Automated evaluation of probe-based confocal laser endomicroscopy in the lung.
Probe-based confocal endomicroscopy provides real time videos of autoflourescent elastin structures within the alveoli. With it, multiple changes in the elastin structure due to different diffuse parenchymal lung diseases have previously been described. However, these evaluations have mainly relied on qualitative evaluation by the examiner and manually selected parts post-examination. To develop a fully automatic method for quantifying structural properties of the imaged alveoli elastin and to perform a preliminary assessment of their diagnostic potential. 46 patients underwent probe-based confocal endomicroscopy, of which 38 were divided into 4 groups categorizing different diffuse parenchymal lung diseases. 8 patients were imaged in representative healthy lung areas and used as control group. Alveolar elastin structures were automatically segmented with a trained machine learning algorithm and subsequently evaluated with two methods developed for quantifying the local thickness and structural connectivity. The automatic segmentation algorithm performed generally well and all 4 patient groups showed statistically significant differences with median elastin thickness, standard deviation of thickness and connectivity compared to the control group. Alveoli elastin structures can be quantified based on their structural connectivity and thickness statistics with a fully-automated algorithm and initial results highlight its potential for distinguishing parenchymal lung diseases from normal alveoli.


Effect of exercise and oral contraceptive agents on fibrinolytic potential in trained females.
It has been shown that physical exercise increases blood fibrinolytic potential, primarily by inducing a release of extrinsic plasminogen activator from the vessel wall. Synthetic estrogens have also been reported to influence fibrinolytic activity. The effect of exercise and the possible additional effect of oral contraceptive agents (OCA) on the fibronolytic system were studied in 20 competitive female rowers. Ten females used OCA (users), and 10 others did not (nonusers). All participants were subjected to standardized exhaustive exercise. Preexercise data revealed higher factor XII, total plasminogen, and free plasminogen levels together with a significantly lower C1-inactivator level in the group of users. No differences were observed in prekallikrein, high-molecular-weight kininogen, alpha 2-antiplasmin, alpha 2-macroglobulin, antithrombin III, and histidine-rich glycoprotein plasma levels. The factor XII-dependent fibrinolytic activator activity and the extrinsic (tissue-type) plasminogen activator were significantly higher; however, the urokinase-like fibrinolytic activator activity was significantly lower. These observations suggest a greater susceptibility to activation of the fibrinolytic pathways during OCA medication. Exercise resulted in a decrease of all factors under study but an increase in all fibrinolytic activities. No differences were observed between the two groups in the percentages of change that occurred with exercise.


Economic Benefits of Diagnostic Testing in Livestock: Anaplasmosis in Cattle.
Anaplasmosis is a costly livestock disease that persists across the United States and the world. While the traditional control options of feed additives, vaccination, and post-infection antibiotic treatments exist, the highly infectious, often asymptomatic onset of anaplasmosis in cattle makes the optimal combination of disease control measures uncertain. Reducing the infection uncertainty through early detection may help producer management decisions and reduce the economic impact of anaplasmosis. To address this, we calculate the costs of applying a range of anaplasmosis control decisions for a representative cow-calf producer in the United States and extend existing analyses to incorporate early detection through diagnostic testing. We use parameters from extant literature, including for mortality, morbidity, and treatment costs to populate a stochastic, dynamic model. Updating the cost estimates finds that production losses account for the majority of anaplasmosis costs, following previous empirical estimates. Using these estimates in our decision model, the outcomes suggest that diagnostic testing with preventative treatments is the optimal herd management strategy. By further framing our findings in the context of three anaplasmosis infection regions in the United States (endemic, disease free, non-endemic buffer), we show that additional considerations exist, which can make sub-optimal control strategies competitive. Our analysis provides an initial exploration of the economic feasibility of diagnostic testing, while helping to assess the burden of anaplasmosis more accurately.


Combined Effects of Prone Positioning and Airway Pressure Release Ventilation on Oxygenation in Patients with COVID-19 ARDS.
Coronavirus disease 2019 (COVID-19) can cause acute respiratory distress syndrome (ARDS). Invasive mechanical ventilation (IMV) support and prone positioning are essential treatments for severe COVID-19 ARDS. We aimed to determine the combined effect of prone position and airway pressure release ventilation (APRV) modes on oxygen improvement in mechanically-ventilated patients with COVID-19. This prospective observational study included 40 eligible patients (13 female, 27 male). Of 40 patients, 23 (57.5%) were ventilated with APRV and 17 (42.5%) were ventilated with controlled modes. A prone position was applied when the PaO2/FiO2 ratio <150 mmHg despite IMV in COVID-19 ARDS. The numbers of patients who completed the first, second, and third prone were 40, 25, and 15, respectively. Incident barotrauma events were diagnosed by both clinical findings and radiological images. After the second prone, the PaO2/FiO2 ratio of the APRV group was higher compared to the PaO2/FiO2 ratio of the control group [189 (150-237)] vs. 127 (100-146) mmHg, respectively, (P=0.025). Similarly, after the third prone, the PaO2/FiO2 ratio of the APRV group was higher compared to the PaO2/FiO2 ratio of the control group [194 (132-263)] vs. 83 (71-136) mmHg, respectively, (P=0.021). Barotrauma events were detected in 13.0% of the patients in the APRV group and 11.8% of the patients in the control group (P=1000). The 28-day mortality was not different in the APRV group than in the control group (73.9% vs. 70.6%, respectively, P=1000). Using the APRV mode during prone positioning improves oxygenation, especially in the second and third prone positions, without increasing the risk of barotrauma. However, no benefit on mortality was detected.


[Rectal pain and tenesmi caused by a large appendix mucocele].
A 60 year-old man presented with rectal pain and tenesmi. On digital rectal examination a pelvic mass was detected. Computed tomography and MR scans showed a 14 × 5.5 × 5 cm large cystic process located in the right side of the pelvis with no clear indication of its origin. Explorative laparotomy revealed a large appendix mucocele based on a cystadenoma. The mucocele was resected in healthy tissue and without perforation. Needle aspiration or drainage should be avoided in cystic intraabdominal processes of unknown origin, and careful operative handling is imperative, because spillage of mucin may result in later development of pseudomyxoma peritonei. Careful digital rectal examination may detect important pelvic pathology.


The effect of albendazole treatment on seizure outcomes in patients with symptomatic neurocysticercosis.
Randomized controlled trials have found an inconsistent effect of anthelmintic treatment on long-term seizure outcomes in neurocysticercosis. The objective of this study was to further explore the effect of albendazole treatment on long-term seizure outcomes and to determine if there is evidence for a differential effect by seizure type. In this trial, 178 patients with active or transitional neurocysticercosis cysts and new-onset symptoms were randomized to 8 days of treatment with albendazole (n=88) or placebo (n=90), both with prednisone, and followed for 24 months. We used negative binomial regression and logistic regression models to determine the effect of albendazole on the number of seizures and probability of recurrent or new-onset seizures, respectively, over follow-up. Treatment with albendazole was associated with a reduction in the number of seizures during 24 months of follow-up, but this was only significant for generalized seizures during months 1-12 (unadjusted rate ratio [RR] 0.19; 95% CI: 0.04-0.91) and months 1-24 (unadjusted RR 0.06; 95% CI: 0.01-0.57). We did not detect a significant effect of albendazole on reducing the number of focal seizures or on the probability of having a seizure, regardless of seizure type or time period. Albendazole treatment may be associated with some symptomatic improvement; however, this association seems to be specific to generalized seizures. Future research is needed to identify strategies to better reduce long-term seizure burden in patients with neurocysticercosis.


Use of dietary supplements in pregnant women in relation to sociodemographic factors - a report from The Environmental Determinants of Diabetes in the Young (TEDDY) study.
The aim of the present study was to examine the prevalence and associated factors of dietary supplement use, particularly supplements containing vitamin D and fatty acids, in pregnant women enrolled in a multi-national study. The Environmental Determinants of Diabetes in the Young (TEDDY) study is a prospective longitudinal cohort study. Maternal dietary supplement use was self-reported through questionnaires at month 3 to 4 postpartum. Six clinical research centres; three in the USA (Colorado, Georgia/Florida and Washington) and three in Europe (Sweden, Finland and Germany). Mothers (n 7326) to infants screened for high-risk HLA-DQ genotypes of type 1 diabetes. Ninety-two per cent of the 7326 women used one or more types of supplement during pregnancy. Vitamin D supplements were taken by 65% of the women, with the highest proportion of users in the USA (80.5 %). Overall, 16% of the women reported taking fatty acid supplements and a growing trend was seen in all countries between 2004 and 2010 (P,0.0001). The use was more common in Germany (32 %) and the USA (24 %) compared with Finland (8.5%) and Sweden (7.0 %). Being pregnant with the first child was a strong predictor for any supplement use in all countries. Low maternal age (<25 years), higher education, BMI<=25.0 kg/m2 and smoking during pregnancy were factors associated with supplement use in some but not all countries. The majority of the women used dietary supplements during pregnancy. The use was associated with sociodemographic and behavioural factors, such as parity, maternal age, education, BMI and maternal smoking.


