Dimethylation of eEF1A at Lysine 55 Plays a Key Role in the Regulation of eEF1A2 on Malignant Cell Functions of Acute Myeloid Leukemia.
This study aimed to explore whether eukaryotic translation elongation factor 1 alpha 2 affected cell proliferation, migration, and apoptosis via regulating the dimethylation of eukaryotic translation elongation factor 1 alpha at lysine 55 in acute myeloid leukemia. The expressions of eukaryotic translation elongation factor 1 alpha 2 and dimethylation of eukaryotic translation elongation factor 1 alpha at lysine 55 in acute myeloid leukemia cell lines and human normal bone marrow mononuclear cells (as control) were assessed. Control CRISPR-Cas9 lentivirus, eukaryotic translation elongation factor 1 alpha 2 knockout CRISPR-Cas9 lentivirus, vector plasmid, eukaryotic translation elongation factor 1 alpha 2 wild type overexpression plasmid, and eukaryotic translation elongation factor 1 alpha 2 with a K55R substitution overexpression plasmid were transfected into AML-193 and Kasumi-1 cells combined or alone, and were accordingly divided into 4 groups (Sgcontrol + vector group, SgeEF1A2 + vector group, SgeEF1A2 + eEF1A2WT group, and SgeEFIA2 + eEF1A2K55R group). Eukaryotic translation elongation factor 1 alpha 2 and dimethylation of eukaryotic translation elongation factor 1 alpha at lysine 55 expressions were higher in AML-193, Kasumi-1, and KG-1 cell lines compared to the control. In AML-193 and Kasumi-1 cells, the knockout and compensated experiments revealed that eukaryotic translation elongation factor 1 alpha 2 promoted cell proliferation and migration but repressed apoptosis. Additionally, the knockout of eukaryotic translation elongation factor 1 alpha 2 decreased dimethylation of eukaryotic translation elongation factor 1 alpha at lysine 55 expression, meanwhile, eukaryotic translation elongation factor 1 alpha 2 wild type overexpression enhanced while eukaryotic translation elongation factor 1 alpha 2 with a K55R substitution overexpression did not influence the dimethylation of eukaryotic translation elongation factor 1 alpha at lysine 55 expression. Furthermore, eukaryotic translation elongation factor 1 alpha 2 wild type overexpression promoted cell proliferation, enhanced migration, and decreased apoptosis, but eukaryotic translation elongation factor 1 alpha 2 with a K55R substitution overexpression did not influence these cellular functions in AML-193 and Kasumi-1 cells, suggesting the implication of dimethylation of eukaryotic translation elongation factor 1 alpha at lysine 55 in eukaryotic translation elongation factor 1 alpha 2 mediated oncogenesis of acute myeloid leukemia. Eukaryotic translation elongation factor 1 alpha 2 and its dimethylated product may serve as therapeutic targets, and these findings may provide support for exploring novel strategies in acute myeloid leukemia treatment.


[Morphologic changes of gastric mucosa in Helicobacter pylori infection in patients with peptic ulcer disease and chronic gastritis].
The present study was carried out to value the histological method of detection of Helicobacter pylori in gastric mucosa and to analyse the pathological changes of the Helicobacter pylori-positive patients with duodenal and gastric ulcer and with chronic gastritis. The studies population consisted of 150 patients, 76 F and 74 M. The samples were taken during gastroscopy from the prepyloric region and gastric body. After routine histological procedure they were stained HE and by Giemsa method for detection of H.pylori. The latter was identified in 56.6% of of the patients with duodenal ulcer, in 54.8% of those with gastric ulcer, in 17.6% of the patients after partial gastrectomy and in 46.9% of patients suffering from chronic gastritis. The increased density of H.pylori (third grade) was detected in the cases with clinical diagnosis of duodenal and gastric ulcer and erythematous endoscopic gastritis and was correlated with the histological picture of chronic active gastritis, which was characterized by diffuse mixed cellular exudation, lesion of the superficial epithelial, mucosal erosions and by foveolar hyperplasia. We suggest that this morphological picture corresponded to the active phase of Helicobacter infection. We conclude, that the histological method by Giemsas staining is effective especially in the acute phase of H.pylori infection and represents high diagnostic value and may be used in monitoring during the antibacterial (anti H.pylori) treatment. There is the correlation between massive density of H.pylori observed in gastric mucosa and chronic active gastritis.


Examining the effects of alexithymia on the relation between posttraumatic stress disorder and over-reporting.
Empirical examinations of the relation between posttraumatic stress disorder (PTSD) and objective measures of symptom over-reporting may be useful for identification of mechanisms of this previously observed relation. The present study examined the moderating effect of alexithymia, defined as a deficit in the ability to identify and describe emotions, on the relation between PTSD and over-reporting. Seventy-five veterans diagnosed with PTSD were recruited from an outpatient Veterans Affairs facility and the community. Participants were administered the Miller Forensic Assessment of Symptoms Test, along with the Toronto Alexithymia Scale and PTSD Checklist within a larger study of behavioral and physiological correlates of PTSD. Hierarchical linear regression analyses showed a significant moderating effect of alexithymia, such that the relation between PTSD symptom severity and over-reporting was only significant in the presence of elevated alexithymia. Evaluation of the subscales of the Toronto Alexithymia Scale showed that the effect was greatest for the Difficulty Describing Emotions subscale. Alexithymia should be considered as a potential mechanism contributing to the over-reporting phenomena observed in the assessment and treatment of PTSD, and warrants further study. Brief interventions to improve understanding and description of emotional experiences may help to improve accuracy of symptom report. (PsycINFO Database Record


Dual-specificity Phosphatase 9 protects against Cardiac Hypertrophy by targeting ASK1.
The functions of dual-specificity phosphatase 9 (DUSP9) in hepatic steatosis and metabolic disturbance during nonalcoholic fatty liver disease were discussed in our prior study. However, its roles in the pathophysiology of pressure overload-induced cardiac hypertrophy remain to be illustrated. This study attempted to uncover the potential contributions and underpinning mechanisms of DUSP9 in cardiac hypertrophy. Utilizing the gain-and-loss-of-functional approaches of DUSP9 the cardiac phenotypes arising from the pathological, echocardiographic, and molecular analysis were quantified. The results showed increased levels of DUSP9 in hypertrophic mice heart and angiotensin II treated cardiomyocytes. In accordance with the results of cellular hypertrophy in response to angiotensin II, cardiac hypertrophy exaggeration, fibrosis, and malfunction triggered by pressure overload was evident in the case of cardiac-specific conditional knockout of DUSP9. In contrast, transgenic mice hearts with DUSP9 overexpression portrayed restoration of the hypertrophic phenotypes. Further explorations of molecular mechanisms indicated the direct interaction of DUSP9 with ASK1, which further repressed p38 and JNK signaling pathways. Moreover, blocking ASK1 with ASK1-specific inhibitor compensated the pro-hypertrophic effects induced by DUSP9 deficiency in cardiomyocytes. The main findings of this study suggest the potential of DUSP9 in alleviating cardiac hypertrophy at least partially by repressing ASK1, thereby looks promising as a prospective target against cardiac hypertrophy.


Next-generation sequencing technology for the diagnosis of Pneumocystis pneumonia in an immunocompetent female: A case report.
Pneumocystis pneumonia (PCP) is a serious fungal infection usually seen in patients with human immunodeficiency virus, and it is more frequently found and has a high fatality rate in immunocompromised people. Surprisingly, it rarely occurs in immunocompetent patients. However, the clinical diagnosis of this pathogen is made more difficult by the difficulty of obtaining accurate microbiological evidence with routine tests. This case reports a PCP patient with normal immune function who was diagnosed through next-generation sequencing (NGS). A 23-year-old female who had no special disease in the past was admitted to the hospital with a persistent fever and cough. Based on the initial examination results, the patient was diagnosed with bipulmonary pneumonia, and empirical broad-spectrum antibiotic therapy was administered. However, due to the undetermined etiology, the patient's condition continued to worsen. She was transferred to the intensive care unit because of acute respiratory failure. After the diagnosis of Pneumocystis jirovecii infection through NGS in bronchoalveolar lavage fluid and treatment with trimethoprim/sulfamethoxazole and caspofungin, the patient gradually recovered and had a good prognosis. This case emphasizes that, for patients with normal immune function the possibility of PCP infection, although rare, cannot be ignored. NGS plays an important role in the diagnosis of refractory interstitial pneumonia and acute respiratory failure.


A Chemically Modified Curcumin (CMC 2.24) Inhibits Nuclear Factor κB Activation and Inflammatory Bone Loss in Murine Models of LPS-Induced Experimental Periodontitis and Diabetes-Associated Natural Periodontitis.
The purpose of this study was to assess the effect of a novel chemically modified curcumin (CMC 2.24) on NF-κB and MAPK signaling and inflammatory cytokine production in two experimental models of periodontal disease in rats. Experimental model I: Periodontitis was induced by repeated injections of LPS into the gingiva (3×/week, 3 weeks); control rats received vehicle injections. CMC 2.24, or the vehicle, was administered by daily oral gavage for 4 weeks. Experimental model II: Diabetes was induced in adult male rats by streptozotocin injection; periodontal breakdown then results as a complication of uncontrolled hyperglycemia. Non-diabetic rats served as controls. CMC 2.24, or the vehicle, was administered by oral gavage daily for 3 weeks to the diabetics. Hemimaxillae and gingival tissues were harvested, and bone loss was assessed radiographically. Gingival tissues were pooled according to the experimental conditions and processed for the analysis of matrix metalloproteinases (MMPs) and bone-resorptive cytokines. Activation of p38 MAPK and NF-κB signaling pathways was assessed by western blot. Both LPS and diabetes induced an inflammatory process in the gingival tissues associated with excessive alveolar bone resorption and increased activation of p65 (NF-κB) and p38 MAPK. In both models, the administration of CMC 2.24 produced a marked reduction of inflammatory cytokines and MMPs in the gingival tissues, decreased bone loss, and decreased activation of p65 (NF-κB) and p38 MAPK. Inhibition of these cell signaling pathways by this novel tri-ketonic curcuminoid (natural curcumin is di-ketonic) may play a role in its therapeutic efficacy in locally and systemically associated periodontitis.


