[Nutritional counseling for children under two years of age: opportunities and obstacles as strategic challenges].
This article sought to conduct a critical analysis of the state of the art of the scientific production in counseling practices in nutrition for infants under two years of age, discussing the opportunities and obstacles in terms of strategic challenges for implementing healthy nutrition practices. It is a bibliographic study, with the core interest being counseling on infant nutrition. The majority (58.1 %) of the articles analyzed was published between 2008 and 2011. The educative actions of counseling on infant feeding were directed at health professionals and caregivers to children under two years of age. As opportunities in the practice of nutritional counseling the following stand out: an improvement in the performance of health professionals, the adoption of healthy eating practices by caregivers, and an improvement in the nutritional status of the children. As obstacles, the following stand out: a lack of training of caregivers and professionals regarding complementary healthy feeding and difficulties regarding the practice of counseling by a health care professional. This evidence reaffirms the importance of incorporating counseling guidance on infant feeding, as permanent practices in policies for child health care.


Site-directed mutagenesis of firefly luciferase: implication of conserved residue(s) in bioluminescence emission spectra among firefly luciferases.
The bioluminescence colours of firefly luciferases are determined by assay conditions and luciferase structure. Owing to red light having lower energy than green light and being less absorbed by biological tissues, red-emitting luciferases have been considered as useful reporters in imaging technology. A set of red-emitting mutants of Lampyris turkestanicus (Iranian firefly) luciferase has been made by site-directed mutagenesis. Among different beetle luciferases, those from Phrixothrix (railroad worm) emit either green or red bioluminescence colours naturally. By substitution of three specific amino acids using site-specific mutagenesis in a green-emitting luciferase (from L. turkestanicus), the colour of emitted light was changed to red concomitant with decreasing decay rate. Different specific mutations (H245N, S284T and H431Y) led to changes in the bioluminescence colour. Meanwhile, the luciferase reaction took place with relative retention of its basic kinetic properties such as K(m) and relative activity. Structural comparison of the native and mutant luciferases using intrinsic fluorescence, far-UV CD spectra and homology modelling revealed a significant conformational change in mutant forms. A change in the colour of emitted light indicates the critical role of these conserved residues in bioluminescence colour determination among firefly luciferases. Relatively high specific activity and emission of red light might make these mutants suitable as reporters for the study of gene expression and bioluminescence imaging.


[Mo2(CN)11]:5- A detailed description of ligand-field spectra and magnetic properties by first-principles calculations.
An ab initio multiconfigurational approach has been used to calculate the ligand-field spectrum and magnetic properties of the title cyano-bridged dinuclear molybdenum complex. The rather large magnetic coupling parameter J for a single cyano bridge, as derived experimentally for this complex by susceptibility measurements, is confirmed to a high degree of accuracy by our CASPT2 calculations. Its electronic structure is rationalized in terms of spin-spin coupling between the two constituent hexacyano-monomolybdate complexes. An in-depth analysis on the basis of Anderson's kinetic exchange theory provides a qualitative picture of the calculated CASSCF antiferromagnetic ground-state eigenvector in the Mo dimer. Dynamic electron correlations as incorporated into our first-principles calculations by means of the CASPT2 method are essential to obtain quantitative agreement between theory and experiment.


Cumulative effects of successive restorative procedures on anterior crown flexure: intact versus veneered incisors.
When successive restorative procedures (e.g., porcelain veneers, interdental resin composite restorations, and endodontic treatment) are carried out on the same tooth, significant effects on crown flexure can be expected. Dentin-bonded porcelain veneers (experimental group) were assessed in vitro using functional and cyclic thermal loads. They were compared to natural teeth (control group) with respect to 2 parameters: coronal flexure (investigated using experimental strain gauges) and morphology of the tooth-restoration interface (scanning electron microscopic evaluation). For both veneered and natural teeth, crown deformation was recorded at 5 sequential experimental steps: intact tooth (baseline), Class III cavities, Class III resin composite restorations, endodontic treatment, and endodontic restoration (without posts). No significant differences in crown flexure were found between natural and veneered incisors when compared across experimental steps. The main effect for experimental steps was highly significant. When averaged across all specimens (natural and veneered teeth), the endodontic treatment step resulted in the highest crown flexure (1.55x the baseline value). The unrestored Class III cavities and the endodontic restoration were next highest (1.30x and 1.28x the baseline value, respectively). The lowest crown flexures were found after restoration of the Class III cavities (1.13x the baseline value). No measurable microleakage or gaps were detected at the ceramic-resin, resin-enamel, or resin-dentin interfaces (Optibond FL, Kerr). Each subsequent reduction in tooth structure resulted in a substantial increase in crown flexibility, even after restoration. Endodontic procedures were responsible for most of the loss in crown stiffness. Extensive proximal cutting and restorations seemed to minimally affect crown flexure. Porcelain veneers showed perfect biomimetic behavior, because cumulated restoration procedures had the same effect on natural and veneered incisors.


[Self care in adolescents].
The purpose of this work is to offer an analysis on the adolescence as stage of the life with some specific characteristics due to the transformations that happen so much at biological level as cognitive and psychosocial. During this period, the adolescent develops their autonomy and by so much starts their independence of the parents, some and other will experience situations that them will make be felt insecure in their/its performances until each one assume their new paper in the familiar environment and before itself. Parents and adolescent can need support and advice to obtain that this stage, in conflicting occasions, is developed normally, so that the adolescent become a capable adult about caring whether same and about others, with a life style that favor the integrated operation and the continued development. The family nurse can lend them the support and advice that need, since occupies a privileged place in the health equipment to guarantee the continuity of the assistance to the familiar group from the birth until the maturity. Basing us on the theory of the self-care developed by D. Orem, we will check the specific requirements of self-care of this stage of the vital cycle, of great transcendency for the step to the adult age.


Selective inhibition of peripheral cholinergic transmission in the cat produced by AF64A.
The neuropharmacologic effects of ethylcholine aziridinium ion, AF64A, were studied in cats, using various physiological techniques, to ascertain its synaptic site of action and to determine whether it may act as a cholinergic specific neurotoxin in vivo. Nictitating membrane contractions elicited by preganglionic nerve stimulation (1-16 Hz) were diminished in a dose-dependent manner after injection of AF64A into the carotid artery. Contractions due to injection of l-norepinephrine, tetramethylammonium or acetylcholine were not changed. Postganglionic action potentials from the superior cervical ganglion evoked by preganglionic stimulation were also abolished by AF64A, whereas the postganglionic firing produced by tetramethylammonium was unchanged. Neither the nictitating membrane nor ganglionic responses on the contralateral side of the animal were affected by AF64A treatment. In the same animals, twitch tension in the tongue produced by stimulation of the ipsilateral hypoglossal nerve (1-16 Hz) was gradually reduced and in most experiments completely blocked by AF64A. Repetitive stimulation of either the autonomic or somatic nerves at high frequencies (greater than 10 Hz) magnified and accelerated the onset of neurotoxic effects of AF64A. The suppression of ganglionic and neuromuscular transmission by AF64A was irreversible during the course of the experiments (12-18 hr). From these results, we can conclude that AF64A produces in the peripheral nervous system a longlasting inhibition of cholinergic transmission, without interfering with adrenergic transmission. Moreover, because AF64A did not block the postganglionic responses elicited by cholinergic nicotinic and muscarinic agonists, the inhibitory effects of AF64A must be mediated by a presynaptic action on cholinergic nerve terminals.


A pilot study into bosentan (Tracleer®) as an immunomodulating agent in patients with Behçet's disease.
Behçet's disease (BD) is an auto-inflammatory vasculitis characterized by aphthous oro-genital ulcers, inflammatory skin changes and uveitis. Treatment is mainly immunosuppressive. Interestingly, elevated endotheline-1 (ET-1) levels suggest a possible beneficial effect of treatment with an ET-1 receptor antagonist. The aim of our study was to investigate the possible beneficial effect of the ET-1 inhibitor bosentan. We performed a prospective double-blind placebo controlled pilot study into the effect and safety of bosentan in BD patients. Disease activity was measured using the Behçet Disease Current Activity Form. The primary objective of the study was to determine whether bosentan is therapeutically effective in patients with BD. Secondary endpoints were safety, tapering of medication and the effect of bosentan on possible disease activity markers such as ET-1, circulating endothelial cells (CECs), soluble interleukin-2 receptor (sIL2R) and cytokine levels. Ten patients were randomized to either bosentan or placebo. Overall, no effect on disease activity was observed, although one patient responded clinically and continued treatment after the study period. Despite one SAE, bosentan seems safe to use. No effect on tapering of medication, CECs, sIL2R and cytokine levels was found. In the bosentan group, ET-1 levels were elevated during the treatment period, with no correlation with disease activity. Although this is a small pilot study, bosentan appears to be safe in BD patients. One patient had a durable and significant clinical response. Our observations should be confirmed and extended in a larger patient cohort to be of significant impact in the treatment options for BD.


[Early pregnancy: revision of the Dutch terminology for clinical and ultrasound findings].
The nomenclature used to describe findings during early pregnancy in The Netherlands needs to be revised. Various terms, like 'abortion' and 'miscarriage', are used to describe the same phenomenon, which is confusing for both patients and doctors. In addition, the meaning of some terms, like 'missed abortion', has changed over time. In accordance with the revision of the European nomenclature in the English language by the Special Interest Group for Early Pregnancy of the European Society for Human Reproduction and Embryology (ESHRE), a revision of the nomenclature in the Dutch language is needed as well. An unambiguous Dutch terminology pertaining to early pregnancy is recommended that corresponds to the English terminology; this includes the Dutch terms 'embryo' [embryo], 'foetus' [foetus], 'biochemische zwangerschap' [biochemical pregnancy], 'zwangerschap met onbekende lokalisatie' [pregnancy of unknown location], 'miskraam' [miscarriage], 'lege vruchtzak' [empty sac], 'gestopte hart-activiteit' [fetal loss], 'herhaalde miskraam' [recurrent miscarriage], 'extra-uteriene zwangerschap' [ectopic pregnancy], and 'trofoblast-ziekte' [gestational trophoblastic disease], because these are based on well-defined clinical and ultrasonographic concepts.