Effects of corticosterone and dietary changes in the hen on ovarian function, plasma LH and steroids and the response to exogenous LH-RH.
Ovarian regression was induced in hens by infusing 30 micrograms corticosterone/h, feeding diets deficient in Ca2+ or Na+ and by withdrawal of food and water. The weight of the ovary was most severely reduced by the corticosterone infusion. The total number of normal ovarian follicles weighing greater than 0.012 g was not altered by any of the treatments. However, the number of large yolk-filled follicles decreased while the numbers of smaller follicles and atretic follicles increased when ovarian regression was induced by dietary changes or hormone infusion as compared to normally fed or solvent-infused hens. These experimental treatments resulted in decreases in plasma concentrations of LH, progesterone and oestradiol, and increases in the plasma levels of corticosterone. These changes were immediate except for the low sodium diet with which there was a delay of about 6 days. When fasted birds were fed oats and given water, plasma LH and oestradiol, but not progesterone, increased. The infusion of corticosterone did not affect the ability of the pituitary gland to secrete LH after an injection of LH-RH, but this response was reduced or eliminated by the other experimental treatments. It is concluded that the regression of the ovary induced by these experimental treatments is a consequence of the reduction in the secretion of LH, which may itself be caused by increased plasma levels of corticosterone. It also appears that recruitment of follicles in the maturational stage which precedes entry into the hierarchy of large yolky follicles was unaffected by all of the methods of inducing ovarian regression which were studied.


Arrhythmias, sudden cardiac death and incapacitation of pilots.
Inflight medical emergencies occur at a rate of 20 to 100 per million passengers, with a death rate of 0.1 to 1 per million. Cardiac, neurologic, and respiratory complaints comprise the more serious emergencies, as defined by aircraft diversion or use of ground-based medical assistance. In this paper, we review changes seen in the resting electrocardiogram in normal individuals exposed to high altitude, alongside important implications for patients with heart diseases in high altitude exposures and the possible effects of high altitude to permanent cardiac pacemakers. Arrhythmias in pilots and public safety are revisited together with the guidelines of the Joint Aviation Requirements (JAR) in Europe. The situation of Military flights is also discussed. Physicians ought to become knowledgeable about the specific medical job standards for their patients when asked to render opinions regarding fitness to fly. A database must be established to obtain prospective data that defines the risk of accidents in patients who are or were being treated for arrhythmias. Current guidelines could then be updated and revised as appropriate.


Volumetric changes in the upper airway after bimaxillary surgery for skeletal class III malocclusions: a case series study using 3-dimensional cone-beam computed tomography.
Postsurgical changes of the airway have become a great point of interest and often have been reported to be a predisposing factor for obstructive sleep apnea after mandibular setback surgery. The purpose of this study was to evaluate the 3-dimensional volumetric changes in the upper airway space of patients who underwent bimaxillary surgery to correct Class III malocclusions. This study was performed retrospectively in a group of patients who underwent bimaxillary surgery for Class III malocclusion and had full cone-beam computed tomographic (CBCT) images taken before surgery and 1 day, 3 months, and 6 months after surgery. The upper and lower parts of the airway volume and the diameters of the airway were measured from 2 different levels. Presurgical measurements and the amount of surgical correction were evaluated for their effect on airway volume. Data analyses were performed by analysis of variance and multiple stepwise regression analysis. The subjects included 21 patients (6 men and 15 women; mean age, 22.7 yrs). The surgeries were Le Fort I impaction (5.27 ± 2.58 mm impaction from the posterior nasal spine) and mandibular setback surgery (9.20 ± 4.60 mm set back from the pogonion). No statistically significant differences were found in the total airway volume for all time points. In contrast, the volume of the upper part showed an increase (12.35%) and the lower part showed a decrease (14.07%), with a statistically significant difference 6 months after surgery (P < .05). Predictor variables affecting the upper and lower parts of the airway volume were presurgical A point to Nasion-perpendicular (A to N-perp) and vertical surgical correction of the pogonion and the posterior nasal spine (P < .05). Bimaxillary surgery for the correction of Class III malocclusion affected the morphology by increasing the upper part and decreasing the lower part of the airway, but not the total volume.


Polymorphism of IL-10 gene promoter region: association with T cell proliferative responses after acellular pertussis vaccination in adults.
Individual variation in immune responses is always encountered after vaccination. This phenomenon is also seen after acellular pertussis vaccination. The aim of this present study was to investigate whether single nucleotide polymorphisms (SNPs) in the IL-10 gene promoter region (rs1800890, rs1800896, rs1800871), IL-12B (rs2546890), IL-12RB1 (rs372889), IL-17A (rs2275913), and IL-23R (rs11209026) affect the immune responses after acellular pertussis vaccination. The T cell proliferative response was evaluated in 38 Finnish young adults who received a second booster dose of a vaccine combination of diphtheria, tetanus, and acellular pertussis, 10 years after the previous booster. The response was evaluated with a proliferation assay in which vaccine antigens pertussis toxin (PT), filamentous hemagglutinin (FHA), and pertactin (PRN) were used for the stimulation, before and 1 month after the second vaccination. Specific proliferation of peripheral blood mononuclear cells against pertussis antigens was affected by IL-10 SNP in the promoter region at position -1082 (A>G, rs1800896). One month after the vaccination, subjects with the AA and AG genotypes had a significantly higher T cell proliferative response against PT and FHA compared to those with the GG genotype. Subjects with the GG genotype had the lowest responses. As a conclusion, our preliminary results indicate that IL-10 SNP -1082 might play an important role in T cell-mediated immune responses after acellular pertussis vaccination.


Using the natural evolution of a rotavirus-specific human monoclonal antibody to predict the complex topography of a viral antigenic site.
Understanding the interaction between viral proteins and neutralizing antibodies at atomic resolution is hindered by a lack of experimentally solved complexes. Progress in computational docking has led to the prediction of increasingly high-quality model antibody-antigen complexes. The accuracy of atomic-level docking predictions is improved when integrated with experimental information and expert knowledge. Binding affinity data associated with somatic mutations of a rotavirus-specific human adult antibody (RV6-26) are used to filter potential docking orientations of an antibody homology model with respect to the rotavirus VP6 crystal structure. The antibody structure is used to probe the VP6 trimer for candidate interface residues. Three conformational epitopes are proposed. These epitopes are candidate antigenic regions for site-directed mutagenesis of VP6, which will help further elucidate antigenic function. A pseudo-atomic resolution RV6-26 antibody-VP6 complex is proposed consistent with current experimental information. The use of mutagenesis constraints in docking calculations allows for the identification of a small number of alternative arrangements of the antigen-antibody interface. The mutagenesis information from the natural evolution of a neutralizing antibody can be used to discriminate between residue-scale models and create distance constraints for atomic-resolution docking. The integration of binding affinity data or other information with computation may be an advantageous approach to assist peptide engineering or therapeutic antibody design.


Description of the pupa of Climaciella brunnea (Say) (Mantispidae, Mantispinae) and a key to known pupae of mantispids from North America.
Most works about Climaciella brunnea (Say) deal with predation and parasitoidism on wolf spiders (Lycosidae), or records of populations that mimic vespid wasps (Polistes spp.). Knowledge on immature mantispids is scarce, particularly about pupae. Currently, a key by Hoffman and Brushwein provides information on larvae and pupae of several North American species. Herein, the pupa of C. brunnea is described for the first time, on the basis of a specimen from central Mexico, and its position in the available key is proposed,. The pupa of Plega yucatanae Parker Stange from southeastern Mexico is also incorporated to the key on the basis of a published description.