Calcium uptake and membrane trafficking in response to PTH or 25(OH)D3 in polarized intestinal epithelial cells.
Cell culture techniques providing retention of the polarized enterocyte morphology has allowed, for the first time, comparison of parathyroid hormone (PTH)- and 25-hydroxyvitamin D(3) [25(OH)D(3)]-induced (45)Ca uptake with membrane trafficking events discerned using confocal microscopy. Treatment of cells with 65 pM bPTH(1-34) promoted enhanced (45)Ca uptake between 1 and 10 min after peptide. The protein kinase A (PKA) antagonist, RpcAMP inhibited hormone-mediated uptake. At the microscopic level, cells labeled with the endocytic tracking dye FM1-43 revealed increased punctate staining 50-550s after hormone. Pretreatment of cells with RpcAMP abolished this pattern of staining. The calcium indicator dye fluo-3 AM revealed faint punctate labeling in controls, with increased bands of punctate labeling in the apical region of the cells after peptide hormone, and ultimately the basal region. Parallel studies conducted with the metabolite 25(OH)D(3) resulted in a slower stimulation of (45)Ca uptake 5-10 min after steroid, which was also inhibited by preincubation with RpcAMP. Cells labeled with FM1-43 and then treated with steroid showed no change in distribution of fluorescence during the 10 min incubation period. Confocal microscopy with fluo-3 revealed intense apical fluorescence--that after steroid --streamed to a perinuclear position, and ultimately the basal area. Uniformly diffuse staining, which would indicate cytoplasmic calcium transport, was observed only in controls. Membrane trafficking and compartmentalized calcium appear to be integral to agonist mediated cation transport.


Delayed extended "midthenar" flap for reconstruction of total fingertip avulsion injury and a proposal of ideal postoperative immobilization for a palmar flap.
Avulsion injury through the base of the distal phalanx of the ring finger with a large projecting tip of exposed bone was treated with delayed extended midthenar flap. The midthenar flap adding delay procedure at the first stage of a consecutive 2-stage operation could provide extended flap length to wrap the large projecting tip of the exposed bone at the second stage. This flap is useful for the middle and ring fingertip injury because motions of the all thumb joints are not restricted during the immobilization period. To reduce immobilization discomfort between the 2 stages of the palmar flap operation, a "relaxed flexion position" immobilization technique devised by the authors is also introduced.


[New aspects in the pathogenesis and therapy of hyperreflexive rhinopathy].
Hyperreflectory rhinopathy (HR) is a non-specific hyperreactivity of the nasal mucosa. It causes hypersecretion, decreased nasal patency, sneezing and sometimes headache by local reflexes, which are beyond voluntary control. The synonymous name "vasomotor rhinitis" or "vasomotor rhinopathy" is no longer adequate with regard to our present state of knowledge of the autonomous innervation of the human nasal mucosa. Recent pharmacological investigations show the great importance of peptidergic neurons. In our own studies, the presence and the topical effects of the neuropeptide substance-P in human nasal mucosa are examined. A new concept of the autonomous innervation of the human nasal mucosa is presented. Apart from adrenergic and cholinergic neurons, it also includes peptidergic neurons (SP, CGRP, NKA, VIP, PHI, APP, GRP). According to this model, a hypothesis on the pathogenesis of HR is developed. A key position is occupied by the so-called "axon reflex" which is mediated by substance-P immunoreactive nerve fibers. It is released by a chemical, thermal or mechanical irritation. This axon reflex mediates pain, vasodilation and plasma extravasation (neurogenic oedema), hypersecretion such as smooth muscle contraction, and sneezing reflex. Capsaicin (8-methyl-N-vanillyl-6-nonenamid) leads to a selective degeneration of substance-P immunoreactive nerve fibres and desensitisation of its receptors after repeated topical or systemic application, thus blocking the axon reflex. The risk-free application of capsaicin was shown in self-experiments and in volunteers. Our hypothesis was confirmed by the good results of the treatment of a group of volunteer patients who suffered from HR.(ABSTRACT TRUNCATED AT 250 WORDS)


Structural behaviour of synthetic Co2SiO4 at low temperatures.
Synthetic Co2SiO4 has an olivine structure with isolated SiO4 groups (space group Pnma) and shows magnetic ordering below 50 K. Single-crystal neutron diffraction was applied to determine precise crystal structure parameters at low temperatures. No structural phase transition was revealed in the temperature range 2.5-300 K. Lattice parameters were determined by high-resolution X-ray powder diffraction between 15 and 300 K. There is a clear evidence of an anomalous thermal expansion related to the magnetic phase transition which can be attributed to magnetostriction.


[Interleukin-2 and human seminal fluid].
Involvement of the cytokines in male infertility were considered from in vitro studies, however the nature and the role of the mediator susceptible of arising in vivo is not completely elucidated. The purpose of this study is to measure the concentrations of interleukin-2 (IL-2) in normal and pathologic seminal fluid and to analyse their relationship with semen parameters. Our results demonstrate the presence of IL-2 in seminal fluid. Comparatively to normal seminal fluid, significant lower IL-2 concentrations were observed in infected semen samples or with sperm antibodies. No correlation was found between the cytokines values and the sperm parameters.


Spinal cord neuroplasticity following repeated opioid exposure and its relation to pathological pain.
Convincing evidence has accumulated that indicates neuroplastic changes within the spinal cord in response to repeated exposure to opioids. Such neuroplastic changes occur at both cellular and intracellular levels. It has been generally acknowledged that the activation of N-methyl-D-aspartate (NMDA) receptors plays a pivotal role in the development of neuroplastic changes following repeated opioid exposure. Intracellular cascades can also be activated subsequent to NMDA receptor activation. In particular, protein kinase C has been shown to be a key intracellular element that contributes to the behavioral manifestation of neuroplastic changes. Moreover, interactions between NMDA and opioid receptors can lead to potentially irreversible degenerative neuronal changes in the spinal cord in association with the development of opioid tolerance. Interestingly, similar cellular and intracellular changes occur in the spinal cord following peripheral nerve injury. These findings indicate that interactions exist in the spinal cord neural structures between two seemingly unrelated conditions-chronic opioid exposure and a pathological pain state. These observations may help understand mechanisms of chemical intolerance and multiple chemical sensitivity as well as have significant clinical implications in pain management with opioid analgesics.


High self-reactivity drives T-bet and potentiates Treg function in tissue-specific autoimmunity.
T cell receptor (TCR) affinity is a critical factor of Treg lineage commitment, but whether self-reactivity is a determining factor in peripheral Treg function remains unknown. Here, we report that a high degree of self-reactivity is crucial for tissue-specific Treg function in autoimmunity. Based on high expression of CD5, we identified a subset of self-reactive Tregs expressing elevated levels of T-bet, GITR, CTLA-4, and ICOS, which imparted significant protection from autoimmune diabetes. We observed that T-bet expression in Tregs, necessary for control of Th1 autoimmunity, could be induced in an IFNγ-independent fashion and, unlike in conventional T cells (Tconv), was strongly correlated with the strength of TCR signaling. The level of CD5 similarly identified human Tregs with an increased functional profile, suggesting that CD5hi Tregs may constitute an efficacious subpopulation appropriate for use in adoptive Treg therapies for treatment of inflammatory conditions. Overall, this work establishes an instrumental role of high TCR self-reactivity in driving Treg function.


[Microbial production of chondroitin sulfate: a review].
Chondroitin sulfate (CS) is the typical sulfation glycosaminoglycan and widely applied in the industries of pharmaceutical, health products and cosmetic for its peculiar properties. CS is the main component of cartilage proteoglycans in animal and capsular polysaccharide in a few bacteria. CS can be extracted from animal sources and produced via microbial fermentation. In this article, development of chondroitin sulfate by fermentation, biosynthesis and regulating mechanisms of CS in bacteria are described. Furthermore, prospect and tendency of chondroitin sulfate from bacterial fermentation are addressed.


C-terminal Agrin Fragment as a potential marker for sarcopenia caused by degeneration of the neuromuscular junction.
Sarcopenia is considered to be an enormous burden for both the individuals affected and for society at large. A multifactorial aetiology of this geriatric syndrome has been discussed. Amongst other pathomechanisms, the degeneration of the neuromuscular junction (NMJ) may be of major relevance. The intact balance between the pro-synaptic agent agrin and the anti-synaptic agent neurotrypsin ensures a structurally and functionally intact NMJ. Excessive cleavage of the native motoneuron-derived agrin by neurotrypsin into a C-terminal Agrin Fragment (CAF) leads to functional disintegration at the NMJ and may consecutively cause sarcopenia. The present study evaluates the hypothesis that CAF serum concentration is a potential marker for the loss of appendicular lean mass in older adults. It also explores how CAF concentration is influenced by vitamin D supplementation and physical exercise. Serum was taken from 69 (47 female) prefrail community-dwelling older adults participating in a training intervention study to measure the CAF concentration using the Western blot technique. All participants were supplemented orally with vitamin D3 before the training intervention period commenced. Appendicular lean mass (aLM) was evaluated by dual energy X-ray absorptiometry. Multiple linear regression models were used to identify factors significantly associated with CAF concentration. Appendicular lean mass, age and sex were identified as significant explanatory factors for CAF concentration. Gait speed and hand grip strength were not associated with CAF concentration. Male participants showed a strong correlation (r=-0.524) between CAF serum concentration and aLM, whereas this was not the case (r=-0.219) in females. Vitamin D supplementation and physical exercise were significantly associated with a reduction in CAF concentration, especially in participants with initially high CAF concentrations. C-terminal Agrin Fragment could be a potential marker for identifying sarcopenia in a subgroup of affected individuals in the future. The decline of muscle mass seems to be a CAF-associated process in males, whereas the situation in females may be more complex and multifactorial. CAF concentration is reduced by vitamin D supplementation and physical exercise and therefore suggests a potentially positive effect on NMJs. Further prospective studies of sarcopenic patients in addition to muscle biopsy and electromyographical investigations are planned to verify the external validity of the CAF concept.


Hide-and-seek or show-and-tell? Emerging issues of informed consent.
This article reviews key philosophical and legal underpinnings of mental health professionals' obligation to obtain informed consent from consumers of their services. The basic components of informed consent are described, and strategies for clinically and ethically appropriate methods of obtaining informed consent are discussed. Emerging issues in informed consent involving duty to assess and protect against client dangerousness, obligations to third parties, and issues of deception are considered as well. The article proposes that part of the process of obtaining informed consent is the cultivation of a treatment environment that emphasizes beneficence and client autonomy.