Selenoprotein W depletion induces a p53- and p21-dependent delay in cell cycle progression in RWPE-1 prostate epithelial cells.
The anticancer activity of selenium (Se) has been demonstrated in myriad animal and in vitro studies, yet the mechanisms remain obscure. The main form of Se in animal tissues is selenocysteine in selenoproteins, but the relative importance of selenoproteins versus smaller Se compounds in cancer protection is unresolved. Selenoprotein W (SEPW1) is a highly conserved protein ubiquitously expressed in animals, bacteria, and archaea. SEPW1 depletion causes a delay in cell cycle progression at the G1/S transition of the cell cycle in breast and prostate epithelial cells. Tumor suppressor protein p53 is a master regulator of cell cycle progression and is the most frequently mutated gene in human cancers. p53 was increased in SEPW1 silenced cells and was inversely correlated with SEPW1 mRNA in cell lines with altered SEPW1 expression. Silencing SEPW1 decreased ubiquitination of p53 and increased p53 half-life. SEPW1 silencing increased p21(Cip1/WAF1/CDKN1A), while p27 (Kip1/CDKN1B) levels were unaffected. G1-phase arrest from SEPW1 knockdown was abolished by silencing p53 or p21. Cell cycle arrest from SEPW1 silencing was not associated with activation of ATM or phosphorylation of Ser-15 in p53, suggesting the DNA damage response pathway was not involved. Silencing GPX1 had no effect on cell cycle, suggesting that G1-phase arrest from SEPW1 silencing was not due to loss of antioxidant protection. More research is required to identify the function of SEPW1 and how it affects stability of p53.


Cell cycle specific toxicity of the Hoechst 33342 stain in untreated or irradiated murine tumor cells.
Hoechst 33342 stain has been used previously in conjunction with fluorescence activated cell sorting to separate cell populations into the various phases of the cell cycle. The present experiments were designed to evaluate the toxicity of this stain in cells derived from solid KHT sarcomas which were untreated or irradiated prior to dissociation. In both untreated and irradiated cells, stain toxicity increased with increasing exposure times but was always greater in treated cells; for example, after stain exposures of 1 h at 10 microM or 2 h at 5 microM, survival was reduced 25- to 45-fold in irradiated cells and 4- to 5-fold in untreated cells. To determine whether this toxicity occurred equally in all phases of the cell cycle or preferentially in a particular phase, cells derived from untreated or irradiated KHT tumors were separated into the various phases of the cell cycle using centrifugal elutriation. Cells in each phase then were divided in two, one-half being stained with 5 microM Hoechst 33342 for 2 h, the other half being handled in an identical manner but not stained. These particular stain exposure conditions were chosen for detailed evaluation because they provided the best DNA histograms. In both untreated and irradiated cells the cytotoxic effects of Hoechst 33342 were found to be significantly greater in cells in the S phase than in cells in G1 and G2-M phases of the cell cycle. This toxicity, particularly its cell cycle specificity, suggests a potentially severe limitation for the use of Hoechst 33342 in combination with fluorescence activated cell sorting in studies aimed at determining treatment efficacies in various phases of the cell cycle.


Osteomalacia revealing celiac disease and primary biliary cirrhosis-related Fanconi syndrome in a patient with systemic sclerosis.
Systemic sclerosis (SSc) may affect the gastrointestinal tract and cause very rarely malabsorption syndrome related to bacterial overgrowth. Malabsorption syndrome may be responsible for weight loss, diarrhea, osteomalacia, and iron and vitamins deficiency. We report the case of a SSc patient who developed osteomalacia caused by the combination of two exceptional conditions in the setting of SSc: celiac disease (CD) and primary biliary cirrhosis (PBC)-related Fanconi syndrome. Oral prednisone with angiotensin-converting enzyme inhibitors, was initiated because of active lesions of tubulitis, and led to the complete regression of bone pains, and by the improvement of renal function and regression of the features of proximal tubulopathy. Thus, in the presence of vitamin deficiencies in a patient with SSc, together with a search for malabsorption syndrome secondary to bacterial overgrowth, CD and/or PBC-associated Fanconi syndrome should be investigated.


Gemcitabine and vinorelbine in recurrent head and neck cancer: pharmacokinetic and clinical results.
To evaluate the pharmacokinetic parameters, efficacy and toxicity of a combination of gemcitabine (GEM) and vinorelbine (VNB) in recurrent heavily pre-treated squamous cell head and neck carcinoma. Twenty-four patients previously treated with concomitant chemo-radiotherapy (n = 13), surgery plus radiotherapy (n = 10) and surgery + concomitant chemo-radiotherapy (n = 1) were enrolled; 7 patients had received one or more courses of palliative chemotherapy. Twenty patients had a local-regional recurrence and 4 patients had metastases. The doses were 1200 mg/m2 for GEM and 30 mg/m2 for VNB on days 1 and 8 every 21 days; a maximum of 6 cycles was allowed. Pharmacokinetic investigations were performed on 9 patients receiving GEM and VNB. As a PK control, a second group of 5 patients was given GEM as single agent, at the same doses and with the same i.v. infusion length. Twenty-four patients received a total of 135 cycles (median per patient, 5). Neutropenia was the most frequent side-effect (92% of patients; grade 3-4 in 50%). The overall response rate was 25% which included 1 of 24 complete responses (4%) and 4 of 24 partial responses (21%). Responses were observed only in patients with good prognostic characteristics. The median response duration was 5.5 months (2-16 months) and the overall median survival was 9 months (range, 2-25+). Vinorelbine serum levels showed no evidence of any pharmacokinetic interaction with GEM; most of all, no rebound in VNB disposition can be produced by GEM pre-administration. The GEM-VNB combination has a palliative role in patients with favourable characteristics; the results seem no better than those observed with VNB alone. Moreover, this drug association does not alter the pharmacokinetic profile of both drugs, also compared to GEM monotherapy.


Effects of Whole-Body Cryostimulation (-90°C) on Somnolence and Psychological Well-Being in an Older Patient with Restless Legs Syndrome.
We aimed at evaluating the feasibility and effects of intense (i.e. -90°C) whole-body cryostimulation (WBC) on somnolence and psychological well-being in an older-adult patient diagnosed with restless legs syndrome (RLS). An interrupted time series approach was used in which the efficacy of cryostimulation was evaluated by measuring self-reported fatigue, wake time sleepiness, and well-being several times prior to, during, and after exposure to treatment (i.e. daily 3-min sessions of intense WBC). No adverse event occurred. Reported levels of sleepiness decreased immediately following the beginning of the treatment phase. In the same time, self-reported well-being significantly increased. Effects sizes were of large magnitude. In summary, the present study demonstrated that daily exposure to extremely cold air in an enclosed space for 2 weeks was feasible and effective in promoting physical and psychological states in an older patient with sleep disturbances.


Erectile function in SRY positive 46,XX males with normal phenotype.
The 46,XX male syndrome is a rare disorder of sex development and has two different forms, depending on the sex-determining region's presence on the Y chromosome (SRY) gene. The SRY positive 46,XX males are usually diagnosed during infertility workup. We evaluated the erectile function of 46,XX SRY positive males and compared it with healthy males. Ten patients with azoospermia and 46,XX SRY positive disorder who referred to a urology clinic with infertility were analyzed retrospectively. Controls were chosen from healthy males at similar ages. The physical examination was performed, and serum hormones were obtained at admission. The clinical assessment of erectile dysfunction was evaluated by the International Index of Erectile Function (IIEF) questionnaire. There was no statistically significant difference between the two groups in terms of age, serum prolactin, luteinizing hormone (LH) levels and IIEF scores (P >0.05). In 46,XX males, serum follicle-stimulating hormone (FSH) levels were significantly higher, and total testosterone levels and testicular volumes were found to be significantly lower when compared to controls (p <0.001, p <0.05, p <0.01, respectively). This study indicates that these males' erectile function is similar to those of 46,XY males.


Evaluation of Radiation-induced Pleural Effusions after Radiotherapy to Support Development of Animal Models of Radiation Pneumonitis.
Not all animal models develop radiation-induced pleural effusions (RIPEs) as a form of radiation-induced lung injury (RILI). Such effusions are also not well characterized in humans. The purpose of this study is to identify occurrences of RIPE in humans, provide justification for development of relevant animal models, and further characterize its risk factors in cancer patients. We also aim to identify dose thresholds for cardiopulmonary toxicity in humans to shed light on possible pathogenic mechanisms for RIPEs. We carried out a retrospective review of medical records of 96 cancer patients receiving thoracic irradiation (TRT) at our institution. Fifty-three (53%) patients developed a new pleural effusion post TRT; 18 (19%) had RIPE; and 67% developed RIPE ipsilateral to the site irradiated. None developed "contralateral only" effusions. Median time to development was 6 mo (IQR; 4-8 mo). Of 18, 8 patients (44%) had concomitant asymptomatic (radiographic only) or symptomatic radiation pneumonitis and pericardial effusion. Dosimetric factors, including combined and ipsilateral mean lung dose (MLD), were significantly associated with increased risk of RIPE. Angiotensin converting enzyme inhibition, steroids, or concurrent chemotherapy did not modify incidence of RIPE. Our results substantiate the occurrence and incidence of RIPEs in humans. In cancer patients, a median time to development of effusions around 6 mo also supports the onset of RIPEs concurrent with radiation pneumonitis. Future work needs to include large populations of cancer survivors in whom delayed RIPEs can be tracked and correlated with cardiovascular changes in the context of injury to multiple organs.


[Techniques for neurovascular bundle preservation during retropubic radical prostatectomy].
Although radical prostatectomy is a curative therapy that has proven effective in many of our patients with prostate cancer, it is still associated with significant morbidity, which includes postoperative erectile dysfunction among its leading exponents. Potency after the intervention may be influenced by many factors, among which, presence of adequate erectile function before surgery, patient's age, stage of disease at the time of treatment, surgeon's experience and, of course, interpersonal anatomical variations may be pointed out. In recent decades, the exact knowledge of the neuroanatomy of the male pelvis has become very important, for both the student of human anatomy and the pelvic surgeon. Therefore, the anatomical nerve sparing techniques have led to fewer complications related to the injury of these structures. This article presents a brief description of the neuroanatomical substrate of the neurovascular bundles along with a detailed compilation of the different surgical techniques for their preservation during radical retropubic prostatectomy.