When the brain hurts the heart: status epilepticus inducing tako-tsubo cardiomyopathy.
Tako-tsubo cardiomyopathy (TTC) is a transient myocardial dysfunction mainly affecting the left ventricle, mimicking an acute coronary syndrome. This condition can be precipitated either by psychological/physical stressful events or by a number of medical conditions among which are seizures and status epilepticus (SE). The evolution is mostly favourable but sometimes TTC can evolve into life-threatening conditions. We searched for cases of TTC among all consecutive SE episodes observed at our department during the period 2013-2018. In addition, we searched MEDLINE (accessed through PubMed from inception to August 31, 2018) to identify reports of patients with TTC associated with an SE episode. Three TTC cases among 392 SE episodes were identified. Adding our cases to those previously reported, overall, we identified 45 cases of TTC induced by SE. The majority were females of around 60 years of age experiencing a first episode of SE with prominent motor phenomena, mostly in the context of remote aetiology. The most frequent presenting symptom was mild hypotension but cases with a severe presentation were also reported. The overall evolution was positive in all cases but some severe complications such as pulmonary oedema, cardiogenic shock, ventricular fibrillation, and a giant apical thrombus were also reported (19%). TTC may be a rare potentially life-threatening consequence of SE. It is frequently unrecognized, and therefore underdiagnosed. Clinicians dealing with SE should be aware of this entity.


Factor Structure of the Foreign Language Classroom Anxiety Scale: Comment on Park (2014).
By employing both exploratory and confirmatory factor analysis, Park has provided an important contribution to understanding the underlying constructs of the Foreign Language Classroom Anxiety Scale. To Park's concerns about previous research on the components of the measure, this article adds the necessity of considering the specific learner populations and learning contexts where foreign language anxiety (FLA) is being examined since the components of FLA likely vary in different learner populations, especially with respect to cultural and proficiency differences. It is particularly important to consider that FLA has different triggers and manifestations in different cultures.


Muscle contraction-blood flow interactions during upright knee extension exercise in humans.
To test for evidence of a muscle pump effect during steady-state upright submaximal knee extension exercise, seven male subjects performed seven discontinuous, incremental exercise stages (3 min/stage) at 40 contractions/min, at work rates ranging to 60-75% peak aerobic work rate. Cardiac cycle-averaged muscle blood flow (MBF) responses and contraction-averaged blood flow responses were calculated from continuous Doppler sonography of the femoral artery. Net contribution of the muscle pump was estimated by the difference between mean exercise blood flow (MBFM) and early recovery blood flow (MBFR). MBFM rose in proportion with increases in power output with no significant difference between the two methods of calculating MBF. For stages 1 and 5, MBFM was greater than MBFR; for all others, MBFM was similar to MBFR. For the lighter work rates (stages 1-4), there was no significant difference between exercise and early recovery mean arterial pressure (MAP). During stages 5-7, MAP was significantly higher during exercise and fell significantly early in recovery. From these results we conclude that 1) at the lightest work rate, the muscle pump had a net positive effect on MBFM, 2) during steady-state moderate exercise (stages 2-4) the net effect of rhythmic muscle contraction was neutral (i.e., the impedance due to muscle contraction was exactly offset by the potential enhancement during relaxation), and 3) at the three higher work rates tested (stages 5-7), any enhancement to flow during relaxation was insufficient to fully compensate for the contraction-induced impedance to muscle perfusion. This necessitated a higher MAP to achieve the MBFM.


Co-spray-dried mannitol-ciprofloxacin dry powder inhaler formulation for cystic fibrosis and chronic obstructive pulmonary disease.
The aim of this study was to assess the potential of delivering a combination therapy, containing mannitol (a sugar alcohol with osmotic characteristics), and ciprofloxacin hydrochloride (an antibacterial fluoroquinolone), as a dry powder inhaler (DPI) formulation for inhalation. Single and combination powders were produced by spray drying ciprofloxacin and mannitol, from aqueous solution, at different ratios and under controlled conditions, as to obtain similar particle size distributions. Each formulation was characterised using laser diffraction, scanning electron microscopy, differential scanning calorimetry, dynamic vapour sorption, X-ray powder diffraction, and colloidal force microscopy. The in vitro aerosol performance of each formulation was studied using an Aerolizer DPI device and a multi-stage liquid impinger (analysed using high performance liquid chromatography). In addition, a disk diffusion test was performed to assess the in vitro antimicrobial activity of each formulation and starting materials. All formulations had similar particle size distributions, however, the morphology, thermal properties and moisture sorption was dependent on the relative percentages of each component. In general, the combination formulation containing 50% (w/w) mannitol appeared to have the best aerosol performance, good stability and lowest particle cohesion (as measured by colloid probe microscopy). Furthermore, of the formulations tested, mannitol did not appear to alter the effectiveness of the ciprofloxacin antimicrobial activity to Staphylococcus aureus, Pseudomonas aeruginosa and Streptococcus pyogenes. The combination of co-spray-dried mannitol and ciprofloxacin from a DPI is an attractive approach to promote mucous clearance in the respiratory tract while simultaneously treating local chronic infection, such as chronic obstructive pulmonary disease and cystic fibrosis.


[Opioid receptors and their selective ligands].
Opioid receptors (micro, delta, and kappa) belong to a large family of G protein-coupled receptors and play an important physiological role. Stimulation of these receptors triggers analgesic effects and affects the function of gastrointestinal tract. The discovery of opioid peptides, which are endogenous ligands of opioid receptors, including delta-selective enkephalins, kappa-selective dynorphins, and micro-selective endomorphins, initiated their structure-activity relationship studies. For the last 30 years, hundreds of analogs of opioid peptides have been synthesized in an effort to obtain the compounds more active, selective, and resistant to biodegradation than the endogenous ligands. Different unnatural amino acids, as well as cyclisation procedures, leading to conformationaly restricted analogs, were employed. All these modifications resulted in obtaining very selective agonists and antagonists with high affinity at micro-, dlta-, and kappa-opioid receptors, which are extremely useful tools in further studies on the pharmacology of opioid receptors in a mammalian organism.


[Demographic Ageing in Russia at the Present Stage].
In the article data ofthe article describes thefindings of modern demographic indicators in Russia andforeign countriesfor the period 2010-2013. As a result of monitoring was defined Russia's status by demographics such as: proportion of population aged 60 years and older; total (men and women) life expectancy at birth; the rate of natural increase of population; mortality; fertility rate; the coefficient of migration growth and the dependency ratio. This article was presented analogy age-sex structure disclosed negative and positive aspects of the ageing of the population in Russia and foreign countries.


Early revascularization of membranous inlay bone graft in canine mandible model.
The early revascularization of a membranous inlay bone graft in a canine mandible was investigated using bone scans and histological examinations. Eight 5-month-old mongrel dogs were used as the subjects. The inlay bone graft, a 1- x 2-cm critically sized bone, was completely separated from the lower border of the canine mandible and then refixed to the original site. Bone scans and histological examinations were performed in the first, second, third, and fourth postoperative weeks. The bone scan in the first postoperative week revealed radioisotope uptake on the margin of the grafted bone with an isotope count that was 21% of the normal bone uptake. In the second postoperative week, the radioisotope uptake in the grafted bone increased to 52% of the normal bone uptake. In the third and fourth postoperative weeks, the isotope uptakes were 111% and 124%, respectively. The histological findings in the first postoperative week showed an absence of osteoblastic activity and 6 viable blood vessels in a field magnified x 200, which was the equivalent of 25% of the vessels of the normal bone. In the second postoperative week, osteoblastic activity was noted, and the number of viable blood vessels increased to 15, that is, 63% of the vessels of the normal bone. In the third postoperative week, there was an increase in osteoblastic activity, and the number of viable blood vessels was 21, that is, 88% of the vessels of the normal bone. In the fourth postoperative week, there was a marked increase in osteoblastic activity with the number of vessels reaching 23, that is, 96% of the normal bone. In summary, revascularization of the membranous inlay bone graft began within the first week after the bone graft and thereafter gradually increased. In the third postoperative week, revascularization returned to a near-normal value compared with the value of the adjacent normal mandibular bone.


Lymphoepithelioma-like gastric carcinoma: an unusual consequence of Epstein-Barr virus infection in an HIV-infected woman.
Lymphoepithelioma-like-gastric carcinoma (LEL-GC) is an Epstein-Barr virus (EBV)-associated neoplasm of the stomach reported to have a better prognosis than conventional gastric adenocarcinoma. Unlike other EBV-associated malignancies, particularly lymphoproliferative disorders and undifferentiated nasopharyngeal carcinoma, for which risk has been shown to increase in human immunodeficiency virus (HIV) infection, LEL-GC remains rare; only one HIV-infected patient with LEL-GC has been reported previously. We describe an aggressive case of EBV-associated LEL-GC in a woman co-infected with HIV 1 and hepatitis C virus. In situ hybridization of an endoscopic biopsy specimen for EBV-encoded small RNA confirmed the presence of this agent exclusively in the gastric cancer cells. Our patient had recently started antiretroviral therapy, suggesting that immune reconstitution may have been a factor in presentation of this tumour.