Recovery of humic-reducing bacteria from a diversity of environments.
To evaluate which microorganisms might be responsible for microbial reduction of humic substances in sedimentary environments, humic-reducing bacteria were isolated from a variety of sediment types. These included lake sediments, pristine and contaminated wetland sediments, and marine sediments. In each of the sediment types, all of the humic reducers recovered with acetate as the electron donor and the humic substance analog, 2,6-anthraquinone disulfonate (AQDS), as the electron acceptor were members of the family Geobacteraceae. This was true whether the AQDS-reducing bacteria were enriched prior to isolation on solid media or were recovered from the highest positive dilutions of sediments in liquid media. All of the isolates tested not only conserved energy to support growth from acetate oxidation coupled to AQDS reduction but also could oxidize acetate with highly purified soil humic acids as the sole electron acceptor. All of the isolates tested were also able to grow with Fe(III) serving as the sole electron acceptor. This is consistent with previous studies that have suggested that the capacity for Fe(III) reduction is a common feature of all members of the Geobacteraceae. These studies demonstrate that the potential for microbial humic substance reduction can be found in a wide variety of sediment types and suggest that Geobacteraceae species might be important humic-reducing organisms in sediments.


Probing the Antiallergic and Anti-inflammatory Activity of Biflavonoids and Dihydroflavonols from Dietes bicolor.
Dietes bicolor (Iridaceae) is an ornamental plant used by African local healers to treat diarrhea and dysentery. A new dihydroflavonol, (2R,3R)-3,5,7-trihydroxy-8-methoxyflavanone (1); two known dihydroflavonols, trans-3-hydroxy-5-methoxy-6,7-methylenedioxyflavanone (2) and trans-3-hydroxy-5,7-dimethoxyflavanone (3); the known isoflavone orobol 7,3'-di-O-methyl ether (4); the known biflavones lanaroflavone (5), robustaflavone (6), and amentoflavone (7); and β-sitosterol (8) were isolated from the CH2Cl2 fraction of D. bicolor leaves. The extract showed potent activity in antiallergic and anti-inflammatory assays. The structures of the isolates were identified by spectroscopic and spectrometric methods. Compounds 6 and 7 (400 μM) exhibited antiallergic activity by inhibiting antigen-induced β-hexosaminidase release at 45.7% and 46.3%, respectively. Moreover, 6 and 7 exerted anti-inflammatory activity as demonstrated by the inhibition of superoxide anion generation with an IC50 value of 1.0 μM as well as the inhibition of elastase release with IC50 values of 0.45 and 0.75 μM, respectively. The anti-inflammatory activity was further explained by the virtual docking of the isolated compounds to the binding sites in the human neutrophil elastase (HNE) crystal structure using Discovery Studio 2.5. It was concluded that the biflavonoids bind directly to HNE and inhibit its enzymatic activity based on the CDOCKER algorithm. The data provided evidence for the potential use of D. bicolor against certain diseases related to allergy and inflammation.


Low S-adenosylmethionine concentrations found in patients with severe inflammatory bowel disease.
S-adenosylmethionine is a methyl donor in many cellular reactions including detoxification of constantly produced hydrogen sulphide in the colon. A reduced capacity to detoxify hydrogen sulphide may be implicated in the pathogenesis of inflammatory bowel disease. S-adenosylmethionine could be low if this assumption is correct. We compared S-adenosylmethionine concentrations in whole blood in patients with severe and moderate inflammatory bowel disease with healthy reference persons. S-adenosylmethionine concentrations in whole blood were measured using high-pressure liquid chromatography. Patients with Crohn's disease (n=21), ulcerative colitis (n=7) and healthy age-matched reference persons (or controls) (n=17) were studied. S-adenosylmethionine concentrations were significantly decreased in patients with severe inflammatory bowel disease (mean 1.10 mg/l) as compared to patients with moderate Crohn's disease and ulcerative colitis (mean 1.83 mg/l) and reference persons (mean 1.84 mg/l). Statistically significant inverse correlations were found between S-adenosylmethionine concentration and activity index (p<0.01 and R2=0.86) as well as Crohn's disease activity index (p<0.01 and R2=0.50) scores. Low concentrations of S-adenosylmethionine were found in patients with severe inflammatory bowel disease. Future studies will show whether S-adenosylmethionine is a marker for disease activity and a possible tool for investigation of sulphur toxicity as a causative mechanism in inflammatory bowel disease.


Comparison of Nutech Functional Score with European Stroke Scale for Patients with Cerebrovascular Accident Treated with Human Embryonic Stem Cells: NFS for CVA Patients Treated with hESCs.
Stem cell therapy is a promising modality for treatment of patients with chronic cerebrovascular accident (CVA) in whom treatment other than physiotherapy or occupational therapy does not address the repair or recovery of the lost function. In this study, the author aimed at evaluating CVA patients treated with human embryonic stem cell (hESC) therapy and comparing their study outcomes with globally accepted European Stroke Scale (ESS) to that with novel scoring system, Nutech functional score (NFS), a 21-point positional and directional scoring system for assessing patients with CVA. Patients diagnosed with CVA were assessed with NFS and ESS before and after hESC therapy. NFS assessed the patients in the direction of 1-5 (bad to good), where 5 was considered as the highest possible grade (HPG). The findings were obtained for the patients who scored HPG, and had shown improvement by at least one grade. Overall, 66.7% of patients scored HPG level on the NFS scale and about 62.5% of the patients scored HPG according to the ESS scale. Approximately, 52.2% patients showed an improvement of 100% (by at least one grade) on NFS scale. None of the patients showed 100% improvement in the alteration of the score by at least one grade when scored with ESS. NFS and ESS scores show that a large population of CVA patients was benefitted with hESC therapy. NFS was found to give more convincing results than ESS, and overcomes the shortcomings of ESS.


Design and Usability of a Heart Failure mHealth System: A Pilot Study.
Despite the advances in mobile health (mHealth) systems, little is known about patients' and providers' experiences using a new mHealth system design. This study aimed to understand challenges and provide design considerations for a personalized mHealth system that could effectively support heart failure (HF) patients after they transition into the home environment. Following exploratory interviews with nurses and preventive care physicians, an mHealth system was developed. Patients were asked to measure their weight, blood pressure, and blood glucose (if they had diabetes). They were also instructed to enter symptoms, view notifications, and read messages on a mobile app that we developed. A Bluetooth-enabled weight scale, blood pressure monitor, glucometer, and mobile phone was provided after an introductory orientation and training session. HF nurses used a dashboard to view daily measurements for each patient and received text and email alerts when risk was indicated. Observations of usage, cases of deterioration, readmissions, and metrics related to system usability and quality of life outcomes were used to determine overall effectiveness of the system, whereas focus group sessions with patients were conducted to elicit participants' feedback on the system's design. A total of 8 patients with HF participated over a 6-month period. Overall, the mean users' satisfaction with the system ranked 73%, which was above average. Quality of life improvement was 3.6. Patients and nurses used the system on a regular basis and were able to successfully identify and manage 8 health deteriorations, of which 5 were completely managed remotely. Focus groups revealed that, on one hand, the system was beneficial and helped patients with: recording and tracking readings; receiving encouragement and reassurance from nurses; spotting and solving problems; learning from past experiences; and communication. On the other hand, findings also highlighted design issues and recommendations for future systems such as the need to communicate via other media, personalize symptom questions and messages, integrate other health tracking technologies, and provide additional methods to analyze and visualize their data. Understanding users' experiences provides important design considerations that could complement existing design recommendations from the literature, and, when combined with physician and nurse requirements, have the potential to yield a feasible telehealth system that is effective in supporting HF self-care. Future studies will include these guidelines and use a larger sample size to validate the outcomes.


mRNA profiling identifies low levels of phosphatases dual‐specific phosphatase‐7 (DUSP7) and cell division cycle‐25B (CDC25B) in patients with early arthritis.
Phosphotyrosine phosphatases (PTPs) control phosphorylation levels and, consequently, regulate the output of intracellular signalling networks in health and disease. Despite the high number of PTPs expressed in CD4 T cells and their involvement in autoimmunity, information about the expression profile of PTPs in these cells has not been obtained in patients diagnosed with autoimmune diseases. Here, we compare the expression profile of PTPs in CD4 T cells of healthy volunteers and patients submitted to an early arthritis clinic, due to suspicion of rheumatoid arthritis, an autoimmune disease mediated by CD4 T cells. We found lower transcript levels of the mitogen-activated protein kinase (MAPK) phosphatase dual-specific phosphatase-7 (DUSP7) and the cell division cycle-25B (CDC25B) in T cells of patients. While the low expression level of DUSP7 was restricted to patients with positive rheumatoid factor and anti-citrullinated protein antibodies, the altered expression of CDC25B correlated with the activity of the disease. Low levels of CDC25B might contribute to the progression of the autoimmune arthritis and/or might be consequence of the inflammatory environment in the active disease. The possible role of DUSP7 and CDC25B as biomarkers of the disease in clinical protocols is discussed.


Video-assisted thoracoscopic surgery atrial clipping for atrial fibrillation.
The majority of thrombi that arise due to atrial fibrillation occur in the left atrial appendage. Eliminating this cul-de-sac within the left atrium reduces the risk of stroke in these patients. We present a unique case of left atrial appendage occlusion performed via video-assisted thoracoscopic surgery, using an Atriclip to occlude the left atrial appendage in a patient with atrial fibrillation in whom anticoagulation was contraindicated due to a history of recurrent upper gastrointestinal bleeding.


Outline of the report on cardiovascular diseases in China, 2014.
The risk factors for cardiovascular diseases (CVDs) are more prevalent in the Chinese population, and therefore, increase the incidence of CVD. In general, CVD morbidity and mortality will remain an upward trend in the next 10 years. Cardiovascular disease is the leading cause of death in China, which accounts for >40% of deaths from any cause. The burden of CVD is substantial and has become an important public health issue. Measures for the prevention and treatment of CVD in China should be further enforced without delay. Since 2005, the National Center for Cardiovascular Diseases has organized experts of cardiology, neurology, nephrology, diabetes, epidemiology, community health, health economics, biostatistics, and other related fields to compile the annual Report on Cardiovascular Diseases in China. The report aims to provide a timely review of the trend of the epidemic of CVD and to assess the progress of prevention and control of CVD. We present an abstract from the Report on Cardiovascular Diseases in China (2014), including trends in CVD, morbidity and mortality of major CVD, up-to-date assessment of risk factors, as well as health resources for CVD, and a profile of medical expenditure, with the aim of providing evidence for decision making in CVD prevention and control programmes in China, and of delivering the most authoritative information on CVD prevention and control for all citizens.