A review of Iliad and Quick Medical Reference for primary care providers. Two diagnostic computer programs.
Two diagnostic computer programs, Iliad (Applied Informatics Inc, Salt Lake City, Utah) and Quick Medical Reference (Camdat Corp, Pittsburgh, Pa), are commercially available. Both describe a large subset of internal medicine diseases. These programs, also called knowledge bases, have different structures and diagnostic methods and emphasize somewhat different experimental observations regarding problem solving and expert diagnostic behavior. Quick Medical Reference has more diverse reference features and emphasizes diagnostic competitors (diseases that explain similar features in a case). Iliad has better developed teaching features and emphasizes a probabilistic approach. The histories and methods of these programs place various constraints on their diagnostic capabilities, especially from a primary care perspective. Although neither program is currently likely to fill the role of an expert diagnostic consultant for a family practitioner, both have potential as unique reference and teaching tools in primary care.


Deep learning-based concrete defects classification and detection using semantic segmentation.
Visual damage detection of infrastructure using deep learning (DL)-based computational approaches can facilitate a potential solution to reduce subjectivity yet increase the accuracy of the damage diagnoses and accessibility in a structural health monitoring (SHM) system. However, despite remarkable advances with DL-based SHM, the most significant challenges to achieving the real-time implication are the limited available defects image databases and the selection of DL networks depth. To address these challenges, this research has created a diverse dataset with concrete crack (4087) and spalling (1100) images and used it for damage condition assessment by applying convolutional neural network (CNN) algorithms. CNN-classifier models are used to identify different types of defects and semantic segmentation for labeling the defect patterns within an image. Three CNN-based models-Visual Geometry Group (VGG)19, ResNet50, and InceptionV3 are incorporated as CNN-classifiers. For semantic segmentation, two encoder-decoder models, U-Net and pyramid scene parsing network architecture are developed based on four backbone models, including VGG19, ResNet50, InceptionV3, and EfficientNetB3. The CNN-classifier models are analyzed on two optimizers-stochastic gradient descent (SGD), root mean square propagation (RMSprop), and learning rates-0.1, 0.001, and 0.0001. However, the CNN-segmentation models are analyzed for SGD and adaptive moment estimation, trained with three different learning rates-0.1, 0.01, and 0.0001, and evaluated based on accuracy, intersection over union, precision, recall, and F1-score. InceptionV3 achieves the best performance for defects classification with an accuracy of 91.98% using the RMSprop optimizer. For crack segmentation, EfficientNetB3-based U-Net, and for spalling segmentation, IncenptionV3-based U-Net model outperformed all other algorithms, with an F1-score of 95.66 and 89.43%, respectively.


The Influence of Dimethyl Sulfoxide as Electrolyte Additive on Anodic Dissolution of Alkaline Zinc-Air Flow Battery.
The present work describes the effects of dimethyl sulfoxide (DMSO) in KOH aqueous electrolyte on the performance of a zinc-air flow battery. Aqueous electrolytes containing 7 M KOH and (0 to 20)% v/v DMSO were studied revealing a critical role of DMSO on the dissolution and deposition of zinc. The anodic zinc dissolution process was studied via cyclic voltammetry, Tafel polarization and electrochemical impedance spectroscopy (EIS). The presence of DMSO showed improved zinc dissolution performance with the highest peak of zinc dissolution being the electrolyte containing 5% v/v DMSO. Tafel analysis demonstrated a significant decrease in polarization resistance and an increase in corrosion rate due to the introduction of DMSO to the electrolyte. This suggests that DMSO has the ability to suspend zinc oxide in the electrolyte, thus preventing passivation of the zinc surface. EIS results revealed that by adding DMSO to the electrolyte, charge transfer resistance increased. This is attributed to the formation of passive layers having arisen from DMSO adsorption, the formation of zincate ions in the vicinity of the zinc surface, and the deposition of discharged products. A difference in Nyquist plots was observed for 20% v/v DMSO/KOH and 0% v/v DMSO/KOH electrolytes implying non-Debye relaxation behavior taking place due to the surface effects. The electrolytes were implemented in a zinc-air flow battery. Maximum power densities of 130 mW/cm2 (5% v/v DMSO) and 125 mW/cm2 (20% v/v DMSO) were obtained and were observed to be about 43% and 28% higher than that of the DMSO-free electrolyte. Results indicated that when 20% v/v DMSO was added to KOH solution, there was 67% zinc utilization efficiency (550 mAh/g) which provided 20% improvement in discharge capacity. Further, the battery with 20% v/v DMSO demonstrated excellent cyclability. Overall, DMSO shows great promise for enhancement of zinc dissolution/deposition in zinc-air batteries.


Continuous 500,000-year climate record from vein calcite in devils hole, nevada.
Oxygen-18 (delta(18)O) variations in a 36-centimeter-long core (DH-11) of vein calcite from Devils Hole, Nevada, yield an uninterrupted 500,000-year paleotemperature record that closely mimics all major features in the Vostok (Antarctica) paleotemperature and marine delta(18)O ice-volume records. The chronology for this continental record is based on 21 replicated mass-spectrometric uranium-series dates. Between the middle and latest Pleistocene, the duration of the last four glacial cycles recorded in the calcite increased from 80,000 to 130,000 years; this variation suggests that major climate changes were aperiodic. The timing of specific climatic events indicates that orbitally controlled variations in solar insolation were not a major factor in triggering deglaciations. Interglacial climates lasted about 20,000 years. Collectively, these observations are inconsistent with the Milankovitch hypothesis for the origin of the Pleistocene glacial cycles but they are consistent with the thesis that these cycles originated from internal nonlinear feedbacks within the atmosphere-ice sheet-ocean system.


Facility assessment of the best functioning facility providing emergency obstetric care in the selected blocks of Varanasi, Uttar Pradesh.
UP has the 2nd highest MMR which is 197 compared with national average of 113 (RGI-SRS-2016-2018).Although institutional deliveries in India has been increased from 78.9% (NFHS-4) to 89% (NFHS-5) [ UP from 67.8% to 83.4%] but still we are far away from SDG -3 target. It reflects that there may be increase in crude coverage but not in effective coverage. It is a cross sectional study conducted in May - June 2017. Out of 8 blocks of rural Varanasi, 4 blocks were selected randomly. Best functioning facility for EmOC services in each selected block were assessed using Facility Gap Assessment Schedule of IPHS. None of the facility met the recommended standard for BEmOC .Tracking of drop out of ANC and PNC services, use of Partograph, treatment of abortion-related complications, were not found at all the 4 facility. Blood grouping and RH typing was also not functional at 2 of the 4 centers. Caesarean section and availability of blood bank were also lacking in CHC (FRU). If condition of best functioning facility in a block is not according to the recommendation then how can we expect to provide a good maternal health service to public.


Diagnosis of late-onset congenital adrenal hyperplasia in clinical practice: current evaluation.
Aim of the study was to investigate the need to perform the adrenocorticotropic hormone (ACTH) stimulation test by recognizing the importance of a second look at basal serum 17-alpha hydroxyprogesterone (17-OHP) levels and calculating new serum 17-OHP cut-off level. A total of 142 patients who had hyperandrogenism symptoms and had basal serum 17-OHP levels of higher than 1.3 ng/mL were scheduled to have an ACTH stimulation test performed. Prior to ACTH stimulation, 17-OHP levels were recorded and as second-look levels. Patients were divided to two groups, late-onset congenital adrenal hyperplasia (LOCAH) (25/142), non-LOCAH (117/142). There were statistically significant results related to cycle length and menstrual irregularity between two groups (P=0.042, P=0.041, respectively). In the LOCAH group, basal serum 17 OHP levels were higher than non-LOCAH (P=0.001). When basal serum 17-OHP levels were measured a second time, the need for performing the ACTH stimulation test was decreased. According to cut-off levels of 1.3 ng/mL, 100% of patients needing to take the second serum 17-OHP decreased to 83.1%, a cut-off level of 2 ng/mL decreased numbers from 74.65% to 35.92% and for 2.25 ng/mL 58.42% of patients was decreased to 26.77%. In this study we established 2.25 ng/ml is a superior cut-off level for 17-OHP, its sensitivity is 84% and specifity is 50.4%. The incidence of LOCAH is 1.35% among the patients with hyperandrogenism symptoms. We found a single measurement of serum 17-OHP level can be unreliable. Second 17-OHP test reduces the need of performing the ACTH stimulation test by approximately 30%.


Purification, characterization, and gene cloning of purine nucleosidase from Ochrobactrum anthropi.
A bacterium, Ochrobactrum anthropi, produced a large amount of a nucleosidase when cultivated with purine nucleosides. The nucleosidase was purified to homogeneity. The enzyme has a molecular weight of about 170,000 and consists of four identical subunits. It specifically catalyzes the irreversible N-riboside hydrolysis of purine nucleosides, the K(m) values being 11.8 to 56.3 microM. The optimal activity temperature and pH were 50 degrees C and pH 4.5 to 6.5, respectively. Pyrimidine nucleosides, purine and pyrimidine nucleotides, NAD, NADP, and nicotinamide mononucleotide are not hydrolyzed by the enzyme. The purine nucleoside hydrolyzing activity of the enzyme was inhibited (mixed inhibition) by pyrimidine nucleosides, with K(i) and K(i)' values of 0.455 to 11.2 microM. Metal ion chelators inhibited activity, and the addition of Zn(2+) or Co(2+) restored activity. A 1.5-kb DNA fragment, which contains the open reading frame encoding the nucleosidase, was cloned, sequenced, and expressed in Escherichia coli. The deduced 363-amino-acid sequence including a 22-residue leader peptide is in agreement with the enzyme molecular mass and the amino acid sequences of NH(2)-terminal and internal peptides, and the enzyme is homologous to known nucleosidases from protozoan parasites. The amino acid residues forming the catalytic site and involved in binding with metal ions are well conserved in these nucleosidases.


The umbilical cord, preeclampsia and the VEGF family.
The VEGF family has been identified as abnormal in preeclampsia (PE). Hypertensive disorders of pregnancy (HDP) are major contributors to maternal and neonatal morbidity and mortality worldwide; likewise, umbilical cord anatomical abnormalities (UCAA) are linked to poor neonatal outcomes. Based on the relationship described between PE and UCAA and the role of the VEGF family in PE, this study explored VEGF expression in placental and UC tissued from patients with PE and with UCAA. We performed an observational, analytical study on placentas, comparing protein and mRNA expression in four groups: patients with PE, patients with UC abnormalities, patients with both, and patients with none of them. Using immunohistochemistry, we studied VEGF A, VEGF R1 (FLT1), MMP1, and PLGF. With quantitative reverse transcription polymerase chain reaction we described mRNA expression of PLGF, VEGF and sFLT1, and sFLT1/PLGF ratio. Forty newborns were included. Sixty-seven percent of mothers and 45% of newborns developed no complications. Immunohistochemistry was performed on UC and placental disc paraffin-embedded tissue; in the latter, the mRNA of the VEGF family was also measured. Statistically significant differences were observed among different expressions in both HDP and UCAA groups. Interestingly, the UCAA group exhibited lower levels of sFLT1 and VEGF-A in comparison with other groups, with significant P-value for sFLT1 (P=0000.1). The origin of UCAA abnormalities and their relation with HDP are still unknown. VEGF family alterations could be involved in both. This study provides the first approach related to molecules linked to UCAA.