Blood culture-positive infections in patients with alcoholic hepatitis.
Acute alcoholic hepatitis (AH) is a life-threatening disease and its course is often determined by infections. However, the pattern of pathogens has not been studied. We examined the microbiological pathogens that caused blood-borne infection in patients with AH. We included 32 AH patients without infection at inclusion. Patients were followed for 1 month and their infection status was recorded based on clinical records, radiologic exams and cultures of different secreta. Nine patients (28%) developed blood culture-positive infections. The agents were of heterogeneous aetiology and came from various sites of infection. Candida species accounted for three of these infections (33%). Five patients (16%) died, two of which had positive blood cultures. A high fraction was invasively infected by a heterogeneous spectrum of microbes including yeasts and commensal bacteria. This may reflect the severe immune impairment of AH and suggests thorough infection screening and an immediate broad-spectrum antibiotic approach if infection is suspected.


Analysis of serine proteases from marine sponges by 2-D zymography.
Proteolytic activities isolated from the marine demosponges Geodia cydonium and Suberites domuncula were analyzed by 2-D zymography, a technique that combines IEF and zymography. After purification, a 200 kDa proteolytically active protein band was obtained from G. cydonium when analyzed in gelatin copolymerized 1-D zymograms. The enzymatic activity was quantified using alpha-N-benzoyl-D-arginine p-nitroanilide (BAPNA) as a substrate and corresponded to a serine protease. The protease activity was resistant to urea and SDS. DTT and 2-mercaptoethanol (2-ME) did not significantly change the protease activity, but induced a shift in molecular mass of the proteolytic band to lower M(r) values as detected by zymography. Under mild denaturing conditions, lower M(r) bands (<200 kDa) were identified in 1-D zymograms, suggesting that the protease is composed of subunits which retain the catalytic activity. After 2-D zymography, the protease from G. cydonium revealed a pI of 8.0 and an M(r) shift from 200 to 66 kDa. To contrast these results, a cytosolic sample from S. domuncula was analyzed. The proteolytic activity of this sponge after 2-D zymography corresponded to an M(r) of 40 kDa and a pI of 4.0. The biological function of both sponge proteases is not yet known. This study demonstrates that mild denaturing conditions required for IEF may alter the interpretation of the 2-D zymography, and care must be taken during sample preparation.


Remifentanil infusion in association with fentanyl-propofol anaesthesia in patients undergoing cardiac surgery: effects on morphine requirement and postoperative analgesia.
We have prospectively assessed the effects of remifentanil on morphine requirement in the first hour after emerging from general anaesthesia after elective coronary artery bypass surgery and in the first 12 h postoperatively, and pain and agitation scores in the first hour after emerging from general anaesthesia. Twenty patients undergoing off-pump coronary artery bypass surgery, receiving standardized propofol-fentanyl-based anaesthesia, randomly received infusions of either remifentanil 0.1 microg kg(-1) min(-1) (Group R, n=10) or saline (Group S, n=10), each infused at 0.12 ml kg(-1) h(-1). Propofol and trial drug infusion were continued into the postoperative period until the patients were ready to be woken up. Postoperative analgesia was provided with morphine infusion commenced immediately after operation, and was additionally nurse controlled on the basis of a visual analogue scale (VAS) score (0-10). Agitation score was recorded using a VAS of 0-3. In the first hour after discontinuing propofol and trial infusion, morphine requirements were significantly higher in the remifentanil group (8.15 (sd 3.59) mg) compared with the saline group (3.29 (2.36) mg) (P<0.01). There was no difference in the total morphine given during the period after stopping propofol or in the total requirement in the first 12 h postoperatively. There was no significant difference in either pain scores or agitation scores between the two groups. Use of remifentanil is associated with increased opioid requirement in the first hour after it has been discontinued.


Generating ultrashort coherent soft X-ray radiation in storage rings using angular-modulated electron beams.
A technique is proposed to generate ultrashort coherent soft x-ray radiation in storage rings using angular-modulated electron beams. In the scheme a laser operating in the TEM01 mode is first used to modulate the angular distribution of the electron beam in an undulator. After passing through a special beam line with nonzero transfer matrix element R54, the angular modulation is converted to density modulation which contains considerable high harmonic components of the laser. It is found that the harmonic number can be 1 or 2 orders of magnitude higher than the standard coherent harmonic generation method which relies on beam energy modulation. The technique has the potential of generating femtosecond coherent soft x-ray radiation directly from an infrared seed laser and may open new research opportunities for ultrafast sciences in storage rings.


Cell polarity protein CRB3 is an independent favorable prognostic factor for clear cell renal cell carcinoma.
Epithelial cells possess apical‑basal polarity and loss of epithelial cell polarity contributes to tumorigenesis and cancer progression. The Crumbs (CRB) polarity protein plays a crucial role in epithelial polarity maintenance, apical membrane formation, and tissue morphogenesis. Although evidence is increasing on involvement of deregulated polarity proteins in cancers, little is currently known about the roles of the CRB (Drosophila), especially the roles of CRB3, a homolog of the CRB, in clear cell renal cell carcinoma (ccRCC). Studies have shown that CRB3 may act as a tumor suppressor in non‑human mammalian cells; the study here was aimed to examine the expression status of CRB3 in ccRCC and the relationships between CRB3 expression and clinicopathologic parameters of ccRCC patients. Our results showed that CRB3 was weakly expressed in ccRCC tissues, but strongly expressed in adjacent normal kidney tissues. Patients with loss of CRB3 expression showed a significantly shorter overall survival (OS) than patients with positive CRB3 expression. Our results suggested that CRB3 may be an independent favorable prognostic factor for patients with ccRCC. We also found that overexpression of CRB3 restrained invasion and migration of 786‑O cells and loss of CRB3 expression promoted invasion and migration of human embryonic kidney 293T (HEK 293T) cells. This finding may explain why the negative CRB3 expression was associated with poor prognosis in human ccRCC. Altogether, our data demonstrated that CRB3 may be used as a new independent favorable prognostic factor for human ccRCC.


Effect of cross-reactivity of alpha-fetoprotein monoclonal antibody on quantitation of serum AFP and radioimmunodetection of hepatocellular carcinoma.
Murine hybridomas were generated by immunizing mice with purified alpha-fetoprotein (AFP) to determine the cross-reactivity of AFP antibodies with human serum albumin (HSA). Eighteen out of 23 hybridoma products were cross-reactive with HSA. All 23 monoclonal antibodies (MAbs) could be subgrouped into 3 groups by their binding affinity to AFP and HSA, and one MAb from each group, IIB3, IIIB1 and IIIC6, were chosen for this study. The affinity to AFP was highest for IIB3 followed by IIIC6 and IIIB1, and that to HSA was in the order IIIB1 greater than IIB3. Immunohistochemical staining revealed no cross-reactions of these antibodies with HSA at the tissue level. IIB3 was the most efficient in the quantitative assays, indicating high affinity and specificity (non-cross-reactivity) of the antibodies, both of which are prerequisites for in vitro assays. Scans with good tumor localization were obtained from mice which received 131I-labeled IIB3 or IIIC6 after xenografting of the human hepatocellular carcinoma. The images obtained with IIB3 were inhibited in the presence of either AFP or HSA in the circulation. These findings indicated that the use of cross-reactive and/or high-affinity antibodies against AFP provides an inhibitory factor for tumor localization. MAb IIIC6, because of its high specificity and moderate affinity to AFP, seems to be the best candidate for immunoscintigraphy of the tumors in future studies.


Cost per responder analysis in patients with secondary hyperparathyroidism on dialysis treated with cinacalcet.
Growing financial pressure on US dialysis providers requires economic efficiency considerations. The objective of this study was to examine short-term economic efficiencies of a cinacalcet-based treatment approach for secondary hyperparathyroidism. This study retrospectively assessed cost per biochemical response of the OPTIMA trial. OPTIMA was conducted in end-stage renal disease patients to compare biochemical control in patients receiving cinacalcet in addition to vitamin D sterols and phosphate binders vs patients receiving vitamin D sterol and phosphate binders alone. It explored three laboratory measurement response definitions from baseline to week 23: (1) decreases in parathyroid hormone (PTH) ≥30%; (2) PTH ≤ 300 pg/ml; and (3) PTH ≤ 300 pg/mL, calcium <9.5 mg/dL and phosphorus <5.5 mg/dL. Medication use and costs were measured to calculate average costs and incremental cost per responder. Stratification by lower and higher baseline PTH assessed cost per response by disease severity. There were 38-77% more responders with cinacalcet vs control, depending on response definition. Mean (SD) per patient total medication costs were $5423 ($3698) for cinacalcet and $2633 ($2334) for control, leading to a mean difference of $2790 over 23 weeks. When response was defined as a decrease in PTH ≥ 30% from baseline, the average cost per responder was $11,266 for control vs $7027 for cinacalcet. The incremental cost per incremental responder ranged from $5186-$9168. Across all response measures, cost per responder was lower in patients with lower baseline PTH. Representing a more efficient allocation of economic resources over the short-term, cinacalcet-based treatment algorithm led to a lower cost per biochemical response, particularly in patients with lower disease severity, vs vitamin D sterols and phosphate binders alone. These findings should be interpreted alongside the study limitation of converting international trial-based medication utilization into US costs.


