hirschsprung disease (HD) were investigated. A retrospective analysis of 96 HD patients (experimental group) admitted to Xuzhou Children's Hospital, Xuzhou Medical University from June 2015 to June 2017 was performed. According to the samples, the experimental group was divided into the stenosis group, the transition group and the expansion group. Forty-seven children with colostomy due to intestinal obstruction were selected as the control group. The expression levels of BMP-4 and Smad1 proteins were detected by immunohistochemical staining. The expression levels of BMP-4 and Smad mRNA were detected by real-time quantitative PCR (RT-qPCR), and were quantified and compared. Via immunohistochemistry, BMP-4 and Smad1 proteins were detected in the samples of different parts of HD patients and children with intestinal obstruction. The positive expression levels of BMP-4 and Smad1 proteins in the transition group were decreased compared with those in the expansion and control groups (P<0.05), and the positive expression levels of BMP-4 and Smad1 proteins in the stenosis group were decreased compared with those in the transition, expansion, and control groups (P<0.05). Also, the gene expression levels of BMP-4 and Smad1 in the transition and stenosis groups were successively decreased, and the differences were statistically significant (P<0.05). In conclusion, the expression of BMP-4 and Smad1 in the intestinal plexus of HD lesions was significantly reduced, indicating that BMP-4 and Smad1 are closely related to the occurrence of HD, and it is suspected that they have a certain influence on the intestinal development of congenital digestive tract malformations.


Polymer nanowire elastic moduli measured with digital pulsed force mode AFM.
The mechanical bending behavior of polymer nanowires-polypyrrole and poly(3,4-ethylene dioxythiophene-co-styrene sulfonate)-produced by template molding were measured using a new innovation in atomic force microscopy (AFM). Digital pulsed force mode (DPFM) was used to image and simultaneously perform three-point bend tests along nanowires spanning microchannels in silicon. The bending profiles were analyzed for apparent elastic moduli variations along the suspended length of individually isolated nanowires and compared to classic beam deflection models for various geometric and boundary conditions. The elastic moduli calculated from these AFM data are 2-7 times that expected for bulk polymer values (approximately 1-3 GPa), demonstrating an apparent strengthening of nanostructured polymer even for diameters greater than 100 nm--the accepted boundary for nanoscience. Furthermore, detailed analysis of deflection data versus loading location demonstrates the experimental dependence on test geometry and inherent errors in relying solely on midpoint bending measurements or any single loading configuration for nanomechanical testing as well as the significant contribution of nanoindentation effects.


Promoting effect of Zn in high-loading Zn/Ni-SiO2 catalysts for selective hydrogen evolution from methylcyclohexane.
The dehydrogenation of methylcyclohexane to toluene was investigated over high-loading monometallic Ni-SiO2 and bimetallic Zn/Ni-SiO2 catalysts. The catalysts were prepared by the impregnation coupled with the advantageous heterophase sol-gel technique. Their performance was tested in a fixed-bed flow reactor at 250-350 °C, 0.1 MPa pressure, equimolar ratio H2/Ar (24 nL h-1 in total), and a methylcyclohexane feed rate of 12 mL h-1. Information regarding the structure of Ni-Zn catalysts was obtained by N2 and CO adsorption, temperature-programmed reduction, high-resolution transmission electron microscopy, X-ray photoelectron spectroscopy, in situ X-ray diffraction, and in situ X-ray absorption spectroscopy. The results have shown that the addition of zinc leads to the hindrance of Ni reducibility along with weakening the Ni interaction with the silica matrix. This behavior particularly indicated the formation of solid oxide nickel-zinc solutions. The catalytic properties of Zn-modified catalysts in the dehydrogenation of methylcyclohexane appeared significantly superior to their Ni-Cu counterparts. For example, the selectivity of Zn/Ni-SiO2 catalysts toward toluene formation increased markedly with a decrease in the Ni : Zn mass ratio, achieving 97% at 350 °C over the sample with Ni : Zn = 80 : 20. This is attributed to the promoting geometric and electronic effects arising from the formation of bimetallic Ni-Zn solid solutions. Moreover, a deeper reduction of zinc and a more efficient formation of solid bimetallic solutions are observed after the catalytic tests.


Integrated Backward Second-Harmonic Generation through Optically Induced Quasi-Phase-Matching.
Quasi-phase-matching for efficient backward second-harmonic generation requires sub-μm poling periods, a nontrivial fabrication feat. For the first time, we report integrated first-order quasiphase-matched backward second-harmonic generation enabled by seeded all-optical poling. The self-organized grating inscription circumvents all fabrication challenges. We compare backward and forward processes and explain how grating period influences the conversion efficiency. These results showcase unique properties of the coherent photogalvanic effect and how it can bring new nonlinear functionalities to integrated photonics.


Anatomical Considerations and Procedure-Specific Aspects Important in Preventing Operative Morbidity during Transanal Total Mesorectal Excision.
As transanal total mesorectal excision (taTME) becomes increasingly utilized, its technical challenges and potential pitfalls have become more clearly appreciated. This chapter explores the differences in how anatomy presents itself from the taTME vantage point as compared with traditional approaches to taTME, and how special problems unique to taTME pose a new set of operative challenges. Morbidity related, specifically, to the technique of taTME is also delineated with particular focus on male urethral injury.


Interference of poly(ethylene glycol)-lipid analogues with cationic-lipid-mediated delivery of oligonucleotides; role of lipid exchangeability and non-lamellar transitions.
Cationic liposomes are applied to transfer oligonucleotides (ODNs) into cells to regulate gene expression for gene therapeutic or cell biological purposes. In vivo, poly(ethylene glycol) (PEG)-lipid derivatives are employed to stabilize and prolong the circulation lifetime of nucleic acid-containing particles, and to improve targeting strategies. In this study, we have studied the effects of PEG-lipid analogues, i.e. PEG coupled to either phosphatidylethanolamine (PE) or ceramide, on cationic-lipid-DNA complex ('lipoplex') assembly and the mechanism of cationic-lipid-mediated delivery of ODNs in vitro. Inclusion of 10 mol% PEG-PE in ODN lipoplexes inhibited their internalization in Chinese hamster ovary cells by more than 70%. The intracellular fraction remained entrapped in the endosomal/lysosomal pathway, and no release of ODNs was apparent. Similar observations were made for complexes prepared from liposomes that contained PEG-ceramides. Interestingly, delivery resumed when lipoplexes had been externally coated with PEG-ceramides. In this case, the kinetics of delivery were dependent on the length of the ceramide acyl chain, consistent with a requirement for the PEG-lipid to dissociate from the complex. Moreover, although the chemical nature of the PEG-ceramides distinctly affected the net internalization of the complexes, impediment of delivery was largely related to an inhibitory effect of the PEG-lipid on the release of ODNs from the endosomal compartment. Cryo-electron microscopy and small-angle X-ray scattering revealed that the PEG-lipids stabilize the lamellar phase of the lipoplexes, while their acyl-chain-length-dependent transfer from the complex enables adaptation of the hexagonal phase. Within the endosomal compartment, this transition appears to be instrumental in causing the dissociation and cytosolic release of the ODNs for their nuclear homing.


Oromandibular Dystonia - A Systematic Review.
Oromandibular dystonia (OMD) is a clinical problem which is commonly encountered in the practice of movement disorders. OMD results from a variety of genetic and acquired etiologies and can occur as an isolated manifestation, or as part of an isolated generalized or a combined dystonia syndrome. There are only very few systematic reviews on this condition which often causes significant disability. We review here the etiology, clinical features, diagnostic approach and management of OMD.


Purification and characterisation of two porcine hypothalamic fractions with LH-releasing activity: evidence for a single LH and FSH-releasing hormone.
Two fractions with LH releasing activity were obtained from porcine hypothalami by Enzmann et al. (1972). These fractions were designated E 57 and E 203. By comparison with pure natural LH-RH and synthetic LH-RH, E 203 was identified as purified natural LH-RH. E 57, which has only 25% of the activity of E 203, was further evaluated for its biological and immunological activity. E 57 has the same ratio of LH/FSH-releasing activity as natural LH-RH. The binding to a specific LH-RH antiserum is identical with that of natural LH-RH and E 203. It is concluded, that E 57 is an artifact of isolation derived from natural LH-RH. These findings can be considered as additional evidence for the identity of the hypothalamic LH and FSH releasing hormone, since both gonadotrophins are stimulated by the two hypothalamic fractions.


Acute porphyrias: diagnosis and management.
To summarize recent information about acute porphyrias and to provide clinicians with a practical diagnostic and management approach, we reviewed the pertinent literature and our clinical experience. The acute porphyrias are characterized by recurrent attacks of abdominal pain with or without additional manifestations of autonomic dysfunction or neuropsychiatric symptoms. On the basis of the potential of these disorders to affect the skin, they are further subdivided into neuroporphyrias and neurocutaneous porphyrias. During acute attacks, acute porphyria is always associated with increased levels of urinary porphyrin precursors. Between attacks, patients with neurocutaneous porphyrias may have normal urinary porphyrins; therefore, stool porphyrins, which are invariably increased, are the most helpful. Latent disease can be detected by the measurement of either urinary and stool porphyrins or cellular enzyme activity. Specific intravenous therapy with hematin has resulted in biochemical remissions, but its clinical benefit remains controversial. Measurement of urinary and stool porphyrins or porphyrin precursors is critical for the diagnosis of clinically overt acute porphyria. Enzyme assays are helpful in supporting the diagnosis but are best used to identify family members with latent disease. Preventive measures and supportive therapy are the mainstays of current management of patients with porphyria.


Confidence of IRB/REC Members in Their Assessments of Human Research Risk: A Study of IRB/REC Decision Making in Action.
Understanding how institutional review boards/research ethics committees (IRBs/RECs) perform risk/benefit assessment is important to help improve their function. In environmental ethics, uncertainty about potential outcomes and the precautionary principle play important roles in regulatory oversight but have received little attention in the context of human research ethics. We carried out an empirical study to gain insight into uncertainty by asking IRB/REC members about confidence in their risk assessments immediately after discussion of new protocols under review. Based on 12 meetings carried out by four IRBs/RECs over a 6-month period, we found a robust, inverse relationship between risk and confidence. As risk increased, confidence decreased. We detected different patterns of consensus between different IRBs/RECs and their members. Our study introduces a novel and relatively easy to implement approach to begin to understand IRB/REC decision making in real time that can be used within or across institutions.