Synthetic Circulating Cell-free DNA as Quality Control Materials for Somatic Mutation Detection in Liquid Biopsy for Cancer.
Detection of somatic genomic alterations in tumor-derived cell-free DNA (cfDNA) in the plasma is challenging owing to the low concentrations of cfDNA, variable detection methods, and complex workflows. Moreover, no proper quality control materials are available currently. We developed a set of synthetic cfDNA quality control materials (SCQCMs) containing spike-in cfDNA on the basis of micrococcal nuclease digestion carrying somatic mutations as simulated cfDNA and matched genomic DNA as genetic background to emulate paired tumor-normal samples in real clinical tests. Site-directed mutagenesis DNA that contained 1500-2000 bases with single-nucleotide variants or indels and genomic DNA from CRISPR/Cas9 edited cells with EML4-ALK rearrangements was fragmented, quantified, and added into micrococcal nuclease-digested DNA derived from HEK293T cells. To prove their suitability, the SCQCMs were compared with patient-derived plasma samples and validated in a collaborative study that encompassed 11 laboratories. The results of SCQCM analysis by next-generation sequencing showed strong agreement with those of patient-derived plasma samples, including the size profile of cfDNA and the quality control metrics of the sequencing data. More than 95% of laboratories correctly detected the SCQCMs with EGFR T790M, L858R, KRAS G12D, and a deletion in exon 19, as well as with EML4-ALK variant 2. The SCQCMs were successfully applied in a broad range of settings, methodologies, and informatics techniques. We conclude that SCQCMs can be used as optimal quality controls in test performance assessments for circulating tumor DNA somatic mutation detection.


Linking of global intensive care (LOGIC): An international benchmarking in critical care initiative.
Benchmarking is a common and effective method for measuring and analyzing ICU performance. With the existence of national registries, objective information can now be obtained to allow benchmarking of ICU care within and between countries. The present manuscript briefly describes the current status of benchmarking in healthcare and critical care and presents the LOGIC project, an initiative to promote international benchmarking for intensive care units. Currently 13 registries have joined LOGIC. We showed large differences in the utilization of ICU as well as resources and in outcomes. Despite the need for careful interpretation of differences due to variation in definitions and limited risk adjustment, LOGIC is a growing worldwide initiative that allows access to insightful epidemiologic data from ICUs in multiple databases and registries.


[Primary myelofibrosis--pathogenesis, diagnosis and treatment].
Primary myelofibrosis is a malignant myeloproliferative disease. It is characterised by proliferation of megakaryocytes in the bone marrow, dysregulated cytokine production and reactive fibrosis that causes bone marrow failure. The purpose of this article is to provide an up-to-date presentation of the pathophysiology, diagnostics and treatment of the disease. The article is based on the authors' own experience and on a selection of articles identified through many years of experience of treating patients with myelofibrosis. The molecular mechanisms that trigger the disease remain unidentified, but mutations in two genes (JAK2 and MPL) occur in 70% of patients and result in increased production of haematopoietic cells. Diagnosis is based on clinical examination, bone marrow histology and molecular biological examinations. The clinical course of primary myelofibrosis varies. Life expectancy depends on a number of factors and is severely decreased by high-risk disease. Allogeneic stem cell transplantation is the only treatment with a curative potential, but only a minority of patients are eligible for it. If transplantation is not possible, therapy is symptomatic. JAK2-inhibitors are new drugs that counteract cytokine production and cell proliferation. Ruxolitinib is the first approved JAK2 inhibitor and has proved effective on symptoms and quality of life. Medical inhibition of the JAK2 gene and associated JAK-STAT signalling pathway is a step forward in treatment. However, stem cell transplantation remains the only potentially curative treatment for myelofibrosis.


An evaluation of SOX2 and hTERC gene amplifications as screening markers in oral and oropharyngeal squamous cell carcinomas.
Oral and oropharyngeal squamous cell carcinomas (OSCC) are among the most common cancers. The poor survival rate among oral cancer patients can be attributed to several factors, one of them being lack of early detection. A key approach to this problem would be to detect potentially malignant lesion at their early stage. Using the FISH technique, oral brush cytology slides can be an easy and rapid screening approach for malignant cell detection. The present study was designed to detect hTERC and SOX2 amplifications in OSSC exfoliative tumor cells and evaluate whether those two gene amplifications might serve as a supportive biomarker in early detection and diagnosis of oral and oropharyngeal SCC. Brush biopsies were collected from exophytic and exulcerated oral and oropharyngeal lesions of the oral cavity of 71 patients and 22 healthy controls. FISH techniques using a TERC-specific DNA probe and a SOX2 DNA specific probe both combined with a centromere 3-specific control probe was performed on the cytology slides. A 100 squamous epithelial cell nuclei of the smears per slide were analysed. As abnormal FISH pattern were considered amplified and polyploid patterns.From 71 brush biopsies of oropharynx and other locations in oral cavity analysed by FISH 49 were considered to be abnormal (69%). The over representation of polyploidy and/or TERC/SOX2 amplification in tumour samples was statistically significant when compared to controls (p = 0.01). SOX2 and TERC gene amplifications are common in all squamous cell carcinomas and their detection in early stages could be crucial for early detection and more accurate prognosis. Our study strongly suggests that early detection by FISH on cytobrushed samples could be a possible non-invasive screening method even before a tissue biopsy is performed.


Genital human papillomavirus infection.
Over the past few decades, epidemiology and natural history studies have led to improved understanding of human papillomavirus (HPV) infection and to promising prevention strategies. HPV infection is the cause of anogenital warts and cervical cancer, as well as a proportion of other anogenital and head and neck cancers. Data from clinical trials have resulted in recommendations that support the use of an HPV test in the context of cervical cancer screening and management. Prophylactic HPV vaccine trials have demonstrated high efficacy, and an HPV vaccine that prevents cervical cancer precursors, cervical cancer, and anogenital warts caused by HPV types 6, 11, 16, and 18 was licensed for use in girls and women aged 9-26 years by the US Food and Drug Administration (FDA) in June 2006. In this article, we review genital HPV for the clinician, with a primary focus on the prevalence of HPV infection in the United States.


Testing concordance of instrumental variable effects in generalized linear models with application to Mendelian randomization.
Instrumental variable regression is one way to overcome unmeasured confounding and estimate causal effect in observational studies. Built on structural mean models, there has been considerable work recently developed for consistent estimation of causal relative risk and causal odds ratio. Such models can sometimes suffer from identification issues for weak instruments. This hampered the applicability of Mendelian randomization analysis in genetic epidemiology. When there are multiple genetic variants available as instrumental variables, and causal effect is defined in a generalized linear model in the presence of unmeasured confounders, we propose to test concordance between instrumental variable effects on the intermediate exposure and instrumental variable effects on the disease outcome, as a means to test the causal effect. We show that a class of generalized least squares estimators provide valid and consistent tests of causality. For causal effect of a continuous exposure on a dichotomous outcome in logistic models, the proposed estimators are shown to be asymptotically conservative. When the disease outcome is rare, such estimators are consistent because of the log-linear approximation of the logistic function. Optimality of such estimators relative to the well-known two-stage least squares estimator and the double-logistic structural mean model is further discussed.


Patient characteristics upon initial presentation to chiropractic teaching clinics: A descriptive study conducted at one university.
Objective : The purpose of this study was to compare demographics and chief complaints of the new patient population at our institution's fee-for-service clinics to the patient population of practicing chiropractors in the United States. We also compared the prevalence of obesity and hypertension to reference standards for the adult population. Methods : Patient data were obtained from the electronic health records. All records identified as new patients during October 2013 were included. Variables of interest were clinic site, patient demographics, blood pressure, body mass index (BMI), chief complaint, and ICD-9 codes. Descriptive statistics were computed and compared to reference standards from previous reports. Results : During October 2013, there were 224 new patients that entered the clinics. The average patient was a 31- to 50-year-old white male. Our clinic patients differed from those seen by US chiropractors in the distribution of all demographic variables. For adult patients, 31.4% were overweight, 29% were obese, and 8% stage 1 or 2 hypertension. Conclusion : New patients in the fee-for-service teaching clinics appear to be dissimilar to those of US practicing chiropractors in several important demographics, characteristics, and types of complaints. The new patients had lower levels of overweight, obesity, and hypertension compared to US reference standards.


Growth-defense tradeoffs in plants: a balancing act to optimize fitness.
Growth-defense tradeoffs are thought to occur in plants due to resource restrictions, which demand prioritization towards either growth or defense, depending on external and internal factors. These tradeoffs have profound implications in agriculture and natural ecosystems, as both processes are vital for plant survival, reproduction, and, ultimately, plant fitness. While many of the molecular mechanisms underlying growth and defense tradeoffs remain to be elucidated, hormone crosstalk has emerged as a major player in regulating tradeoffs needed to achieve a balance. In this review, we cover recent advances in understanding growth-defense tradeoffs in plants as well as what is known regarding the underlying molecular mechanisms. Specifically, we address evidence supporting the growth-defense tradeoff concept, as well as known interactions between defense signaling and growth signaling. Understanding the molecular basis of these tradeoffs in plants should provide a foundation for the development of breeding strategies that optimize the growth-defense balance to maximize crop yield to meet rising global food and biofuel demands.