[The influence of smoking on the development of various subtypes of pulmonary carcinoma].
The influence of smoking on the development of pulmonary carcinoma of various histological subtypes was analyzed in 244 primary and 84 metastatic lung cancer patients. The new WHO classification was used for histological diagnosis. The relative risk was calculated by the odds ratio. It was found that all the squamous, small-and large-cell carcinomas were derived from smokers with relative risks of 27.8, 7.5 and 5.6, respectively. On the other hand, adenocarcinoma had very little, if any, relationship with smoking, the relative risk being 1.5 in men and 1.3 in women. The high-risk group for squamous cell carcinoma in men was defined with a regression line of y (duration of smoking) on x (number of cigarettes per day) as y = -0.24 x + 48.6.


Mean and variance quality control for multiple correlated levels of replicated control samples.
Mean and variance rules for quality control are more powerful than rules based on individual values. An algorithm for applying such rules is described that controls type I errors (false alarms), while allowing for multiple levels of quality control samples, correlation between levels, small numbers of preliminary values, replication of samples and autocorrelation arising from random effects. Based on ANOVA and empirical approximations for small samples, the algorithm maintains a low per-batch probability of type I errors. Three statistics are computed, z(m), z(b) and z(w), which are shown by simulations to be primarily sensitive to a concordant shift in the quality control values, a discordant shift in the values, and an increase in random variability, respectively. Simulations also show that for a Gaussian distribution of analytical errors, the per-batch probability of a type I error is likely to be within the range 0.0045-0.0071 for two to four levels where there are 20-100 preliminary batches and the inter-level correlations are between zero and 0.8. This partial separation of out-of-control alarms into three components provides more assistance with trouble-shooting than do multivariate quality control schemes based on Hotelling's T2, while retaining comparable power.


Influence of growth rate and nutrient limitation on monobactam production and peptidoglycan synthesis in Pseudomonas aeruginosa.
The effects of growth rate and nutrient limitation on monobactam production, peptidoglycan content and mean cell length in Pseudomonas aeruginosa was studied using continuous culture techniques. All three parameters increased progressively with growth rate, a greater response being shown under carbon limitation compared to that occurring under nitrogen limiting conditions. Interestingly, monobactum production mirrored peptidoglycan synthesis. In addition, the monobactam exhibited a broad range of antibacterial activity and bound preferentially to PBP 1A in the producing organism. Moreover, addition of the monobactam to a growing culture inhibited cell wall synthesis. These results are discussed in relation to the control and regulation of peptidoglycan synthesis.


Complex regional pain syndrome - phenotypic characteristics and potential biomarkers.
Complex regional pain syndrome (CRPS) is a pain condition that usually affects a single limb, often following an injury. The underlying pathophysiology seems to be complex and probably varies between patients. Clinical diagnosis is based on internationally agreed-upon criteria, which consider the reported symptoms, presence of signs and exclusion of alternative causes. Research into CRPS biomarkers to support patient stratification and improve diagnostic certainty is an important scientific focus, and recent progress in this area provides an opportunity for an up-to-date topical review of measurable disease-predictive, diagnostic and prognostic parameters. Clinical and biochemical attributes of CRPS that may aid diagnosis and determination of appropriate treatment are delineated. Findings that predict the development of CRPS and support the diagnosis include trauma-related factors, neurocognitive peculiarities, psychological markers, and local and systemic changes that indicate activation of the immune system. Analysis of signatures of non-coding microRNAs that could predict the treatment response represents a new line of research. Results from the past 5 years of CRPS research indicate that a single marker for CRPS will probably never be found; however, a range of biomarkers might assist in clinical diagnosis and guide prognosis and treatment.


A contradictory GLABRA3 allele helps define gene interactions controlling trichome development in Arabidopsis.
Previously characterized Arabidopsis gl3 mutants have trichomes that are smaller, less branched and undergo fewer rounds of endoreplication than wild-type trichomes. A new gl3 mutant, called gl3-sst, has oddly shaped trichomes that over expand during early development, undergo more endoreduplication and that have a striking nuclear morphology. The mutant nuclei consist of many interconnected lobes; however, only a single set of polytene-like chromosomes reside in the mutant nuclei. The predicted gl3-sst polypeptide has a Leu to Phe substitution (codon 78) within a region responsible for protein-protein interaction. Yeast interaction assays comparing GL3 with gl3-sst proteins show that the mutant protein interaction with GL1 and TTG1 is decreased by 75% and 50%, respectively, but there is no difference in its interaction with TRY. Furthermore, TRY has the ability to prevent the GL1 GL3 interaction and the GL1 gl3-sst interaction is even more sensitive to TRY. Analysis of plants expressing functional GFP-tagged versions of GL1, GL3 and TRY show that the proteins are localized in trichome nuclei. These results have been used to model trichome initiation in terms of protein interactions and threshold levels of activator complex.


Genetic complementation in cystic fibrosis pancreatic cells by somatic cell fusion.
The biochemical defect that underlies the genetic disorder cystic fibrosis (CF) has been proposed to involve an altered regulation of epithelial Cl- permeability by agents such as adenosine 3',5'-cyclic monophosphate (cAMP). We report here the successful complementation of this functional defect achieved by using the technique of somatic cell fusion to introduce the normal CF allele into mutant cells. CF epithelial cells were fused with transfectant mouse fibroblasts that contain the normal human gene. The resulting heterokaryons were examined for restoration of cAMP-activated Cl- transport using an optical assay of Cl- permeability. Our results provide direct evidence for the involvement of the protein product of the normal CF allele in modulating epithelial Cl- permeability.


Inflammatory microcrystals alter the functional phenotype of human osteoblast-like cells in vitro: synergism with IL-1 to overexpress cyclooxygenase-2.
Chronic crystal-associated arthropathies such as gout and pseudogout can lead to local bone destruction. Because osteoblasts, which orchestrate bone remodeling via soluble factors and cell-to-cell interactions, have been described in contact with microcrystals, particularly in uratic foci of gout, we hypothesized that microcrystals of monosodium urate monohydrate (MSUM) and of calcium pyrophosphate dihydrate (CPPD) could alter osteoblastic functions. MSUM and CPPD adhered to human osteoblastic cells (hOB) in vitro and were partly phagocytized as shown by scanning electron microscopy. MSUM and CPPD dose-dependently stimulated the production of PGE(2) in hOB as assessed by enzyme immunoassay, a response that was synergistically enhanced in the presence of IL-1. The mechanism of this synergism was, at least in part, at the level of the expression of cyclooxygenase-2 as evaluated by immunoblot analysis. MSUM and CPPD also stimulated the expression of IL-6 and IL-8 and reduced the 1,25-dihydroxyvitamin D(3)-induced activity of alkaline phosphatase and osteocalcin in hOB (with no synergism with IL-1). MSUM- or CPPD-stimulated expression of IL-6 in hOB pretreated with the selective cyclooxygenase-2 inhibitor NS-398 was increased, unlike that induced by IL-1 alone which was partially reduced. MSUM-, CPPD- or IL-1-induced expression of IL-8 was unchanged by pretreating hOB with NS-398. These results suggest that inflammatory microcrystals alter the normal phenotype of hOB, redirecting them toward reduced bone formation and amplified osteoblast-mediated bone resorption, abnormalities that could play a role in the bone destruction associated with chronic crystal-induced arthritis.


[Study on the relationship between TCM syndrome and gastroscopic feature in children with stomachache].
To explore the characteristic of TCM syndrome in children with stomachache and its relationship with the gastroscopic feature. The relationship between macroscopic syndrome differentiation and microscopic syndrome differentiation in 300 children with stomachache was analyzed by means of clinical syndrome differentiation and gastroscopic examination. The total disease detectable rate of gastroscopic examination was 98.3%. Dampness-heat syndrome and deficiency-cold syndrome were the syndromes that most commonly be detected by either macroscopic or microscopic syndrome differentiation. And the two methods of differentiation showed better coherence in superficial gastritis. The clinical effect of TCM therapy on children stomachache could be improved through integrative Chinese and Western medical approach, by conducting syndrome differentiation referring to the gastroscopic features, which could conduct microscopic guidance to the clinical macroscopic syndrome differentiation.


Hypodontia in an adolescent with the HbSC genotype: a case report.
The haemoglobin SC (HbSC) genotype is a variant form of haemoglobin disorder that often presents less frequent clinical symptoms of sickle cell disease (SCD) than the homozygous state (HbSS). Various dental abnormalities, such as enamel hypoplasia, hypomineralization, midline diastema and delayed eruption of the teeth, have been associated with SCD. This report presents the case of an adolescent patient with HbSC who had hypodontia, transposition of a permanent canine, retained primary teeth, multiple carious lesions and poor oral hygiene complicated by sickle cell crisis. This report focuses on the need for preventive care in such patients to reduce functional and aesthetic complications.


FTO Gene Polymorphisms and Their Roles in Acromegaly.
The major causes of both morbidity and mortality in patients with acromegaly are cardiovascular diseases (CVDs). The polymorphisms of the fat mass and obesity-associated gene (FTO) are associated with obesity, as well as with an increased risk of CVDs. The aim of the study was to determine the relationship of risk alleles of four FTO gene polymorphisms with selected parameters of lipid and glucose metabolism as well as with IGF-1 and GH levels in the group of patients with acromegaly compared to the control group. The study group consisted of 104 patients with acromegaly and 64 healthy subjects constituting the control group. In the whole acromegaly group, the data reveal that the homozygous for risk allele carriers (rs1421085, rs9930506, rs9939609) as well as carriers of only one risk allele have lower IGF-1 concentrations. In the well-controlled acromegaly group, the homozygous for three risk allele carriers of FTO gene polymorphisms have lower HDL cholesterol concentration (rs1121980, rs1421085, rs993609). In the cured acromegaly group, homozygous risk allele carriers rs9930506 tend to have higher levels of total cholesterol and LDL cholesterol. These associations are not observed in the control group. Conclusion: there is an association between FTO gene polymorphisms and the metabolism of lipids, suggesting that the FTO gene may be associated with higher CVD risk in patients with acromegaly. In addition, there is an association between FTO gene polymorphisms and IGF-1, implying that FTO gene may influence/modify IGF-1 synthesis. Further investigation on a larger scale is required to provide more precise evidence.