Chicks produced in the Magellanic penguin (Spheniscus magellanicus) after cloacal insemination of frozen-thawed semen.
The in vitro and in vivo functionality of cryopreserved spermatozoa was examined over two breeding seasons in a zoological colony of Magellanic penguins (Spheniscus magellanicus). Frozen-thawed semen was inseminated into five anesthetized females, over a total of eight egg production cycles, with a different male used for each artificial insemination (AI) within each season. Females were maintained within the colony in cordoned nest sites to prevent copulation with their paired male, and were inseminated every 3-10 days until the first oviposition. Semen frozen from seven males using a straw method retained 39.8%, 25.7%, 74.0%, and 52.1% of its initial total motility, progressive motility, average path velocity, and plasma membrane integrity, respectively. Normal morphology of motile cells was reduced (P < 0.05) during freeze-thawing from 76.7% immediately prior to freezing to 65.3% post-thawing. Conceptive females received 1.6 ± 0.2 inseminations before the first oviposition, with 19.2 ± 1.6 × 10(6) motile, morphologically normal spermatozoa per insemination. Overall fertility was 53.3% (8/15 eggs), hatchability was 50.0% (4/8), and genetic analyses confirmed that all embryos and hatchlings were sired by the AI male. Fertile eggs were laid at 4.0-12.1 days after AI, indicating that frozen-thawed spermatozoa resided in the female reproductive tract for up to ∼7.2 days prior to fertilization. Results demonstrate that frozen-thawed Magellanic penguin spermatozoa are fully functional in vivo and support the use of genome banking and AI as tools for managing the sustainability of zoological penguin populations. Zoo Biol. 35:326-338, 2016. © 2016 Wiley Periodicals, Inc.


Danhong injection in the treatment of chronic stable angina: study protocol for a randomized controlled trial.
Chronic stable angina is a leading cause of death worldwide. Danhong injection, a complementary alternative medicine for chronic stable angina, has been demonstrated to be effective in numerous studies and is widely prescribed to patients. However, the methodological quality of most prior studies was found to be, in general, low. Therefore, we designed this randomized controlled trial to evaluate the efficacy and safety of using Danhong injection to treat chronic stable angina. This is a randomized multicentre, double-blind, placebo-controlled, adaptive clinical trial. A total of 870 patients meeting the eligibility criteria will be randomly assigned into either the Danhong injection or the placebo group in a 2:1 ratio. Participants will then undergo a 2-week treatment regimen and a 76-day follow-up period. Because this is an adaptive trial, two interim analyses are prospectively planned. These will be performed after one-third and two-thirds of the patients, respectively, have completed the trial. Based on the results of these interim analyses, a data monitoring committee will determine how to modify aspects of the study without undermining the validity and integrity of the trial. The primary outcome measure is the proportion of patients who show a clinically significant change, which is defined as at least a 20-point improvement in angina frequency score on the Seattle Angina Questionnaire, which will be administered on day 30. Other secondary efficacy and safety outcomes will also be assessed. This trial will provide high-quality evidence regarding the use of Danhong injection to treat chronic stable angina. ClinicalTrials.gov: NCT01681316 .


Are anomalous invasion speeds robust to demographic stochasticity?
Two important issues for conservation are the range expansion of species as a result of climate change and the invasion of exotic species. Being able to predict the rate at which species spread is key for successful management. In deterministic models, the invasion speed of a polymorphic population can be faster than that of any of the component phenotypes, and these "anomalous" invasion speeds persist even when the mutation rate between phenotypes is vanishingly small. Here we investigate whether the same phenomenon is observed in a model with demographic stochasticity. The model that we use is discrete in time and space and we carry out numerical simulations to determine the invasion speed of a population that has two morphs which differ in their dispersal abilities. We find that anomalous speeds are observed in the stochastic model, but only when the carrying capacity of the population is large or the mutation rate between morphs is high enough. These results suggest that only species with large population sizes, such as many insect species, may be able to invade faster if they are polymorphic than if there is only a single morph present in the population.


Stroke in Melbourne. A study of the relationship between a teaching hospital and the community.
A prospective study of stroke admissions to the Royal Melbourne Hospital was planned to provide baseline information before the introduction of Medibank. One hundred and seventy patients admitted consecutively over a six-month period were followed until either three months had elapsed since admission, or death had supervened. Although over half the cases (57%) came from areas close to the hospital, the remainder come from a wide area embracing most of suburban Melbourne. Of the cases expected from the local areas, less than one-third were admitted to the Royal Melbourne Hospital. Expected demand for admission to hospital from the north-west region was calculated and the dependence of the community upon the private sector demonstrated.


Preoperative serum thyroglobulin concentration is an independent predictive factor of malignancy in follicular neoplasms of the thyroid gland.
A distinction between a benign follicular neoplasm (FN) and a malignant FN based entirely on cytologic examination of fine-needle aspiration biopsy is not possible. The aim of this retrospective study was to find predictive factors of carcinoma in patients with FN. A chart review of 388 patients (314 females, 74 males; mean age 50 years, range 9-81 years) with FN, who were surgically treated between 1988 and 2009, was performed. Predictive factors for malignancy were identified by the chi-squared test and multivariate logistic regression. The histopathological diagnoses were carcinoma, adenoma, and benign goiter in 127 (33%), 126 (32%), and 135 (35%) patients, respectively. The independent predictors of malignancy as shown by multivariate logistic regression were age of patients (OR 1.88, P = 0.008), solitary tumor (OR 1.72, P = 0.037), and Tg concentration (OR 2.36, P = 0.001). Carcinoma was more common in patients younger than 45 years, with solitary tumor and with preoperative serum Tg concentration more than 400 ng/ml. The independent predictors of malignancy in FN were age of patients, solitary tumor, and preoperative Tg concentration. The determination of Tg concentration may be useful in the patients with FN in order to decrease the number of completion thyroidectomies.


Percutaneous pediculoplasty for traumatic pedicle fracture. A technical case report.
The objective of this case is to illustrate a technique for performing fluoroscopically guided percutaneous pediculoplasty in the setting of traumatic or non-neoplastic pedicle fractures. Pediculoplasty has been described in the literature as a complimentary technique performed during vertebroplasty. In this case, isolated pediculoplasty is demonstrated using existing vertebroplasty equipment, which may be utilized as a primary intervention for pedicle fractures in patients who are poor surgical candidates.


Ontogeny of neuroepithelial bodies: correlations with mitogenesis and innervation.
This paper summarizes current knowledge and advances speculation about the formation of the neuroendocrine system of mammalian lungs (comprising uninnervated solitary and clustered small-granule cells and innervated neuroepithelial bodies). It relates the initial appearance of neuroendocrine cells to regulation of mitotic activity in the epithelium during the development of the lung and pays special attention to the later in growth of nerves that converts some of them into neuroepithelial bodies, structures considered ideally adapted to function as chemoreceptors. A few original observations from ongoing immunohistochemical, electron microscopic, and analytical studies have been included here and there to point the discussion. The neuroendocrine cells are derived from undifferentiated precursors present in the endodermal pulmonary epithelium. At an early pseudoglandular stage of lung development these precursors begin to differentiate into neuroendocrine small-granule cells, commencing in the larynx and upper trachea, and expanding centrifugally into pulmonary airways almost as rapidly as these are laid down. Subsequently many of the intrapulmonary small-granule cell clusters become innervated. This event, the delayed appearance of small-granule cells synthesizing other than the dominant peptides and amines (calcitonin gene-related peptide and serotonin in rodents, gastrin-releasing peptide and serotonin in human beings), and other regional adjustments yield the population distribution present in the lungs of adults. Neuroendocrine cell precursors normally differentiate into typical serotonin- or peptide-synthesizing small-granule cells without requiring direct contact by nerves, and dissociated cells from a previously innervated population continue to exhibit physiological characteristics of oxygen sensors despite the loss of contact with nerves. Development of the innervation occurs in stages. Small-granule cell clusters are reached first by ganglion cells derived from pulmonary neuroblasts and later on by processes of extrinsic sensory nerves. The latter not only convey information to the central nervous system but also serve in a variety of ways to extend the neuroepithelial bodies' sphere of influence within the lung itself.


The nature and distribution of gonadotropin-releasing hormones in brains and plasma of ranid frogs.
Highly specific antisera and reversed-phase HPLC were used to examine the nature and distribution of gonadotropin-releasing hormone (GnRH) in the brains and plasma of three species of Rana frog previously reported to vary in the types and brain distributions of GnRH. The three species examined, Rana pipiens, Rana esculenta, and Rana ridibunda, exhibit differences in total concentration and relative concentrations of GnRH in brain areas, but all three contain the mammalian and chicken-II forms of GnRH (mGnRH and cGnRH-II). Both forms were found in the telencephalon and diencephalon, but mGnRH is consistently the most abundant form in the preoptic-hypothalamic area (e.g., ratios of mGnRH:cGnRH-II > 3:1 in hypothalamus-median eminence), whereas, cGnRH-II is the most abundant in the telencephalon and the only form measured in the cerebellum and medulla. The total content of cGnRH-II in the whole brain is about 1.5-2 times higher than that of mGnRH, due to the larger size of the areas outside the preoptic-hypothalamic area. These general patterns were the same for adults and juveniles. We found no evidence of a third form of GnRH corresponding to salmon GnRH in hypothalamus or other brain areas of R. esculenta as previously reported. These analyses also revealed the presence of both forms of GnRH in plasma draining the hypothalamic area, but the concentration of cGnRH-II is relatively higher than that in the corresponding hypothalamic tissue, suggesting differentially greater release or slower degradation of this form. These results do not support the conclusion that the ranid frogs are highly variable in the nature and distribution of GnRH in the brain, although they suggest that both forms of GnRH are potentially involved in the direct regulation of pituitary gonadotropes.