Rechargeable Mg-Ion Full Battery System with High Capacity and High Rate.
Thanks to the low cost, free dendritic hazards, and high volumetric capacity, magnesium (Mg)-ion batteries have attracted increasing attention as alternative energy storage devices to lithium-ion batteries. Despite the successful development of electrode materials, the real-life application potential of Mg-ion full battery systems (MIFBSs) is largely hindered by the lack of suitable electrode couples and hence low diffusion kinetics, which induce low specific capacity, poor rate performance, and low working voltage. Herein, we report an aqueous rechargeable MIFBS by employing copper hexacyanoferrate (CuHCF) as the cathode and 3,4,9,10-perylene-tetracarboxylic acid diimide (PTCDI) as the anode in 1 moL L-1 MgCl2 electrolyte. The combination of PTCDI//CuHCF allows efficient redox and convenient intercalation/deintercalation of Mg2+ at the electrodes while facilitating a fast transfer of Mg2+ between the two electrodes. As a result, the system delivers a high capacity of ∼120.3 mAh g-1 at a current density of 0.5 A g-1 after 200 operation cycles with a broadened voltage range (0-1.95 V) and maintains a capacity of ∼97.8 mAh g-1 at 2.0 A g-1 after 1000 cycles. Even at a high current density of 5.0 A g-1, the battery provides a steady capacity of ∼81.4 mAh g-1 over 5000 cycles. Moreover, after being applied at 11.0 A g-1, the system can deliver a capacity of ∼126.5 mAh g-1 at 0.5 A g-1. This work emphasizes the great promise of developing suitable electrode couples for aqueous MIFBSs to achieve high capacity and high rate.


Nasal carriers are more likely to acquire exogenous Staphylococcus aureus strains than non-carriers.
We performed a prospective observational study in a clinical setting to test the hypothesis that prior colonization by a Staphylococcus aureus strain would protect, by colonization interference or other processes, against de novo colonization and, hence, possible endo-infections by newly acquired S. aureus strains. Three hundred and six patients hospitalized for >7 days were enrolled. For every patient, four nasal swabs (days 1, 3, 5, and 7) were taken, and patients were identified as carriers when a positive nasal culture for S. aureus was obtained on day 1 of hospitalization. For all patients who acquired methicillin-resistant S. aureus (MRSA) or methicillin-susceptible S. aureus via colonization and/or infection during hospitalization, strains were collected. We note that our study may suffer from false-negative cultures, local problems with infection control and hospital hygiene, or staphylococcal carriage at alternative anatomical sites. Among all patients, 22% were prior carriers of S. aureus, including 1.9% whom carried MRSA upon admission. The overall nasal staphylococcal carriage rate among dermatology patients was significantly higher than that among neurosurgery patients (n = 25 (55.5%) vs. n = 42 (16.1%), p 0.005). This conclusion held when the carriage definition included individuals who were nasal culture positive on day 1 and day 3 of hospitalization (p 0.0001). All MRSA carriers were dermatology patients. There was significantly less S. aureus acquisition among non-carriers than among carriers during hospitalization (p 0.005). The mean number of days spent in the hospital before experiencing MRSA acquisition in nasal carriers was 5.1, which was significantly lower than the score among non-carriers (22 days, p 0.012). In conclusion, we found that nasal carriage of S. aureus predisposes to rather than protects against staphylococcal acquisition in the nose, thereby refuting our null hypothesis.


Glucose-lowering drugs in patients with chronic kidney disease: a narrative review on pharmacokinetic properties.
The achievement of a good glycaemic control is one of the cornerstones for preventing and delaying progression of microvascular and macrovascular complications in patients with both diabetes and chronic kidney disease (CKD). As for other drugs, the presence of an impaired renal function may significantly affect pharmacokinetics of the majority of glucose-lowering agents, thus exposing diabetic CKD patients to a higher risk of side effects, mainly hypoglycaemic episodes. As a consequence, a reduction in dosing and/or frequency of administration is necessary to keep a satisfactory efficacy/safety profile. In this review, we aim to summarize the pharmacology of the most widely used glucose-lowering agents, discuss whether and how it is altered by a reduced renal function, and the recommendations that can be made for their use in patients with different degrees of CKD.


Essential role of STAT3 for embryonic stem cell pluripotency.
Propagation of mouse embryonic stem (ES) cells in vitro requires exogenous leukemia inhibitory factor (LIF) or related cytokines. Potential downstream effectors of the LIF signal in ES cells include kinases of the Src, Jak, and mitogen-activated protein families and the signal transducer and transcriptional activator STAT3. Activation of nuclear STAT3 and the ability of ES cells to grow as undifferentiated clones were monitored during LIF withdrawal. A correlation was found between levels of STAT3 activity and maintenance of an undifferentiated phenotype at clonal density. In contrast, variation in STAT3 activity did not affect cell proliferation. The requirement for STAT3 was analyzed by targeted mutagenesis in ES cell lines exhibiting different degrees of LIF dependency. An insertional mutation was devised that abrogated Stat3 gene expression but could be reversed by Cre recombination-mediated excision. ES cells heterozygous for the Stat3 mutation could be isolated only from E14 cells, the line least dependent on LIF for self-renewal. Targeted clones isolated from other ES cell lines were invariably trisomic for chromosome 11, which carries the Stat3 locus, and retained normal levels of activated STAT3. Cre-regulated reduction of Stat3 gene copy number in targeted, euploid E14 clones resulted in dose-dependent losses of STAT3 activity and the efficiency of self-renewal without commensurate changes in cell cycle progression. These results demonstrate an essential role for a critical amount of STAT3 in the maintenance of an undifferentiated ES cell phenotype.


A contemporary assessment of nocturia: definition, epidemiology, pathophysiology, and management--a systematic review and meta-analysis.
Nocturia is a common urologic symptom that has been covered in a variety of reported studies in the literature but is not specifically covered in current guidelines. To comprehensively review the literature pertaining to the definition, etiologies, and consequences of nocturia and assess the evidence supporting the use of conservative medical and interventional therapy. A literature search was conducted using the keyword nocturia, restricted to articles in the English language, after 2000 and before April 2012, in PubMed/Medline, Embase, Scopus, Web of Science, and Cochrane Library databases. Regarding treatment modalities, studies were included only if nocturia was a primary end point and if the studies were designed as randomized controlled trials without limit of date. When suitable, a meta-analysis was conducted. Papers covering treatment options for nocturia specifically related to nonurologic conditions were excluded. Nocturia is still defined as the symptom of wakening from sleep once or more often to void. The prevalence is high in both genders and increases with age. Frequency-volume charts, which are the pivotal tool of clinical assessment, detect 24-h polyuria, nocturnal polyuria (NP), or reduced nocturnal bladder capacity and help to target specific nonurologic etiologies. Nocturia is a morbid condition that significantly affects quality of life and increases mortality. Besides behavioral measures, validated treatment options include oral desmopressin, which is superior to placebo in treating NP. While the level of evidence for desmopressin is high, limited data support the use of α1-blockers and antimuscarinics; however, only rarely has nocturia been a primary end point when studying these drug classes, and studies have not consistently controlled for the effect of NP. Our knowledge of nocturia, its etiology, and its management has substantially improved in recent years. The evidence available on the management of nocturia remains limited; contributory factors include (1) the complexity of associated conditions, (2) the underuse of objective evaluation tools, and (3) the lack of specific focus on nocturia in clinical trials.


Severe hyponatraemia peripartum associated with omeprazole therapy.
Hyponatraemia is the most commonly encountered electrolyte abnormality in pregnancy and may be associated with adverse maternal and neonatal outcomes. Rapid onset, severe hyponatraemia has been reported with proton-pump inhibitor therapy in non-pregnant individuals. Gastro-oesophageal reflux is very common during pregnancy, and proton-pump inhibitors are available without a prescription in many countries. A case of severe maternal hyponatraemia in the setting of recent omeprazole therapy is presented. Health professionals should be aware of this complication given the availability of proton-pump inhibitors without prescription and high rates of gestational gastro-oesophageal reflux.


Cancer stem cells CD133 and CD24 in colorectal cancers in Northern Iran.
We aimed to study the expression of CD24 and CD133 in colorectal cancer and normal adjacent tissues to assess a relationship between these markers and clinic-pathological characteristics and patient's survival. Cancer stem cells are a group of tumor cells that have regeneration and multi-order differentiation capabilities. Expression of CD24 and CD133 was studied in a paraffin block of colorectal cancer and normal tissues near tumors with the immuneohistochemical method in patients who were referred to Imam Khomeini Hospital in Sari. A total of 50 samples (25 males and 25 females) with a mean age of 67.57±13.9 years old with range 28-93 years, included 3 mucinous carcinoma and 47 adenocarcinoma. Expression of CD133 marker was negative in 29 cases and positive in 21 cases. Expression of CD24 in tissue near tumor cells was found in 30% of available samples. The relationship between expressing CD24 with treatment (surgery and chemotherapy) was significant and its relationship with patient's survival was insignificant statistically. However, there was a clear difference as mean survival age of patients based on CD24 expression was 26.64±18.15 for negative cases and 41.75±28.76 months for positive cases. CD24 and CD133 expressions and their co-expression with other clinic-pathological factors were not significant. During this study, the relationship between CD24 and treatment type was significant. To confirm this result, various studies with high sample numbers and other stem cell markers are recommended.


Mass spectrometric characterization of the affinity-purified human 26S proteasome complex.
The 26S proteasome is a multisubunit complex responsible for degradation of ubiquitinated substrates, which plays a critical role in regulating various biological processes. To fully understand the function and regulation of the proteasome complex, an important step is to elucidate its subunit composition and posttranslational modifications. Toward this goal, a new affinity purification strategy has been developed using a derivative of the HB tag for rapid isolation of the human 26S proteasome complex for subsequent proteomic analysis. The purification of the complex is achieved from stable 293 cell lines expressing a HB-tagged proteasome subunit and by high-affinity streptavidin binding with TEV cleavage elution. The complete composition of the 26S proteasome complex, including recently assigned new subunits, is identified by LC-MS/MS. In addition, all known proteasome activator proteins and components involved in the ubiquitin-proteasome degradation pathway are identified. Aside from the subunit composition, the N-terminal modification and phosphorylation of the proteasome subunits have been characterized. Twelve novel phosphorylation sites from eight subunits have been identified, and N-terminal modifications are determined for 25 subunits, 12 of which have not been previously reported in mammals. We also observe different N-terminal processing of subunit Rpn2, which results in identification of two different N-termini of the protein. This work presents the first comprehensive characterization of the human 26S proteasome complex by affinity purification and tandem mass spectrometry. The detailed proteomic profiling obtained here is significant to future studies aiming at a complete understanding of the structure-function relationship of the human 26S proteasome complex.