Targeted expression of tetanus toxin: a new tool to study the neurobiology of behavior.
Over the past few decades, the explosion of molecular genetic knowledge, particularly in the fruit fly Drosophila melanogaster, has led to the identification of a large number of genes, which, when mutated, directly or indirectly affect fly behavior. Beyond the genetic and molecular characterization of genes and their associated molecular pathways, recent advances in molecular genetics also have allowed the development of new tools dedicated more directly to the dissection of the neural bases for various behaviors. In particular, the conjunction of the development of two techniques--the enhancer-trap detection system and the targeted gene expression system, based on the yeast GAL4 transcription factor--has led to the development of the binary enhancer-trap P[GAL4] expression system, which allows the selective activation of any cloned gene in a wide variety of tissue- and cell-specific patterns. Thus, this development, in addition to allowing the anatomical characterization of neuronal circuitry, also allows, via the expression of tetanus toxin light chain (known to specifically block synaptic transmission), an investigation of the role of specific neurons in certain behaviors. Using this system of "toxigenetics," several forms of behavior--from those mediated by sensory systems, such as olfaction, mechanoreception, and vision, to those mediated by higher brain function, such as learning, memory and locomotion--have been studied. These studies aim to map neuronal circuitry underlying specific behaviors and thereby unravel relevant neurophysiological mechanisms. The advantage of this approach is that it is noninvasive and permits the investigation of behavior in the free moving animal. We review a number of behavioral studies that have successfully employed this toxigenetic approach, and we hope to persuade the reader that transgenic tetanus toxin light chain is a useful and appropriate tool for the armory of neuroethologists.


Extracolonic manifestations of familial polyposis coli.
The original concept of familial polyposis coli (FPC) as only a genetically determined premalignant disease of the colon changed in the 1950's, with the description of Gardner's syndrome. The extracolonic manifestations of osteoma and epidermoid cyst have since been shown to be only a small part of the spectrum of both benign and malignant extracolonic manifestations of the disease. The modern concept of this condition is that FPC is a genetically determined generalized growth disorder that gives rise to tumors in various parts of the body.


A Strategy for Screening and Confirmation of HTLV-1/2 Infections in Low-Endemic Areas.
Serological tests have been widely used for detecting human T-cell lymphotropic virus type 1/2 (HTLV-1/2) antibodies in the endemic areas, but their performance in low-risk populations is rarely reported. The aim of this study was to evaluate the performance of four HTLV-1/2 screening assays and to discuss a strategy for diagnosis of HTLV-1/2 infection in a non-endemic area. At the present study, 1546 specimens repeatedly reactive (RR) by one screening ELISA were collected from blood centers/banks from January 2016 to April 2019. Avioq-ELISA, Murex-ELISA, Roche-ECLIA and Fujirebio-CLIA were independently performed on each plasma sample and compared to WB and LIA confirmatory tests. Positive or indeterminate specimens with blood available were quantified by qPCR. The results showed that 48 samples were finally confirmed as HTLV-1 positive, 13 were HTLV positive, 151 were indeterminate, and 387 were negative. All the WB-positive samples were also LIA-positive. Roche-ECLIA showed the highest sensitivity that was able to detect 91.8% positives and combined with the Murex-ELISA would significantly increase the positive detection rate (98.4%). In addition, LIA yield more indeterminate and HTLV-untyped results than WB (152 vs. 27), but was able to resolve infection status of some individuals with an indeterminate WB. Besides, 3 WB indeterminate and 1 LIA-untyped samples were confirmed as HTLV-1 positive by qPCR. Based on these findings, we put forward a proper test strategy for HTLV-1/2 diagnosis in low-prevalence areas. If possible, the Roche-ECLIA with the highest sensitivity is suggested as a second screening assay in primary labs. If not, all RR specimens are recommended to be firstly retested by Roche-ECLIA and Murex-ELISA in the reference lab. Secondly, samples reactive to any one of the two tests were quantified by qPCR, and then the NAT-negatives were furtherly submitted to LIA for confirmation. Thereby, the cost can be reduced and the diagnostic accuracy would be improved.


Pedagogical preparation of nursing professors for professional secondary technical education.
To understand how the nursing professor pedagogically prepares for teaching in professional secondary technical education in the light of the Pedagogical Content Knowledge of Lee S. Shulman. This is a qualitative and descriptive study. Two pedagogical workshops were held to collect data, with the participation of six professors from two technical schools. The categories Pedagogical preparation for the teaching exercise and Knowledge Base for Teaching have emerged. The results revealed the difference between the real and the ideal in the perception of the participants, when their preparation for the teaching exercise is situated between the learning with colleagues and the mastery over the Content Knowledge. Professor training is necessary and is an alternative to qualify teaching in professional secondary technical education.


SNP-RFLPing 2: an updated and integrated PCR-RFLP tool for SNP genotyping.
PCR-restriction fragment length polymorphism (RFLP) assay is a cost-effective method for SNP genotyping and mutation detection, but the manual mining for restriction enzyme sites is challenging and cumbersome. Three years after we constructed SNP-RFLPing, a freely accessible database and analysis tool for restriction enzyme mining of SNPs, significant improvements over the 2006 version have been made and incorporated into the latest version, SNP-RFLPing 2. The primary aim of SNP-RFLPing 2 is to provide comprehensive PCR-RFLP information with multiple functionality about SNPs, such as SNP retrieval to multiple species, different polymorphism types (bi-allelic, tri-allelic, tetra-allelic or indels), gene-centric searching, HapMap tagSNPs, gene ontology-based searching, miRNAs, and SNP500Cancer. The RFLP restriction enzymes and the corresponding PCR primers for the natural and mutagenic types of each SNP are simultaneously analyzed. All the RFLP restriction enzyme prices are also provided to aid selection. Furthermore, the previously encountered updating problems for most SNP related databases are resolved by an on-line retrieval system. The user interfaces for functional SNP analyses have been substantially improved and integrated. SNP-RFLPing 2 offers a new and user-friendly interface for RFLP genotyping that can be used in association studies and is freely available at http://bio.kuas.edu.tw/snp-rflping2.


Detection of gastric Helicobacter spp. in stool samples of dogs with gastritis.
The aim of this study was to determine the prevalence and identify the species of gastric Helicobacter in the stool of dogs with gastritis. The study was carried out on thirty dogs of different breeds, of both genders and of various ages, diagnosed with gastritis. Helicobacter spp. was detected in stool samples using the nested-PCR method. Helicobacter bacteria were identified in stool samples from seven (23.3%) dogs. Helicobacter heilmannii was found to be the most common species of gastric Helicobacter. Helicobacter salomonis was identified much less frequently, while Helicobacter felis, Helicobacter pylori and Helicobacter bizzozeronii were not detected in any of the samples.


Non-AIDS-defining cancers in New Orleans.
Non-AIDS-defining cancers in HIV-infected patients in the highly active antiretroviral therapy era have increased. To our knowledge a comprehensive review of non-AIDS-related malignancies in New Orleans has not yet been conducted. Databases from main institutions in New Orleans were queried retrospectively for the years 2001 to 2011. The International Classification of Diseases, Ninth Revision codes were used to search for HIV infection and cancer comorbidity. A total of 16 patients were diagnosed with lung cancer (mean age 50 years) with 81% of the patients presenting with advanced stages. In all, 20 (mean age 47 years) were diagnosed with anal cancer, and 35% presented in late stages. In all, 14 patients (mean age 42 years) were diagnosed with Hodgkin Lymphoma, and 64% were diagnosed at late stage. A total of 5 women (mean age 44 years) were diagnosed with breast cancer with 40% of them presenting at late stage. Malignancies were diagnosed at late stages in the majority of the cases, presented with worse outcomes, and had higher recurrence rates. The role of HIV and other viruses (Epstein Barr virus, human papillomavirus) and the potential mechanisms or pathways of oncogene activation also need to be clarified.


Gene repair of an Usher syndrome causing mutation by zinc-finger nuclease mediated homologous recombination.
Human Usher syndrome (USH) is the most frequent cause of inherited deaf-blindness. It is clinically and genetically heterogeneous, assigned to three clinical types of which the most severe type is USH1. No effective treatment for the ophthalmic component of USH exists. Gene augmentation is an attractive strategy for hereditary retinal diseases. However, several USH genes, like USH1C, are expressed in various isoforms, hampering gene augmentation. As an alternative treatment strategy, we applied the zinc-finger nuclease (ZFN) technology for targeted gene repair of an USH1C, causing mutation by homologous recombination. We designed ZFNs customized for the p.R31X nonsense mutation in Ush1c. We evaluated ZFNs for DNA cleavage capability and analyzed ZFNs biocompatibilities by XTT assays. We demonstrated ZFNs mediated gene repair on genomic level by digestion assays and DNA sequencing, and on protein level by indirect immunofluorescence and Western blot analyses. The specifically designed ZFNs did not show cytotoxic effects in a p.R31X cell line. We demonstrated that ZFN induced cleavage of their target sequence. We showed that simultaneous application of ZFN and rescue DNA induced gene repair of the disease-causing mutation on the genomic level, resulting in recovery of protein expression. In our present study, we analyzed for the first time ZFN-activated gene repair of an USH gene. The data highlight the ability of ZFNs to induce targeted homologous recombination and mediate gene repair in USH. We provide further evidence that the ZFN technology holds great potential to recover disease-causing mutations in inherited retinal disorders.


Effects of vitamin A deficiency and repletion on rat glucagon secretion.
To determine whether vitamin A is involved in pancreatic alpha cell function, we tested for (a) effects of vitamin A deficiency on glucagon release from perifused islets and perfused pancreases, and (b) the presence of cytosolic retinol-binding proteins (CRBP) and retinoic acid-binding proteins (CRABP), in the glucagon-secreting alpha cell line, ln-R1-G9. Arginine 19 mM plus glucose 2.8 mM-stimulated glucagon secretion was markedly impaired in islets and pancreases of vitamin A-deficient rats or rats that had at some time been cycled through vitamin A deficiency (ever A-def) despite repletion with retinoids for 2-4 weeks. Insulin secretion was impaired likewise. Repletion starting early in the development of vitamin A deficiency and for a longer period of time (18 or 60 days) did not restore glucagon secretion, but did normalize insulin secretion. CRBP and CRABP were present in ln-R1-G9 cells. We conclude that (a) vitamin A deficiency is associated with a defect in glucagon secretion; (b) The defect in secretion occurs early in the course of vitamin A deficiency; (c) The defect persists despite repletion; and (d) The requirement of vitamin A for secretion and the presence of CRBP and CRABP in glucagon-secreting cells support a physiologic role for vitamin A at the alpha cell level.


