Optimal intensity and type of leg exercise training for people with chronic obstructive pulmonary disease.
Intensity of exercise is considered a key determinant of training response, however, no systematic review has investigated the effects of different levels of training intensity on exercise capacity, functional exercise capacity and health-related quality of life (HRQoL) in people with chronic obstructive pulmonary disease (COPD). As type of training (continuous or interval) may also affect training response, the effects of the type of training in COPD also require investigation. To determine the effects of training intensity (higher versus lower) or type (continuous versus interval training) on primary outcomes in exercise capacity and secondary outcomes in symptoms and HRQoL for people with COPD. We searched for studies in any language from the Cochrane Airways Group Specialised Register, CENTRAL, MEDLINE, EMBASE, CINAHL, AMED, PsycINFO and PubMed. Searches were current as of June 2011. We included randomised controlled trials comparing higher training intensity to lower training intensity or comparing continuous training to interval training in people with COPD. We excluded studies that compared exercise training with no exercise training. We pooled results of comparable groups of studies and calculated the treatment effect and 95% confidence intervals (CI) using a random-effects model. We made two separate comparisons of effects between: 1) higher and lower training intensity; 2) continuous and interval training. We contacted authors of missing data. We analysed three included studies (231 participants) for comparisons between higher and lower-intensity training and eight included studies (367 participants) for comparisons between continuous and interval training. Primary outcomes were outcomes at peak exercise (peak work rate, peak oxygen consumption, peak minute ventilation and lactate threshold), at isowork or isotime, endurance time on a constant work rate test and functional exercise capacity (six-minute walk distance). When comparing higher versus lower-intensity training, the pooled primary outcomes were endurance time and six-minute walk distance. There were no significant differences in endurance time improvement (mean difference (MD) 1.07 minutes; 95% CI -1.53 to 3.67) and six-minute walk distance improvement (MD 2.8 metres; 95% CI -10.1 to 15.6) following higher or lower-intensity training. However, heterogeneity of the endurance time results between studies was significant. When comparing continuous and interval training, there were no significant differences in any of the primary outcomes, except for oxygen consumption at isotime (MD 0.08; 95% CI 0.01 to 0.16) but the treatment effect was not considered clinically important. According to the GRADE system, studies were of low to moderate quality. Comparisons between the higher and lower training intensity were limited due to the small number of included studies and participants. Consequently, there are insufficient data to draw any conclusions on exercise capacity, symptoms and HRQoL for this comparison. For comparisons between continuous and interval training, both appear to be equally effective in improving exercise capacity, symptoms and HRQoL.


Association of prostate cancer with human papillomavirus infections: a case-control study.
The highly oncogenic human papillomavirus (HPV) is associated with numerous cancer types. While the role of viruses in the development of certain cancers is well established, the association between HPV infections and prostate cancer remains a subject of ongoing debate. This study aimed to investigate a potential association of prostate cancer with HPV infections utilizing a case-control study. We extracted data from the Taiwan Longitudinal Health Insurance Database 2010. We retrieved 5137 patients with prostate cancer as cases and a 3:1 ratio of propensity score-matched patients without prostate cancer (15,411 patients) as controls. Multiple logistic regression analyses were carried out to scrutinize the association of prostate cancer with HPV infections while taking into account age, monthly income category, geographic location and urbanization level of the patient's residence as well as hyperlipidemia, diabetes, hypertension and chronic prostatitis, tobacco use disorder, and alcohol abuse/alcohol dependence syndrome. The data indicate that out of all sampled patients, 1812 (8.8%) had a prior diagnosis of HPV infections before the index date. Among cases and matched controls, HPV infections were diagnosed in 743 (14.5%) and 1069 (6.9%) patients, respectively. The results from the chi-square test demonstrate that individuals with prostate cancer exhibited a significantly higher incidence rate of HPV infections than their control counterparts (p < 0.001). Furthermore, in comparison to controls, individuals with a history of HPV infections had an adjusted odds ratio of 2.321 (95% CI: 2.097~2.568) for developing prostate cancer. Notably, individuals diagnosed with chronic prostatitis were also more likely to be subsequently diagnosed with prostate cancer (adjusted odds ratio=1.586; 95% CI = 1.338~1.879), which aligns with expectations in this context. We found prostate cancer to be significantly associated with HPV infections, contributing to the mounting body of evidence indicating a plausible connection between the two.


Locoregional recurrence (non hepatic abdominal recurrence) of rectal cancer.
Thirty percent of deaths are related to locoreional recurrence. All patients with nonhepatic abdominal recurrence (NHAR) were considered as having locoregional failure. The aims of this study are firstly to retrospectively evaluate the results of potentially curative resection and palliative treatment modalities for a group of 25 patients with NHAR from rectal cancer. The second aim is to determine the effectiveness of R1 resection in these patients in terms of survival. In this study we have followed 25 patients with NHAR of which 10 were able to undergo potentially curative salvage resection, whilst the remaining 15 had either a palliative (R2) or no resection. The goals of treatment for recurrent rectal cancer are palliation of symptoms, a good quality of life, and if possible, cure with a low rate of treatment--related complications. Indications for salvage surgery depend on several factors including the extent of disease, the presence of concomitant illness and the surgeons experience. Systemic disease, systemic disease with peritoneal implants, multiple hepatic metastases, or extensive pelvic involvement preclude surgical treatment for cure. Curative and noncurative surgical procedures were performed width acceptable complications in the series presented hereThe mean survival for the group undergoing R0 resection was 50 months versus 55 months for the group undergoing R1 resection (not significant). Mean survival were 7,3 and 6 months in the groups undergoing R2, NR and NS respectively. The 5-year survival for the 10 patients who had potentially curative resection was 30 per cent versus 0 per cent for 15 patients who had non-curative procedures (p = 0.001). There was 1 post-operative 30 day mortality in the series of 19 patients who underwent surgery. Five patients (6 per cent) developed one or more post-operative complications. Two of them required reoperation.


Genetic identification of inherited cystic kidney diseases for implementing precision medicine: a study protocol for a 3-year prospective multicenter cohort study.
Inherited cystic kidney disease is a spectrum of disorders in which clusters of renal cysts develop as the result of genetic mutation. The exact methods and pipelines for defining genetic mutations of inherited cystic kidney disease are not clear at this point. This 3-year, prospective, multicenter, cohort study was designed to set up a cohort of Korean patients with inherited cystic kidney disease, establish a customized genetic analysis pipeline for each disease subtype, and identify modifying genes associated with the severity of the disease phenotype. From May 2020 to May 2022, we aim to recruit 800 patients and their family members to identify pathogenic mutations. Patients with more than 3 renal cysts in both kidneys are eligible to be enrolled. Cases of simple renal cysts and acquired cystic kidney disease that involve cyst formation as the result of renal failure will be excluded from this study. Demographic, laboratory, and imaging data as well as family pedigree will be collected at baseline. Renal function and changes in total kidney volume will be monitored during the follow-up period. Genetic identification of each case of inherited cystic kidney disease will be performed using a targeted gene panel of cystogenesis-related genes, whole exome sequencing (WES) and/or family segregation studies. Genotype-phenotype correlation analysis will be performed to elucidate the genetic effect on the severity of the disease phenotype. This is the first nationwide cohort study on patients with inherited cystic kidney disease in Korea. We will build a multicenter cohort to describe the clinical characteristics of Korean patients with inherited cystic kidney disease, elucidate the genotype of each disease, and demonstrate the genetic effects on the severity of the disease phenotype. This cohort study was retrospectively registered at the Clinical Research Information Service ( KCT0005580 ) operated by the Korean Center for Disease Control and Prevention on November 5th, 2020.


ResUHUrge: A Low Cost and Fully Functional Ventilator Indicated for Application in COVID-19 Patients.
Although the cure for the SARS-CoV-2 virus (COVID-19) will come in the form of pharmaceutical solutions and/or a vaccine, one of the only ways to face it at present is to guarantee the best quality of health for patients, so that they can overcome the disease on their own. Therefore, and considering that COVID-19 generally causes damage to the respiratory system (in the form of lung infection), it is essential to ensure the best pulmonary ventilation for the patient. However, depending on the severity of the disease and the health condition of the patient, the situation can become critical when the patient has respiratory distress or becomes unable to breathe on his/her own. In that case, the ventilator becomes the lifeline of the patient. This device must keep patients stable until, on their own or with the help of medications, they manage to overcome the lung infection. However, with thousands or hundreds of thousands of infected patients, no country has enough ventilators. If this situation has become critical in the Global North, it has turned disastrous in developing countries, where ventilators are even more scarce. This article shows the race against time of a multidisciplinary research team at the University of Huelva, UHU, southwest of Spain, to develop an inexpensive, multifunctional, and easy-to-manufacture ventilator, which has been named ResUHUrge. The device meets all medical requirements and is developed with open-source hardware and software.


0° vs. 180° CT localiser: The effect of vertical off-centring, phantom positioning and tube voltage on dose optimisation in multidetector computed tomography.
Patient positioning is an essential consideration for the optimisation of radiation dose during CT examinations. The study objectives seek to explore the effects of vertical off-centring, localiser direction (0° and 180°), and phantom positioning (supine and prone) on radiation dose, using three different tube voltages in multidetector computed tomography (MDCT) imaging. The trunk of a PBU-60 anthropomorphic phantom was imaged using a Discovery CT750 HD - 128 slice (GE Healthcare). Images employing 0° and 180° localisers were acquired in supine and prone orientation for each combination of vertical off-centring (±100, ±60 and ±30 mm) and different tube voltages (80, 120 and 140 kVp), using the system's automatic tube current modulation (ATCM) function. The displayed volume CT dose index (CTDIvol ) and dose length product (DLP) were recorded. With incremental table off-centring of ±100 mm, the dose at 120 kVp in the supine position ranged from 63% to 196% (0° localiser) and from 66% to 191% (180° localiser) as compared to iso-centre. While in the prone position, the dose ranged from 62% to 195% (0° localiser); and 62% to 193% (180° localiser), with a notable dose increase at higher tube voltages. Dose variation and vertical off-centring showed a significant relationship for both 0° and 180° localisers (r = 0.94 and 0.96, respectively, P < 0.001). The CTDIvol variation between supine and prone phantom positions at ±100 mm off-centring was 0.22 mGy (2.9%), and 0.19 mGy (2.3%) when the 0° and 180 ° localisers were utilised, respectively. Phantom off-centring and localiser direction evidenced large dose variation. It is recommended that the 0° localiser is employed during CT examinations, in order to minimise the potential additional radiation dose which may result from off-centring and the use of lower tube voltages where clinically appropriate.