Deep learning-based automated diagnosis of fungal keratitis with in vivo confocal microscopy images.
The aim of this study was to develop an intelligent system based on a deep learning algorithm for automatically diagnosing fungal keratitis (FK) in in vivo confocal microscopy (IVCM) images. A total of 2,088 IVCM images were included in the training dataset. The positive group consisted of 688 images with fungal hyphae, and the negative group included 1,400 images without fungal hyphae. A total of 535 images in the testing dataset were not included in the training dataset. Deep Residual Learning for Image Recognition (ResNet) was used to build the intelligent system for diagnosing FK automatically. The system was verified by external validation in the testing dataset using the area under the receiver operating characteristic curve (AUC), accuracy, specificity and sensitivity. In the testing dataset, 515 images were diagnosed correctly and 20 were misdiagnosed (including 6 with fungal hyphae and 14 without). The system achieved an AUC of 0.9875 with an accuracy of 0.9626 in detecting fungal hyphae. The sensitivity of the system was 0.9186, with a specificity of 0.9834. When 349 diabetic patients were included in the training dataset, 501 images were diagnosed correctly and thirty-four were misdiagnosed (including 4 with fungal hyphae and 30 without). The AUC of the system was 0.9769. The accuracy, specificity and sensitivity were 0.9364, 0.9889 and 0.8256, respectively. The intelligent system based on a deep learning algorithm exhibited satisfactory diagnostic performance and effectively classified FK in various IVCM images. The context of this deep learning automated diagnostic system can be extended to other types of keratitis.


Treatment of primary and secondary spontaneous pneumothorax using videothoracoscopy.
Our aim is to assess the results of surgical treatment for spontaneous pneumothorax (SP) using video-assisted thoracoscopic surgery (VATS) and to determine whether this technique is equally effective for primary SP (PSP) and secondary SP (SSP). A prospective study was performed on 54 patients with persistent and recurrent SP (42 PSPs and 12 SSPs). Mean operating time, time before drainage removal, and hospital stay were significantly longer with SSP (67.1 vs. 48.1 minutes; 5.4 vs. 3.6 days; and 7.7 vs. 5.3 days; p < 0.01, respectively). There was one conversion due to bleeding (PSP). Postoperative complications were greater with SSP (33% vs. 12%; p = 0.09). Although these were mild in all cases, the most common was air leak (25 vs. 5%; p = 0.06). The SP persisted in three cases [2 (5%) with PSP, and 1 (8%) with SSP] and recurred in 2 (1 in each group; 2% and 8%, respectively). Follow-up averaged 24 months. VATS is effective in the surgical treatment of PSP and SSP, although the technical difficulty, morbidity rate, hospital stay, and incidence of persistences and recurrences are greater in the latter than in the former.


The natural history of aortic valve stenosis.
Despite different aetiologies, acquired aortic stenosis is a self-maintaining, slowly progressive process with good long-term prognosis. In 142 patients with mild stenosis, there was clinical progression within 10 years of the initial diagnosis in only 12% of patients. Twenty-five years after the diagnosis had been established, the severity of aortic stenosis was clinically unchanged in 38%, while 25% of patients had moderate stenosis and 35% had undergone valve replacement. Progression of moderate aortic stenosis was more rapid: the average time interval between the manifestation of moderate aortic stenosis and surgery was 13.4 years. Age at the onset of initial symptoms was related to aetiology: 39 +/- 18 years with rheumatic aortic stenoses, 48 +/- 6 years in patients with bicuspid valves who had no history of rheumatic fever, infective endocarditis or myocarditis, and 66 +/- 12 years in degenerative, calcific stenoses of tricuspid aortic valves. Patients with haemodynamically severe stenosis who had refused the recommended operation (n = 55) had an overall poor prognosis: mean survival averaged 23 +/- 5 months and the five-year probability of survival was 18 +/- 7%. All these patients died within 12 years of observation. Mean survival after the occurrence of angina pectoris was 45 +/- 13 months, after syncope 27 +/- 15 months, and after first occurrence of left heart failure 11 +/- 10 months.


Prognostic impact of peritonitis in hemodialysis patients: A national-wide longitudinal study in Taiwan.
Peritonitis has been independently associated with increased morbidity and mortality in peritoneal dialysis patients. However, there are few reports on peritonitis in hemodialysis patients. We aim at investigating both the risk profiles and prognostic impact of peritonitis in hemodialysis patients. This nation-wide longitudinal study uses claims data obtained from the Taiwan National Health Insurance Research Database. A total of 80,733 incident hemodialysis patients of age ≥ 20 years without a history of peritonitis were identified between January 1, 1998 and December 31, 2009. Predictors of peritonitis events were estimated using Cox proportional hazard models. Time-dependent Cox proportional hazard models were used to estimate hazard ratio for mortality attributed to peritonitis exposure. Of 80,733 incident hemodialysis patients over a 13-year study period, peritonitis was diagnosed in 935 (1.16%), yielding an incidence rate of 2.91 per 1000 person-years. Female gender, liver cirrhosis and polycystic kidney disease were three of the most significant factors for peritonitis in both non-diabetic and diabetic hemodialysis patients. The cumulative survival rate of patients with peritonitis was 38.8% at 1 year and 10.1% at 5 years. A time-dependent Cox multivariate analysis showed that peritonitis had significantly increased hazard ratio for all cause mortality. Additionally, the risk of mortality remained significantly higher for non-diabetic hemodialysis patients that experienced peritonitis. The risk of peritonitis in hemodialysis patients is higher in female gender, liver cirrhosis and polycystic kidney disease. Although peritonitis is a rare condition, it is associated with significantly poorer outcome in hemodialysis patients.


Development of a reliable and accurate algorithm to quantify the tumor immune stroma (QTiS) across tumor types.
The tumor microenvironment plays an important role in the tumor biology. Overall survival of tumor patients after resection is influenced by tumor-infiltrating lymphocytes (TILs) as a component of the tumor stroma. However, it is not clear how to assess TILs in the tumor stroma due to heterogeneous methods in different cancer types. Therefore, we present a novel Quantification of the Tumor immune Stroma (QTiS) Algorithm to reliably and accurately quantify cells in the tumor stroma. Immunohistochemical staining of CD3 and CD8 cells in sections of metastatic colorectal cancer (mCRC), ovarian cancer (OvCa), hepatocellular carcinoma (HCC), and pancreatic ductal adenocarcinoma (PDAC), alltogether N = 80, was performed. Hot spots of infiltrating immune cells are reported in the literature. Reliability of the hot spot identification of TILs was examined by two blinded observers. Accuracy was tested in 1 and 3 hot spots using computed counting methods (ZEN 2 software counting (ZC), ImageJ software with subjective threshold (ISC) and ImageJ with color deconvolution (IAC)) and compared to manual counting. All tumor types investigated showed an accumulation of TILs in the tumor stroma (peri- and intratumoral). Reliability between observers indicated a high level consistency. Accuracy for CD8+/CD3+ ratio and absolute cell count required 1 and 3 hot spots, respectively. ISC was found to be the best for paraffin sections, whereas IAC was ideal for frozen sections. ImageJ software is cost-effective and yielded the best results. In conclusion, an algorithm for quantification of tumoral stroma could be established. With this QTiS Algorithm counting of tumor stromal cells is reliable, accurate, and cost-effective.


Panel-based genetic diagnostic testing for inherited eye diseases is highly accurate and reproducible, and more sensitive for variant detection, than exome sequencing.
Next-generation sequencing-based methods are being adopted broadly for genetic diagnostic testing, but the performance characteristics of these techniques with regard to test accuracy and reproducibility have not been fully defined. We developed a targeted enrichment and next-generation sequencing approach for genetic diagnostic testing of patients with inherited eye disorders, including inherited retinal degenerations, optic atrophy, and glaucoma. In preparation for providing this genetic eye disease (GEDi) test on a CLIA-certified basis, we performed experiments to measure the sensitivity, specificity, and reproducibility, as well as the clinical sensitivity, of the test. The GEDi test is highly reproducible and accurate, with sensitivity and specificity of 97.9 and 100%, respectively, for single-nucleotide variant detection. The sensitivity for variant detection was notably better than the 88.3% achieved by whole-exome sequencing using the same metrics, because of better coverage of targeted genes in the GEDi test as compared with a commercially available exome capture set. Prospective testing of 192 patients with inherited retinal degenerations indicated that the clinical sensitivity of the GEDi test is high, with a diagnostic rate of 51%. Based on quantified performance metrics, the data suggest that selective targeted enrichment is preferable to whole-exome sequencing for genetic diagnostic testing.


Lumbar Cryptococcal Osteomyelitis Mimicking Metastatic Tumor.
Cryptococcus is an encapsulated, yeast-like fungus that rarely causes infection in immunocompetent patients. We present the case report of a 66-year-old female patient with a history of rectal cancer with an isolated lumbar vertebral cryptococcosis proven by biopsy performed during operation. The patient was not an immunocompromised host and did not have any other risk factors except the history of cured rectal cancer. The presumptive diagnosis based on imaging studies was metastatic spine cancer, so operation was performed. However, cryptococcal osteomyelitis was diagnosed in the pathologic examination. This case report emphasizes that we should be aware that lumbar cryptococcosis can be a rare cause of mimicking lesions with metastatic cancer.


Nitric Oxide Function in an Echinoderm.
In vertebrates, nitric oxide (NO) is synthesized from L-arginine by NO synthase (NOS) and regulates relaxation of smooth muscle by activating the cyclic-GMP (cGMP) generating enzyme soluble guanylyl cyclase (SGC). Here we show that the NO-cGMP pathway mediates relaxation of the cardiac stomach in the starfish Asterias rubens. The NO-donors hydroxylamine, S-nitrosoglutathione (SNOG) and S-nitroso-N-acetylpenicillamine (SNAP) and the NOS substrate L-arginine cause relaxation of the cardiac stomach. The relaxing effect of SNAP is blocked by the SGC inhibitor 1H-[1,2,4]-oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), and the relaxing effect of L-arginine is inhibited by ODQ and the NOS inhibitor Nw-monomethyl-L-arginine (L-NMMA). ODQ and methylene blue also cause contraction, which may be due to inhibition of the relaxing action of NO produced by cells in the cardiac stomach. These results suggest that NO is synthesized in the cardiac stomach and regulates relaxation by activating SGC. NO-cGMP-mediated relaxation of the cardiac stomach may be important during feeding in starfish where the relaxed stomach is everted through an oral opening and over the digestible parts of prey. The discovery of NO-cGMP-mediated relaxation in an echinoderm demonstrates that regulation of smooth muscle tone by this signaling pathway also occurs in animals other than vertebrates.