Artefact reduction with alternative cuff configurations.
In nerve cuff electrode recordings of neural signals, the pick-up of interfering signals can be reduced by choosing appropriate cuff configurations. In the traditionally used tripolar configuration, short circuiting of the end electrodes is expected to reduce the field inside the cuff from interfering signals. A model study suggests that moving the end electrodes toward the center of the cuff reduces the pick-up of interfering signals. In this paper, these properties are studied in more detail using a rabbit model. In addition, a new cuff configuration is suggested, which has an additional set of short circuited end electrodes. The total improvement of signal-to-noise ratio in the new configuration as compared with the traditionally used tripolar configuration was 73% for muscle signals and 127% for the stimulus pulse.


Differential effectiveness of depression disease management for rural and urban primary care patients.
Federally qualified health centers across the country are adopting depression disease management programs following federally mandated training; however, little is known about the relative effectiveness of depression disease management in rural versus urban patient populations. To explore whether a depression disease management program has a comparable impact on clinical outcomes over 2 years in patients treated in rural and urban primary care practices and whether the impact is mediated by receiving evidence-based care (antidepressant medication and specialty care counseling). A preplanned secondary analysis was conducted in a consecutively sampled cohort of 479 depressed primary care patients recruited from 12 practices in 10 states across the country participating in the Quality Enhancement for Strategic Teaming study. Depression disease management improved the mental health status of urban patients over 18 months but not rural patients. Effects were not mediated by antidepressant medication or specialty care counseling in urban or rural patients. Depression disease management appears to improve clinical outcomes in urban but not rural patients. Because these programs compete for scarce resources, health care organizations interested in delivering depression disease management to rural populations need to advocate for programs whose clinical effectiveness has been demonstrated for rural residents.


DNA-microarray technology: comparison of methodological factors of recent technique towards gene expression profiling.
DNA-microarray technology is a powerful tool to explore the potentials of functional genomics. Almost a decade back, it was not so easy to trace the molecular pathway process in the pathogenesis of diseases. Today DNA-microarray technology allows identifying candidate genes that are implicated in the development and progression of diseases. Because this technology is new, it remains a challenge to perform DNA-microarray experiments in the laboratory. All the technical factors associated with DNA-microarray experiments have a strong impact on the results and therefore, all parts/steps of the protocol, in particular sample preparation (i.e., RNA isolation, RNA preamplification, and cDNA labeling), hybridization, washing, and scanning settings need to be checked and optimized to obtain reliable results from DNA-microarray experiments.


7-Deaza-2'-deoxy-7-propynylguanosine.
The title compound, C14H16N4O4, adopts the anti conformation at the glycosylic bond [chi -117.1 (5) degrees]. The sugar pucker of the 2'-deoxyribofuranosyl moiety is C2'-endo-C3'-exo, 2T3 (S-type). The orientation of the exocyclic C4'-C5' bond is +sc (gauche). The propynyl group is linear and coplanar with the nucleobase moiety. The structure of the compound is stabilized by several hydrogen bonds (N-H...O and O-H...O), leading to the formation of a multi-layered network. The nucleobases, as well as the propynyl groups, are stacked. This stacking might cause the extraordinary stability of DNA duplexes containing this compound.


Natural Leishmania sp. reservoirs and phlebotomine sandfly food source identification in Ibitipoca State Park, Minas Gerais, Brazil.
Leishmania spp are distributed throughout the world and different species are associated with varying degrees of disease severity. However, leishmaniasis is thought to be confined to areas of the world where its insect vectors, sandflies, are present. Phlebotomine sandflies obtain blood meals from a variety of wild and domestic animals and sometimes from humans. These vectors transmit Leishmania spp, the aetiological agent of leishmaniasis. Identification of sandfly blood meals has generally been performed using serological methods, although a few studies have used molecular procedures in artificially fed insects. In this study, cytochrome b gene (cytB) polymerase chain reaction (PCR) was performed in DNA samples isolated from 38 engorged Psychodopygus lloydi and the expected 359 bp fragment was identified from all of the samples. The amplified product was digested using restriction enzymes and analysed for restriction fragment length polymorphisms (RFLPs). We identified food sources for 23 females; 34.8% yielded a primate-specific banding profile and 26.1% and 39.1% showed banding patterns specific to birds or mixed restriction profiles (rodent/marsupial, human/bird, rodent/marsupial/human), respectively. The food sources of 15 flies could not be identified. Two female P. lloydi were determined to be infected by Leishmania using internal transcribed spacer 1 and heat shock protein 70 kDa PCR-RFLP. The two female sandflies, both of which fed on rodents/marsupials, were further characterised as infected with Leishmania (Viannia) braziliensis. These results constitute an important step towards applying methodologies based on cytB amplification as a tool for identifying the food sources of female sandflies.


Adaptation of the Test of Performance Strategies Competition Subscale to Spanish.
In recent years, there has been a growing interest in the conceptualisation and assessment of athletes' psychological skills and the study of their impact on sports performance. The aim of this study was to adapt the Test of Performance Strategies 3 competition subscale to the Spanish context. The items included in the original test in English were translated using a double-back method, and the test was completed by a sample of 1,003 Spanish athletes of both sexes. Analyses of the factorial validity, reliability and invariance of the measurement model were carried out. Favourable evidence was obtained for a measurement model comprising 36 items grouped in 9 factors, similar to the original model. Model fit was reasonable for both individual parameters and overall. Reliability rates were satisfactory for the overall test and for each factor. Evidence was also favourable for sex-based measurement model invariance. The adaptation is satisfactory and fit for use by sports psychology researchers and professionals in assessing the psychological skills employed by athletes in competition.


Prevalence of night sweats in primary care patients: an OKPRN and TAFP-Net collaborative study.
To estimate the prevalence and factors associated with night sweats among adult primary care patients. This was a cross-sectional study. Adult patients in 2 primary care practice-based research networks (PBRNs) during 1 week in the summer and 1 week in the winter in the years 2000 and 2001. We measured the prevalence of pure night sweats and night and day sweats in all patients and subgroups defined by age and sex, clinical variables associated with night sweats, and the frequency, severity, and rate of reporting. Of the 2267 patients who participated, 41% reported experiencing night sweats within the last month, including 23% with pure night sweats and an additional 18% with day and night sweats. The prevalence of night sweats in both men and women was highest in the group aged 41 years to 55 years. In multivariate analyses, factors associated with pure night sweats in women were hot flashes and panic attacks; in men, sleep problems. Variables associated with night and day sweats in women were increased weight, hot flashes, sleep disturbances, use of antihistamines, selective serotonin reuptake inhibitors (SSRIs), and other (non-SSRI, non-tricyclic) antidepressants; in men, increased weight, hot flashes, and greater alcohol use. A majority of patients had not reported their night sweats to their physicians, even when frequent and severe. Night sweats are common and under-reported. Pure night sweats and night and day sweats may have different causes. With regard to the etiologies of pure night sweats, panic attacks and sleep disorders need further investigation.


[Clinical Functionality of Dispersive OVDs: Improvement of One of the Properties of 3% Hyaluronic Acid and 4% Chondroitin Sulfate Combination].
Ophthalmic viscosurgical devices (OVDs) are mainly divided into two general categories: cohesive and dispersive. Dispersive OVDs such as the 3% hyaluronic acid and 4% chondroitin sulfate (HA/CS) combination have excellent adhesion to ocular tissues and protect the corneal endothelium to a greater extent than cohesive OVDs. Herein, we summarize our recent findings regarding one of the properties of the HA/CS combination related to clinical performance. (i) The room temperature stability of OVDs and needle clogging by OVDs remain clinical issues. We demonstrated that adding d-sorbitol to the HA/CS combination preserved its viscosity, which was equivalent after 2 year-storage at room temperature to the viscosity of HA/CS combination stored under refrigeration for 2 years without d-sorbitol. Besides, the HA/CS combination with d-sorbitol could be used repeatedly without cleaning or replacing the needle, suggesting that the addition of d-sorbitol prevents drying and solidification of the OVD on the needle. (ii) Although it can be inferred from numerous studies that the tissue adhesion of OVDs influences their retention by the eye, little is known about the physical properties of OVDs that contribute to intraocular retention. To address this issue, we compared two types of adhesive forces, detachment force and repulsive force, for each OVD. Compared with other dispersive OVDs, the HA/CS combination showed higher values for both adhesive forces. These results suggest that adhesive forces may be used as an index of dispersive OVD retention in the eye.