WY7 is a newly identified promoter from the rubber powdery mildew pathogen that regulates exogenous gene expression in both monocots and dicots.
Promoters are very important for transcriptional regulation and gene expression, and have become invaluable tools for genetic engineering. Owing to the characteristics of obligate biotrophs, molecular research into obligate biotrophic fungi is seriously lagging behind, and very few of their endogenous promoters have been developed. In this study, a WY7 fragment was predicted in the genome of Oidium heveae Steinmann using PromoterScan. Its promoter function was verified with transient transformations (Agrobacterium tumefaciens-mediated transformation, ATMT) in Nicotiana tabacum cv. Xanthi nc. The analysis of the transcription range showed that WY7 could regulate GUS expression in both monocots (Zea mays Linn and Oryza sativa L. spp. Japonica cv. Nipponbare) and dicots (N. tabacum and Hylocereus undulates Britt). The results of the quantitative detection showed that the GUS transient expression levels when regulated by WY7 was more than 11.7 times that of the CaMV 35S promoter in dicots (N. tabacum) and 5.13 times that of the ACT1 promoter in monocots (O. sativa). GUS staining was not detected in the T1 generation of the WY7-GUS transgenic N. tabacum. This showed that WY7 is an inducible promoter. The cis elements of WY7 were predicted using PlantCARE, and further experiments indicated that WY7 was a low temperature- and salt-inducible promoter. Soluble proteins produced by WY7-hpa1Xoo transgenic tobacco elicited hypersensitive responses (HR) in N. tabacum leaves. N. tabacum transformed with pBI121-WY7-hpa1Xoo exhibited enhanced resistance to the tobacco mosaic virus (TMV). The WY7 promoter has a lot of potential as a tool for plant genetic engineering. Further in-depth studies will help to better understand the transcriptional regulation mechanisms of O. heveae.


[The characteristics of exploratory behavior in Wistar rats in a permanent inhomogeneous magnetic field].
The influence of weak perturbations (up to 300 microT) in the natural magnetic field (MF) (induced by the introduction of three constant magnets) on the problem task solving in complex maze was studied in two groups of Wistar rats. Under the action of inhomogeneous magnetic field there were no spontaneous change of the orienting for the explorative activity independently of the individual in the presence of the organization of goal-directed behaviour. Instead of the expected explorative activity, the stable and deep locomotor depression developed after orienting in a new environment. Such a phenomenon was manifested only against the background of informational loading. Nevertheless, under these conditions a short external stimulations resulted in even faster rat learning than in the control animals. It is suggested that the MF characteristics play a crucial role in the transformation of the orienting (unconditioned activity) into the explorative (cognitive) activity.


Temporary Mechanical Circulatory Support in Acute Heart Failure.
Cardiogenic shock (CS) is a challenging syndrome, associated with significant morbidity and mortality. Although pharmacological therapies are successful and can successfully control this acute cardiac illness, some patients remain refractory to drugs. Therefore, a more aggressive treatment strategy is needed. Temporary mechanical circulatory support (TCS) can be used to stabilise patients with decompensated heart failure. In the last two decades, the increased use of TCS has led to several kinds of devices becoming available. However, indications for TCS and device selection are part of a complex process. It is necessary to evaluate the severity of CS, any early and prompt haemodynamic resuscitation, prior TCS, specific patient risk factors, technical limitations and adequacy of resources and training, as well as an assessment of whether care would be futile. This article examines options for commonly used TCS devices, including intra-aortic balloon pumps, a pulsatile percutaneous ventricular assist device (the iVAC), veno-arterial extra-corporeal membrane oxygenation and Impella (Abiomed) and TandemHeart (LivaNova) percutaneous ventricular assist device.


The Important Role of STAT3 in Chronic Lymphocytic Leukaemia Biology.
Signal transducer and activator of transcription (STAT) proteins are cytoplasmic transcription factors that transmit the signal of cytokines, hormones and growth factors. STAT proteins control fundamental cellular processes including survival, proliferation and differentiation. Inappropriate activation of STATs might contribute to cellular transformation and leukaemogenesis. About 70% of all solid and haematological tumours exhibit aberrant STAT3 expression and/or activation, highlighting its essential role in tumourigenesis. Aberrant STAT3 activation has been found in several solid tumours and haematologic malignancies. Importantly, constitutive activation of STAT proteins has been found in several leukaemias including acute myeloid leukaemia, acute promyelocytic leukaemia, acute lymphoblastic leukaemia, chronic myeloid leukaemia and chronic lymphocytic leukaemia (CLL). Constitutively activated STAT3 plays an important role in CLL biology. CLL cells harbour constitutive phosphorylation on S727 and acetylation on K685 and transient phosphorylation on Y705 residues. Moreover, STAT3 messenger RNA expression is significantly higher in CLL cells compared to healthy B-lymphocytes. Interestingly, STAT3 inhibition was disclosed as an important by-product of ibrutinib treatment in CLL patients. The purpose of this review is to describe the consequences of STAT3 dysregulation in CLL cells. Here, we discuss aberrantly modified processes by STAT3 activation in CLL cells such as proliferation, apoptosis, B cell receptor signalling, cytokine secretion, immune checkpoint regulation, microRNA regulation, free fatty acid metabolism and electron transport chain in the mitochondria.


Relationship between (8-14C)adenosine transport and growth inhibition in Neurospora crassa strain ad-8.
The kinetic parameters of [8-(14)C]adenosine transport by a general nucleoside uptake system were studied in germinated conidia of the ad 8 strain of Neurospora crassa. The apparent K(m) for adenosine uptake by this system was found to be 6.2 muM. The apparent K(i) values for other nucleosides competing with adenosine for uptake were measured by using Dixon plots. Nucleosides which were efficient competitive inhibitors of adenosine transport were found to inhibit severely the rate of growth of strain ad-8 on adenosine-supplemented medium. Xanthosine and thymidine did not inhibit [8-(14)C]adenosine uptake as severely as other nucleosides, nor did they cause significant inhibition of ad-8 growth rate on adenosine.


Individuation of objects and events: a developmental study.
This study investigates children's ability to use language to guide their choice of individuation criterion in the domains of objects and events. Previous work (Shipley, E. F., & Shepperson, B. (1990). Countable entities: developmental changes. Cognition, 34, 109-136.) has shown that children have a strong bias to use a spatio-temporal individuation strategy when counting objects and that children will ignore a conflicting linguistic description in favor of this spatio-temporal bias. Experiment 1 asked children (3-, 4-, and 5-year-olds) and adults to count objects and events under different linguistic descriptions. In the object task, subjects counted pictures of familiar objects split into multiple pieces (as in Shipley, E. F., & Shepperson, B. (1990). Countable entities: developmental changes. Cognition, 34, 109-136.) and described either using an appropriate kind label (e.g. "car") or the general term "thing". In the event task, subjects watched short animated movies consisting of a goal-oriented event achieved via multiple, temporally separated steps. The events were described either with an appropriate telic predicate targeting the goal (e.g. "paint a flower") or with an atelic predicate targeting the steps in the process (e.g. "paint") and the subjects' task was to count the events. Relative to adults, children preferred a spatio-temporal counting strategy in both tasks; there was no difference among the three groups of children. However, children were able to significantly change their counting strategy to follow the linguistic description in the event but not the object task. Experiment 2 extended the object task to include counting of other types of non-spatio-temporal units such as sub-parts of objects and collections. Results showed that children could use the linguistic descriptions to guide their counting strategy for these new items, though they continued to show a bias for a spatio-temporal individuation strategy with the collections. We suggest potential cognitive origins for the spatio-temporal individuation bias and how it interacts with children's developing linguistic knowledge.


Novel actions of next-generation taxanes benefit advanced stages of prostate cancer.
To improve the outcomes of patients with castration-resistant prostate cancer (CRPC), there is an urgent need for more effective therapies and approaches that individualize specific treatments for patients with CRPC. These studies compared the novel taxane cabazitaxel with the previous generation docetaxel, and aimed to determine which tumors are most likely to respond. Cabazitaxel and docetaxel were compared via in vitro modeling to determine the molecular mechanism, biochemical and cell biologic impact, and cell proliferation, which was further assessed ex vivo in human tumor explants. Isogenic pairs of RB knockdown and control cells were interrogated in vitro and in xenograft tumors for cabazitaxel response. The data herein show that (i) cabazitaxel exerts stronger cytostatic and cytotoxic response compared with docetaxel, especially in CRPC; (ii) cabazitaxel induces aberrant mitosis, leading to pyknotic and multinucleated cells; (iii) taxanes do not act through the androgen receptor (AR); (iv) gene-expression profiling reveals distinct molecular actions for cabazitaxel; and (v) tumors that have progressed to castration resistance via loss of RB show enhanced sensitivity to cabazitaxel. Cabazitaxel not only induces improved cytostatic and cytotoxic effects, but also affects distinct molecular pathways, compared with docetaxel, which could underlie its efficacy after docetaxel treatment has failed in patients with CRPC. Finally, RB is identified as the first potential biomarker that could define the therapeutic response to taxanes in metastatic CRPC. This would suggest that loss of RB function induces sensitization to taxanes, which could benefit up to 50% of CRPC cases.


Possibilities of Controlling the Quantum States of Hole Qubits in an Ultrathin Germanium Layer Using a Magnetic Substrate: Results from ab Initio Calculations.
Using density functional theory in the noncollinear approximation, the behavior of quantum states of hole qubits in a Ge/Co:ZnO system was studied in this work. A detailed analysis of the electronic structure and the distribution of total charge density and hole states was carried out. It was shown that in the presence of holes, the energetically more favorable quantum state is the state |0˃, in contrast to the state |1˃ when there is no hole in the system. The favorability of hole states was found to be dependent on the polarity of the applied electric field.


Assessing the skill of yes/no predictions.
Should healthy, middle-aged women receive precautionary mammograms? Should trauma surgeons use the popular TRISS score to predict the likelihood of patient survival? These are examples of questions confronting us when we decide whether to use a yes/no prediction. In order to trust a prediction we must show that it is more valuable than would be our best guess of the future in the absence of the prediction. Calculating value means identifying our loss should the prediction err and examining the past performance of the prediction with respect to that loss. A statistical test to do this is developed. Predictions that pass this test are said to have skill. Only skillful predictions should be used. Graphical and numerical methods to identify skill will be demonstrated. The usefulness of mammograms is explored.