Two novel mutations of thiazide-sensitive Na-Cl cotrans porter (TSC) gene in two sporadic Japanese patients with Gitelman syndrome.
Gitelman syndrome is a renal disorder characterized by hypokalemia, hypomagnesemia, metabolic alkalosis and hypocalciuria due to the defective tubular reabsorption of magnesium and potassium. This disease is caused by mutations of thiazide-sensitive Na-Cl cotransporter (TSC) gene. Gitelman syndrome is usually distinguished from Bartter syndrome by the presence of both hypomagnesemia and hypocalciuria. However, a phenotypic overlap is sometimes observed. We encountered two sporadic Japanese patients with Gitelman syndrome and analyzed their TSC gene. These patients were diagnosed as Gitelman syndrome by the typical clinical findings and biochemical abnormalities, such as mild muscular weakness, periodic paralysis, tetany, metabolic alkalosis, hypomagnesemia and hypocalciuria. In patient 1, a novel two base deletion (del TG at nucleotide 731 and 732) in exon 5 and a two base deletion (del TT at nucleotide 2543 and 2544) in exon 21 previously reported in a Japanese patient were identified. The patient 2 had a missense mutation (L623P), that was also identified in Japanese patients, and a novel in-frame 18 base insertion in exon 6 as a heterozygous state. Family analysis of two patients confirmed an autosomal recessive inheritance. In conclusion, we add two new mutations of the TSC gene in Japanese patients with Gitelman syndrome. Because the differential diagnosis between Bartter syndrome and Gitelman syndrome is sometimes difficult, molecular analysis would be a useful diagnostic tool, particularly in unusual cases with phenotypic overlapping.


Reading the operation of an acid/base-controllable molecular switch by naked eye.
We report a pH-controllable molecular switch whose switching can be monitored by the naked eye; this system involves the formation of a complex between a [2]rotaxane--featuring dibenzylammonium and 4,4'-bipyridinium stations--and a TTF-side-walled molecular clip.


New drug discovery: extraordinary opportunities in an uncertain time.
The way in which new medicines are discovered has irreversibly changed and the future sustainability of the enterprise is characterized by an unprecedented period of uncertainty. Herein, we convey that these changes provide unprecedented opportunities for many different players within the private and public sectors to work together and develop new models that ensure the sustainability of activities that have had an extraordinary impact; in terms of promoting public health and driving economic value. Specific examples of experiments are provided to demonstrate some of the new thinking that will be needed to ensure continuation of new drug discovery.


The Biological Effects of Pharmaceuticals in the Marine Environment.
Environmental pharmaceuticals represent a threat of emerging concern for marine ecosystems. Widely distributed and bioaccumulated, these contaminants could provoke adverse effects on aquatic organisms through modes of action like those reported for target species. In contrast to pharmacological uses, organisms in field conditions are exposed to complex mixtures of compounds with similar, different, or even opposing therapeutic effects. This review summarizes current knowledge of the main cellular pathways modulated by the most common classes of environmental pharmaceuticals occurring in marine ecosystems and accumulated by nontarget species-including nonsteroidal anti-inflammatory drugs, psychiatric drugs, cardiovascular and lipid regulator agents, steroidal hormones, and antibiotics-and describes an intricate network of possible interactions with both synergistic and antagonistic effects on the same cellular targets and metabolic pathways. This complexity reveals the intrinsic limits of the single-chemical approach to predict the long-term consequences and future impact of pharmaceuticals at organismal, population, and community levels.


Efficacy and outcome of an open access endoscopy service.
In a retrospective questionnaire-study, we have attempted to elucidate how open access endoscopy influences management of dyspeptic patients, with special focus on young patients (less than 40 year), since the proportion of examinations with findings not requiring medical treatment are consistently reported higher in this age group. During a one-year period, 436 patients referred for open access endoscopy and their general practitioners completed questionnaires giving details of medical treatment, consultation rate for dyspepsia and global assessment before and 6-18 months after the endoscopy. Relevant changes in medical treatment (stopped in patients with no or minor abnormalities or started in patients with major abnormalities) was found for 27% of the patients, irrespective of age group. The result of the endoscopy provided reassurance for 70% of the patients with no or minor abnormalities. Reassurance was coupled with a lower consultation rate and with fewer symptoms. Altogether, 83% of the young patients with no or minor abnormalities had a positive outcome of the endoscopy. As the endoscopy service introduced relevant and lasting prescription habits and reduced consultation rates at general practitioners, also for younger dyspeptic patients with no or minor abnormalities, the strategy generally proposed of a trial with H2-receptor antagonists before considering referral for endoscopy should be subjected to formal clinical trial evaluating all relevant levels of efficacy.


In vitro desensitization of isolated nephron segments to vasopressin.
Recent studies have demonstrated that in vivo administration of 1-deamino-8-D-arginine-vasopressin, an analog of arginine-8-vasopressin, induces homologous desensitization to vasopressin in the thick ascending limb of the loop of Henle. Desensitization has been documented by a decreased physiological response to vasopressin in vivo and by a reduced cAMP accumulation in the cortical thick ascending limb (CTAL). By measuring cAMP content in single isolated medullary thick ascending limbs (MTALs), we now report that desensitization can occur all along the thick ascending limb and, more importantly, that it can also be induced in vitro. In a first series of experiments, we observed that 1 hr after in vivo injection of 1-deamino-8-D-arginine-vasopressin, MTALs were desensitized by 80% to vasopressin, whereas the effects of the other hormones acting on the same cyclase pool (glucagon, calcitonin) were fully maintained. In a second set of experiments, desensitization was induced in vitro by vasopressin, the natural hormone. A 60-min preincubation of MTALs with vasopressin caused a marked (up to 86%) and highly reproducible desensitization. The process was dose and time dependent. The apparent Ka for desensitization was 0.2 nM, and the half-maximal effect was obtained within 20 min. The desensitization induced in vitro by vasopressin was again essentially homologous in nature, with 80% of the maximal stimulation of cAMP accumulation being obtained in the presence of glucagon. Desensitization to vasopressin was observed in the presence and absence of indomethacin, indicating that it is independent of prostaglandin synthesis. It is concluded that (i) vasopressin and its analog 1-deamino-8-D-arginine-vasopressin cause marked desensitization in the CTAL and MTAL and (ii) the low vasopressin concentrations required to induce desensitization and the rapid onset of the process suggest that it has a physiological significance.


Labrador sea water spreading and the Atlantic meridional overturning circulation.
In 1982, Talley and McCartney used the low potential vorticity signature of Labrador Sea Water (LSW) to make the first North Atlantic maps of its properties. Forty years later, our understanding of LSW variability, spreading time scales and importance has deepened. In this review and synthesis article, I showcase recent observational advances in our understanding of how LSW spreads from its formation regions into the Deep Western Boundary Current and southward into the subtropical North Atlantic. I reconcile the fact that decadal variability in LSW formation is reflected in the Deep Western Boundary Current with the fact that LSW formation does not control subpolar overturning strength and discuss hypothesized connections between LSW spreading and decadal Atlantic Meridional Overturning Circulation variability. Ultimately, LSW spreading is of fundamental interest because it is a significant pathway for dissolved gasses such as oxygen and carbon dioxide into the deep ocean. We should hence prioritize adding dissolved gas measurements to standard hydrographic and circulation observations, particularly at targeted western boundary locations. This article is part of a discussion meeting issue 'Atlantic overturning: new observations and challenges'.


[Psychological disorders in mental handicap--prevalence and psychopathologic characteristics].
This review of studies on the prevalence of mental disorders in adults with mental retardation comes to the following principal conclusions: Mainly due to research problems and psychiatry's disinterest there continues to be limited epidemiological findings; compared with people of normal intelligence the prevalence rate of mental disorders is about 50% higher in mild mental retardation and 2-3 fold higher in severe mental retardation; substance abuse or -dependence as well as neurotic disturbances may be less prevalent while self-injurious behaviour seems to be rather specific for subjects with mental retardation. As far as factor analytic studies are concerned the two syndromes of irritability and lethargy may play a major role in the psychopathology of severe mental retardation.


Global Gray Water Footprint and Water Pollution Levels Related to Anthropogenic Nitrogen Loads to Fresh Water.
This is the first global assessment of nitrogen-related water pollution in river basins with a specification of the pollution by economic sector, and by crop for the agricultural sector. At a spatial resolution of 5 by 5 arc minute, we estimate anthropogenic nitrogen (N) loads to freshwater, calculate the resultant gray water footprints (GWFs), and relate the GWFs per river basin to runoff to calculate the N-related water pollution level (WPL) per catchment. The accumulated global GWF related to anthropogenic N loads in the period 2002-2010 was 13×10(12) m3/y. China contributed about 45% to the global total. Three quarters of the GWF related to N loads came from diffuse sources (agriculture), 23% from domestic point sources and 2% from industrial point sources. Among the crops, production of cereals had the largest contribution to the N-related GWF (18%), followed by vegetables (15%) and oil crops (11%). The river basins with WPL>1 (where the N load exceeds the basin's assimilation capacity), cover about 17% of the global land area, contribute about 9% of the global river discharge, and provide residence to 48% of the global population.