Antioxidant protective effects of mushroom metabolites.
Global industrialization and agricultural development are related to the release of various pollutants into the environment including huge amounts of free radicals, which are associated with the occurrence of various destructive diseases and disorders. Due to different rates of metabolic activity and oxygen consumption, organisms, organs and tissues have distinct protective antioxidant systems and defence mechanisms. Although numerous synthetic antioxidants can improve defence system capacity, because of their toxic and mutagenic effects as well as rising awareness of a healthy lifestyle, preference is given to natural antioxidants. Therefore, studies of metabolites of various fungal species and their activity are currently increasing. Antioxidant features of numerous compounds isolated from mushrooms, such as phenolic compounds, vitamins, polysaccharides, peptides, proteins, organic acids, carotenoids, alkaloids, and nucleotides, have been reported. Chemical composition and antioxidant potential of mushrooms are highly-dependent on species, habitat, phase of life cycle (mycelium, young or mature fruiting body), method of processing, extraction solvent, and extract dose. Although around only 5% of fungal species have been well studied and thousands more species have potential benefit for mankind, reports on the antioxidant potentials of fungi are already numerous and these are the subject of this review.


Dynamics at the serine loop underlie differential affinity of cryptochromes for CLOCK:BMAL1 to control circadian timing.
Mammalian circadian rhythms are generated by a transcription-based feedback loop in which CLOCK:BMAL1 drives transcription of its repressors (PER1/2, CRY1/2), which ultimately interact with CLOCK:BMAL1 to close the feedback loop with ~24 hr periodicity. Here we pinpoint a key difference between CRY1 and CRY2 that underlies their differential strengths as transcriptional repressors. Both cryptochromes bind the BMAL1 transactivation domain similarly to sequester it from coactivators and repress CLOCK:BMAL1 activity. However, we find that CRY1 is recruited with much higher affinity to the PAS domain core of CLOCK:BMAL1, allowing it to serve as a stronger repressor that lengthens circadian period. We discovered a dynamic serine-rich loop adjacent to the secondary pocket in the photolyase homology region (PHR) domain that regulates differential binding of cryptochromes to the PAS domain core of CLOCK:BMAL1. Notably, binding of the co-repressor PER2 remodels the serine loop of CRY2, making it more CRY1-like and enhancing its affinity for CLOCK:BMAL1.


Which factors affect the choice of the inhaler in chronic obstructive respiratory diseases?
Inhalation is the preferred route of drug administration in chronic respiratory diseases because it optimises delivery of the active compounds to the targeted site and minimises side effects from systemic distribution. The choice of a device should be made after careful evaluation of the patient's clinical condition (degree of airway obstruction, comorbidities), as well as their ability to coordinate the inhalation manoeuvre and to generate sufficient inspiratory flow. These patient factors must be aligned with the specific advantages and limitations of each inhaler when making this important choice. Finally, adherence to treatment is not the responsibility of the patient alone, but should be shared also by clinicians. Clinicians have access to a wide selection of pressurised metered dose inhalers (pMDIs) and dry powder inhalers (DPIs) that can be used effectively when matched to the needs of individual patients; this should be perceived as an opportunity rather than a limitation.


A prospective study of asymptomatic bacteriuria in sexually active young women.
Asymptomatic bacteriuria is common in young women, but little is known about its pathogenesis, natural history, risk factors, and temporal association with symptomatic urinary tract infection. We prospectively evaluated 796 sexually active, nonpregnant women from 18 through 40 years of age over a period of six months for the occurrence of asymptomatic bacteriuria (defined as at least 10(5) colony-forming units of urinary tract pathogens per milliliter). The women were patients at either a university student health center or a health maintenance organization. Periodic urine cultures were taken, daily diaries were kept, and regularly scheduled interviews were performed. Escherichia coli strains were tested for hemolysin, the papG genotype, and the ribosomal RNA type. The prevalence of asymptomatic bacteriuria (the proportion of urine cultures with bacteriuria in asymptomatic women) was 5 percent (95 percent confidence interval, 4 percent to 6 percent) among women in the university group and 6 percent (95 percent confidence interval, 5 percent to 8 percent) among women in the health-maintenance-organization group. Persistent asymptomatic bacteriuria with the same E. coli strain was rare. Symptomatic urinary tract infection developed within one week after 8 percent of occasions on which a culture showed asymptomatic bacteriuria, as compared with 1 percent of occasions when asymptomatic bacteriuria was not found (P<0.001). Asymptomatic bacteriuria was associated with the same risk factors as for symptomatic urinary tract infection, particularly the use of a diaphragm plus spermicide and sexual intercourse. Asymptomatic bacteriuria in young women is common but rarely persists. It is a strong predictor of subsequent symptomatic urinary tract infection.


Spinal Cord Stimulators: An Analysis of the Adverse Events Reported to the Australian Therapeutic Goods Administration.
Spinal cord stimulators are used to treat intractable pain. Placebo-controlled trials of spinal cord stimulators typically involve short-term treatment and follow-up, so long-term safety and efficacy are unclear. The aim of the study was to describe the adverse events relating to spinal cord stimulators reported to the Therapeutic Goods Administration of Australia between July 2012 and January 2019. Adverse events were coded by seriousness, severity, body system affected, type of event, action taken, and attribution of fault. Data on the number of stimulators implanted and removed were sourced from the Admitted Patient Care Minimum Data Set. Five hundred twenty adverse events were reported for spinal cord stimulators. Most events were rated as severe (79%) or life-threatening (13%). Device malfunction was the most common event (56.5%). The most common action taken in response to an adverse event was surgical intervention with or without antibiotics (80%). The ratio of removals to implants was 4 per every 10 implanted. Spinal cords stimulators have the potential for serious harm, and each year in Australia, many are removed. In view of the low certainty evidence of their long-term safety and effectiveness, our results raise questions about their role in providing long-term management of intractable pain.


AraC protein, regulation of the l-arabinose operon in Escherichia coli, and the light switch mechanism of AraC action.
This review covers the physiological aspects of regulation of the arabinose operon in Escherichia coli and the physical and regulatory properties of the operon's controlling gene, araC. It also describes the light switch mechanism as an explanation for many of the protein's properties. Although many thousands of homologs of AraC exist and regulate many diverse operons in response to many different inducers or physiological states, homologs that regulate arabinose-catabolizing genes in response to arabinose were identified. The sequence similarities among them are discussed in light of the known structure of the dimerization and DNA-binding domains of AraC.


Long term trends and spatial distribution of animal bite injuries and deaths due to human rabies infection in Uganda, 2001-2015.
In the absence of accurate data on trends and the burden of human rabies infection in developing countries, animal bite injuries provide useful information to bridge that gap. Rabies is one of the most deadly infectious diseases, with a case fatality rate approaching 100%. Despite availability of effective prevention and control strategies, rabies still kills 50,000 to 60,000 people worldwide annually, the majority of whom are in the developing world. We describe trends and geographical distribution of animal bite injuries (a proxy of potential exposure to rabies) and deaths due to suspected human rabies in Uganda from 2001 to 2015. We used 2001-2015 surveillance data on suspected animal bite injuries, collected from health facilities in Uganda. To describe annual trends, line graphs were used and linear regression tested significance of observed trends at P<0.05. We used maps to describe geographical distribution of animal bites by district. A total of 208,720 cases of animal bite injuries were reported. Of these, 27% were in Central, 22% in Eastern, 27% in Northern and 23% in Western regions. Out of 48,720 animal bites between 2013 and 2015, 59% were suffered by males and 81% were persons aged above 5 years. Between 2001 and 2015, the overall incidence (per 100,000 population) of animal bites was 58 in Uganda, 76 in Northern, 58 in Central, 53 in Western and 50 in Eastern region. From 2001 to 2015, the annual incidence (per 100,000 population) increased from 21 to 47 (P = 0.02) in Central, 27 to 34 (P = 0.04) in Eastern, 23 to 70 (P = 0.01) in Northern and 16 to 46 (P = 0.001) in Western region. A total of 486 suspected human rabies deaths were reported, of which 29% were reported from Eastern, 28% from Central, 27% from Northern and 17% from Western region. Animal bite injuries, a potential exposure to rabies infection, and mortality attributed to rabies infection are public health challenges affecting all regions of Uganda. Eliminating rabies requires strengthening of rabies prevention and control strategies at all levels of the health sector. These strategies should utilize the "One Health" approach with strategic focus on strengthening rabies surveillance, controlling rabies in dogs and ensuring availability of post exposure prophylaxis at lower health facilities.


GibbsCluster: unsupervised clustering and alignment of peptide sequences.
Receptor interactions with short linear peptide fragments (ligands) are at the base of many biological signaling processes. Conserved and information-rich amino acid patterns, commonly called sequence motifs, shape and regulate these interactions. Because of the properties of a receptor-ligand system or of the assay used to interrogate it, experimental data often contain multiple sequence motifs. GibbsCluster is a powerful tool for unsupervised motif discovery because it can simultaneously cluster and align peptide data. The GibbsCluster 2.0 presented here is an improved version incorporating insertion and deletions accounting for variations in motif length in the peptide input. In basic terms, the program takes as input a set of peptide sequences and clusters them into meaningful groups. It returns the optimal number of clusters it identified, together with the sequence alignment and sequence motif characterizing each cluster. Several parameters are available to customize cluster analysis, including adjustable penalties for small clusters and overlapping groups and a trash cluster to remove outliers. As an example application, we used the server to deconvolute multiple specificities in large-scale peptidome data generated by mass spectrometry. The server is available at http://www.cbs.dtu.dk/services/GibbsCluster-2.0.


Strong-field ionization and dissociation studies on small early transition metal carbide clusters via time-of-flight mass spectrometry.
In this work, we report experimental results from the strong-field ionization and subsequent Coulomb explosion of narrow distributions of small (<40 atoms) heteronuclear clusters composed of transition metal (Ti, V, Cr, Nb, or Ta) and carbon atoms. Analysis of the resulting multiply charged ions was performed through time-of-flight mass spectrometry, and evidence regarding ionization dynamics was obtained. The data reveal the presence of enhanced ionization during exposure to the ultrashort (∼100 fs) pulse resulting in the formation of ions possessing significantly higher charge states than those predicted from atomic species. Regardless of the transition metal species, we observe the absorption of similar amounts of energy from the external field, as indicated by the maximum observed charge states in each experiment. These results are compared to our previously reported study on the strong-field ionization of transition metal oxide clusters. We observe identical maximum observable charge states for each of the transition metal species resulting from both metal oxide and metal carbide clusters.