Thalidomide and tuberculosis.
The anti-inflammatory and immunomodulatory effects of thalidomide have led scientists to explore its clinical therapeutic values. Thalidomide is now being considered as an adjuvant treatment for tuberculosis. This literature review examines the drug's mechanism of action and clinical applications. Thalidomide affects cytokine production and T lymphocyte proliferation. It appears that thalidomide suppresses TNF-alpha production by macrophages and thereby reduces inflammatory response. Thalidomide elevates the IFN-gamma level and modulates several other cytokines as well, noteworthily IL-2 and IL-12. Thalidomide costimulates T lymphocytes, with greater effect on CD8+ than on CD4+ T cells. This finding is important, since CD8+ T cells have been shown to be contributory to the protective immune response to Mycobacterium tuberculosis infection. The clinical application of thalidomide as part of standard tuberculosis therapy is inconclusive amid variability among reports. However, thalidomide has been shown to be an effective adjuvant for tuberculosis patients complicated with severe inflammatory reaction or wasting conditions.


Antecedent hypercortisolemia is not primarily responsible for generating hypoglycemia-associated autonomic failure.
Hypoglycemia-associated autonomic failure (HAAF) occurs commonly in patients with longstanding diabetes, placing affected patients at increased risk for severe hypoglycemia. Previous studies have suggested that hypoglycemia-induced hypercortisolemia may be responsible for blunting subsequent sympathoadrenal responses to hypoglycemia; however, this view remains highly controversial. In this work, we sought to better define the role of antecedent hypercortisolemia in generating HAAF, using two complimentary experimental models in nondiabetic human subjects: 1) antecedent hydrocortisone infusions (simulating physiologic cortisol responses to hypoglycemia) and 2) antecedent hypoglycemia, with and without concurrent blockade of endogenous cortisol production using oral metyrapone. Our results showed no effect of antecedent hypercortisolemia on epinephrine responses to subsequent hypoglycemia (area under the curve/time 280 +/- 53 vs. 337 +/- 57 pg/ml, P = 0.16). Of particular importance, selective blockade of endogenous cortisol production during antecedent hypoglycemia had no effect on subsequent counterregulatory responses to hypoglycemia. Compared with epinephrine responses following antecedent euglycemia (area under the curve/time 312 +/- 38 pg/ml), epinephrine responses were comparably blunted following antecedent hypoglycemia, regardless of whether concurrent metyrapone blockade was employed (198 +/- 28 vs. 192 +/- 28 pg/ml, P = NS). Similar results were obtained for glucagon and ACTH levels. Considered together, these observations provide strong evidence that hypoglycemia-induced hypercortisolemia is not primarily responsible for the generation of HAAF.


Nandrolone decanoate relieves joint pain in hypogonadal men: a novel prospective pilot study and review of the literature.
Testosterone is an archetypal androgenic-anabolic steroid (AAS), while its exogenous administration is considered to be the gold standard for the treatment of male hypogonadism. The benefits are not due to its intrinsic nature alone but are due to the result of its interactions with the androgen receptor (AR). As the management of hypogonadism continues to advance into the modern era, it would be preferable for modern andrologists to have multiple tools at their disposal to influence AR activity. Nandrolone, or 19-nortestosterone, is one such compound. In the following review of the literature, we examine the history, pharmacology, and clinical applications of this medication. We also present the results of our novel pilot study examining the favorable effects of nandrolone on joint pain for hypogonadal men.


A role for A-to-I RNA editing in temperature adaptation.
A-to-I RNA editing can recode mRNAs, giving organisms the option to express diverse, functionally distinct protein isoforms. Here, we propose that RNA editing is inherently geared for temperature adaptation because it tends to recode to smaller, less stabilizing amino acids. Studies on how editing affects protein function support this idea.


Tests of the roles of two diffusible substances in long-term potentiation: evidence for nitric oxide as a possible early retrograde messenger.
Although long-term potentiation (LTP) in the CA1 region of the hippocampus is initiated postsynaptically by the influx of Ca2+ through N-methyl-D-aspartate receptor channels, the maintenance of LTP seems to be at least in part presynaptic. This suggests that the postsynaptic cell releases a retrograde messenger to activate the presynaptic terminals. It is likely that this messenger is membrane-permeant and reaches the presynaptic neuron by diffusion. We therefore have investigated two major membrane-permeant candidate retrograde messengers, arachidonic acid and nitric oxide (NO). Consistent with arachidonic acid or a lipoxygenase metabolite being a retrograde messenger, the phospholipase A2 and lipoxygenase inhibitor nordihydroguaiaretic acid blocked LTP in the guinea pig CA1 region in vitro. However, arachidonic acid (up to 100 microM) did not reliably produce activity-independent LTP, and activity-dependent potentiation by arachidonic acid was blocked by DL-aminophosphonovaleric acid. Since nordihydroguaiaretic acid also interferes with signal transduction involving NO, we next examined whether inhibitors of NO synthase block LTP. NG-Nitro-L-arginine blocked LTP when given in the bath, and this inhibition was partially overcome by high concentrations of L-arginine, suggesting that the inhibitor is specific to NO synthase. NG-Nitro-L-arginine and NG-methyl-L-arginine (but not NG-methyl-D-arginine) also blocked LTP when injected intracellularly, indicating that NO synthase is located in the postsynaptic cell. The NO, in turn, seems to be released into the extracellular space, since bathing the slice with hemoglobin, a protein that binds NO and is not taken up by cells, also blocked LTP. Moreover, NO enhances spontaneous presynaptic release of transmitter from hippocampal neurons in dissociated cell culture. These data favor the idea that NO might be a retrograde messenger in LTP.


Nociceptin (1-13)NH2 inhibits stimulated calcitonin-gene-related-peptide release from primary cultures of rat trigeminal ganglia neurones.
In this work we have developed and characterized primary cultures of neonatal rat trigeminal ganglia neurones; calcitonin-gene-related-peptide (CGRP) released from cells was taken as a marker of neuronal function. A significant and consistent increase in CGRP secretion was elicited by non-specific (56 mm KCl or veratridine) or specific (capsaicin) depolarizing stimuli. This paradigm was subsequently used to investigate the effects of nociceptin, an opioid-like peptide involved in central and peripheral control of nociception. We found that the nociceptin analogue nociceptin (1-13)NH2 (NOC) did not affect baseline CGRP release, but it reduced in a concentration-dependent manner CGRP release induced by all tested stimuli. NOC-induced reduction was statistically significant from 0.01 nm onward and achieved maximal effects at 10 nm. Such effects of NOC were seemingly mediated by the activation of specific ORL1 receptors, as a well-known nociceptin antagonist, N(Phe1)nociceptin (1-13)NH2, was able to completely revert NOC inhibition of capsaicin-stimulated CGRP release.


Serum concentrations of prostate-specific antigen measured using immune extraction, trypsin digestion, and tandem mass spectrometry quantification of LSEPAELTDAVK peptide.
Prostate-specific antigen (PSA) is a 34-kDa glycoprotein with chymotrypsin-like enzyme activity that circulates both in free forms and complexed to various enzyme inhibitors including antichymotrypsin and α2-macroglobulin. Prostate-specific antigen bound to α2-macroglobulin is not detected by commercial PSA immunoassays. To develop a mass spectrometry assay that detects the same forms of PSA as the immunoassays, which could serve as a reference for harmonizing PSA immunoassays. Prostate-specific antigen was immune extracted from serum, trypsin was digested, and the LSEPAELTDAVK peptide was quantitated on an API 5000 spectrometer. Calibrators were made by adding 10% free and 90% antichymotrypsin-bound PSA to female sera. The assay was standardized to the World Health Organization 96/670 reference standard. Validation of clinical utility and comparisons with 2 immunoassays (Roche cobas and Beckman Access) were performed using frozen sera aliquots from 100 men undergoing prostate biopsy (50 negative, 50 with cancer) and 5 serial samples collected over time from 5 men with advanced prostate cancer. The antibody extraction efficiency was greater than 99%. The assay has an analytic range from 1.2 to 76 ng/mL, with precision ranging from 8.6% at 1.5 ng/mL to 5.4% at 27 ng/mL. The mass spectrometry assay correlated well with 2 immunoassays. All 3 assays showed statistically equivalent separation of prostate cancer from benign disease using receiver operating characteristic curve analysis. This mass spectrometry assay can reliably measure PSA concentrations in human serum and could serve as a reference standard for harmonizing PSA immunoassays.


A prospective study of chlamydial, mycoplasmal, and viral infections in a neonatal intensive care unit.
In a prospective study of non-bacterial infection in a neonatal intensive care unit in north west London, Chlamydia trachomatis infection was identified in 4 of 280 babies (1.4%) and was the most common cause of neonatal ophthalmia. One of the four developed pneumonitis. Ureaplasma urealyticum was found to colonise the nasopharynx in 53 of 235 babies (22.6%), with Mycoplasma hominis present in 6 of 235 babies (2.6%). There was a statistically significant association between U urealyticum colonisation and preterm birth or prolonged rupture of membranes. Colonisation occurred more commonly in babies with apnoea. Viral infection was detected in 16 of 280 babies (5.7%). Rotavirus was identified in 5 of 170 babies (2.9%) and was associated with necrotising enterocolitis in two infants and with bloody diarrhoea in another. Respiratory syncytial virus, which was identified in 4 of 280 babies (1.4%), was not associated with lower respiratory tract infection.


Chronic striatal cholinergic interneuron excitation induces clinically-relevant dystonic behavior in mice.
Dystonia is common, debilitating, often medically refractory, and difficult to diagnose. The gold standard for both clinical and mouse model dystonia evaluation is subjective assessment, ideally by expert consensus. However, this subjectivity makes translational quantification of clinically-relevant dystonia metrics across species nearly impossible. Many mouse models of genetic dystonias display abnormal striatal cholinergic interneuron excitation, but few display subjectively dystonic features. Therefore, whether striatal cholinergic interneuron pathology causes dystonia remains unknown. To address these critical limitations, we first demonstrate that objectively quantifiable leg adduction variability correlates with leg dystonia severity in people. We then show that chemogenetic excitation of striatal cholinergic interneurons in mice causes comparable leg adduction variability in mice. This clinically-relevant dystonic behavior in mice does not occur with acute excitation, but rather develops after 14 days of ongoing striatal cholinergic interneuron excitation. This requirement for prolonged excitation recapitulates the clinically observed phenomena of a delay between an inciting brain injury and subsequent dystonia manifestation and demonstrates a causative link between chronic striatal cholinergic interneuron excitation and clinically-relevant dystonic behavior in mice. Therefore, these results support targeting striatal ChIs for dystonia drug development and suggests early treatment in the window following injury but prior to dystonia onset. Chronic excitation of dorsal striatal cholinergic interneuron causes clinically-relevant dystonic phenotypes in mice.