Oxidant stress stimulates anion secretion from the human airway epithelial cell line Calu-3: implications for cystic fibrosis lung disease.
Exposure to reactive oxygen species (ROS) is associated with tissue damage in the lung and may be a common element in the pathogenesis of all inflammatory lung diseases. Exposure to the ROS hydrogen peroxide (H2O2) evoked a rapid increase in transepithelial anion secretion across monolayers of the human submucosal gland serous cell line Calu-3. This increase was almost entirely abolished by the addition of diphenylamine-2-carboxylate (DPC), implicating the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel in the response. The response was also reduced by inhibitors of basolateral K+ channels. Studies of electrically isolated apical and basolateral membranes revealed that H2O2 stimulated both apical Cl- and basolateral K+ conductances (G(Cl) and G(K)). Apical G(Cl) was sensitive to DPC, but unaffected by 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), suggesting that CFTR is the major anion conduction pathway mediating the response to H2O2. Additionally, H2O2 had no effect on G(Cl) in the presence of the adenylate cyclase inhibitor SQ22536 or following maximal stimulation of G(Cl) with forskolin, implicating the cAMP-dependent protein kinase pathway in the apical response to H2O2. Basolateral G(K) was reduced by the K+ channel inhibitors clotrimazole and clofilium, indicating roles for KCNN4 and KCNQ1 in the H2O2-stimulated response. We propose that ROS-stimulated anion secretion from serous cells plays an important role in keeping the airways clear from damaging radicals that could potentially initiate tissue destruction. Our finding that this response is CFTR dependent suggests that an important host defence mechanism would be dysfunctional in the cystic fibrosis (CF) lung. Loss of this compensatory protective mechanism could expose the CF lung to ROS for extended periods, which could be important in the pathogenesis of CF lung disease.


Enzymatic synthesis of esters using an immobilized lipase.
Various esters were synthesized in nearly anhydrous hexane from alcohols and carboxylic acids using a lipase from Candida cylindracea. The enzyme was immobilized on a nylon support and protein loadings as high as 10 mg/g were obtained. The activity of the immobilized enzyme was maximum in a range of temperatures from 25 to 37 degrees C. Ethylpropionate was formed from ethanol and propionic acid at a rate of 0.017 mol/h g immobilized protein. Different esters were formed at comparable rates and equilibrium conversions could generally be approached in less than 10 h in a batch reaction system. The immobilized lipase catalyst was quite stable and retained about one third of the initial activity after repeated experiments during the course of 72 days. A stirred tank continuous flow reactor was used successfully for the continuous production of esters.


Visible-Light-Induced Palladium-Catalyzed Selective Defluoroarylation of Trifluoromethylarenes with Arylboronic Acids.
Selective functionalization of inactive C(sp3)-F bonds to prepare medicinally interesting aryldifluoromethylated compounds remains challenging. One promising route is the transition-metal-catalyzed cross-coupling through oxidative addition of the C(sp3)-F bond in trifluoromethylarenes (ArCF3), which are ideal precursors for this process due to their ready availability and low cost. Here, we report an unprecedented excited-state palladium catalysis strategy for selective defluoroarylation of trifluoromethylarenes with arylboronic acids. This visible-light-induced palladium-catalyzed cross-coupling proceeds under mild reaction conditions and allows transformation of a variety of arylboronic acids and ArCF3. Preliminary mechanistic studies reveal that the oxidative addition of the C(sp3)-F bond in ArCF3 to excited-state palladium(0) via a single electron transfer pathway is responsible for the C(sp3)-F bond activation.


Comparison of axial lengths in occludable angle and angle-closure glaucoma--the Bhaktapur Glaucoma Study.
To compare the anterior chamber depth (ACD) and axial length of eyes in a population-based sample among normal, occludable angle, and primary angle-closure glaucoma (PACG) groups. Totally, 3979 subjects from a population-based glaucoma prevalence study underwent complete ocular examination including four mirror gonioscopy. An angle in which >270 degrees of the posterior trabecular meshwork could not be seen was considered occludable. PACG was considered when there were features of primary angle closure (eye with an occludable angle, peripheral anterior synechiae, increased intraocular pressure, iris whorling, lens opacities, or excessive pigment deposition on the trabecular surface) and optic disc with glaucomatous damage. A handheld ultrasonic biometer was used to measure the ACD and the axial length of subjects. The ACD and axial length were measured on all subjects with occludable angles (n = 86), PACG (n = 17) and a random subgroup of normal subjects (n = 685). The measurements were compared among the groups and statistically analyzed using independent t-test, one-way analysis of variance, χ2 test, and logistic regression. The mean age among occludable angle and angle-closure glaucoma groups was 59.64 ± 9.70 and 71.23 ± 8.5 years, respectively, which was significantly higher (p < 0.001) than the mean of 54.03 ± 10.90 years among normal subjects. In the occludable angle group, the ACD (2.55 ± 0.69) was significantly different (p < 0.001) than the normal group (2.85 ± 0.39). The axial length (22.08 ± 0.83) in the occludable angle group was also considerably different (p < 0.001) when compared with the normal group (22.62 ± 0.90). In PACG group, the ACD and axial length were 2.48 ± 0.22 and 22.02 ± 0.59, respectively. The risk of having an occludable angle decreased with per unit millimeter increase in ACD (odds ratio, 0.24; 95% confidence intervals, 0.12-0.50) and axial length (odds ratio, 0.49; 95% confidence intervals, 0.36-0.67) after adjusting the effect of age and sex. The eyes in Nepalese population with occludable angle and angle-closure glaucoma appear to have significantly shallower anterior chambers and shorter axial lengths when compared with the normal group.


Human macrophage colony-stimulating factor induces the differentiation of trophoblast.
When human cytotrophoblastic cells in the early stage of pregnancy were cultured in a serum-free medium in the presence of human macrophage colony-stimulating factor (M-CSF), the cytotrophoblastic cells fused and formed a typical syncytiotrophoblast which had a dense distribution of microvilli revealed under an electron microscope. On the other hand, cytotrophoblasts incubated with anti-M-CSF antibody showed hardly any syncytiotrophoblast formation. Following this finding, we studied the differentiation of chorionic cells from the viewpoint of hormone secretion. When cytotrophoblasts were incubated in the presence of M-CSF, the supernatant of the culture showed an increase in human chorionic gonadotropin and human placental lactogen levels in proportion to the concentration of M-CSF added. When cytotrophoblasts were incubated in the presence of anti-M-CSF antibody or anti-fms antibody, human chorionic gonadotropin and human placental lactogen secretion were suppressed. Thus, M-CSF was morphologically and endocrinologically found to induce the differentiation of chorionic cells.


Non-linear conduction due to depinning of charge order domains in Fe3O2BO3.
The oxyborate Fe3O2BO3 presents a charge density wave (CDW) transition close to room temperature. As we show here, this is associated with a well defined anomaly in the specific heat. Below this transition, when applying in a single crystal of Fe3O2BO3 a DC voltage above a temperature dependent threshold, a high current is liberated in this material. We study the conduction in single crystals of Fe3O2BO3 with voltage applied parallel and perpendicular to the crystallographic c axis direction. The observed currents are attributed to the depinning of charge ordered domains above a threshold voltage V T2 that gives rise to a collective conduction due to coherent domains. Compliance limited DC data shows that above a lower threshold voltage depinning is smooth and follows a power law scaling. Similar depinning with power law scaling is also revealed in the AC conductivity.


Interleukin-6 and B-type natriuretic peptide are independent predictors for worsening of heart failure in patients with progressive congestive heart failure.
Neurohormones and cytokines have been associated with poor prognosis in patients with congestive heart failure. However, direct comparisons between them are rare and the best predictor for worsening of heart failure remains to be elucidated. The aim of the study was to identify independent predictors for worsening of heart failure. We studied 100 patients with congestive heart failure (LVEF <or=25%). Serum levels of interleukin-6 (IL-6), tumor necrosis factor (TNF)-alpha, aminoterminal atrial natriuretic peptide (N-ANP), B-type natriuretic peptide (BNP), aminoterminal B-type natriuretic peptide (N-BNP), norepinephrine (NE) and big endothelin (big ET) were measured at entry into the study. Univariate and stepwise multivariate regression analyses were performed to compare variables and clinical parameters. The end-point was defined as worsening of heart failure. During follow-up (mean 378 days), 30 patients reached the defined end-point. Univariate risk factors of worsening of heart failure were IL-6 (p = 0.0001), log BNP (p = 0.0001), log N-ANP (p = 0.0259), big ET (p = 0.0032), log N-BNP (p = 0.0495), NE (p = 0.0013) and LVEF (p = 0.0498). Multivariate regression analysis identified IL-6 as the strongest independent predictive value for worsening of heart failure (p = 0.0001), followed by log BNP (p = 0.0002). Mean IL-6 levels were 1.76 +/- 1.73 in stable patients and 4.62 +/- 7.98 pg/ml in patients who developed worsening of heart failure. Using a cut-off value of 1.66 pg/ml for IL-6 Kaplan-Meier freedom from worsening, heart failure was significantly higher in patients below this cut-off point compared with those above (p = 0.0002). IL-6 and BNP independently predict worsening of heart failure. A single measurement could help to identify patients at high risk for worsening of heart failure and these individuals should be monitored more closely.


Occupational biohazards: a review.
There are at least 193 important biological agents that show infectious, allergenic, toxic, or carcinogenic activities in the working population. These agents are viruses, bacteria, fungi, plant substances, invertebrate animals (mostly arthropods), and substances derived from vertebrate animals. At least 20 large occupational groups are exposed to these biohazards. The risk is greatest among health care and laboratory workers who are threatened by human pathogens and among agricultural workers who are at risk from dust-borne biological allergens and toxins and by parasitic worms in warm climates. There is growing evidence that biohazards are also important risk factors for many other professions, including woodworkers, workers of textile plants, sewage and compost workers, miners and renovators. Some suggestions for research and prevention for reducing the occupational risks from biohazards are discussed.


Enhancing marketing recruitment strategies: administrator tenure and nursing expenditures.
When recruiting nurses, long-term care facilities require an ability to identify salient organizational characteristics that are attractive to potential nursing services candidates vis-à-vis their competitors. The findings of this study suggest that information on administrative tenure can be utilized to attract applicants by appealing to criteria within their high-involvement job search activities. High-involvement applicants proactively seek recruitment content that provides essential job attributes that match their needs and skills and are more apt to be a higher quality candidate. Based upon the study's findings, managers are offered marketing strategy recommendations for tailoring recruiting messages that appeal to high-involvement job seekers.