Intravitreal ranibizumab therapy for retinal capillary hemangioblastoma related to von Hippel-Lindau disease.
To evaluate the effect of intravitreal ranibizumab on retinal capillary hemangioblastomas (RCHs) associated with von Hippel-Lindau (VHL) disease that are not amenable or responsive to standard therapy. Prospective, noncomparative, interventional case series. Five patients with VHL-associated RCH with exudative changes and visual loss. Monthly intravitreal injections of ranibizumab (0.5 mg) were given over a course of 6 months for a total of 7 injections, with additional injections considered until week 52. The final study visit was designated as 8 weeks after the final study injection. The primary outcome was the change in best-corrected visual acuity (BCVA) of >/=15 letters at the final visit compared with baseline. Secondary outcomes included change in lesion size, exudation as assessed clinically and by fluorescein angiography, change in retinal thickness as evaluated by optical coherence tomography, and adverse event assessments. Patients received an average of 10.0+/-3.1 injections over an average period of 47+/-14 weeks, including follow-up. Mean change in BCVA was a decrease of 9+/-20 letters, with 1 patient gaining >/=15 letters, and 2 patients losing >/=15 letters. Changes in both lesion size and exudation were variable. Intravitreal ranibizumab, delivered as monotherapy every 4 weeks, had minimal beneficial effects on most VHL-related RCHs. Possible treatment efficacy was demonstrated in the patient with the smallest lesion with less exudation. Future prospective studies are needed to determine the potential role of an antiangiogenic agent, possibly in combination with other therapies for the treatment of such advanced ocular tumors associated with VHL.


Interventional chest radiology.
The authors review the various interventional radiology techniques currently used in 1996 by a Medico-Radio-Surgical team. CT guided needle biopsy has an important place in the diagnostic approach to parenchymal as well as mediastinal tumours. But CT guidance allows also routine drainage of thoracic collections and sometimes thoracic sympatholysis. Superior vena cava and tracheobronchial stenting are palliative treatments as the percutaneous aspergilloma treatment. Embolization of bronchial and thoracic systemic arteries are also palliative but effective therapeutic procedures as well as vasoocclusion for arterio-venous fistulae.


Adenosine receptor agonists: from basic medicinal chemistry to clinical development.
Adenosine is a physiological nucleoside which acts as an autocoid and activates G protein-coupled membrane receptors, designated A(1), A(2A), A(2B) and A(3). Adenosine plays an important role in many (patho)physiological conditions in the CNS as well as in peripheral organs and tissues. Adenosine receptors are present on virtually every cell. However, receptor subtype distribution and densities vary greatly. Adenosine itself is used as a therapeutic agent for the treatment of supraventricular paroxysmal tachycardia and arrhythmias and as a vasodilatatory agent in cardiac imaging. During the past 20 years, a number of selective agonists for A(1), A(2A) and A(3) adenosine receptors have been developed, all of them structurally derived from adenosine. Several such compounds are currently undergoing clinical trials for the treatment of cardiovascular diseases (A(1)and A(2A)), pain (A(1)), wound healing (A(2A)), diabetic foot ulcers (A(2A)), colorectal cancer (A(3)) and rheumatoid arthritis (A(3)). Clinical evaluation of some A(1) and A(2A) adenosine receptor agonists has been discontinued. Major problems include side effects due to the wide distribution of adenosine receptors; low brain penetration, which is important for the targeting of CNS diseases; short half-lifes of compounds; or a lack of effects, in some cases perhaps due to receptor desensitisation or to low receptor density in the targeted tissue. Partial agonists, inhibitors of adenosine metabolism (adenosine kinase and deaminase inhibitors) or allosteric activators of adenosine receptors may be advantageous for certain indications, as they may exhibit fewer side effects.


Role of interstitial potassium.
Interstitial potassium concentration, [K+], is modulated during muscle activity due to a number of different mechanisms: diffusion and active transport of K+ in combination with water fluxes. The relative significance of the various mechanisms for muscle function is quantified. The effect of interstitial [K+] locally on the single muscle fiber is discussed along with its effect on the cardiovascular and respiratory systems and its role in motor control. It is concluded that K+ may play a significant role in the prevention as well as the development of fatigue.


Application of SNPs to improve yield of Pisum sativum L. (pea).
Nanotechnology opens an enormous scope of novel application in the fields of biotechnology and agricultural industries, because nanoparticles (NPs) have unique physicochemical properties, i.e. high surface area, high reactivity, tunable pore size and particle morphology. Present study was carried out to determine the role of silver NPs (SNPs) to improve yield of Pisum sativum L. SNPs (10-100 nm) were synthesised by green method using extract of Berberis lycium Royle. Pea seeds were soaked and seedling were foliage sprayed by 0, 30, 60 and 90 ppm SNPs. The experiment was arranged as split-split plot randomised complete block design with three replicates. The application of SNPs enhanced significantly number of seeds pod-1, number of pods plant-1, hundred seed weight, biological yield and green pod yield over control. The highest yield was found when 60 ppm SNPs were applied. However, exposure to 90 ppm SNPs, the yield of the pea decreased significantly as compared with 30 and 60 ppm. This research shows that SNPs have definite ability to improve growth and yield of crops. Nevertheless, a comprehensive experimentation is needed to establish the most appropriate concentration, size and mode of application of SNPs for higher growth and maximum yield of pea.


Gene order evolution and paleopolyploidy in hemiascomycete yeasts.
The wealth of comparative genomics data from yeast species allows the molecular evolution of these eukaryotes to be studied in great detail. We used "proximity plots" to visually compare chromosomal gene order information from 14 hemiascomycetes, including the recent Génolevures survey, to Saccharomyces cerevisiae. Contrary to the original reports, we find that the Génolevures data strongly support the hypothesis that S. cerevisiae is a degenerate polyploid. Using gene order information alone, 70% of the S. cerevisiae genome can be mapped into "sister" regions that tile together with almost no overlap. This map confirms and extends the map of sister regions that we constructed previously by using duplicated genes, an independent source of information. Combining gene order and gene duplication data assigns essentially the whole genome into sister regions, the largest gap being only 36 genes long. The 16 centromere regions of S. cerevisiae form eight pairs, indicating that an ancestor with eight chromosomes underwent complete doubling; alternatives such as segmental duplications can be ruled out. Gene arrangements in Kluyveromyces lactis and four other species agree quantitatively with what would be expected if they diverged from S. cerevisiae before its polyploidization. In contrast, Saccharomyces exiguus, Saccharomyces servazzii, and Candida glabrata show higher levels of gene adjacency conservation, and more cases of imperfect conservation, suggesting that they split from the S. cerevisiae lineage after polyploidization. This finding is confirmed by sequences around the C. glabrata TRP1 and IPP1 loci, which show that it contains sister regions derived from the same duplication event as that of S. cerevisiae.


Comparison of health-related quality of life and prostate-associated symptoms after primary and salvage cryotherapy for prostate cancer.
Recent advances in cryosurgery of the prostate have led to the ability to treat tumors successfully with decreased morbidity. The patients' perspectives of this relatively new technique, however, have not yet been addressed. The purpose of this study was to compare health related quality of life (QoL) as well as prostate-associated symptoms in patients after primary and salvage cryoablation for clinically localized prostate cancer using a self-administered questionnaire. A total of 131 consecutive patients who underwent cryoablation of the prostate between 1997 and 2001 were included in this confidential mailing study. The patients were either (a) patients with localized prostate cancer with contraindications for radical surgery, including patients refusing other forms of therapy, or (b) had locally recurrent prostate cancer after failure of radiation therapy. All patients received 3 months of neoadjuvant androgen deprivation therapy prior to cryosurgery and were surgically treated by the same surgeon using an argon-based system. We used the EORTC QLQ-C30, a commonly used, multidimensional instrument together with a supplementing, prostate-cancer-specific module. Eighty-one of the 131 patients (response rate 62%) returned the questionnaires. The two groups were comparable regarding age (mean age 72.8 vs 70.1 for the primary and the salvage group, respectively; p=0.22). The overall QoL scores were high in both groups. Primary cryotherapy patients fared significantly better regarding physical (p=0.005) and social (p=0.024) functioning compared with salvage cryotherapy patients. The most prominent prostate-related symptom in both patient groups was sexual dysfunction, followed by urinary symptoms, which were significantly more severe in the salvage group (p=0.001). Incontinence rates were 5.9 and 10% in the primary and the salvage group, respectively. Severe erectile dysfunction was reported in 86 and 90% of the primary and the salvage group, respectively. The present study demonstrates that, in selected patients, cryotherapy is a treatment option which has a functional outcome comparable to traditionally used prostate cancer treatments. More information regarding QoL is necessary for appropriate patient counseling and individual decision-making in the presence of various treatment alternatives.


An open-label trial of levetiracetam in severe myoclonic epilepsy of infancy.
To conduct an open-label, add-on trial on safety and efficacy of levetiracetam in severe myoclonic epilepsy of infancy (SMEI). SMEI patients were recruited from different centers according to the following criteria: age > or =3 years; at least four tonic-clonic seizures/month during the last 8 weeks; previous use of at least two drugs. Levetiracetam was orally administrated at starting dose of approximately 10 mg/kg/day up to 50 to 60 mg/kg/day in two doses. Treatment period included a 5- to 6-week up-titration phase and a 12-week evaluation phase. Efficacy variables were responder rate by seizure type and reduction of the mean number per week of each seizure type. Analysis was performed using Fisher exact and Wilcoxon tests. Twenty-eight patients (mean age: 9.4 +/- 5.6 years) entered the study. Sixteen (57.1%) showed SCN1A mutations. Mean number of concomitant drugs was 2.5. Mean levetiracetam dose achieved was 2,016 mg/day. Twenty-three (82.1%) completed the trial. Responders were 64.2% for tonic-clonic, 60% for myoclonic, 60% for focal, and 44.4% for absence seizures. Number per week of tonic-clonic (median: 3 vs 1; p = 0.0001), myoclonic (median: 21 vs 3; p = 0.002), and focal seizures (median: 7.5 vs 3; p = 0.031) was significantly decreased compared to baseline. Levetiracetam effect was not related to age at onset and duration of epilepsy, genetic status, and concomitant therapy. Levetiracetam was well tolerated by subjects who completed the study. To date, follow-up ranges 6 to 36 months (mean, 16.2 +/- 13.4). Levetiracetam add-on is effective and well tolerated in severe myoclonic epilepsy of infancy. Placebo-controlled studies should confirm these findings.