Malignant Transformation of an HNF1a-Inactivated Hepatocellular Adenoma to Hepatocellular Carcinoma.
Hepatocellular adenomas (HCA) are rare benign tumors of the liver, occurring predominantly in females using oral contraceptives. Our case describes a 66-year-old woman presenting with a palpable mass in her upper abdomen. Contrast-enhanced computed tomography and magnetic resonance imaging showed a large exophytic mass protruding from the caudal border of liver segments IV and V, without visible metastases. Laparoscopic resection of the tumor and gallbladder was performed. Histopathological examination showed a hepatocellular carcinoma with areas of HNF1a-HCA (H-HCA). This case shows that malignant transformation is possible in H-HCA. We present our preoperative decision-making process, as well as the role of imaging techniques in this rare case.


Role of flanking sequences and phosphorylation in the recognition of the simian-virus-40 large T-antigen nuclear localization sequences by importin-alpha.
The nuclear import of simian-virus-40 large T-antigen (tumour antigen) is enhanced via phosphorylation by the protein kinase CK2 at Ser112 in the vicinity of the NLS (nuclear localization sequence). To determine the structural basis of the effect of the sequences flanking the basic cluster KKKRK, and the effect of phosphorylation on the recognition of the NLS by the nuclear import factor importin-alpha (Impalpha), we co-crystallized non-autoinhibited Impalpha with peptides corresponding to the phosphorylated and non-phosphorylated forms of the NLS, and determined the crystal structures of the complexes. The structures show that the amino acids N-terminally flanking the basic cluster make specific contacts with the receptor that are distinct from the interactions between bipartite NLSs and Impalpha. We confirm the important role of flanking sequences using binding assays. Unexpectedly, the regions of the peptides containing the phosphorylation site do not make specific contacts with the receptor. Binding assays confirm that phosphorylation does not increase the affinity of the T-antigen NLS to Impalpha. We conclude that the sequences flanking the basic clusters in NLSs play a crucial role in nuclear import by modulating the recognition of the NLS by Impalpha, whereas phosphorylation of the T-antigen enhances nuclear import by a mechanism that does not involve a direct interaction of the phosphorylated residue with Impalpha.


After being challenged by a video game problem, sleep increases the chance to solve it.
In the past years many studies have demonstrated the role of sleep on memory consolidation. It is known that sleeping after learning a declarative or non-declarative task, is better than remaining awake. Furthermore, there are reports of a possible role for dreams in consolidation of declarative memories. Other studies have reported the effect of naps on memory consolidation. With similar protocols, another set of studies indicated that sleep has a role in creativity and problem-solving. Here we hypothesised that sleep can increase the likelihood of solving problems. After struggling to solve a video game problem, subjects who took a nap (n = 14) were almost twice as likely to solve it when compared to the wake control group (n = 15). It is interesting to note that, in the nap group 9 out 14 subjects engaged in slow-wave sleep (SWS) and all solved the problem. Surprisingly, we did not find a significant involvement of Rapid Eye Movement (REM) sleep in this task. Slow-wave sleep is believed to be crucial for the transfer of memory-related information to the neocortex and implement intentions. Sleep can benefit problem-solving through the generalisation of newly encoded information and abstraction of the gist. In conclusion, our results indicate that sleep, even a nap, can potentiate the solution of problems that involve logical reasoning. Thus, sleep's function seems to go beyond memory consolidation to include managing of everyday-life events.


(4'-Acet-yloxy-1,3,1'-trioxo-1,3,4,4a,4b,5,6,7,9,9a-deca-hydro-spiro-[indene-2,9'-pyrano[4,3-a]pyrrolizin]-3'-yl)methyl acetate.
In the title compound, C23H23NO8, the dihedral angle between the five- and six-membered rings of the indene-dione moiety is 3.09 (13)°. The mean plane of the five-membered ring (which has a flat envelope conformation with the spiro C atom as the flap) is inclined to the mean plane of the central five-membered ring of the pyrrolizine unit by 76.48 (12)°. This central ring has a twist conformation on the N-C(spiro) bond. The outer ring of the pyrrolizine unit has an envelope conformation with the N atom as the flap. The mean planes of these two fused rings are inclined to one another by 65.28 (15)°. The pyran ring has a screw-boat conformation and its mean plane makes a dihedral angle of 29.50 (11)° with the mean plane of the central five-membered ring of the pyrrolizine unit. In the crystal, mol-ecules are linked via C-H⋯O hydrogen bonds, forming two-dimensional networks lying parallel to the ab plane.


Enhanced catalytic activity of polyethylenedioxythiophene towards tri-iodide reduction in DSSCs via 1-dimensional alignment using hollow carbon nanofibers.
Here, we report a highly conducting 1-dimensionally (1-D) aligned polyethylenedioxythiophene (PEDOT) along the inner and outer surfaces of a hollow carbon nanofiber (CNF) and its application as a counter electrode in a dye sensitized solar cell (DSSC). The hybrid material (CP-25) displays a conversion efficiency of 7.16% compared to 7.30% for the standard Pt counter electrode, 4.48% for bulk PEDOT and 5.56% for CNF. The enhanced conversion efficiency of CP-25 is attributed to the accomplishment of high conductivity and surface area of PEDOT through the 1-D alignment compared to its bulk counterpart. Reduced charge transfer resistance and high conductivity of CP-25 could be proven by cyclic voltammetry, impedance analysis and Tafel experiments. Further, through a long-term stability test involving efficiency profiling for 20 days, it is observed that CP-25 possesses excellent durability compared to the bulk PEDOT.


X-ray crystallographic validation of structure predictions used in computational design for protein stabilization.
Protein engineering aimed at enhancing enzyme stability is increasingly supported by computational methods for calculation of mutant folding energies and for the design of disulfide bonds. To examine the accuracy of mutant structure predictions underlying these computational methods, crystal structures of thermostable limonene epoxide hydrolase variants obtained by computational library design were determined. Four different predicted effects indeed contributed to the obtained stabilization: (i) enhanced interactions between a flexible loop close to the N-terminus and the rest of the protein; (ii) improved interactions at the dimer interface; (iii) removal of unsatisfied hydrogen bonding groups; and (iv) introduction of additional positively charged groups at the surface. The structures of an eightfold and an elevenfold mutant showed that most mutations introduced the intended stabilizing interactions, and side-chain conformations were correctly predicted for 72-88% of the point mutations. However, mutations that introduced a disulfide bond in a flexible region had a larger influence on the backbone conformation than predicted. The enzyme active sites were unaltered, in agreement with the observed preservation of catalytic activities. The structures also revealed how a c-Myc tag, which was introduced for facile detection and purification, can reduce access to the active site and thereby lower the catalytic activity. Finally, sequence analysis showed that comprehensive mutant energy calculations discovered stabilizing mutations that are not proposed by the consensus or B-FIT methods.


Natural infectivity of Anopheles species from the Pacific and Atlantic Regions of Colombia.
Malaria is an important public health problem in Colombia. Among the major vectors in Colombia, Anopheles albimanus is recognized for its importance on the Pacific Coast where it is the predominant species; it is also found in the Atlantic Coast, although its vectorial role in this region is not clear. We examined the occurrence of An. albimanus in four localities of the Pacific and three of the Atlantic Coast. Morphological identification of problematic specimens was confirmed by a molecular assay. All identified mosquitoes at these sites, including An. albimanus, were also tested for malaria parasite infection. From 12,189 anophelines collected, 6370 were from the Pacific Coast, and corresponded to 99% An. albimanus, 0.8% Anopheles neivai, and three other species at <0.2%. From the Atlantic Coast we identified 5819 specimens with 61% An. albimanus, 36% Anopheles triannulatus s.l. and five other species at <2%. In both coasts, species present at lower percentages included several incriminated as vectors in neighboring countries. Six Pacific Coast specimens were infected with malaria parasites: four An. albimanus, two with Plasmodium vivax VK247, one with P. vivax VK210 and one with Plasmodium falciparum; two An. neivai with P. falciparum. Our data support the continued predominance of An. albimanus in the Pacific Coast, and demonstrate that this species is the most abundant in the Atlantic Coast as well.


Interventional pulmonology: a new medical specialty.
Interventional pulmonology (IP) is the newest chapter in respiratory medicine. IP includes both diagnostic and therapeutic methods. Nanotechnology, in both instrumental engineering and optical imaging, will further advance this competitive discipline towards cell diagnosis and therapy as part of the future's personalized medicine.


Amyloid-beta peptide and tau protein crosstalk in Alzheimer's disease.
Alzheimer's disease is a neurodegenerative disease that accounts for most of the 50-million dementia cases worldwide in 2018. A large amount of evidence supports the amyloid cascade hypothesis, which states that amyloid-beta accumulation triggers tau hyperphosphorylation and aggregation in form of neurofibrillary tangles, and these aggregates lead to inflammation, synaptic impairment, neuronal loss, and thus to cognitive decline and behavioral abnormalities. The poor correlation found between cognitive decline and amyloid plaques, have led the scientific community to question whether amyloid-beta accumulation is actually triggering neurodegeneration in Alzheimer's disease. The occurrence of tau neurofibrillary tangles better correlates to neuronal loss and clinical symptoms and, although amyloid-beta may initiate the cascade of events, tau impairment is likely the effector molecule of neurodegeneration. Recently, it has been shown that amyloid-beta and tau cooperatively work to impair transcription of genes involved in synaptic function and, more importantly, that downregulation of tau partially reverses transcriptional perturbations. Despite mounting evidence points to an interplay between amyloid-beta and tau, some factors could independently affect both pathologies. Thus, the dual pathway hypothesis, which states that there are common upstream triggers causing both amyloid-beta and tau abnormalities has been proposed. Among others, the immune system seems to be strongly involved in amyloid-beta and tau pathologies. Other factors, as the apolipoprotein E ε4 isoform has been suggested to act as a link between amyloid-beta and tau hyperphosphorylation. Interestingly, amyloid-beta-immunotherapy reduces not only amyloid-beta but also tau levels in animal models and in clinical trials. Likewise, it has been shown that tau-immunotherapy also reduces amyloid-beta levels. Thus, even though amyloid-beta immunotherapy is more advanced than tau-immunotherapy, combined amyloid-beta and tau-directed therapies at early stages of the disease have recently been proposed as a strategy to stop the progression of Alzheimer's disease.