ESOS: a European study on the orthopaedic status of patients with haemophilia and inhibitors.
Published studies highlight the paucity of data relating to orthopaedic health and quality of life in haemophilia patients with inhibitors. A European Study on the Orthopaedic Status of Patients with Haemophilia and Inhibitors (ESOS) has been set up to address this lack of data. This cross-sectional retrospective study aims to enrol 400 patients from 50 sites in nine European countries. The primary outcome measure of the study is the burden of orthopaedic complications in patients with severe haemophilia and inhibitors. The data that will be generated should demonstrate the high level of arthropathy in haemophilia patients with inhibitors and show the need to focus on improving orthopaedic health in this rare, but particularly vulnerable, patient population.


Delayed Intraparenchymal Hematoma Following Diagnostic Lumbar Puncture.
Lumbar puncture is a safe and commonly performed procedure, with an overall complication rate of 0.1% to 0.5%. Well-known contraindications to lumbar puncture are an intracranial tumor, noncommunicating hydrocephalus, coagulopathy, and ruptured aneurysm with subarachnoid hemorrhage. We report a case of a young man with epilepsy who, after a lumbar puncture performed for research purposes, presented with an intracerebral hematoma and neurological deficits. To the best of our knowledge, post-tap intraparenchymal hematoma is extremely rare and only 1 case has been reported previously. In consideration, all patients undergoing a lumbar puncture should be informed about this possible rare complication, even in the absence of documented hemorrhagic risk factors.


Single staged carotid and coronary arteries surgery. Indications and results.
The frequency of associated carotid and coronary stenosis is estimated to be 40% of cases including asymptomatic patients. The risk of death by myocardial infarction during carotid surgery is in the neighborhood of 1%. With Doppler and ultrasonic imaging, it is now possible to recognize carotid stenosis in 11% of these cases. Depending on the extent of the clinical manifestations caused by one or the other of these lesions, there are several therapeutic options: surgery of the carotid with special protection of the myocardium during and after surgery or simultaneous carotid and coronary surgery in high risk cases. Between 1979 and 1986, simultaneous surgery was performed in 18 cases in our department. During the same period, 1194 carotid endarterectomies and 505 coronary bypasses were performed. These 18 cases included 13 males and 5 females with a mean age of 62 years (range: 43 to 78 years). The procedure performed in these cases were 17 carotid endarterectomies, one bypass of the innominate artery and an average of 2 aorto-coronary bypasses per patient. One death from myocardial infarction occurred in the immediate postoperative period and another patient died suddenly 8 months later. The evolution of arterial disease was evaluated in the other patients. Simultaneous carotid and coronary artery surgery is indicated in high risk carotid and coronary patients. Perusal of the results in the literature allows a better appreciation of the indications and risks of this type of surgery. Our series indicates the necessity for a careful preoperative workup in these patients and the need for more accurate screening to obtain better results.


[Essential hypertension: psychosomatic aspects].
Essential hypertension is a heterogenous disease made up of several still unspecified subgroups. According to the dual theory of hypertension, acquired and genetic factors can lead to vascular muscle defects and increased sympathetic drive, which, either both or each alone, can lead to an increased vascular resistance. Acquired factors can either help a predisposition to become a manifest disease, act by itself to cause a disease or prevent a genetic trait from becoming manifest. Among the acquired factors, we differentiate those lying inside the personality, which means they are early acquired in life and those lying outside of the individual such as job-stress, noise exposure etc. How the external stressors are perceived depends upon the personality make up and the social support system. Within the presented model the Greek word 'crisis' plays an important role, since it characterizes a state in which a decision has to be made. In a situation in which a decision cannot be made in reality or tried out in fantasy life, the organism will activate the autonomic nervous system in a planless fashion, alternating very rapidly between its two branches, with concomitant symptoms of trembling, piloerection, salivation, vomiting, urination, defecation etc. In case this persists for a longer period of time the reciprocity between the two systems may break down, the organism seems to be psychologically immobilized. In the beginning of stress exposure, increased sympathetic nervous system activity is seen as an adaptive response, thereby enabling the individual to cope, but if the attempts to cope meet with constant failure, the stress becomes unbearable. The attitude of giving up which then develops is accompanied by a low level functioning of the sympathetic nervous system. This goes along with an adaptation for the baroreceptor depressor mechanism. As a consequence, any kind of additional stress would not lead to a relieving counterregulation. This might be a small stone within the mosaic of essential hypertension.


[Conservative therapy of supraventricular tachycardias].
The conservative management of supraventricular tachycardias is briefly surveyed and described. For exact treatment a description of different ways of origin of tachycardias such as focal activity or reentry-mechanisms is given. A diagram at the end shows the site of influence of different antiarrhythmic drugs in the atria and the AV-node as well as the accessory pathways in preexcitation. A list of antiarrhythmics according to their classification of Vaughan Williams and of Tournboul completes this survey.


Association of Myopia and Intraocular Pressure With Retinal Detachment in European Descent Participants of the UK Biobank Cohort: A Mendelian Randomization Study.
Rhegmatogenous retinal detachment is a potentially sight-threatening condition. The role of myopia or intraocular pressure (IOP) in retinal detachment remains unclear. To determine if myopia or IOP is associated with retinal detachment risk using genetic data. Observational analyses and 2-sample mendelian randomization were used to evaluate the associations between myopia, IOP, and retinal detachment risk in European descent participants from the UK Biobank (UKBB) cohort (n = 405 692). For retinal detachment, a genome-wide association study on 4257 cases and 39 181 controls in the UKBB was conducted. Genetic variants associated with mean spherical equivalent (MSE) refractive error (n = 95 827) and IOP (n = 101 939) were derived using independent participants from the retinal detachment genome-wide association study. Recruitment to the UKBB occurred between 2006 and 2010, and data analysis occurred from February 2019 to March 2020. The odds ratio (OR) of retinal detachment caused by per-unit increases in MSE refractive error (in diopters [D]) and IOP (in mm Hg). Of the 405 692 participants in the UKBB cohort, the mean (SD) age was 56.87 (7.96) years, the mean (SD) MSE was -0.31 (2.65) D, the mean (SD) corneal-compensated IOP was 16.05 (3.49) mm Hg, and 4253 participants (1.0%) had retinal detachment. Genetic analyses of the 4257 cases and 39 181 controls identified 2 novel retinal detachment genes: COL22A1 (lead single-nucleotide variant rs11992725; P = 4.8 × 10-10) and FAT3 (lead single-nucleotide variant rs10765568; P = 1.2 × 10-15). Genetically assessed MSE refractive error was negatively associated with retinal detachment (per-unit [D] increase in MSE refractive error: OR, 0.72; 95% CI, 0.69-0.76; P = 3.8 × 10-44). For each 6-D decrease in MSE refractive error (representing the move of refractive error from emmetropia to high myopia), retinal detachment risk increased 7.2-fold (95% CI, 5.19-9.27). For per-unit (mm Hg) genetically assessed increase in IOP, the risk of retinal detachment increased by 8% (OR, 1.08; 95% CI, 1.03-1.14; P = .001). This study provides genetic support for the assertion that myopia and IOP are associated with the risk of retinal detachment and that myopia prevention efforts may help prevent retinal detachment.


Brassinosteroids are involved in response of cucumber (Cucumis sativus) to iron deficiency.
Brassinosteroids (BR) are a class of plant polyhydroxysteroids with diverse functions in plant growth and development. However, there is little information on the role of BRs played in the response to nutrient deficiency. To evaluate the role of BR in the response of plants to iron (Fe) deficiency, the effect of 24-epibrassinolide (EBR) on ferric reductase (FRO) activity, acidification of the rhizosphere and Fe content in cucumber (Cucumis sativus) seedlings under Fe-deficient (1 µm FeEDTA) and Fe-sufficient (50 µm FeEDTA) conditions were investigated. There was a significant increase in FRO activity upon exposure of cucumber seedlings to an Fe-deficient medium, and the Fe deficiency-induced increase in FRO activity was substantially suppressed by EBR. In contrast, application of EBR to Fe-sufficient seedlings stimulated FRO activity. Ethylene production evoked by Fe deficiency was suppressed by EBR, while EBR induced ethylene production from Fe-sufficient seedlings. Fe contents in shoots were reduced by treatment with EBR, while Fe contents in roots were markedly increased under both Fe-deficient and Fe-sufficient conditions. The reductions in Fe contents of shoots were independent of chlorophyll (CHL) contents under Fe-sufficient conditions, but they were positively correlated with CHL contents under Fe-deficient conditions. At the transcriptional level, transcripts encoding FRO (CsFRO1) and Fe transporter (CsIRT1) were increased upon exposure to the Fe-deficient medium, and the increases in transcripts were reversed by EBR. The results demonstrate that BRs are likely to play a negative role in regulating Fe-deficiency-induced FRO, expressions of CsFRO1 and CsIRT1, as well as Fe translocation from roots to shoots.