The deformation of cancer cells through narrow micropores holds the potential to regulate genes that impact cancer malignancy.
Surgery, radiation, hormonal therapy, chemotherapy, and immunotherapy are standard treatment strategies for metastatic breast cancer. However, the heterogeneous nature of the disease poses challenges and continues to make it life-threatening. It is crucial to elucidate further the underlying signaling pathways to improve treatment efficacy. Our study established two triple-negative breast cancer cell lines (TW-1 and TW-2) that were physically deformed using 3 μm pores to investigate the relationship between cancer cell deformation and metastasis within a heterogeneous population. The physical transformation of TW-1 and TW-2 cells significantly affected their growth and migration speed, as evidenced by wound healing assays for collective cell migration and microchannel assays for single-cell migration. We conducted bulk RNA sequencing to gain insights into the genes influenced by physical deformation. Additionally, we evaluated the effects of trametinib resistance on breast cancer cell metastasis by assessing cell viability and migration rates. Interestingly, TW-1 and TW-2 cells exhibited resistance to trametinib treatment. We observed a significant upregulation of GABRA-3, a protein commonly expressed in malignant breast cancer, and the critical transcription factor Myc in TW-1 and TW-2 cells compared to the control group (Ori). However, we did not observe a significant difference in Myc expression between TW-1 and TW-2 cells. In contrast, in the trametinib-resistant cell lines (TW-1-Tra and TW-2-Tra), we found increased expression of OCT4 and SOX2 rather than GABRA-3 or Myc. These findings highlight the differential expression patterns of these genes in our study, suggesting their potential role in cancer cell deformation and drug resistance. Our study presents a potential in vitro model for metastatic and drug-resistant breast cancer cells. By investigating the correlation between cancer cell deformation and metastasis, we contribute to understanding breast cancer heterogeneity and lay the groundwork for developing improved treatment strategies.


MiR-223 as a Regulator and Therapeutic Target in Liver Diseases.
MicroRNAs (miRNAs) are endogenous non-coding single-stranded small molecule RNAs consisting of 20-24 nucleotides that are highly conserved in species evolution. Expression of miRNAs is strictly tissue-specific, and it is chronological in fungi and plants, as well as in animals. MiR-223 has been shown to play a key role in innate immunity, and dysregulation of its expression contributes to the pathogenesis of multiple inflammatory diseases, and cancers. In this article the biosynthesis and functions of miR-223 in innate immunity are reviewed, and the role of miR-223 in liver physiopathology and therapeutic prospects are highlighted.


RGS9: a regulator of G-protein signalling with specific expression in rat and mouse striatum.
A clone of the regulator of G-protein signalling, RGS9, was isolated from a rat striatum-minus-cerebellum-minus-hippocampus subtracted library generated by directional tag polymerase chain reaction subtraction. The full-length cDNA clone encodes a 444 amino acid protein containing an 118 amino acid RGS domain, which corresponds to an evolutionarily conserved domain that is present in all members of the RGS family of proteins. Outside of the homology domain, RGS9 shows more extended similarity to human RGS6 and RGS7, rat RGS12, and the C. elegans protein EGL-10. During embryonic and early postnatal stages of development, two RGS9 transcripts of approximately 1.4 Kb and 1.8 Kb were detected in whole brain. After postnatal day 10, accumulation of the larger transcript increased progressively until adulthood at the expense of the smaller transcript, which was undetectable in the adult. In adult rat brain, the 1.8-Kb RGS9 transcript was detected in the striatum but not in other brain regions or peripheral tissues. In situ hybridization in rat and mouse demonstrates that RGS9 mRNA is expressed predominantly in medium-sized, spiny neurons of the neostriatum and in neurons of the nucleus accumbens and olfactory tubercle. Relatively strong signals were also detected in some hypothalamic nuclei. Its selective expression suggests that RGS9 may play an important role in modulation of the complex signalling pathways of the basal ganglia.


Isolation of cancer stem cells from adult glioblastoma multiforme.
Glioblastoma multiforme (GBM) is the most common adult primary brain tumor and is comprised of a heterogeneous population of cells. It is unclear which cells within the tumor mass are responsible for tumor initiation and maintenance. In this study, we report that brain tumor stem cells can be identified from adult GBMs. These tumor stem cells form neurospheres, possess the capacity for self-renewal, express genes associated with neural stem cells (NSCs), generate daughter cells of different phenotypes from one mother cell, and differentiate into the phenotypically diverse populations of cells similar to those present in the initial GBM. Having a distinguishing feature from normal NSCs, these tumor stem cells can reform spheres even after the induction of differentiation. Furthermore, only these tumor stem cells were able to form tumors and generate both neurons and glial cells after in vivo implantation into nude mice. The identification of tumor stem cells within adult GBM may represent a major step forward in understanding the origin and maintenance of GBM and lead to the identification and testing of new therapeutic targets.


The stress response: implications for the clinical development of hsp90 inhibitors.
In their role as molecular chaperones, heat shock proteins serve as central integrators of protein homeostasis within cells. As part of this function, they guide the folding, assembly, intracellular disposition and proteolytic turnover of many key regulators of cell growth, differentiation and survival. Not surprisingly then, heat shock proteins are over expressed in many types of cancer, and induction of the stress response may actually be required for cells to tolerate the genetic disarray characteristic of malignant transformation. Regulation of heat shock protein levels via the stress response is complex, but recent data indicate that the molecular chaperone Hsp90 plays a key role. Specifically, Hsp90 inhibitors alter the multi-chaperone complexes associated with Heat Shock Factor 1 (HSF1), the dominant transcription factor controlling induction of the stress response, and stimulate HSF1-activated heat shock gene expression. Induction of this heat shock response has now emerged as an important consideration in the further clinical development of Hsp90 inhibitors for several reasons. First, tumors in which the stress response is compromised appear particularly sensitive to Hsp90 inhibition. Second, induction of the stress response by Hsp90 inhibitors provides a sensitive pharmacodynamic endpoint with which to monitor drug action in individual patients. Third, Hsp90 inhibitors display important therapeutic interactions with both conventional DNA-targeted chemotherapeutics and newer molecularly targeted agents. These interactions are, at least in part, due to modulation of the stress response by these drugs. Lastly, stress response induction by Hsp90 inhibitors may have therapeutic benefits in non-neoplastic disorders such as heart disease, stroke and neurodegenerative diseases. These benefits are just beginning to be explored.


Prediction of amino acid residues participated in substrate recognition by cytochrome P450 subfamilies with broad substrate specificity.
Cytochromes P450 comprise a large superfamily and several of their isoforms play a crucial role in metabolism of xenobiotics, including drugs. Although these enzymes demonstrate broad and cross-substrate specificity, different cytochrome P450 subfamilies exhibit certain selectivity for some types of substrates. Analysis of amino acid residues of the active sites of six cytochrome subfamilies (CYP1А, CYP2А, CYP2С, CYP2D, CYP2E and CYP3А) enables to define subfamily-specific patterns that consist of four residues. These residues are located on the periphery of the active sites of these cytochromes. We suggest that they can form a primary binding site at the entrance to the active site, defining cytochrome substrate recognition.


Antioxidant and cancer cell proliferation inhibition effect of citrus pectin-oligosaccharide prepared by irradiation.
Pectin was dissolved in deionized distilled water (2%, vol/vol) and irradiated at 20 kGy using a Co-60 gamma ray irradiator. The resulting solution was dialyzed and lyophilized. The samples were separated into three groups to estimate their antioxidant and cancer cell proliferation effects: non-irradiated (0 kGy), irradiated (20 kGy), and dialyzed (20 kGy-F, mol wt <10,000) samples. Antioxidant properties of each treatment was tested by a beta-carotene-linoleic acid bleaching assay and electron donating ability and compared for antioxidant index, which indicated that the activity was higher in the order of 20 kGy-F > 20 kGy > 0 kGy. Spleen cell survival effect of the irradiated pectin (20 kGy) and dialyzed (20 kGy-F) samples was higher than the non-irradiated control (0 kGy). The pectins inhibited growth of the cancer cell in the order of 20 kGy- F > 20 kGy > 0 kGy. The Ames test revealed that none of the fractions was mutagenic, and there was no indication of a dose-dependent response for any of the samples. These results suggest that a functional pectin oligosaccharide can be produced by irradiation for the food industry without any chemical treatment.


Deciphering the dual emission in the photoluminescence of Au14Cd(SR)12: A theoretical study using TDDFT and TDDFT + TB.
Determining excited state processes for small nanoclusters, specifically gold, aids in our ability to fine-tune luminescent materials and optical devices. Using TDDFT and TDDFT + TB, we present a detailed theoretical explanation for the dual emission peaks displayed in Au14Cd(S-Adm)12 (Adm = adamantane). As dual emission is relatively rare, we decipher whether the mechanism originates from two different excited states or from two different minima on the same excited state surface. This unique mechanism, which proposes that the dual emission results from two minima on the first excited state, stems from geometrical changes in the bi-tetrahedron core during the emission process.


Identification of Bacterial Drug-Resistant Cells by the Convolutional Neural Network in Transmission Electron Microscope Images.
The emergence of bacteria that are resistant to antibiotics is common in areas where antibiotics are used widely. The current standard procedure for detecting bacterial drug resistance is based on bacterial growth under antibiotic treatments. Here we describe the morphological changes in enoxacin-resistant Escherichia coli cells and the computational method used to identify these resistant cells in transmission electron microscopy (TEM) images without using antibiotics. Our approach was to create patches from TEM images of enoxacin-sensitive and enoxacin-resistant E. coli strains, use a convolutional neural network for patch classification, and identify the strains on the basis of the classification results. The proposed method was highly accurate in classifying cells, achieving an accuracy rate of 0.94. Using a gradient-weighted class activation mapping to visualize the region of interest, enoxacin-resistant and enoxacin-sensitive cells were characterized by comparing differences in the envelope. Moreover, Pearson's correlation coefficients suggested that four genes, including lpp, the gene encoding the major outer membrane lipoprotein, were strongly associated with the image features of enoxacin-resistant cells.