Effect of the finite pixel boundary on the angular emission characteristics of top-emitting organic light-emitting diodes.
We numerically investigate the effect of the pixel boundary on the angular emission characteristics of top-emitting organic light-emitting diodes (OLEDs) using the finite element method. A three-dimensional OLED structure has the square pixel boundary, which is surrounded by the pixel definition layer. The angular emission characteristics based on the Poynting vectors are calculated in various positions of a Hertz dipole emitter within the pixel boundary. When the dipole emitter is located near the center of the square pixel, the angular emission characteristics have a symmetric forward-directed pattern, which is similar to the angular emission pattern calculated by the thin-film-based optical model. When the position of the dipole emitter is close to the pixel boundary, the angular emission pattern becomes asymmetric because the optical reflections from the pixel boundary in the horizontal direction affect the emission pattern of the dipole emitter. The total angular emission characteristics of the top-emitting OLED are obtained by summing the individual angular emission pattern of the whole dipole emitters, which are assumed to be uniformly distributed in the two-dimensional emission plane. The asymmetrical angular emission characteristics of the dipole emitters near the pixel boundary contribute to narrowing the total angular emission pattern.


Alcohol-induced miR-27a regulates differentiation and M2 macrophage polarization of normal human monocytes.
Alcohol abuse is a leading cause of liver disease characterized by liver inflammation, fatty liver, alcoholic hepatitis, or liver cirrhosis. Immunomodulatory effects of alcohol on monocytes and macrophages contribute to alcoholic liver disease. Alcohol use, an independent risk factor for progression of hepatitis C virus (HCV) infection-mediated liver disease, impairs host defense and alters cytokine production and monocyte/macrophage activation. We hypothesized that alcohol and HCV have synergistic effects on the phenotype and function of monocytes. Our data show that acute alcohol binge drinking in healthy volunteers results in increased frequency of CD16(+) and CD68(+) and M2-type (CD206(+), dendritic cell [DC]-SIGN(+)-expressing and IL-10-secreting) circulating CD14(+) monocytes. Expression of HCV-induced CD68 and M2 markers (CD206 and DC-SIGN) in normal monocytes was further enhanced in the presence of alcohol. The levels of microRNA (miR)-27a was significantly upregulated in monocytes cultured in the presence of alcohol or alcohol and HCV as compared with HCV alone. The functional role of miR-27a in macrophage polarization was demonstrated by transfecting monocytes with an miR-27a inhibitor that resulted in reduced alcohol- and HCV- mediated monocyte activation (CD14 and CD68 expression), polarization (CD206 and DC-SIGN expression), and IL-10 secretion. Overexpression of miR-27a in monocytes enhanced IL-10 secretion via activation of the ERK signaling pathway. We found that miR-27a promoted ERK phosphorylation by downregulating the expression of ERK inhibitor sprouty2 in monocytes. Thus, we identified that sprouty2 is a target of miR-27a in human monocytes. In summary, our study demonstrates the regulatory role of miR-27a in alcohol-induced monocyte activation and polarization.


Pooling of animal experimental data reveals influence of study design and publication bias.
The extensive neuroprotective literature describing the efficacy of candidate drugs in focal ischemia has yet to lead to the development of effective stroke treatments. Ideally, the choice of drugs taken forward to clinical trial should be based on an unbiased assessment of all available data. Such an assessment might include not only the efficacy of a drug but also the in vivo characteristics and limits--in terms of time window, dose, species, and model of ischemia used--to that efficacy. To our knowledge, such assessments have not been made. Nicotinamide is a candidate neuroprotective drug with efficacy in experimental stroke, but the limits to and characteristics of that efficacy have not been fully described. Systematic review and modified meta-analysis of studies of experimental stroke describing the efficacy of nicotinamide. The search strategy ensured ascertainment of studies published in full and those published in abstract only. DerSimonian and Laird random effects meta-analysis was used to account for heterogeneity between studies. Nicotinamide improved outcome by 0.287 (95% confidence interval 0.227 to 0.347); it was more effective in temporary ischemia models, after intravenous administration, in animals without comorbidities, and in studies published in full rather than in abstract. Studies scoring highly on a quality measure gave more precise estimates of the global effect. Meta-analysis provides an effective technique for the aggregation of data from experimental stroke studies. We propose new standards for reporting such studies and a systematic approach to aggregating data from the neuroprotective literature.


Measuring spike pattern reliability with the Lempel-Ziv-distance.
Spike train distance measures serve two purposes: to measure neuronal firing reliability, and to provide a metric with which spike trains can be classified. We introduce a novel spike train distance based on the Lempel-Ziv complexity that does not require the choice of arbitrary analysis parameters, is easy to implement, and computationally cheap. We determine firing reliability in vivo by calculating the deviation of the mean distance of spike trains obtained from multiple presentations of an identical stimulus from a Poisson reference. Using both the Lempel-Ziv-distance (LZ-distance) and a distance focussing on coincident firing, the pattern and timing reliability of neuronal firing is determined for spike data obtained along the visual information processing pathway of macaque monkey (LGN, simple and complex cells of V1, and area MT). In combination with the sequential superparamagnetic clustering algorithm, we show that the LZ-distance groups together spike trains with similar but not necessarily synchronized firing patterns. For both applications, we show how the LZ-distance gives additional insights, as it adds a new perspective on the problem of firing reliability determination and allows neuron classifications in cases, where other distance measures fail.


Deregulation of Long Intergenic Non-coding RNAs in CD4+ T Cells of Lamina Propria in Crohn's Disease Through Transcriptome Profiling.
The aetiology of Crohn's disease [CD] involves immune dysregulation in a genetically susceptible individual. Genome-wide association studies [GWAS] have identified 200 loci associated with CD, ulcerative colitis, or both, most of which fall within non-coding DNA regions. Long non-coding RNAs [lncRNAs] regulate gene expression by diverse mechanisms and have been associated with disease activity in inflammatory bowel disease. However, disease-associated lncRNAs have not been characterised in pathogenic immune cell populations. Terminal ileal samples were obtained from 22 CD patients and 13 controls. RNA from lamina propria CD4+ T cells was sequenced and long intergenic non-coding RNAs [lincRNAs] were detected. Overall expression patterns, differential expression [DE], and pathway and gene enrichment analyses were performed. Knockdown of novel lincRNAs XLOC_000261 and XLOC_000014 was performed. Expression of Th1 or Th17-associated transcription factors, T-bet and RORγt, respectively, was assessed by flow cytometry. A total of 6402 lincRNAs were expressed, 960 of which were novel. Unsupervised clustering and principal component analysis showed that the lincRNA expression discriminated patients from controls. A total of 1792 lincRNAs were DE, and 295 [79 novel; 216 known] mapped to 267 of 5727 DE protein-coding genes. The novel lincRNAs were enriched in inflammatory and Notch signalling pathways [p <0.05]. Furthermore, DE lincRNAs in CD patients were more frequently found in DNA regions with known inflammatory bowel disease [IBD]-associated loci. The novel lincRNA XLOC_000261 negatively regulated RORγt expression in Th17 cells. We describe a novel set of DE lincRNAs in CD-associated CD4+ cells and demonstrate that novel lincRNA XLOC_000261 appears to negatively regulate RORγt protein expression in Th17 cells.


Effects of an educational compliance enhancement programme and therapeutic drug monitoring on treatment adherence in depressed patients managed by general practitioners.
Medication non-adherence is a major obstacle in the treatment of affective disorders. The primary objective of this study was to evaluate two different interventions to improve adherence to antidepressant drugs. Secondary objectives included response to treatment, relation between adherence and response, patient satisfaction and tolerability. A randomized controlled design was used to assess the effect of a patient educational compliance enhancing programme (CP) and therapeutic drug monitoring in 1031 major depressed patients treated with sertraline for 24 weeks and managed by their general practitioner. Adherence was measured by questioning, measurable serum levels of sertraline and desmethylsertraline, appointments kept and a composite index including all three methods. Treatment adherence was found in 37-70% of patients, depending on the method used. Neither of the interventions resulted in a significant increase in adherence rate. However, significantly more patients in the CP group had responded at week 24 compared to patients in the control group. Overall, significantly more adherent patients responded to treatment compared to non-adherent patients, regardless of method used to determine adherence. This large study demonstrates that treatment response increases when using an educational compliance programme and that a strong relationship between treatment adherence and response exists.


Coactivation by OCA-B: definition of critical regions and synergism with general cofactors.
Molecular dissection of the B-cell-specific transcription coactivator OCA-B has revealed distinct regions important, respectively, for recruitment to immunoglobulin promoters through interaction with octamer-bound Oct-1 and for subsequent coactivator function. Further analysis of general coactivator requirements showed that selective removal of PC4 from the essential USA fraction severely impairs Oct-1 and OCA-B function in a cell-free system reconstituted with partially purified factors. Full activity can be restored by the combined action of recombinant PC4 and the PC4-depleted USA fraction, thus suggesting a joint requirement for PC4 and another, USA-derived component(s) for optimal function of Oct-1/OCA-B in the reconstituted system. Indeed, USA-derived PC2 was found to act synergistically with PC4 in reproducing the function of intact USA in the assay system. Consistent with the requirement for PC4 in the reconstituted system, OCA-B was found to interact directly with PC4. Surprisingly, however, removal of PC4 from the unfractionated nuclear extract has no detrimental effect on OCA-B/Oct-1-dependent transcription. These results lead to a general model for the synergistic function of activation domains in Oct-1 and OCA-B (mediated by the combined action of the multiple USA components) and, further, suggest a functional redundancy in general coactivators.