Neurobehavioural and Analgesic Properties of Ethanol Bark Extract of Terminalia ivorensis A Chev. (Combrataceae) in Mice.
Terminalia ivorensis A. Chev (Combretaceae) is a medicinal plant used in folk medicine in the management of pain, rheumatic condition, gastroenteritis and as a tranquilizer in psychotic disorder. We evaluated neurobehavioural and analgesic properties of the ethanol bark extract of T. ivorensis (EBETI). Effects of EBETI (1.25, 2.5, and 5 mg/kg) on novelty-induced behaviours were determined using open field test. Anxiolytic effect of EBETI (1.25, 2.5, and 5 mg/kg) was assessed using hole-board and elevated-plus maze paradigms. Analgesic property of EBETI (2.5, 5, and 10 mg/kg) was evaluated using acetic acid induced writhing, formalin and tail immersion tests. The extract was administered once intraperitoneally. The LD50 of EBETI was 173 mg/kg. EBETI (1.25, 2.5, and 5 mg/kg) significantly reduced rearing (142.3±1.6, 83.5±1.9, 39.3±1.5) and grooming (33.8±3.4, 28.4±3.0, 18.2±1.7) as compared with controls (180.5±4.9; 52.4±5.2). Treatment with EBETI (1.25, 2.5, and 5 mg/kg) significantly reduced head-dipping on hole-board (9.4±2.3, 6.2±1.9, 5.4±2.9) as compared with control (26.8±1.9). However, there was no anxiolytic effect on EPM. EBETI (2.5, 5, and 10 mg/kg) significantly inhibited abdominal constriction in writhing assay (21.8.0±2.4, 12.2±1.6, 5.8±2.1) as compared with control (35.0±1.7). Inhibition of neurogenic and inflammatory phases of formalin test was notice. However, the extract could not alter response to thermal stimulus in tail immersion test. EBETI is sedative and has analgesic effect, thus supporting its folkloric use in pain management and as a tranquilizer in psychosis.


Nonpeptidic inhibitors of human leukocyte elastase. 3. Design, synthesis, X-ray crystallographic analysis, and structure-activity relationships for a series of orally active 3-amino-6-phenylpyridin-2-one trifluoromethyl ketones.
A series of nonpeptidic inhibitors of human leukocyte elastase (HLE) is reported. These trifluoromethyl ketone-based inhibitors contain a 3-amino-6-phenylpyridone group as a central template. The effect of varying the N-3 substituent in these inhibitors on in vitro potency, physical properties, and oral activity in a hamster based, HLE-induced lung damage model is described. The variety of substituents at this position that have little effect on in vitro potency supports the idea that this region of the molecule does not interact strongly with the enzyme. One exception to this generality is 13k, which is substituted with a (4-acetamidophenyl)sulfonyl group. This compound has a K(i) of 0.7 nM and is, in vitro, the most potent inhibitor in the series. In contrast, variation of the N-3 substituent was found to have a dramatic effect on activity after oral administration. Several analogs, including the parent amine, 7, formamide, 2u, benzyl sulfamide, 13e, and benzyl sulfonamide, 13f, show significant activity when administered at an oral dose of 2.5 mg/kg. Support for the modeling-based design concepts was obtained through in vitro SAR results and X-ray crystallographic analysis of the complex between 13d and porcine pancreatic elastase (PPE), a closely related enzyme.


What is the evidence for stress resistance and slowed aging?
Stress resistance is thought to contribute to slowed-aging, although cause and effect between the two is controversial. On October 30, 2015 researchers gathered at the Front Range Consortium on Stress Resistance and Slowed Aging in Fort Collins, CO, to discuss what the current evidence is that stress resistance imparts slowed aging. Included in that discussion was defining stress resistance, distinguishing if there are key stresses to which resistance imparts slowed aging, what models aid in our understanding of stress resistance and aging, and how to translate that knowledge into slowed aging treatment. The following article is a brief summary of that discussion and recommendations for moving forward.


Hepatic regeneration and functional recovery following partial liver resection in an experimental model of hepatic steatosis treated with omega-3 fatty acids.
Omega-3 fatty acids (FAs) have been shown to reduce experimental hepatic steatosis and protect the liver from ischaemia-reperfusion injury. The aim of this study was to examine the effects of omega-3 FAs on regeneration of steatotic liver. Steatosis was induced in rats by a 3-week methionine/choline-deficient diet, which was continued for an additional 2 weeks in conjunction with oral administration of omega-3 FAs or saline solution. Steatosis was graded histologically and quantified by proton magnetic resonance spectroscopy ((1) H-MRS) before and after the diet/treatment. Liver function was determined by (99m) Tc-labelled mebrofenin hepatobiliary scintigraphy (HBS). In separate experiments, the hepatic regenerative capacity and functional recovery of omega-3 FA-treated, saline-treated or non-steatotic (control) rats were investigated 1, 2, 3 and 5 days after partial (70 per cent) liver resection by measurement of liver weight change and hepatocyte proliferation (Ki-67) and HBS. Severe steatosis (over 66 per cent) in the saline group was reduced by omega-3 FAs to mild steatosis (less than 33 per cent), and hepatic fat content as assessed by (1) H-MRS decreased 2·2-fold. (99m) Tc-mebrofenin uptake in the saline group was more than 50 per cent lower than in the control group, confirming the functional effects of steatosis. (99m) Tc-mebrofenin uptake and regenerated liver mass were significantly greater in the omega-3 group compared with the saline group on days 1 and 3. The posthepatectomy proliferation peak response was delayed until day 2 in saline-treated rats, compared with day 1 in the omega-3 and control groups. Omega-3 FAs effectively reduced severe hepatic steatosis, which was associated with improved liver regeneration and functional recovery following partial hepatectomy.


Flip-flop phenomenon in systemic sclerosis on fluorodeoxyglucose positron emission tomography/computed tomography.
Systemic sclerosis (SSc) is a rare autoimmune disease, which may affect multiple organ systems. Fluorine-18-fluorodeoxyglucose (18F-FDG) positron emission tomography/computed tomography (PET/CT) can demonstrate the degree and anatomical extent of involvement in the entire body and coexisting malignancies in connective tissue diseases. We present a case of SSc with an increased 18F-FDG uptake in the cutaneous and subcutaneous tissues even higher than the neighboring skeletal muscles ("flip-flop phenomenon," that is, an increased 18F-FDG uptake in the skin but a decreased 18F-FDG uptake in the skeletal muscles).


Performance Assessment of a Trypanosoma cruzi Chimeric Antigen in Multiplex Liquid Microarray Assays.
Diagnosing chronic Chagas disease (CD) requires antibody-antigen detection methods, which are traditionally based on enzymatic assay techniques whose performance depend on the type and quality of antigen used. Previously, 4 recombinant chimeric proteins from the Instituto de Biologia Molecular do Paraná (IBMP-8.1 to 8.4) comprising immuno-dominant regions of diverse Trypanosoma cruzi antigens showed excellent diagnostic performance in enzyme-linked immunosorbent assays. Considering that next-generation platforms offer improved CD diagnostic accuracy with different T. cruzi-specific recombinant antigens, we assessed the performance of these chimeras in liquid microarrays (LMAs). The chimeric proteins were expressed in Escherichia coli and purified by chromatography. Sera from 653 chagasic and 680 healthy individuals were used to assess the performance of these chimeras in detecting specific anti-T. cruzi antibodies. Accuracies ranged from 98.1 to 99.3%, and diagnostic odds ratio values were 3,548 for IBMP-8.3, 4,826 for IBMP-8.1, 7,882 for IBMP-8.2, and 25,000 for IBMP-8.4. A separate sera bank (851 samples) was employed to assess cross-reactivity with other tropical diseases. Leishmania, a pathogen with high similarity to T. cruzi, showed cross-reactivity rates ranging from 0 to 2.17%. Inconclusive results were negligible (0 to 0.71%). Bland-Altman and Deming regression analysis based on 200 randomly selected CD-positive and negative samples demonstrated interchangeability with respect to CD diagnostic performance in both singleplex and multiplex assays. Our results suggested that these chimeras can potentially replace antigens currently used in commercially available assay kits. Moreover, the use of multiplex platforms, such as LMA assays employing 2 or more IBMP antigens, would abrogate the need for 2 different testing techniques when diagnosing CD.


A deep learning-based framework for retinal fundus image enhancement.
Low-quality fundus images with complex degredation can cause costly re-examinations of patients or inaccurate clinical diagnosis. This study aims to create an automatic fundus macular image enhancement framework to improve low-quality fundus images and remove complex image degradation. We propose a new deep learning-based model that automatically enhances low-quality retinal fundus images that suffer from complex degradation. We collected a dataset, comprising 1068 pairs of high-quality (HQ) and low-quality (LQ) fundus images from the Kangbuk Samsung Hospital's health screening program and ophthalmology department from 2017 to 2019. Then, we used these dataset to develop data augmentation methods to simulate major aspects of retinal image degradation and to propose a customized convolutional neural network (CNN) architecture to enhance LQ images, depending on the nature of the degradation. Peak signal-to-noise ratio (PSNR), structural similarity index measure (SSIM), r-value (linear index of fuzziness), and proportion of ungradable fundus photographs before and after the enhancement process are calculated to assess the performance of proposed model. A comparative evaluation is conducted on an external database and four different open-source databases. The results of the evaluation on the external test dataset showed an significant increase in PSNR and SSIM compared with the original LQ images. Moreover, PSNR and SSIM increased by over 4 dB and 0.04, respectively compared with the previous state-of-the-art methods (P < 0.05). The proportion of ungradable fundus photographs decreased from 42.6% to 26.4% (P = 0.012). Our enhancement process improves LQ fundus images that suffer from complex degradation significantly. Moreover our customized CNN achieved improved performance over the existing state-of-the-art methods. Overall, our framework can have a clinical impact on reducing re-examinations and improving the accuracy of diagnosis.


Ab initio quantum mechanical calculations of the variation of the 1H and 13C nuclear magnetic shielding constants in proline as a function of the angle psi.
The variation of the nuclear magnetic shielding constant of the different protons and carbons of trans HCO-L-Pro-NH2 with the value of the angle psi is calculated by a non-empirical method for three conformations of the proline ring. The results concerning the CH protons show that the chemical shift of the alpha, beta and gamma endo hydrogens can vary by more than 1 ppm when psi goes from -30 degrees to 180 degrees. The theoretical variation of the chemical shift difference between alpha and gamma or beta and gamma carbons is found to be sensitive to the puckering of the proline ring. For the second of these differences the theoretical results are in agreement with Siemion's relation only for a limited range of molecular conformations. Additional calculations show that the variations of the proton shifts with the value of psi are due to the magnetic anisotropy of the proline carbonyl group and to the polarization of the CH bonds by the multipolar charge distribution carried by this carbonyl. The results are discussed in relation to experiment and the possibility of using 1H and 13C chemical shifts for the determination of the value of the torsion angle about the C alpha C' bond.