The anxiogenic drug yohimbine activates central viscerosensory circuits in rats.
Systemic administration of the alpha(2)-adrenoceptor antagonist yohimbine (YO) activates the HPA stress axis and promotes anxiety in humans and experimental animals. We propose that visceral malaise contributes to the stressful and anxiogenic effects of systemic YO and that YO recruits brainstem noradrenergic (NA) and peptidergic neurons that relay viscerosensory signals to the hypothalamus and limbic forebrain. To begin testing these hypotheses, the present study explored dose-related effects of YO on food intake, conditioned flavor avoidance (CFA), and Fos immunolabeling in rats. Systemic YO (5.0 mg/kg BW, i.p.) inhibited food intake, supported CFA, and increased Fos immunolabeling in identified NA neurons in the ventrolateral medulla, nucleus of the solitary tract, and locus coeruleus. YO also increased Fos in the majority of corticotropin releasing hormone-positive neurons in the paraventricular nucleus of the hypothalamus. YO administered at 1.0 mg/kg BW did not inhibit food intake, did not support CFA, and did not increase Fos immunolabeling. Retrograde neural tracing demonstrated that neurons activated by YO at 5.0 mg/kg BW included medullary and pontine neurons that project to the central nucleus of the amygdala and to the lateral bed nucleus of the stria terminalis, the latter region receiving comparatively greater input by Fos-positive neurons. We conclude that YO produces anorexigenic and aversive effects that correlate with activation of brainstem viscerosensory inputs to the limbic forebrain. These findings invite continued investigation of how central viscerosensory signaling pathways interact with hypothalamic and limbic regions to influence interrelated physiological and behavioral components of anxiety, stress, and visceral malaise.


The Toll Signaling Pathway in the Chinese Oak Silkworm, Antheraea pernyi: Innate Immune Responses to Different Microorganisms.
The Toll pathway is one of the most important signaling pathways regulating insect innate immunity. Spatzle is a key protein that functions as a Toll receptor ligand to trigger Toll-dependent expression of immunity-related genes. In this study, a novel spatzle gene (ApSPZ) from the Chinese oak silkworm Antheraea pernyi was identified. The ApSPZ cDNA is 1065 nucleotides with an open reading frame (ORF) of 777 bp encoding a protein of 258 amino acids. The protein has an estimated molecular weight of 29.71 kDa and an isoelectric point (PI) of 8.53. ApSPZ is a nuclear and secretory protein with no conserved domains or membrane helices and shares 40% amino acid identity with SPZ from Manduca sexta. Phylogenetic analysis indicated that ApSPZ might be a new member of the Spatzle type 1 family, which belongs to the Spatzle superfamily. The expression patterns of several genes involved in the Toll pathway were examined at different developmental stages and various tissues in 5th instar larvae. The examined targets included A. pernyi spatzle, GNBP, MyD88, Tolloid, cactus and dorsalA. The RT-PCR results showed that these genes were predominantly expressed in immune-responsive fat body tissue, indicating that the genes play a crucial role in A. pernyi innate immunity. Moreover, A. pernyi infection with the fungus Nosema pernyi and the gram-positive bacterium Enterococcus pernyi, but not the gram-negative bacterium Escherichia coli, activated the Toll signaling pathway. These results represent the first study of the Toll pathway in A. pernyi, which provides insight into the A. pernyi innate immune system.


Ultrastructure of rat pituitary gonadotrophs after injection of synthetic luteinizing hormone releasing factor (LRF).
The effects of intracarotid injection of synthetic luteinizing hormone releasing factor (LRF) on the ultrastructure of pituitary gonadotrophs are reported. Chronically ovariectomized rats of the Sprague-Dawley strain pretreated with oestradiol and progesterone were sacrificed at zero time and at 30, 120 min after injection of 1 mug of synthetic LRF. A careful observation was made for evidence of granule extrusion from gonadotrophs following intracarotid injection of LRF, but no signs of granule extrusion were demonstrated. However, granule migration to the periphery of the cell was observed 30 min after LRF injection and the distribution of secretory granules recovered 2 hours after injection. The result of ultrastructural observations suggests that LRF plays an important role in releasing and synthesizing gonadotrophins in pituitary gonadotrophs. Further investigations should elucidate more clearly the morphologic aspect of mode of LRF action in pituitary gonadotrophs.


An RNA-dependent UMP-incorporating activity is associated with the small subunit of cytoplasmic ribosomes in bloodstream forms of Trypanosoma brucei.
Trypanosoma brucei cytoplasm contained a UMP-incorporating activity which was dependent upon the presence of endogenous RNA, stimulated by exogenous RNA and precipitable by ammonium sulphate. The activity cosedimented with ribosomes, and after ribosome dissociation using EDTA was mainly associated with the small (31S) subunit. Ribosome-associated activity was selective for UTP as a substrate and greatly stimulated by the presence of at least one other nucleoside triphosphate.


Drosophila Pax6 promotes development of the entire eye-antennal disc, thereby ensuring proper adult head formation.
Paired box 6 (Pax6) is considered to be the master control gene for eye development in all seeing animals studied so far. In vertebrates, it is required not only for lens/retina formation but also for the development of the CNS, olfactory system, and pancreas. Although Pax6 plays important roles in cell differentiation, proliferation, and patterning during the development of these systems, the underlying mechanism remains poorly understood. In the fruit fly, Drosophila melanogaster, Pax6 also functions in a range of tissues, including the eye and brain. In this report, we describe the function of Pax6 in Drosophila eye-antennal disc development. Previous studies have suggested that the two fly Pax6 genes, eyeless (ey) and twin of eyeless (toy), initiate eye specification, whereas eyegone (eyg) and the Notch (N) pathway independently regulate cell proliferation. Here, we show that Pax6 controls eye progenitor cell survival and proliferation through the activation of teashirt (tsh) and eyg, thereby indicating that Pax6 initiates both eye specification and proliferation. Although simultaneous loss of ey and toy during early eye-antennal disc development disrupts the development of all head structures derived from the eye-antennal disc, overexpression of N or tsh in the absence of Pax6 rescues only antennal and head epidermis development. Furthermore, overexpression of tsh induces a homeotic transformation of the fly head into thoracic structures. Taking these data together, we demonstrate that Pax6 promotes development of the entire eye-antennal disc and that the retinal determination network works to repress alternative tissue fates, which ensures proper development of adult head structures.


[The captopril test in nonselected hypertensive patients: absence of value for screening of renovascular patients and for unilateral small kidneys].
The decrease of blood pressure and the increase of plasma renin activity (PRA) after oral administration of captopril was evaluated in 104 consecutive hypertensive out-patients in whom the morphology of the renal arteries and parenchyma was assessed thanks to an intravenous digitalized angiography. Twenty five of these patients were excluded because of a natriuresis less than 50 mmol/24 h or non discontinuation of their treatment; 50 of these patients were classified as essential hypertension, 12 had renovascular disease (10 unilateral stenosis 2 of which significant; 2 bilateral significant stenosis); 9 had unilateral small kidneys (4 significant). The significance was ascertained on the PRA in the renal veins and/or the decrease of hypertension after surgery. The decrease of blood pressure after captopril was not different between the various groups. The increase after captopril of PRA was higher in unilateral significant lesions. However the highest post captopril PRA value was found in the essential hypertension group so that renovascular diseases could not be screened by higher post captopril PRA values. However taking into account that these latter were decreasing with age, a better discrimination of significant unilateral disease was possible in the hypertensive patients above 40 years of age. Furthermore once the diagnosis of unilateral kidney disease is established by radiological investigation, the captopril test allows to predict unilateral hypersecretion of renin with a sensitivity of 100% and a specificity of 78%. the captopril test does not allow to screen the non selected hypertensive patients for an efficient radiological investigation by intravenous digitalized angiography but may help to select the patients with unilateral renal disease for the renal venous PRA evaluation.