Do fragments and glycosylated isoforms of alpha-1-antitrypsin in CSF mirror spinal pathophysiological mechanisms in chronic peripheral neuropathic pain? An exploratory, discovery phase study.
Post-translational modifications (PTMs) generate a tremendous protein diversity from the ~ 20,000 protein-coding genes of the human genome. In chronic pain conditions, exposure to pathological processes in the central nervous system could lead to disease-specific PTMs detectable in the cerebrospinal fluid (CSF). In a previous hypothesis-generating study, we reported that seven out of 260 CSF proteins highly discriminated between neuropathic pain patients and healthy controls: one isoform of angiotensinogen (AG), two isoforms of alpha-1-antitrypsin (AT), three isoforms of haptoglobin (HG), and one isoform of pigment epithelium-derived factor (PEDF). The present study had three aims: (1) To examine the multivariate inter-correlations between all identified isoforms of these seven proteins; (2) Based on the results of the first aim, to characterize PTMs in a subset of interesting proteins; (3) To regress clinical pain data using the 260 proteins as predictors, thereby testing the hypothesis that the above-mentioned seven discriminating proteins and/or the characterized isoforms/fragments of aim (2) would be among the proteins having the highest predictive power for clinical pain data. CSF samples from 11 neuropathic pain patients and 11 healthy controls were used for biochemical analysis of protein isoforms. PTM characterization was performed using enzymatic reaction assay and mass spectrometry. Multivariate data analysis (principal component analysis and orthogonal partial least square regression) was applied on the quantified protein isoforms. We identified 5 isoforms of AG, 18 isoforms of AT, 5 isoforms of HG, and 5 isoforms of PEDF. Fragments and glycosylated isoforms of AT were studied in depth. When regressing the pain intensity data of patients, three isoforms of AT, two isoforms of PEDF, and one isoform of angiotensinogen "reappeared" as major results, i.e., they were major findings both when comparing patients with healthy controls and when regressing pain intensity in patients. Altered levels of fragments and/or glycosylated isoforms of alpha-1-antitrypsin might mirror pathophysiological processes in the spinal cord of neuropathic pain patients. In particular, we suggest that a putative disease-specific combination of the levels of two different N-truncated fragments of alpha-1-antitrypsin might be interesting for future CSF and/or plasma biomarker investigations in chronic neuropathic pain.


Will eradication of Helicobacter pylori infection influence the risk of gastric cancer?
Gastric adenocarcinoma is a disease of high mortality and poor prognosis that is second only to lung cancer as a leading cause of cancer-related deaths worldwide. Although gastric cancer has a multifactorial etiology, infection with Helicobacter pylori is highly associated with its development. New information on bacterial and host genetics and results of epidemiologic studies suggest that better identification of individuals at high risk for gastric malignancy may be possible. Studies suggest that cure of H pylori infection may be associated with retardation of glandular atrophy and intestinal metaplasia but not reversal of dysplasia. Theoretically, it is attractive to believe that eradication of H pylori infection might prevent gastric cancer; however, studies supporting this hypothesis are not yet available. Public policy strategies for the identification of patients at risk for H pylori-related gastric malignancy are likely to be complex, but testing and treating for the infection earlier rather than later in life is anticipated to be the more beneficial approach.


Efficacy and safety of the long-acting C5 inhibitor ravulizumab in patients with atypical hemolytic uremic syndrome triggered by pregnancy: a subgroup analysis.
Atypical hemolytic uremic syndrome (aHUS) triggered by pregnancy is a rare disease caused by dysregulation of the alternative complement pathway that occurs in approximately 1 in 25,000 pregnancies. The 311 phase 3 trial (NCT02949128) showed that ravulizumab, a long-acting C5 inhibitor obtained through selective modifications to eculizumab, is efficacious in inhibiting complement-mediated thrombotic microangiopathy (TMA) in patients with aHUS. In this analysis, we report outcomes in a subgroup of patients from the 311 study who developed TMA postpartum. This was a phase 3, multicenter trial evaluating efficacy and safety of ravulizumab in adults (≥18 years of age) with aHUS naïve to complement inhibitor treatment. The primary endpoint was complete TMA response (simultaneous platelet count normalization [≥150 × 109/L], lactate dehydrogenase normalization [≤246 U/L] and 25% improvement in serum creatinine) through the 183-day initial evaluation period. Additional efficacy endpoints included time to complete TMA response, hematologic normalization, and dialysis requirement status. Eight patients presenting with TMA postpartum (median age of 37.7 [range; 22.1-45.2] years) were diagnosed with aHUS and received ≥1 dose of ravulizumab. Five patients (63%) were on dialysis at baseline. Complete TMA response was achieved in 7/8 patients (87.5%) in a median time of 31.5 days. Hematologic normalization was observed in all patients. All patients on dialysis at baseline discontinued dialysis within 21 days after treatment with ravulizumab. All patients showed continued improvements in the estimated glomerular filtration rate from baseline to Day 183. Three possible treatment-related adverse events were observed in 2 patients (arthralgia and nasopharyngitis [both non-severe]; urinary tract infection). No deaths or meningococcal infections occurred. Treatment with ravulizumab provided immediate and complete C5 inhibition, resulting in rapid clinical and laboratory improvements and complete TMA response through 183 days in patients with aHUS triggered by pregnancy. The safety profile observed in this subset of patients analysed is consistent with the 311 study investigating ravulizumab in patients with aHUS naïve to complement treatment. Clinical trial identifier: NCT02949128 .


[Minimally invasive cytoselective radiation therapy using boron neutron capture reaction].
The cell-killing effect of boron neutron capture therapy (BNCT) is due to the nuclear reaction of two essentially nontoxic species, boron-10 ((10)B) and thermal neutrons, whose destructive effect is well observed in boron-loaded tissues. High accumulation and selective delivery of boron into tumor tissue are the most important requirements to achieve efficient neutron capture therapy of cancers. This review focuses on liposomal boron delivery system (BDS) as a recent promising approach that meet these requirements for BNCT. BDS involves two strategies: (1) encapsulation of boron in the aqueous core of liposomes and (2) accumulation of boron in the liposomal bilayer. In this review, recent development of liposomal boron delivery system is summarized.


Mitochondrial DNA phylogeography of Caiman crocodilus in Mesoamerica and South America.
The Neotropical crocodylian species, Caiman crocodilus, is widely distributed through Mesoamerica, northern South America, and the Amazon basin. Four subspecies are recognized within C. crocodilus, suggesting some geographic variation in morphology. In this study, we utilized mitochondrial DNA (mtDNA) sequence data from 45 individuals of C. crocodilus throughout its range to infer its evolutionary history and population structure, as well as to evaluate genealogical support for subspecies and their geographic distributions. Our molecular phylogenetic results identified five mtDNA haplotype clades with a mean sequence divergence of 3.4%, indicating considerable evolutionary independence among phylogeographic lineages. Our results were also broadly consistent with current subspecific taxonomy, with some important additional findings. First, we found substantial genetic structuring within C. c. fuscus from southern Mesoamerica. Second, though we confirmed the existence of a widespread Amazonian clade, we also discovered a cryptic and divergent mtDNA lineage that was indistinguishable from C. c. crocodilus based on external morphology. Third, we confirm the status of C. c. chiapasius as a distinct evolutionary lineage, and provide evidence that C. c. fuscus may be moving northward and hybridizing with C. c. chiapasius in northern Mesoamerica. Finally, our results parallel previous phylogeographic studies of other organisms that have demonstrated significant genetic structure over shorter geographic distances in Mesoamerica compared with Amazonia. We support conservation efforts for all five independent lineages within C. crocodilus, and highlight the subspecies C. c. chiapasius as a unit of particular conservation concern.


Isolation and characterisation of an anticryptococcal protein in human cerebrospinal fluid.
An earlier study reported that human cerebrospinal fluid (CSF) has fungistatic activity for Cryptococcus neoformans. The present study reports that molecular sieve fractionation of concentrated CSF yielded three protein peaks, one of which (p2) had anticryptococcal activity. On a DEAE-Sephacel anion-exchange column the active molecular sieve peak (p2) gave two peaks that contained anticryptococcal activity. The first (DEAE-1) eluted with 0.1 M NaCl and the second (DEAE-2) eluted with 0.2 M NaCl in buffer. Fungistatic activity of DEAE-1 was reversed by FeCl3. Moreover, FeCl3 reversed inhibition of C. neoformans growth by CSF. In contrast, activity of DEAE-2 was not reversed by FeCl3, indicating that inhibition was produced by an iron-independent mechanism. Immunoblot assays showed that transferrin was present in DEAE-1 but not in DEAE-2, whereas albumin was present in DEAE-2 but not in DEAE-1. On NuPAGE, DEAE-1 protein migrated as a single band corresponding to transferrin and DEAE-2 protein gave a single band corresponding to albumin. In control experiments, human serum albumin subjected to the same isolation protocol acquired anticryptococcal activity similar to that of DEAE-2. Therefore, CSF albumin (DEAE-2) activity was associated with the isolation protocol. These data indicate that transferrin, present in or isolated from CSF, sequesters trace amounts of ferric iron, inhibits growth of C. neoformans and acts as an innate defence mechanism.


Comparative genomic analysis of Brazilian Leptospira kirschneri serogroup Pomona serovar Mozdok.
Leptospira kirschneri is one of the pathogenic species of the Leptospira genus. Human and animal infection from L. kirschneri gained further attention over the last few decades. Here we present the isolation and characterisation of Brazilian L. kirschneri serogroup Pomona serovar Mozdok strain M36/05 and the comparative genomic analysis with Brazilian human strain 61H. The M36/05 strain caused pulmonary hemorrhagic lesions in the hamster model, showing high virulence. The studied genomes presented high symmetrical identity and the in silico multilocus sequence typing analysis resulted in a new allelic profile (ST101) that so far has only been associated with the Brazilian L. kirschneri serogroup Pomona serovar Mozdok strains. Considering the environmental conditions and high genomic similarity observed between strains, we suggest the existence of a Brazilian L. kirschneri serogroup Pomona serovar Mozdok lineage that could represent a high public health risk; further studies are necessary to confirm the lineage significance and distribution.


A novel resistance mechanism to triclosan that suggests horizontal gene transfer and demonstrates a potential selective pressure for reduced biocide susceptibility in clinical strains of Staphylococcus aureus.
The widely used biocide triclosan selectively targets FabI, the NADH-dependent trans-2-enoyl-acyl carrier protein reductase, which is an important target for narrow-spectrum antimicrobial drug development. In relation to the growing concern about biocide resistance, we compared in vitro mutants and clinical isolates of Staphylococcus aureus with reduced triclosan susceptibility. Clinical isolates of S. aureus as well as laboratory-generated mutants were assayed for minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) phenotypes and genotypes related to reduced triclosan susceptibility. A potential epidemiological cut-off (ECOFF) MBC of >4 mg/L was observed for triclosan in clinical isolates of S. aureus. These showed significantly lower MICs and higher MBCs than laboratory mutants. These groups of strains also had few similarities in the triclosan resistance mechanism. Molecular analysis identified novel resistance mechanisms linked to the presence of an additional sh-fabI allele derived from Staphylococcus haemolyticus. The lack of predictive value of in-vitro-selected mutations for clinical isolates indicates that laboratory tests in the present form appear to be of limited value. More importantly, detection of sh-fabI as a novel resistance mechanism with high potential for horizontal gene transfer demonstrates for the first time that a biocide could exert a selective pressure able to drive the spread of a resistance determinant in a human pathogen.