Drug use among Sydney teenagers in 1985 and 1986.
To investigate adolescent drug use behaviour, a random community sample of Sydney teenagers aged 14 to 19 years was interviewed at home in 1985 (N = 996) and again in 1986 (N = 756). Respondents were asked about current use of alcohol, tobacco, marijuana, other illicit drugs, medications and inhalants. Drug use was common: 16 per cent of respondents were heavy drinkers, 28 per cent smoked tobacco, 10 per cent used marijuana and 4 per cent used drugs other than alcohol, tobacco or marijuana. Seventeen per cent were multiple drug users. Drug use was more common among boys than girls, except for tobacco smoking, and increased with age: older males had particularly high prevalences of heavy drinking, tobacco and marijuana use. The prevalence of heavy drinking, tobacco and marijuana use increased by 2-3 per cent over the one year follow-up period. About half of the heavy drinkers and marijuana users, and 80 per cent of tobacco smokers had not changed one year later, which indicates the stability of these behaviours. One-third of eligible teenagers contacted at the first interview declined to participate: it is likely that this study underestimates the prevalence of drug use in the community. Heavy drinking, tobacco smoking and marijuana use remain important target behaviours for adolescent drug use prevention programs.


Evaluation of a multidisciplinary concept of mental demands at work on cognitive functioning in old age.
Previous work has shown that high mental demands are associated with better cognitive functioning in old age. As there is a lack of a general conceptual framework for this association, the aim of the study was to investigate how mental demands and other work-related factors relate to cognitive functioning as a foundation for developing such a framework. An expert panel discussion was conducted with the aim of determining relevant work-related factors, which were then tested in a survey with 346 employees aged 50+ years, who were actively working. Assessment of cognitive functioning comprised complex attention, executive function, learning/memory, language, perceptual-motor, and social cognition. Confirmatory factor analysis was conducted to confirm factor belonging. Associations with cognitive functioning were analyzed using structure equation modelling to confirm associations and to identify additional direct and indirect paths. Only 42.3% (22/52) of the work-related factors and 19.0% (4/21) of the mediating paths suggested by the experts were significant with respect to cognitive functioning. Factor analysis and structural equation modeling indicated that high mental demands are only associated with better cognitive functioning in old age to the extent that they are intellectually stimulating and this effect is embedded in individual capacities and the social context. Based on the panel discussion and the empirical testing, we propose the Conceptual Framework of Social Dependency of Intellectual Stimulation on Cognitive Health. We recommend researchers and workplace health experts to pay attention to the component of this theory when assessing workplace risk.


Liquid biopsy epigenomic profiling for cancer subtyping.
Although circulating tumor DNA (ctDNA) assays are increasingly used to inform clinical decisions in cancer care, they have limited ability to identify the transcriptional programs that govern cancer phenotypes and their dynamic changes during the course of disease. To address these limitations, we developed a method for comprehensive epigenomic profiling of cancer from 1 ml of patient plasma. Using an immunoprecipitation-based approach targeting histone modifications and DNA methylation, we measured 1,268 epigenomic profiles in plasma from 433 individuals with one of 15 cancers. Our assay provided a robust proxy for transcriptional activity, allowing us to infer the expression levels of diagnostic markers and drug targets, measure the activity of therapeutically targetable transcription factors and detect epigenetic mechanisms of resistance. This proof-of-concept study in advanced cancers shows how plasma epigenomic profiling has the potential to unlock clinically actionable information that is currently accessible only via direct tissue sampling.


A year of unprecedented progress in Down syndrome basic research.
The years 2006 and 2007 saw the publication of three new and different approaches to prevention or amelioration of Down syndrome effects on the brain and cognition. We describe the animal model systems that were critical to this progress, review these independent breakthrough studies, and discuss the implications for therapeutic approaches suggested by each.


CARD15 polymorphisms are associated with anti-Saccharomyces cerevisiae antibodies in caucasian Crohn's disease patients.
Carriage of CARD15 gene polymorphisms and the serological marker anti-Saccharomyces cerevisiae antibodies (ASCA) are two markers for Crohn's disease (CD). Similar phenotypes have been associated with both markers. In the present study we analysed whether both markers were associated with each other and, if so, whether this association could be explained by a direct link or by an indirect association with those phenotypes. Therefore, we included 156 consecutive Caucasian CD patients and assessed the prevalence of the three common single nucleotide polymorphisms in the CARD15 gene. Serum samples were analysed for IgA and IgG ASCA by ELISA. CD patients with CARD15 polymorphisms were more frequently ASCA positive (OR 2.7 (1.4-5.2); P = 0.002) and had higher titres for ASCA IgA (P = 0.005) and ASCA IgG (P < 0.001) compared to patients carrying the wild type polymorphisms. Multivariate analysis demonstrated that this association was independent from ileal disease, penetrating disease and stricturing disease, the need for resective bowel surgery, familial cases, smoking habits and early age at onset. Homozygotes or compound heterozygotes for CARD15 polymorphisms had significantly more frequent ASCA positivity compared to single heterozygotes (OR 9.1 (1.1-74.2), P(c) (corrected P-value) = 0.030). These data indicate that there is a significant association between the carriage of CARD15 polymorphisms and ASCA, independent of the described phenotypes. Moreover, ASCA positivity is more frequent in CD patients carrying 2 CARD15 polymorphisms compared to single heterozygotes.


Physiologic effects of forced down-regulation of dnaK and groEL expression in Streptococcus mutans.
Strains of Streptococcus mutans lacking DnaK or GroEL appear not to be isolable. To better distinguish the roles played by these chaperones/chaperonins in the physiology of S. mutans, we created a knockdown strategy to lower the levels of DnaK by over 95% in strain SM12 and the level of GroEL about 80% in strain SM13. Interestingly, GroEL levels were approximately twofold higher in SM12 than in the parent strain, but the levels of DnaK were not altered in the GroEL knockdown strain. Both SM12 and SM13 grew slower than the parent strain, had a strong tendency to aggregate in broth culture, and showed major changes in their proteomes. Compared with the wild-type strain, SM12 and SM13 had impaired biofilm-forming capacities when grown in the presence of glucose. The SM12 strain was impaired in its capacity to grow at 44 degrees C or at pH 5.0 and was more susceptible to H(2)O(2), whereas SM13 behaved like the wild-type strain under these conditions. Phenotypical reversions were noted for both mutants when cells were grown in continuous culture at a low pH, suggesting the occurrence of compensatory mutations. These results demonstrate that DnaK and GroEL differentially affect the expression of key virulence traits, including biofilm formation and acid tolerance, and support that these chaperones have evolved to accommodate unique roles in the context of this organism and its niche.


[Mediastinal sonography in childhood. Results of 310 studies].
Following the performance of 310 mediastinal ultrasonograms over 3 years in children up to an age of 15 years, the authors report on the indication applied, their methods, and the results. Sonography allows an easy approach to upper and anterior mediastinal structures in children. Thus, differentiation between a normal thymus and malignant infiltration, which is difficult on plain films, is possible in almost all cases without further radiological examinations. Except in the hilar area, sonography is sufficient to detect lymphomas. In patients with congenital stridor a vascular etiology, e.g. an aberrant brachiocephalic trunk, can be visualized or excluded.


Early and late auditory information processing show opposing deviations in aniridia.
Aniridia is a congenital disorder, predominantly caused by heterozygous mutations of the PAX6 gene. While ocular defects have been extensively characterized in this population, brain-related anatomical and functional abnormalities are emerging as a prominent feature of the disorder. Individuals with aniridia frequently exhibit auditory processing deficits despite normal audiograms. While previous studies have reported hypoplasia of the anterior commissure and corpus callosum in some of these individuals, the neurophysiological basis of these impairments remains unexplored. This study provides direct assessment of neural activity related to auditory processing in aniridia. Participants were presented with tones designed to elicit an auditory steady-state response (ASSR) at 22 Hz, 40 Hz, and 84 Hz, and infrequent broadband target tones to maintain attention during electroencephalography (EEG) recording. Persons with aniridia showed increased early cortical responses (P50 AEP) in response to all tones, and increased high-frequency oscillatory entrainment (84 Hz ASSR). In contrast, this group showed a decreased cortical integration response (P300 AEP to target tones) and reduced neural entrainment to cortical beta-band stimuli (22 Hz ASSR). Collectively, our results suggest that subcortical and early cortical auditory processing is augmented in aniridia, while functional cortical integration of auditory information is deficient in this population.


Dental fear in adults: a meta-analysis of behavioral interventions.
The aim of this meta-analytic and systematic quantitative approach is to examine the effects of behavioral interventions for dental anxiety and dental phobia. Eighty studies were identified where dental fear treatment with behavioral methods was evaluated. Thirty-eight of 80 met entry criteria and were included in a meta-analysis. The calculated effect sizes (ESs) for self-reported anxiety after intervention indicate positive changes in 36 of the 38 studies and no changes in two. The overall ES = 1.8 (95% CI: 1.6, 1.8). The percent of subjects with post-treatment dental visits in the first 6 months post-treatment varied between 50 and 100%. The overall ES for attendance at dental visits, weighted by sample size, is 1.4 (95% CI: 1.3, 1.6). The homogeneity analysis indicates that the studies cannot be adequately described in one ES. The reported percentage of subjects with a dental visit between 6 months and 4 years post-treatment varied from 48 to 100%. The overall weighted ES for visiting the dentist, adjusted for drop-outs in the studies, is 1.2 (95% CI: 0.99, 1.4). Despite extensive heterogeneity, changes in self-reported anxiety represent medium to large ESs. Patients signing up for a behavioral intervention for dental fear can be expected to report a significant reduction in their fear, and this effect generally seems to be lasting. Mean long-term attendance ( >4 years after treatment) is 77%.