Nest-building behavior of Monk Parakeets and insights into potential mechanisms for reducing damage to utility poles.
The Monk Parakeet (Myiopsitta monachus) commonly uses utility poles as a substrate for building large, bulky nests. These nests often cause fires and electric power outages, creating public safety risks and increasing liability and maintenance costs for electric companies. Previous research has focused on lethal methods and chemical contraception to prevent nesting on utility poles and electrical substations. However, implementation of lethal methods has led to public protests and lawsuits, while chemical contraception may affect other than the targeted species, and must be continually reapplied for effectiveness. One non-lethal alternative, nest removal, is costly and may not be a sustainable measure if Monk Parakeet populations continue to grow. In order to identify cost-effective non-lethal solutions to problems caused by Monk Parakeet nesting, we studied their behavior as they built nests on utility poles. Monk Parakeets initiate nests by attaching sticks at the intersection of the pole and electric lines. We found that parakeets use the electric lines exclusively to gain access to the intersection of lines and pole during nest initiation, and continue to use the lines intensively throughout construction. Monk Parakeets also have more difficulty attaching sticks during the early stages of nest construction than when the nest is nearing completion. These findings suggest that intervention during the earlier stages of nest building, by excluding Monk Parakeets from electric lines adjacent to poles, may be an effective, non-lethal method of reducing or eliminating parakeets nesting on, and damaging, utility poles.


Sphingolipids and mitochondrial function, lessons learned from yeast.
Mitochondrial dysfunction is a hallmark of several neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease, but also of cancer, diabetes and rare diseases such as Wilson's disease (WD) and Niemann Pick type C1 (NPC). Mitochondrial dysfunction underlying human pathologies has often been associated with an aberrant cellular sphingolipid metabolism. Sphingolipids (SLs) are important membrane constituents that also act as signaling molecules. The yeast Saccharomyces cerevisiae has been pivotal in unraveling mammalian SL metabolism, mainly due to the high degree of conservation of SL metabolic pathways. In this review we will first provide a brief overview of the major differences in SL metabolism between yeast and mammalian cells and the use of SL biosynthetic inhibitors to elucidate the contribution of specific parts of the SL metabolic pathway in response to for instance stress. Next, we will discuss recent findings in yeast SL research concerning a crucial signaling role for SLs in orchestrating mitochondrial function, and translate these findings to relevant disease settings such as WD and NPC. In summary, recent research shows that S. cerevisiae is an invaluable model to investigate SLs as signaling molecules in modulating mitochondrial function, but can also be used as a tool to further enhance our current knowledge on SLs and mitochondria in mammalian cells.


Recent evidence of economic evaluation of artificial intelligence in ophthalmology.
Health economic evaluation (HEE) is essential for assessing value of health interventions, including artificial intelligence. Recent approaches, current challenges, and future directions of HEE of artificial intelligence in ophthalmology are reviewed. Majority of recent HEEs of artificial intelligence in ophthalmology were for diabetic retinopathy screening. Two models, one conducted in the rural USA (5-year period) and another in China (35-year period), found artificial intelligence to be more cost-effective than without screening for diabetic retinopathy. Two additional models, which compared artificial intelligence with human screeners in Brazil and Thailand for the lifetime of patients, found artificial intelligence to be more expensive from a healthcare system perspective. In the Thailand analysis, however, artificial intelligence was less expensive when opportunity loss from blindness was included. An artificial intelligence model for screening retinopathy of prematurity was cost-effective in the USA. A model for screening age-related macular degeneration in Japan and another for primary angle close in China did not find artificial intelligence to be cost-effective, compared with no screening. The costs of artificial intelligence varied widely in these models. Like other medical fields, there is limited evidence in assessing the value of artificial intelligence in ophthalmology and more appropriate HEE models are needed.


Adenosine receptor modeling: what does the A2A crystal structure tell us?
For a long time, there have been no experimentally determined structural data for any adenosine receptor (AR) and the only approach available for making structure/function correlations about these proteins has been homology modeling. While the early attempts to model these receptors followed the crystallization of bacteriorhodopsin, the cryo-microscopy studies of bovine and frog rhodopsin, and the modeling of a Calpha-template for the TM helices in the rhodopsin family of GPCRs, the crystallization of bovine rhodopsin by Palczewski was of extreme importance as it first provided the crystal structure of an eukaryotic GPCR to be used as template for more realistic homology models. Since then, rhodopsin-based modeling became the routine approach to develop AR structural models that proved to be useful for interpretation of site-directed mutagenesis data and for molecular docking studies. The recently reported crystal structures of the adrenergic beta1 and beta2 receptors only partially confirmed the structural features showed by bovine rhodopsin, raising a question about which template would have been better for further modeling of ARs. Such question remained actually not-answered, due to the publication in late 2008 of the crystal structure of human adenosine A(2A) receptor (AA(2A)R). Since its publication, this structure has been used for ligands docking analysis and has provided a high similarity template for homology modeling of the other AR subtypes. Still, the AA(2A)R crystal structure allows to verify the hypotheses that were made on the basis of the previously reported homology modeling and molecular docking.


White-to-brown transdifferentiation of omental adipocytes in patients affected by pheochromocytoma.
In all mammals, white adipose tissue (WAT) and brown adipose tissue (BAT) are found together in several fat depots, forming a multi-depot organ. Adrenergic stimulation induces an increase in BAT usually referred to as "browning". This phenomenon is important because of its potential use in curbing obesity and related disorders; thus, understanding its cellular mechanisms in humans may be useful for the development of new therapeutic strategies. Data in rodents have supported the direct transformation of white into brown adipocytes. Biopsies of pure white omental fat were collected from 12 patients affected by the catecholamine-secreting tumor pheochromocytoma (pheo-patients) and compared with biopsies from controls. Half of the omental fat samples from pheo-patients contained uncoupling protein 1 (UCP1)-immunoreactive-(ir) multilocular cells that were often arranged in a BAT-like pattern endowed with noradrenergic fibers and dense capillary network. Many UCP1-ir adipocytes showed the characteristic morphology of paucilocular cells, which we have been described as cytological marker of transdifferentiation. Electron microscopy showed increased mitochondrial density in multi- and paucilocular cells and disclosed the presence of perivascular brown adipocyte precursors. Brown fat genes, such as UCP1, PR domain containing 16 (PRDM16) and β3-adrenoreceptor, were highly expressed in the omentum of pheo-patients and in those cases without visible morphologic re-arrangement. Of note, the brown determinant PRDM16 was detected by immunohistochemistry only in nuclei of multi- and paucilocular adipocytes. Quantitative electron microscopy and immunohistochemistry for Ki67 suggest an unlikely contribution of proliferative events to the phenomenon. The data support the idea that, in adult humans, white adipocytes of pure white fat that are subjected to adrenergic stimulation are able to undergo a process of direct transformation into brown adipocytes. This article is part of a Special Issue entitled Brown and White Fat: From Signaling to Disease.


Descriptions and Correlates of Medication Adherence, Attitudes, and Self-Efficacy in Outpatients With Schizophrenia Spectrum Disorders (SSDs).
The problem of medication adherence in schizophrenia spectrum disorders (SSDs) has challenged researchers and clinicians for decades. Few investigations have examined non-psychiatric adherence in this group. We conducted a descriptive correlational investigation of adherence and related factors in 185 stable outpatients with SSDs. Fifty-seven percent of participants had antipsychotic medication levels within therapeutic range and 42% had levels below therapeutic range. Pill count percentage adherence to antipsychotic medications ranged from 0-100% with a mean of 70% and SD 34.9. Mean non-psychiatric medication adherence ranged from 0 to 100 with a mean of 61% and SD 31.8. The following characteristics were not significantly associated with adherence: age, diagnosis, gender, race, living arrangement, educational level, typical versus atypical antipsychotic medication. Level of symptoms was correlated negatively and significantly with self-reported medication adherence and medication adherence self-efficacy. Our next project will examine the effectiveness of a telephone-delivered intervention designed to support adherence in this group.


Treatment With Balloon Angioplasty of Chronic Portal Vein Thrombosis.
The therapeutic options in portal vein thrombosis cases of young age and low weight, as in this case, are limited. Interventional radiologists also have minimal experience in pediatric patients. There are no reported cases anywhere worldwide, especially in this age group. However, we think that balloon angioplasty can be safely applied in cases in which esophageal variceal bleeding cannot be controlled using traditional treatment.


Quantum melting of valence-bond crystal insulators and novel supersolid phase at commensurate density.
Bosonic and fermionic Hubbard models on the checkerboard lattice are studied numerically for infinite on-site repulsion. At particle density n=1/4 and strong nearest-neighbor repulsion, insulating Valence-Bond crystals (VBC) of resonating particle pairs are stabilized. Their melting into superfluid or metallic phases under increasing hopping is investigated at T=0 K. We identify a novel and unconventional commensurate VBC supersolid region, precursor to the melting of the bosonic crystal. Hardcore bosons (spins) are compared to fermions (electrons), as well as positive to negative (frustrating) hoppings.


Surgery for deep venous incompetence.
Chronic deep venous incompetence (DVI) is caused by incompetent vein valves and/or the blockage of large calibre leg veins, with a range of symptoms including recurrent ulcers, pain and swelling. To establish the effectiveness of various surgical procedures for treating DVI. Trials were identified through the Cochrane Peripheral Vascular Diseases Group's trials register, reference lists of relevant studies, and contact with principal investigators of identified trials and world experts in deep venous surgery. Randomised controlled trials of surgical treatment for patients with DVI. Reviewers extracted data independently. Outcome measures included ambulatory venous pressure (AVP) and venous refill time (VRT). Three trials were included, one trial was excluded. Two trials compared external valvuloplasty using limited anterior plication (LAP) in combination with ligation (L) of incompetent superficial veins (L+LAP) against ligation only (L). The other trial compared external valvuloplasty and ligation (V+L) of incompetent superficial veins against ligation only (L). Trial participants had primary valvular incompetence with mild to moderate symptoms but no venous ulcers.L+LAP produced significant improvement in AVP: the mean difference between L+LAP and L groups was -15 mm Hg (95% confidence interval (CI) -20.9 to -9.0) at one year and -15 mm Hg (95% CI -21 to -8.9) at ten years.AVP values after surgery remained relatively high. Nine of eleven valves repaired remained competent after two years of follow up. No complications occurred. The overall mean score for clinical outcome was +2 (moderate improvement) in the L+LAP group compared with +1 (mild improvement) in the L group. Patients with deteriorating clinical dynamics over the five years preceding surgery had a significantly higher rate of improvement in clinical condition in V+L compared to L (81% versus 51%; p < 0.05) after seven years follow-up. Patients with stable preoperative clinical dynamics demonstrated a similar rate of improvement in both groups (96% versus 90%; p> 0.1). AVPs were not performed. These results indicate that ligation and valvuloplasty may have produced a moderate and sustained improvement for seven to ten years after surgery, in patients with mild to moderate DVI caused by primary valvular incompetence. However, there is insufficient evidence to recommend the treatment to this subgroup of patients, as the trials were small, used different methods of valvuloplasty and different methods of assessment.