Rat model of Achilles tendon disorder. A pilot study.
Three-month-old male rats were subjected 3 times weekly for 1 h to eccentric exercise of one triceps surae muscle (30 stimulations/min) under general anesthesia in order to induce Achilles tendon disorder corresponding to paratenonitis and tendinosis in man. Net muscle work during the sessions ranged between 0.67 and 4.37 mJ (mean 1.72, SD 0.77). After 9 and 13 sessions, respectively, 2 rats started to show gait alterations during the functional test which was performed 2-3 times weekly. These rats were killed after additional sessions which showed a worsening of the limp. The other trained rats and controls did not limp and were killed after 7-11 weeks. Histologic evaluation of the Achilles tendons from the exercised limb showed in the majority of the cases hypervascularization, increased number of nerve filaments and increased immunoreactivity for substance P and calcitonin gene-related peptide. The tendons from the nonstimulated limb looked normal. The distribution of collagen types I and II appeared normal in the tendon and its insertion to the calcaneus. Inflammation of the epi- and paratenon could be provoked in the rat, but tendon changes corresponding to chronic tendinosis did not develop within 11 weeks with the used training regime. The clinical relevance of this model for chronic tendon disease needs to be evaluated further.


New perspectives on anaerobic methane oxidation.
Anaerobic methane oxidation is a globally important but poorly understood process. Four lines of evidence have recently improved our understanding of this process. First, studies of recent marine sediments indicate that a consortium of methanogens and sulphate-reducing bacteria are responsible for anaerobic methane oxidation; a mechanism of 'reverse methanogenesis' was proposed, based on the principle of interspecies hydrogen transfer. Second, studies of known methanogens under low hydrogen and high methane conditions were unable to induce methane oxidation, indicating that 'reverse methanogenesis' is not a widespread process in methanogens. Third, lipid biomarker studies detected isotopically depleted archaeal and bacterial biomarkers from marine methane vents, and indicate that Archaea are the primary consumers of methane. Finally, phylogenetic studies indicate that only specific groups of Archaea and SRB are involved in methane oxidation. This review integrates results from these recent studies to constrain the responsible mechanisms.


Calcium-dependent enhancement of myocardial diastolic tone and energy utilization dissociates systolic work and oxygen consumption during low sodium perfusion.
The relationships and correlations among functional, metabolic, and ionic consequences of low sodium perfusion were studied in isovolumic, retrograde-aortic perfused working rat hearts by 31P nuclear magnetic resonance, oxygen consumption, and atomic absorption spectrometry. Reduction of perfusate sodium from 144 to 74, 51, 39, and 25 mM in four separate groups of hearts via lithium substitution for 15 minutes decreased cell sodium to mean values of 62, 51, 43, and 36 mumol/g dry weight, respectively (P less than 0.001 vs. control of 107). There was a transient rise and then a fall in developed pressure and a decline in phosphocreatine and adenosine triphosphate, all of which were graded and correlated with perfusate sodium (P less than 0.01 for all parameters vs. perfusate sodium). This was accompanied by a 2- to 7-fold elevation of diastolic pressure while oxygen consumption remained near control levels. All parameters except adenosine triphosphate returned toward baseline values when normal perfusate sodium was reintroduced. Although cell calcium as measured by atomic absorption spectrometry did not differ among the groups, the functional and metabolic changes did not occur if the sodium steps were performed in reduced perfusate calcium (0.08 mM). In hearts in which systolic function was obliterated by verapamil, exposure to zero sodium caused a 4-fold increase in oxygen consumption, an increase in diastolic pressure, and a reduction of high energy phosphates. In the presence of ryanodine, a specific inhibitor of sarcoplasmic reticulum calcium release, the metabolic changes did not occur, and the excess oxygen consumption in zero sodium was substantially reduced. Thus, the effect of lowered perfusate sodium in beating hearts, i.e., to dissociate oxygen consumption and systolic function, and to increase diastolic pressure and its effect in arrested hearts to increase oxygen consumption, are calcium dependent, energy consuming, and modulated by sarcoplasmic reticulum calcium cycling.


Importins and exportins as therapeutic targets in cancer.
The nuclear transport proteins, importins and exportins (karyopherin-β proteins), may play an important role in cancer by transporting key mediators of oncogenesis across the nuclear membrane in cancer cells. During nucleocytoplasmic transport of tumor suppressor proteins and cell cycle regulators during the processing of these proteins, aberrant cellular growth signaling and inactivation of apoptosis can occur, both critical to growth and development of tumors. Karyopherin-β proteins bind to these cargo proteins and RanGTP for active transport across the nuclear membrane through the nuclear pore complex. Importins and exportins are overexpressed in multiple tumors including melanoma, pancreatic, breast, colon, gastric, prostate, esophageal, lung cancer, and lymphomas. Furthermore, some of the karyopherin-β proteins such as exportin-1 have been implicated in drug resistance in cancer. Importin and exportin inhibitors are being considered as therapeutic targets against cancer and have shown preclinical anticancer activity. Moreover, synergistic activity has been observed with various chemotherapeutic and targeted agents. However, clinical development of the exportin-1 inhibitor leptomycin B was stopped due to adverse events, including vomiting, anorexia, and dehydration. Selinexor, a selective nuclear export inhibitor, is being tested in multiple clinical trials both as a single agent and in combination with chemotherapy. Selinexor has demonstrated clinical activity in multiple cancers, especially acute myelogenous leukemia and multiple myeloma. The roles of other importin and exportin inhibitors still need to be investigated clinically. Targeting the key mediators of nucleocytoplasmic transport in cancer cells represents a novel strategy in cancer intervention with the potential to significantly affect outcomes.


Why do patients change their general practitioner? A postal questionnaire study of patients in Avon.
The aim of this confidential postal questionnaire study was to determine why patients change their general practitioner. Among 1423 patients who changed their general practitioner without changing their address the reason most commonly given for leaving a general practitioner was distance (41%)--three quarters of these patients mentioned only distance. Just over one third of respondents (35%) mentioned dissatisfaction with the personal care given by the general practitioner and 36% mentioned dissatisfaction with practice organization. Specific patient requirements, such as a woman doctor, were not commonly mentioned. Convenience was the commonest reason given for choosing a new doctor (53%), with recommendation or reputation mentioned by 36% of respondents and positive expectations of service by 37%. The proportion of patients that changed from any one practice was small. This type of survey may prove a useful method for family health services authorities to identify practices with a higher than normal level of patient dissatisfaction.


Hepatic hydatid cyst presenting as a cutaneous fistula.
Hepatic hydatid cysts are usually asymptomatic. Nevertheless, they may rupture, causing anaphylactic shock or fistulation. Cutaneous fistulae caused by ruptured hepatic hydatid cysts are extremely rare. Herein, we report a case of infected cutaneous fistula caused by a ruptured hepatic hydatid cyst. A 57-year-old man presented to Al-Ain Hospital complaining of swelling in his right upper quadrant (RUQ) of 5 months' duration. The abdomen was soft, having a fluctuant tender swelling of 12 cm × 15 cm in the RUQ associated with a pus discharging fistula. The patient was admitted with a provisional diagnosis of abdominal wall abscess with pending sepsis. Surgical incision and drainage were performed under general anesthesia. Initially, around 15 ml of pus was drained, followed by the removal of multiple sized transparent cysts typical of hydatid disease. Postoperative abdominal computed tomography (CT) scan showed multiloculated hepatic cysts in the sixth, seventh, and left lobes with the involvement of the abdominal wall. The patient was treated with oral albendazole 400 mg twice daily for 30 days. Repeated CT scan at 4-month follow-up showed a significant reduction of size of the cysts, indicating proper response to treatment. A cutaneous fistula as a complication of a ruptured hepatic hydatid cyst is extremely rare. Awareness of this complication, especially in endemic areas, and using proper imaging and serological tests are vital for reaching a proper diagnosis.


Iron, microbiota and colorectal cancer.
Iron deficiency and anaemia are common in colorectal cancer. Replacement with oral or intravenous iron effectively treats this deficiency. However, mechanistic and population studies suggest that excess iron promotes colorectal carcinogenesis. Growing research into gut microbiota and dysbiosis suggests one explanation for this association. Iron is growth limiting for many pathogenic bacteria and may promote a shift in the ratio of pathogenic to protective bacteria. This may increase the toxic bacterial metabolites, promoting inflammation and carcinogenesis. This has important implications as we seek to correct anaemia in our patients.


Salivary gland epithelial neoplasms in pediatric population: a single-institute experience with a focus on the histologic spectrum and clinical outcome.
Salivary gland epithelial neoplasms are rare in children and adolescents, with only a handful of large series having been published. A retrospective study was conducted for 57 cases in patients 20 years or younger. The tumors were located in the parotid (n=36), submandibular gland (n=7), and minor salivary glands (n=14). Nineteen (33%) tumors were pleomorphic adenoma, whereas the remaining (67%) were malignant. The histologic types of carcinomas were mucoepidermoid carcinoma (MEC, n=19, 33%), acinic cell carcinoma (n=7, 12%), adenoid cystic carcinoma (n=6, 11%), secretory carcinoma (mammary analogue) (SC, n=4, 7%), and myoepithelial carcinoma (n=2, 4%). Ninety-three percent (13/14) of the minor and 58% (25/43) of the major salivary gland tumors were malignant. A 7-year-old girl (2%) with a high-grade MEC died from her disease because of uncontrollable locoregional recurrence. Seven patients (16%) developed recurrence including 2 distant metastases from adenoid cystic carcinoma and 6 locoregional recurrences (2 pleomorphic adenomas, 1 SC, 1 myoepithelial carcinoma, 1 adenoid cystic carcinoma, and 1 MEC). The following parameters were associated with decreased disease-free survival in malignant tumors: elevated mitotic index of >4/10 high-power fields (log-rank test, P<.001), and advanced American Joint Committee on Cancer pT (P=.029) and pN stage (P<.001). In conclusion, myoepithelial carcinoma and SC can occur in the pediatric population and should be considered in the differential diagnosis. Salivary gland malignancies in children appear to have better clinical outcome, associated with a 10-year recurrence-free survival rate of 74% and a 10-year disease-specific survival of 94%.