Analgesic effect of the electromagnetic resonant frequencies derived from the NMR spectrum of morphine.
Exposure to various types of electromagnetic fields (EMFs) affects pain specificity (nociception) and pain inhibition (analgesia). Previous study of ours has shown that exposure to the resonant spectra derived from biologically active substances' NMR may induce to live targets the same effects as the substances themselves. The purpose of this study is to investigate the potential analgesic effect of the resonant EMFs derived from the NMR spectrum of morphine. Twenty five Wistar rats were divided into five groups: control group; intraperitoneal administration of morphine 10 mg/kg body wt; exposure of rats to resonant EMFs of morphine; exposure of rats to randomly selected non resonant EMFs; and intraperitoneal administration of naloxone and simultaneous exposure of rats to the resonant EMFs of morphine. Tail Flick and Hot Plate tests were performed for estimation of the latency time. Results showed that rats exposed to NMR spectrum of morphine induced a significant increase in latency time at time points (p < 0.05), while exposure to the non resonant random EMFs exerted no effects. Additionally, naloxone administration inhibited the analgesic effects of the NMR spectrum of morphine. Our results indicate that exposure of rats to the resonant EMFs derived from the NMR spectrum of morphine may exert on animals similar analgesic effects to morphine itself.


Identifying signals of suffering by linking verbal and facial cues.
Here, the authors describe microanalytically the two main behavioral states in suffering (enduring and emotional suffering) so that in subsequent research, appropriate comforting responses to ease and relieve suffering can be identified for each behavioral state. Their objectives were to describe the facial expressions of enduring and emotional suffering, and to link them with verbal narrative and thus develop a microanalytic description of each behavioral state. Using Ekman's modified EMFACS, they videotaped interviews with 19 participants and coded co-occurring verbal text and expressions. They also documented differences between each behavioral state and the transitions from enduring to emotional suffering. Enduring and emotional suffering are distinct and identifiable behaviors. These formerly implicit behavioral cues can be used in clinical assessment and research.


Trans-2-Acetoxycyclobutyltrimethylammonium iodide, a cyclobutane analog of "trans-ACTM".
The (+/-) title compound was prepared to evaluate prior observations that certain acetylcholine congeners derived from cyclobutane are devoid of muscarinic effects. It was prepared by a multistep sequence from trans-2-carbomethoxycyclobutyl methyl ketone. In a guinea pig ileum assay, it was 0.02 times as active as AcCh, and in a dog blood pressure assay, it was 0.09 times as active. In these assays, its (+/-)-cyclopropane congener and AcCh were equally active. This is the first cyclobutane-derived AcCh congener possessing significant muscarinic effect.


Slow-pull compared to suction technique for EUS-guided sampling of pancreatic solid lesions: a meta-analysis of randomized controlled trials.
Background and study aims Current ESGE guidelines suggest employing the suction (SU) technique for endoscopic ultrasound (EUS)-guided sampling of pancreatic solid lesions. Nonetheless, recent randomized controlled trials (RCT) have reported that the slow-pull (SP) technique has similar diagnostic accuracy with possibly less blood contamination. However, these results are heterogeneous and limited to small cohorts. The aim of this meta-analysis was to compare adequacy, accuracy, sensitivity and specificity of the SU and SP techniques for EUS-guided sampling of solid pancreatic lesions. Methods A computerized bibliographic search was restricted to RCTs. Pooled effects were calculated using a random-effects model and expressed in terms of pooled sensitivity and specificity and OR (95 % CI) for adequacy and accuracy. Results Overall, seven RCTs were included, for a total of 475 patients (163 lesions sampled with SU, 164 with SP and 148 by both). The adequacy was similar (OR = 0.98) without heterogeneity (I 2 = 0 %), but a high degree of blood contamination was more common with SU than SP (pooled rate 27.6 % vs 19.7 %). A non-significant superiority of SP in terms of pooled accuracy (OR = 0.82; 95 % CI 0.36-1.85) was recorded, with moderate heterogeneity (I 2 = 52.4 %). The SP technique showed a slightly higher pooled sensitivity compared to SU (88.7 % vs 83.4 %), while specificity was similar (97.2 % SP vs 96.9 % SU), with considerable heterogeneity. Conclusion The current meta-analysis reveals non-superiority of SU over SP, while SP results in reduced blood contamination. If the 5 % accuracy difference favouring SP is true, with alfa error = 0.05 and beta = 0.20, a RCT of 982 patients per arm is needed to confirm significance.


An Interactive Ambulatory Nephrology Curriculum for Internal Medicine Interns: Design, Implementation, and Participant Feedback.
While diminishing nephrology fellow recruitment is a known issue, more work is needed to evaluate possible interventions to reverse this trend. We designed and implemented a curriculum to increase exposure to ambulatory nephrology among internal medicine interns. The curriculum focused on key aspects of outpatient nephrology practice, including supervised clinic visits, formal themed didactic content, and an online interactive forum with assigned evidence-based readings and small-group responses to relevant cases. We obtained postcourse surveys from all participating interns. Of the 43 interns who took part in the first year of the ambulatory nephrology curriculum, 100% reported a positive didactic experience and 91% reported a positive interactive online experience. 77% reported an improvement in their familiarity with clinical nephrology practice (median 2-point increase in familiarity score on a 7-point scale, P<0.001 by signed rank testing). Qualitative feedback included praise for the high-yield topics covered by the lectures and energizing teachers. In conclusion, we successfully implemented an ambulatory nephrology curriculum using a framework that integrated formal didactics, interactive online learning, and key clinical components of outpatient nephrology care. Future investigation will evaluate whether early implementation of this curriculum is associated with increased pursuit of nephrology as a career.


Vestibular function and cochlear implant.
The authors illustrate their personal experience relating to 32 patients, aged between 12 and 74 years, undergoing cochlear implant, in whom vestibular reflexes were evaluated before and after surgery. This series did not include cases of areflexia, but only 1 case of reduced vestibular reflexia consequently to surgery. In this case, owing to the probable intervention of central compensation processes, labyrinthine hyporeflexia never became clinically significant. These personal results enable the authors to affirm that preoperative vestibular reflexes do not offer elements able to influence the choice of the ear in which to perform the cochlear implant. In the series of patients reported by the authors, a cochleostomy by removal of the floor of the round window niche, following the suggestions of O'Leary et al., always headed the electrode implant. This contrivance may reduce or eliminate the negative effects on vestibular receptors indirectly caused by the consequent and inevitable alteration of perilymph pressure produced by the implant.


miR-429 regulates alveolar macrophage inflammatory cytokine production and is involved in LPS-induced acute lung injury.
p38 MAPK (mitogen-activated protein kinase) is a critical regulator in lung inflammation. It can be inactivated by DUSP1 (dual-specificity phosphatase 1) which was identified as a putative target of miR-429. miR-429 mimics directly targeted to the 3'-UTR of the gene encoding DUSP1 may result in the translational attenuation of DUSP1. Moreover, the phosphorylation of p38 MAPK was prolonged after miR-429 mimic treatment. Additionally, miR-429 expression was sensitive to LPS (lipopolysaccharide) stimulation and the miR-429 mimics increased the production of pro-inflammatory cytokines. However, anti-miR-429 reduced the LPS-induced production of pro-inflammatory cytokines. These results provide direct evidence that miR-429 is involved in the LPS-induced inflammatory response. In parallel with miR-429, miR-200b and miR-200c, but not miR-200a or miR-141, shared similar effects. In vivo, LPS induced the expression of miR-429, miR-200b and miR-200c in lung. At the same time, inhibiting these miRNAs by anti-miRNAs attenuated the LPS-induced pulmonary inflammatory response and injury. These findings reveal that miR-429 possesses pro-inflammatory activities and may be a potential therapy target for LPS-induced lung injury.


Pharmacological therapy of acute cardiogenic pulmonary oedema in the emergency department.
This paper critically reviews the major drug types that are currently used in the management of acute cardiogenic pulmonary oedema. As decompensated heart failure becomes an increasingly common problem in emergency departments in the developed world, optimization of emergency drug therapy for these critically ill patients is essential. The evidence base for 'routine therapy' in the ED is considered. The review also briefly considers emerging pharmacological therapies that may have an impact on future management of cardiogenic pulmonary oedema.


New thin-film adhesive for sealing full-thickness corneal incisions in rabbits.
To compare the repair of penetrating corneal incisions in an in vivo rabbit model using a laser-activated thin-film adhesive, sutures, or self-seal. The University of Sydney, Camperdown, Australia. Animal studies. Under an operating microscope, 2.0 mm penetrating incisions were created in 162 right eyes. Incisions in one group were repaired with the adhesive, the second group received a single 10-0 nylon suture, and the third group was left to self-seal. Rabbits were killed humanely at predetermined timepoints over 2 weeks, and wound healing was assessed using burst pressure and immunohistological studies. A modified McDonald-Shadduck scoring was used to assess eyes. The mean burst pressure of the adhesive group was significantly higher than the sutured or self-sealed groups at all timepoints within the first 72 hours. At 0 hour, the burst pressure was 98.0 (±17.0) mm Hg, 30.6 (±2.1) mm Hg, and 3.8 (±0.6) mm Hg (P < .00001) for adhesive-treated (n = 5), sutured (n = 5), and self-sealed wounds (n = 5), respectively. These increased to 229.0 (±53.7) mm Hg, 12.4 (±2.9) mm Hg, and 27.3 (±4.0) mm Hg (P = .0011) at 72 hours. The modified McDonald-Shadduck score was significantly higher for eyes repaired using the adhesive than those sutured or left to self-seal for the first 72 hours. On histology and immunofluorescence, adhesive treatment demonstrated better wound approximation and higher myofibroblastic activation than the other groups. The adhesive was efficacious in sealing penetrating corneal incisions and tolerated higher burst pressures than sutures or self-seal. The adhesive was biocompatible in rabbits, and incisions demonstrated a rapid gain in wound strength that sustained over the study period.


Evaluation of x-ray diffraction enhanced imaging in the diagnosis of breast cancer.
The significance of the x-ray diffraction enhanced imaging (DEI) technique in the diagnosis of breast cancer and its feasibility in clinical medical imaging are evaluated. Different massive specimens including normal breast tissues, benign breast tumour tissues and malignant breast tumour tissues are imaged with the DEI method. The images are recorded respectively by CCD or x-ray film at different positions of the rocking curve and processed with a pixel-by-pixel algorithm. The characteristics of the DEI images about the normal and diseased tissues are compared. The rocking curves of a double-crystal diffractometer with various tissues are also studied. The differences in DEI images and their rocking curves are evaluated for early diagnosis of breast cancers.