Evaluating the responsiveness of the fullerton advanced balance scale in patients with lymphedema secondary to breast cancer surgery.
Lymphedema following surgical treatment for breast cancer can impair balance and predispose patients to falling. Fullerton Advanced Balance (FAB) Scale is a reliable and valid tool which can identify persons with different balance levels, but its responsiveness has not been investigated in patients with lymphedema secondary to breast surgery. Thirty women with stage 2 lymphedema secondary to breast cancer treatment received complex decongestive therapy (CDT) for 2 weeks as a routine treatment method. They were evaluated with FAB Scale and Timed Up and Go Test (TUGT), volumetric measurements and circumferential measurements of the upper limbs before and after CDT. A moderate change was found in FAB score after CDT (Cohen's effect size = 0.65). For FAB, the computed standard error of the mean was 0.85 and minimal detectable change was 2.33. Significant improvement in FAB score and TUGT results, and significant reductions in circumferential and volumetric measurements were seen after 2 weeks of CDT. The FAB score change showed a moderate correlation with circumference change and volumetric change (r = -0.41) but a very weak correlation with TUGT change (r = -0.1). The FAB Scale showed acceptable responsiveness in detecting treatment effects in patients with unilateral secondary lymphedema after breast cancer treatment.


Volume Load and Neuromuscular Fatigue During an Acute Bout of Agonist-Antagonist Paired-Set vs. Traditional-Set Training.
The purpose of this study was to investigate the acute effects of performing paired-set (PS) vs. traditional-set (TS) training over 3 consecutive sets, on volume load and electromyographic fatigue parameters of the latissimus dorsi, biceps brachii, pectoralis major, and triceps brachii muscles. Fifteen trained men performed 2 testing protocols (TS and PS) using 10 repetition maximum loads. The TS protocol consisted of 3 sets of bench press (BP) followed by 3 sets of wide-grip seated row (SR). The PS consisted of 3 sets of BP and 3 sets of SR performed in an alternating manner. Volume load was calculated as load × repetitions. The electromyographic signal, time (CRMS) and frequency (Cf5) domain, parameters were recorded during SR. Under the PS protocol, sets of SR were performed immediately after the sets of BP. A 2-minute rest interval between the completion of the set of SR and the subsequent set of BP was implemented (e.g., between PSs). Under the TS protocol, 2-minute rest intervals were implemented between all sets. BP and SR volume loads decreased significantly from set 1 to set 2 and from set 2 to set 3 under both conditions. Volume load was greater for all sets of both exercises under PS as compared with TS. Muscle fatigue indices were greater under PS as compared with TS. In general, these results indicate that as compared with TS, PS produced a greater training volume in less time and may induce greater fatigue and thereby provide an enhanced training stimulus.


Efficiency of the lausanne clinical pathway for proximal femoral fractures.
Purpose/Introduction: The number of hip fractures is rising, due to increases in life expectancy. In such cases, patients are at risk from post-operative complications and subsequently the average length of hospitalization may be extended. In 2011, we established a clinical pathway (CP), a specific model of care for patient-care management, to improve the clinical and economic outcomes of proximal femoral fracture management in elderly patients. The goal was to evaluate the CP using clinical, process, and financial indicators. We included all surgical patients aged 65 and over, admitted to the emergency department with a fracture of the proximal femur following a fall. Assessment parameters included three performance indicators: clinical, process, and financial. The clinical indicators were the presence or absence of acute delirium on the third post-operative day, diagnosis of nosocomial pneumonia, and the number of patients fulfilling at least 75% of their nutritional requirements at the end of the hospitalization period. The process indicator was the time interval between arrival at the emergency department and surgery. The financial indicator was based on the number of days spent in hospital. From 2011 to 2013, 669 patients were included in the CP. We observed that the average length of stay in hospital decreased as soon as the CP was implemented and stabilized afterwards. The goal of 90% of patients undergoing surgery within 48 h of arrival in the emergency department was surpassed in 2013 (93.1%). Furthermore, we observed an improvement in the clinical indicators. The application of a CP allowed an improvement in the qualitative and quantitative efficiency of proximal femoral fracture management in elderly patients, in terms of clinical, process, and financial factors.


Pseudohypoparathyroidism type Ib in 2015.
The term pseudohypoparathryoidism (PHP) refers to a group of rare genetic and epigenetic disorders characterized by resistance to the action of parathyroid hormone (PTH) that activates cAMP signaling in target cells. Together with pseudohypoparathyroidism, Albright hereditary osteodystrophy (AHO) and progressive osseous heteroplasia (POH) represent rare, related and deeply impairing disorders encompassing heterogeneous features, such as brachydactyly, ectopic ossifications, short stature, mental retardation and endocrine deficiencies due to resistance to the action of different hormones. The two main subtypes, PHP-Ia and PHP-Ib, are caused by mutations in GNAS exons 1-13 and methylation defects in the imprinted GNAS cluster respectively, while mutations in the PRKAR1A and PDE4D genes (also involved in mediating cAMP signalling) have been demonstrated in patients with acrodysostosis, a disease of bone formation with characteristics similar to AHO. The molecular overlap among these disorders indicates the need for different classification models and seriously alters our understanding of the mechanisms through which GNAS defects, together with the new recently described defects involving other components of the cAMP signalling cascade, cause AHO-related disorders.


An Extracellular Cell-Attached Pullulanase Confers Branched α-Glucan Utilization in Human Gut Lactobacillus acidophilus.
Of the few predicted extracellular glycan-active enzymes, glycoside hydrolase family 13 subfamily 14 (GH13_14) pullulanases are the most common in human gut lactobacilli. These enzymes share a unique modular organization, not observed in other bacteria, featuring a catalytic module, two starch binding modules, a domain of unknown function, and a C-terminal surface layer association protein (SLAP) domain. Here, we explore the specificity of a representative of this group of pullulanases, Lactobacillus acidophilus Pul13_14 (LaPul13_14), and its role in branched α-glucan metabolism in the well-characterized Lactobacillus acidophilus NCFM, which is widely used as a probiotic. Growth experiments with L. acidophilus NCFM on starch-derived branched substrates revealed a preference for α-glucans with short branches of about two to three glucosyl moieties over amylopectin with longer branches. Cell-attached debranching activity was measurable in the presence of α-glucans but was repressed by glucose. The debranching activity is conferred exclusively by LaPul13_14 and is abolished in a mutant strain lacking a functional LaPul13_14 gene. Hydrolysis kinetics of recombinant LaPul13_14 confirmed the preference for short-branched α-glucan oligomers consistent with the growth data. Curiously, this enzyme displayed the highest catalytic efficiency and the lowest Km reported for a pullulanase. Inhibition kinetics revealed mixed inhibition by β-cyclodextrin, suggesting the presence of additional glucan binding sites besides the active site of the enzyme, which may contribute to the unprecedented substrate affinity. The enzyme also displays high thermostability and higher activity in the acidic pH range, reflecting adaptation to the physiologically challenging conditions in the human gut.IMPORTANCE Starch is one of the most abundant glycans in the human diet. Branched α-1,6-glucans in dietary starch and glycogen are nondegradable by human enzymes and constitute a metabolic resource for the gut microbiota. The role of health-beneficial lactobacilli prevalent in the human small intestine in starch metabolism remains unexplored in contrast to colonic bacterial residents. This study highlights the pivotal role of debranching enzymes in the breakdown of starchy branched α-glucan oligomers (α-limit dextrins) by human gut lactobacilli exemplified by Lactobacillus acidophilus NCFM, which is one of the best-characterized strains used as probiotics. Our data bring novel insight into the metabolic preference of L. acidophilus for α-glucans with short α-1,6-branches. The unprecedented affinity of the debranching enzyme that confers growth on these substrates reflects its adaptation to the nutrient-competitive gut ecological niche and constitutes a potential advantage in cross-feeding from human and bacterial dietary starch metabolism.


Toxoids of Pseudomonas aeruginosa exotoxin-A: photoaffinity inactivation of purified toxin and purified toxin derivatives.
For the preparation of greatly detoxified but highly immunogenic toxoids, two enzymatically active, low-toxicity derivatives of Pseudomonas aeruginosa exotoxin-A were further inactivated by photoaffinity labeling. These derivatives were formed during toxin purification, when a relatively crude toxin preparation was concentrated by ammonium sulfate precipitation and subsequently dialyzed. These derivatives, designated peak-1 protein (PK-1) and peak-2 protein (PK-2) were antigenically indistinguishable from native toxin, but had isoelectric points (5.00 and 4.90, respectively) that were different from that of the native toxin (4.95). Although the enzymatic activities and molecular weights of PK-1 and PK-2 were similar to those of native toxin, their toxicities were greatly reduced (ca. 500-fold). Photoaffinity labeling of fully active toxin-A, purified by a process which limits the formation of these derivatives, decreased its enzymatic activity (ca. 30-fold) and toxicity (ca. 100-fold). Likewise, photoaffinity labeling of purified PK-1 and PK-2 decreased their enzymatic activities and toxicities (ca. 30-fold and 100-fold, respectively) and, thus, yielded toxoids that were ca. 50,000-fold less toxic than unpurified native toxin. These toxoids were irreversibly detoxified and highly immunogenic during 9 months of storage at 4 degrees C.


Emerging investigator series: characterization of silver and silver nanoparticle interactions with zinc finger peptides.
In biological systems, chemical and physical transformations of engineered silver nanomaterials (AgENMs) are mediated, in part, by proteins and other biomolecules. Metalloprotein interactions with AgENMs are also central in understanding toxicity and antimicrobial and resistance mechanisms. Despite their readily available thiolate and amine ligands, zinc finger (ZF) peptides have thus far escaped study in reaction with AgENMs and their Ag(I) oxidative dissolution product. We report spectroscopic studies that characterize AgENM and Ag(I) interactions with two ZF peptides that differ in sequence, but not in metal binding ligands: the ZF consensus peptide CP-CCHC and the C-terminal zinc finger domain of HIV-1 nucleocapsid protein p7 (NCp7_C). Both ZF peptides catalyze AgENM (10 and 40 nm, citrate coated) dissolution and agglomeration, two important AgENM transformations that impact bioreactivity. AgENMs and their oxidative dissolution product, Ag(I)(aq), mediate changes to ZF peptide structure and metalation as well. Spectroscopic titrations of Ag(I) into apo-ZF peptides show an Ag(I)-thiolate charge transfer band, indicative of Ag(I)-ZF binding. Fluorescence studies of the Zn(II)-NCp_7 complex indicate that the Ag(I) also effectively competes with the Zn(II) to drive Zn(II) displacement from the ZFs. Upon interaction with AgENMs, Zn(II) bound ZF peptides show a secondary structural change in circular dichroism spectroscopy toward an apo-like structure. The results suggest that Ag(I) and AgENMs may alter ZF protein function within the cell.