Chiropractic care, including craniosacral therapy, during pregnancy: a static-group comparison of obstetric interventions during labor and delivery.
To determine whether the addition of chiropractic care including craniosacral therapy to a regimen of standard obstetric pregnancy results in fewer obstetric interventions during labor and delivery. Retrospective, case-matched, static-group comparison. The study group was obtained from a college faculty-based clinic and received chiropractic care in addition to their routine obstetrical care. The setting for the comparison group was unkown, but the care rendered was presumed to be primary medical obstetric care only. A consecutive sample of 63 pregnant women who sought chiropractic care within the period under study. The reason for seeking care was not necessarily related to the pregnancy. The sample was primarily between 18 and 35 yr, non-Hispanic caucasian and primiparous. After selection and matching criteria, 35 patients remained in the study group. Chiropractic care and craniosacral therapy delivered during pregnancy vs. unknown care within the same county. Obstetric interventions during labor and delivery as reported by the birth attendant on the certificate of live birth. No statistical differences were detected in the rates of obstetric interventions used during labor or delivery between the two samples. Approximate large-sample 95% confidence intervals are provided. Because of the limitations in the design of the project, this study provides no evidence that the addition of chiropractic care and craniosacral therapy during pregnancy results in any observable benefit or detriment with regard to obstetric interventions used during labor and delivery and that chiropractic care for pregnancy-related neuromusculoskeletal disorders should not complicate labor or delivery.


Multimeric Coactivator Complexes for Steroid/Nuclear Receptors.
Nuclear receptors regulate transcription in direct response to their cognate hormonal ligands. Ligand binding leads to the dissociation of corepressors and the recruitment of coactivators. Many of these factors, acting in large complexes, have emerged as chromatin remodelers through intrinsic histone-modifying activities or through other novel functions. In addition, other ligand-recruited complexes appear to act more directly on the transcriptional apparatus, suggesting that transcriptional regulation by nuclear receptors might involve a process of both chromatin alterations and direct recruitment of key initiation components at regulated promoters.


CRP-binding bacteriophage as a new element of layer-by-layer assembly carbon nanofiber modified electrodes.
Recently, bacteriophage particles have started to be applied as a new biomaterial for developing sensing platforms. They can be used as both a recognition element or/and as building blocks, template/scaffold. In this paper, we studied a bacteriophage selected through phage-display technology. The chosen bacteriophage acted as a building block for creating a carbon nanofiber-based electrode and as a new receptor/binding element that recognizes C-reactive protein (CRP) - one of the markers of inflammatory processes in the human body. The binding efficiency of the selected phage towards CRP is two orders of magnitude higher than in the wild type. We demonstrate that the phage-based sensor is selective against other proteins. Finally, we show that layer-by-layer methods are suitable for deposition of negatively charged phages (wild or CRP-binding) with positively charged carbon nanofibers for electrode surface modification. A three-layered electrode was successfully used for molecular recognition of CRP, and the molecular interactions were studied using electrochemical, biological, and optical methods, including microscopic and spectroscopic analyses.


Structure of the Rhizomucor miehei aspartic proteinase complexed with the inhibitor pepstatin A at 2.7 A resolution.
Crystals of Rhizomucor miehei aspartic proteinase (RMP) complexed with pepstatin A grew in the orthorhombic space group P212121 and were isomorphous to native RMP crystals. The unit-cell dimensions are a = 41.52, b = 50.82, c = 172.71 A. There is one RMP-pepstatin A complex per asymmetric unit. The structure of the RMP-pepstatin A complex has been refined to a crystallographic R value of 19.3% and an Rfree value of 28.0% at 2.7 A resolution. A pepstatin A molecule fits into the large substrate-binding cleft between the two domains of RMP in an extended conformation up to the alanine residue at the P2' position. The dipeptide analogue statine residue at the P3'-P4' position forms an inverse gamma-turn (P3'-P1') with the statine residue at the P1-P1' position and its leucyl side chain binds back into the S1' subsite. The inhibitor interacts with the residues of the substrate-binding pocket by both hydrogen bonds and hydrophobic interactions. The hydroxyl group of the statine residue at the P1-P1' position forms hydrogen bonds with both catalytic aspartate residues (Asp38 and Asp237). This conformation mimics the expected transition state of the enzyme-substrate interaction. The binding of the inhibitor to the enzyme does not produce large distortions of the active site. No domain movement was observed compared with the native enzyme structure. However, the surface-flap region (residues 82-88) undergoes a conformational change, moving toward the inhibitor and becoming rigid owing to the formation of hydrogen bonds with the inhibitor. B-factor calculations of the two domains suggest that the C-terminal domain becomes more rigid in the complex than in the native structure.


[The role of angiography in the diagnosis and therapy of gastrointestinal hemorrhage].
Evaluation of angiography in the diagnosis and treatment of acute and chronic gastrointestinal bleeding. Thirty-seven consecutive patients with clinically suspected gastrointestinal bleeding underwent selective angiography of the visceral arteries. If technically possible, patients with radiologically proven hemorrhage underwent selective embolization with microcoils. Not all angiographically detected bleeding vessels were embolized. Clinical outcome was determined from a review of the medical records. 37 patients underwent 43 angiographies, which demonstrated 25 bleedings considered acute and 18 considered chronic. Seventeen patients had previous intestinal surgery. In 9 of the 37 patients, 12 of the 43 (sensitivity: 28 %) hemorrhages were detected. A postoperative bleeding was found in 6 patients. With one exception, all angiographically positive cases were clinically considered to be acute. Transcatheter embolization of the bleeding vessels with microcoils was performed in 6 patients, with 3 patients having the bleeding stopped permanently and 3 patients requiring further surgical treatment. In 3 patients, embolization was not possible for technical reasons. No complications attributable to angiography were seen. Angiography should be performed in patients with spontaneous and postoperative gastrointestinal bleeding. Transcatheter embolization is an effective and safe therapeutical option in both clinical settings.


The socio-economic status of communities predicts variation in brain serotonergic responsivity.
We reported previously that the socio-economic status (SES) of individuals predicts variation in brain serotonergic responsivity, as assessed by neuropharmacological challenge in an adult community sample, and that this association is qualified by allelic variation in the serotonin transporter gene-linked polymorphic region (5-HTTLPR). Here we examine whether serotonergic responsivity covaries similarly with the SES of communities, as indexed by US Census data in the same study sample. Community SES was defined by levels of income, economic disadvantage, housing costs, and educational attainment of census tracts in which 249 locally recruited study participants (54% male) resided. Serotonergic responsivity was assessed as the baseline-adjusted, peak plasma prolactin (Prl) concentration following acute administration of the serotonin-releasing agent, fenfluramine; tissue for DNA extraction and 5-HTTLPR genotyping was available on 131 participants. Subjects residing in census tracts of lower SES showed a blunted Prl response to fenfluramine (diminished serotonergic responsivity) relative to individuals living in more affluent neighborhoods. When adjusted for personal income and education, SES at the community level continued to predict fenfluramine-stimulated Prl responses and did so independently of 5-HTTLPR genotype. Area-level indices of relative social and economic disadvantage covary with individual differences in brain serotonergic responsivity, and this association is, in part, independent of individually defined SES. These findings may be relevant to reported effects of low community SES on the prevalence of psychiatric disorders or behaviors associated with dysregulation of central serotonergic function, such as depression, impulsive aggression, and suicide.


Osteopathic manipulative treatment for pediatric conditions: a systematic review.
Most osteopaths are trained in pediatric care, and osteopathic manipulative treatment (OMT) is available for many pediatric conditions. The objective of this systematic review was to critically evaluate the effectiveness of OMT as a treatment of pediatric conditions. Eleven databases were searched from their respective inceptions to November 2012. Only randomized clinical trials (RCTs) were included, if they tested OMT against any type of control in pediatric patients. Study quality was critically appraised by using the Cochrane criteria. Seventeen trials met the inclusion criteria. Five RCTs were of high methodological quality. Of those, 1 favored OMT, whereas 4 revealed no effect compared with various control interventions. Replications by independent researchers were available for 2 conditions only, and both failed to confirm the findings of the previous studies. Seven RCTs suggested that OMT leads to a significantly greater reduction in the symptoms of asthma, congenital nasolacrimal duct obstruction (posttreatment), daily weight gain and length of hospital stay, dysfunctional voiding, infantile colic, otitis media, or postural asymmetry compared with various control interventions. Seven RCTs indicated that OMT had no effect on the symptoms of asthma, cerebral palsy, idiopathic scoliosis, obstructive apnea, otitis media, or temporomandibular disorders compared with various control interventions. Three RCTs did not perform between-group comparisons. The majority of the included RCTs did not report the incidence rates of adverse effects. The evidence of the effectiveness of OMT for pediatric conditions remains unproven due to the paucity and low methodological quality of the primary studies.


Dipyridamole stress echocardiography for risk stratification in hypertensive patients with chest pain.
The noninvasive prognostic assessment of coronary artery disease (CAD) in hypertensive patients represents an unresolved task to date. In this study, we investigated the value of dipyridamole stress echocardiography in risk stratification of hypertensive patients with chest pain and unknown CAD. Dipyridamole stress echocardiography was performed in 257 hypertensives (110 men; age, 63+/-9 years) complaining of chest pain and without a history of CAD. No major complications occurred. Four tests were interrupted prematurely because of side effects, with 98. 4% feasibility of test. A positive echocardiographic response was found in 72 patients (27 during the low-dose [</=0.56 mg/kg] and 45 during the high-dose [>0.56 mg/kg]). During the follow-up (32+/-18 months), 27 cardiac events occurred: 3 deaths, 8 infarctions, and 16 cases of unstable angina. Moreover, 27 patients underwent coronary revascularization. At multivariate analysis, the positive echocardiographic result (OR, 5.5; 95% CI, 1.4 to 16.6) was the only predictor of hard cardiac events (death, infarction). Considering spontaneous cardiac events (death, infarction, and unstable angina) as end points, the positive echocardiographic result (OR, 4.2; 95% CI, 1.8 to 9.6) and family history of CAD (OR, 4.2; 95% CI, 1.5 to 6. 9) were independently associated with prognosis. The 3-year survival rates for the negative and the positive populations were, respectively, 97% and 87% (P=0.0019) considering hard cardiac events and 96% and 74% (P=0.0000) considering spontaneous cardiac events. Dipyridamole stress echocardiography is safe, highly feasible, and effective in risk stratification of hypertensives with chest pain and unknown CAD. At present, it represents an attractive option for prognostic assessment of this clinically defined population.