Induction of B cell unresponsiveness to noninherited maternal HLA antigens during fetal life.
Patients who have received many transfusions become highly sensitized and develop antibodies against almost all HLA alloantigens, so that finding a cross-match negative kidney donor is difficult. A survey of those patients showed that 50 percent did not form antibodies against the noninherited maternal HLA antigens. Apart from the obvious clinical implications, the data indicate that a human equivalent of murine neonatal or actively acquired tolerance has now been identified.


Why rejection hurts: a common neural alarm system for physical and social pain.
Numerous languages characterize 'social pain', the feelings resulting from social estrangement, with words typically reserved for describing physical pain ('broken heart', 'broken bones') and perhaps for good reason. It has been suggested that, in mammalian species, the social-attachment system borrowed the computations of the pain system to prevent the potentially harmful consequences of social separation. Mounting evidence from the animal lesion and human neuroimaging literatures suggests that physical and social pain overlap in their underlying neural circuitry and computational processes. We review evidence suggesting that the anterior cingulate cortex plays a key role in the physical-social pain overlap. We also suggest that the physical-social pain circuitry might share components of a broader neural alarm system.


Bilateral Postoperative Cyst after Maxillary Sinus Surgery: Report of a Case and Systematic Review of the Literature.
Purpose. We present a case of a bilateral postoperative maxillary cyst (PMC) and discuss this with a systemic review. Case Report and Literature Review. A 68-year-old female with pain and swelling on the right side of the face. MRI and CT showed a cystic tumors of the right and left maxillary sinus. Radical maxillary surgery via a Caldwell-Luc procedure had been performed 55 years ago and bilateral PMC was diagnosed. The PubMed database was searched for PMC within the last 30 years. Results. Together with the current case, we found 23 reports including 284 patients describing PMC. It was diagnosed at a mean time of 22 years after causal surgery at a mean age of 47 years. Initial symptoms were mostly pain with or without swelling. The main radiological sign was a unilocular radiolucency with a slight preference for the left side. Discussion. PMC is a long-term complication that can occur after maxillary sinus surgery and a second surgical approach is required in order to stop cystic expansion. Therefore, patients' informed consent on this complication as well as a prolonged follow-up is recommended. Simple paranasal ultrasound or paranasal sinus plain radiography may lead to an earlier detection reducing interventional morbidity.


Visibility graphs of ground-level ozone time series: A multifractal analysis.
A recent method based on the concurrence of complex networks and multifractal analyses is applied for the first time to explore ground-level ozone behavior. Ozone time series are converted into complex networks for their posterior analysis. The searched purpose is to check the suitability of this transformation and to see whether some features of these complex networks could constitute a preliminary analysis before the more thorough multifractal formalism. Results show effectively that the exposed transformation stores the original information about the ozone dynamics and gives meaningful knowledge about the time series. Based on these results, the multifractal analysis of the complex networks is performed. Looking at the physical meaning of the multifractal properties (such as fractal dimensions and singularity spectrum), a relationship between those and the degree distribution of the complex networks is found. In addition to all the promising results, this novel connection between time series and complex networks can deal with both stationary and non-stationary time series, overcoming one of the main limitations of multifractal analysis. Therefore, this technique can be regarded as an alternative to give supplementary information within the study of complex signals.


Antigen recognition in the islets changes with progression of autoimmune islet infiltration.
In type 1 diabetes, the pancreatic islets are an important site for therapeutic intervention because immune infiltration of the islets is well established at diagnosis. Therefore, understanding the events that underlie the continued progression of the autoimmune response and islet destruction is critical. Islet infiltration and destruction is an asynchronous process, making it important to analyze the disease process on a single islet basis. To understand how T cell stimulation evolves through the process of islet infiltration, we analyzed the dynamics of T cell movement and interactions within individual islets of spontaneously autoimmune NOD mice. Using both intravital and explanted two-photon islet imaging, we defined a correlation between increased islet infiltration and increased T cell motility. Early T cell arrest was Ag dependent and due, at least in part, to Ag recognition through sustained interactions with CD11c(+) APCs. As islet infiltration progressed, T cell motility became Ag independent, with a loss of T cell arrest and sustained interactions with CD11c(+) APCs. These studies suggest that the autoimmune T cell response in the islets may be temporarily dampened during the course of islet infiltration and disease progression.


Robust detection of oncometabolic aberrations by 1H-13C heteronuclear single quantum correlation in intact biological specimens.
Magnetic resonance (MR) spectroscopy has potential to non-invasively detect metabolites of diagnostic significance for precision oncology. Yet, many metabolites have similar chemical shifts, yielding highly convoluted 1H spectra of intact biological material and limiting diagnostic utility. Here, we show that hydrogen-carbon heteronuclear single quantum correlation (1H-13C HSQC) offers dramatic improvements in sensitivity compared to one-dimensional (1D) 13C NMR and significant signal deconvolution compared to 1D 1H spectra in intact biological settings. Using a standard NMR spectroscope with a cryoprobe but without specialized signal enhancing features such as magic angle spinning, metabolite extractions or 13C-isotopic enrichment, we obtain well-resolved 2D 1H-13C HSQC spectra in live cancer cells, in ex vivo freshly dissected xenografted tumors and resected primary tumors. This method can identify tumors with specific oncometabolite alterations such as IDH mutations by 2-hydroxyglutarate and PGD-deleted tumors by gluconate. Results suggest potential of 1H-13C HSQC as a non-invasive diagnostic in precision oncology.


Detecting mycobacteraemia for diagnosing tuberculosis.
In human immunodeficiency virus (HIV) infected persons with pulmonary tuberculosis (TB), sputum may not always show acid fast bacilli (AFB). Moreover, in most cases of suspected extrapulmonary TB (irrespective of HIV status) mycobacteria-containing material is not readily available for investigation. This study evaluated whether blood culture for Mycobacterium tuberculosis bacteraemia (mycobacteraemia) help in diagnosing TB in such cases. A total of 93 consecutive subjects with a clinical diagnosis of tuberculosis with or without laboratory confirmation, 42 with and 38 without coexisting HIV infection, and 13 patients with HIV infection without clinical evidence of TB were enrolled. Mycobacterial blood cultures were done using lysis centrifugation technique followed by subculturing onto the modified Lowenstein-Jenson medium (LJ-1) and Selective Kirchner's medium followed by subculturing onto the modified Lowenstein-Jenson medium (LJ-2, LJ-3). Of the 15 (16.2%) subjects with evidence of mycobacteremia in 4 (26.7%) blood was the first/ only source of diagnosing TB. Among 80 patients with clinical diagnosis of TB whether supported by laboratory tests or not, 14 (17.5%) had mycobacteraemia. Among the 21 HIV infected patients with laboratory proven TB, 9 (43%) had mycobacteraemia. Blood culture appears to be a useful additional test to diagnose TB in persons with HIV infection. In patients without HIV infection, but with clinical picture compatible with TB, blood culture for mycobacteraemia may occasionally help in the diagnosis. We recommend the use of the lysis centrifugation technique followed by direct smear of the sediment along with inoculation of the sediment into both modified Lowenstein-Jenson medium and the Selective Kirchner's medium with subsequent subculturing onto the modified Lowenstein-Jenson medium for mycobacterial blood culture for detecting mycobacteraemia.


Next generation sequencing in research and diagnostics of ocular birth defects.
Sequence capture enrichment (SCE) strategies and massively parallel next generation sequencing (NGS) are expected to increase the rate of gene discovery for genetically heterogeneous hereditary diseases, but at present, there are very few examples of successful application of these technologic advances in translational research and clinical testing. Our study assessed whether array based target enrichment followed by re-sequencing on the Roche Genome Sequencer FLX (GS FLX) system could be used for novel mutation identification in more than 1000 exons representing 100 candidate genes for ocular birth defects, and as a control, whether these methods could detect two known mutations in the PAX2 gene. We assayed two samples with heterozygous sequence changes in PAX2 that were previously identified by conventional Sanger sequencing. These changes were a c.527G>C (S176T) substitution and a single basepair deletion c.77delG. The nucleotide substitution c.527G>C was easily identified by NGS. A deletion of one base in a long polyG stretch (c.77delG) was not registered initially by the GS Reference Mapper, but was detected in repeated analysis using two different software packages. Different approaches were evaluated for distinguishing false positives (sequencing errors) and benign polymorphisms from potentially pathogenic sequence changes that require further follow-up. Although improvements will be necessary in accuracy, speed, ease of data analysis and cost, our study confirms that NGS can be used in research and diagnostic settings to screen for mutations in hundreds of loci in genetically heterogeneous human diseases.


Performance effects and metabolic consequences of caffeine and caffeinated energy drink consumption on glucose disposal.
This review documents two opposing effects of caffeine and caffeine-containing energy drinks, i.e., their positive effects on athletic performance and their negative impacts on glucose tolerance in the sedentary state. Analysis of studies examining caffeine administration prior to performance-based exercise showed caffeine improved completion time by 3.6%. Similar analyses following consumption of caffeine-containing energy drinks yielded positive, but more varied, benefits, which were likely due to the diverse nature of the studies performed, the highly variable composition of the beverages consumed, and the range of caffeine doses administered. Conversely, analyses of studies administering caffeine prior to either an oral glucose tolerance test or insulin clamp showed a decline in whole-body glucose disposal of ~30%. The consequences of this resistance are unknown, but there may be implications for the development of a number of chronic diseases. Both caffeine-induced performance enhancement and insulin resistance converge with the primary actions of caffeine on skeletal muscle.


Expression of Smad1 and Smad2 during embryogenesis suggests a role in organ development.
Smad proteins are intracellular signalling molecules and putative transcription factors that transduce signals elicited by members of the transforming growth factor beta (TGF-beta) superfamily. By comparing the expression of Smad1 and Smad2 during embryonic development, we show that mRNAs of both Smad isoforms are present in a variety of tissues. The major sites of expression of both Smads can be correlated with the expression domains of several members of the TGF-beta superfamily. Our expression data suggest that Smad proteins are involved in organ development, particularly that of organs arising from mesenchymal-epithelial interactions. A second site of strong expression is the central nervous system. Transcriptional control mediated by Smad1 and Smad2, therefore, may exert an important function in differentiation processes of embryonic development that are controlled by ligands of the TGF-beta superfamily.