[Shortening of the measurement time by 1H-MR turbo spectroscopic imaging of the brain].
Development of a new technique for reduction of measurement time in 1H-MR spectroscopic imaging of the brain. Optimisation of the sequence parameters in volunteer and in patient examinations and comparison to the results obtained with conventional 2 D-SI. Examination of 20 healthy volunteers and 5 patients in a 1.5 T whole-body MR system. In "turbo-spectroscopic imaging" (TSI) sequences, a train of spin-echo signals with different phase encoding is acquired after each 90 degrees excitation. 32 x 32 matrix elements covered a field of view of 20 cm, and additional volume selection was performed by double spin echo excitation. Measurement duration 9 min with acquisition of four phase encoding steps per TR interval, whereas the corresponding 2 D-SI sequence (TR/TE 2000/272 ms) took 30 min. The TSI data sets yielded maps of the regional distribution of metabolite concentrations with a quality comparable to the 2 D-SI results. Signal homogeneity and delineation of brain lesions, however, were superior in conventional spectroscopic imaging. The T2 relaxation of the metabolites required a reduced sampling interval for each phase-encoded echo, and hence the frequency resolution of the corresponding TSI spectra was not always sufficient for separating choline and creatine signals. With measurement durations < 10 min the TSI technique allows in clinical studies a combination with single-voxel MRS for accurate quantification and with a diagnostic MRI within a total examination time of less than one hour.


Using systems thinking and the Intervention Level Framework to analyse public health planning for complex problems: Otitis media in Aboriginal and Torres Strait Islander children.
Middle ear disease (otitis media) is endemic among Aboriginal and Torres Strait Islander children in Australia and represents an important cause of hearing loss. The disease is the result of a mix of biological, environmental and host risk factors that interact in complex, non-linear ways along a dynamic continuum. As such, it is generally recognised that a holistic, systems approach is required to reverse the high rates of otitis media in Aboriginal and Torres Strait Islander children. The objective of this paper is to examine the alignment between efforts designed to address otitis media in Aboriginal and Torres Strait Islander children in Queensland, Australia and core concepts of systems thinking. This paper's overall purpose is to identify which combination of activities, and at which level, hold the potential to facilitate systems changes to better support ear health among Aboriginal and Torres Strait Islander children. We began with a review of documents identified in consultation with stakeholders and an online search. In addition, key informants were invited to participate in an online survey and a face-to-face or phone interview. Qualitative interviews using a semi-structured interview guide were used to explore survey responses in more depth. We also undertook interviews at the community level to elicit a diverse range of views. Ideas, statements or activities reported in the documents and interviews as being performed under the Intervention Level Framework were identified using qualitative thematic and content analysis. A quantitative descriptive analysis was also undertaken, whereby data was extracted into an Excel spreadsheet and coded under the relevant strategic directions and performance indicators of the Framework. Subsequently, we coded activities against the five-level intervention framework developed by Malhi and colleagues, that is: 1) paradigm; 2) goals; 3) system structure; 4) feedback and delays; and 5) structural elements. Overall, twenty documents were reviewed. We examined surveys and interviews with six key informants. Twenty-four individual and 3 group interviews were conducted across central and community level informants. One hundred and four items were coded from the 20 documents and 156 items from interview data. For both data sets, the majority of activities were coded at the structural elements level. The results suggested three key areas where further work is needed to drive sustained improvements: 1) build the governance structures needed for paradigm shift to achieve a multi-sectoral approach; 2) develop shared system level goals; 3) develop system-wide feedback processes. Sustained progress in improving ear health within Aboriginal and Torres Strait Islander children requires a holistic, system-wide approach. To advance such work, governance structures for multi-sectoral collaboration including the development of joint goals and monitoring and feedback are required. Intervening at these higher leverage points could have a profound effect on persistent public health issues.


The OsGSK2 Kinase Integrates Brassinosteroid and Jasmonic Acid Signaling by Interacting with OsJAZ4.
The crosstalk between brassinosteroid (BR) and jasmonic acid (JA) signaling is crucial for plant growth and defense responses. However, the detailed interplay between BRs and JA remains obscure. Here, we found that the rice (Oryza sativa) Glycogen synthase kinase3 (GSK3)-like kinase OsGSK2, a conserved kinase serving as a key suppressor of BR signaling, enhanced antiviral defense and the JA response. We identified a member of the JASMONATE ZIM-domain (JAZ) family, OsJAZ4, as a OsGSK2 substrate and confirmed that OsGSK2 interacted with and phosphorylated OsJAZ4. We demonstrated that OsGSK2 disrupted the OsJAZ4-OsNINJA complex and OsJAZ4-OsJAZ11 dimerization by competitively binding to the ZIM domain, perhaps helping to facilitate the degradation of OsJAZ4 via the 26S proteasome pathway. We also showed that OsJAZ4 negatively modulated JA signaling and antiviral defense and that the BR pathway was involved in modulating the stability of OsJAZ4 protein in an OsCORONATINE INSENSITIVE1-dependent manner. Collectively, these results suggest that OsGSK2 enhances plant antiviral defenses by activating JA signaling as it directly interacts with, phosphorylates, and destabilizes OsJAZ4. Thus, our findings provide a clear link between BR and JA signaling.


Tc-99m HMPAO brain SPECT findings in pediatric viral encephalitis.
Two children were diagnosed with viral encephalitis, due to Epstein-Barr virus infection in one case and to herpes simplex virus infection in the other. Tc-99m HMPAO brain SPECT was arranged to detect changes in regional cerebral blood flow (rCBF) secondary to viral encephalitis. During the acute episode, Tc-99m HMPAO brain SPECT showed that the two cases had increased rCBF. After the acute episode, follow-up brain SPECT was arranged 6 months later. The rCBF in one case was restored to normal on the second brain SPECT, and that in the other case was decreased. The child with normal rCBF in the follow-up brain SPECT had better learning ability and intelligence than the other child with decreased rCBF.


Vulnerable but strong: deaf people challenge established understandings of deafness.
In the last 35 years, hearing majorities have, in variable degree, gradually recognized that deaf people can be strong and able and a common knowledge of deaf people as linguistic minorities is partly embraced in public life. This is by and large a result of a long-time deaf struggle for the recognition of sign language and against paternalistic policies of pity. This article aims at showing how contemporary deaf identities are crafted by balancing vulnerability and empowering forces. In the presentation of life narratives from one deaf Norwegian and one deaf American, different aspects of being deaf are explored. These two stories have emerged from two related deaf research projects. They are deliberately chosen because they illustrate central insights gained within this research. The empowering aspects of being sign language users are in focus, as well as the joys of being connected to a global deaf movement. Difficulties and hardship are also part and parcel of the deaf lives displayed, and strong arguments are put forward against the medical model of deafness in particular. One conclusion is that deaf identities are vulnerable but at the same time can be strongly rewarding.


Characterization of two Ashkenazi Jewish founder mutations in MSH6 gene causing Lynch syndrome.
Founder mutations are an important cause of Lynch syndrome and facilitate genetic testing in specific ethnic populations. Two putative founder mutations in MSH6 were analyzed in 2685 colorectal cancer (CRC) cases, 337 endometrial cancer (EnCa) cases and 3310 healthy controls of Ashkenazi Jewish (AJ) descent from population-based and hospital-based case–control studies in Israel, Canada and the United States. The carriers were haplotyped and the age of the mutations was estimated. MSH6*c.3984_3987dupGTCA was found in 8/2685 CRC cases, 2/337 EnCa cases, and 1/3310 controls, consistent with a high risk of CRC (odds ratio (OR) = 9.9, 95% confidence interval (CI) = 1.2–78.9, p = 0.0079) and a very high risk of EnCa (OR = 19.6, 95% CI = 1.8–217.2, p = 0.0006). MSH6*c.3959_3962delCAAG was identified in 3/2685 CRC cases, 2/337 EnCa cases and no controls. Each mutation was observed on separate conserved haplotypes. MSH6*c.3984_3987dupGTCA and MSH6*c.3959_3962delCAAG probably arose around 585 CE and 685 CE, respectively. No carriers were identified in Sephardi Jews (450 cases and 490 controls). Truncating mutations MSH6*c.3984_3987dupGTCA and MSH6*c.3959_3962delCAAG cause Lynch syndrome and are founder mutations in Ashkenazi Jews. Together with other AJ founder mutations, they contribute substantially to the incidence of CRC and EnCa and are important tools for the early diagnosis and appropriate management of AJ Lynch syndrome patients.


Multiplex Serologic Assessment of Schistosomiasis in Western Kenya: Antibody Responses in Preschool Aged Children as a Measure of Reduced Transmission.
AbstractCurrently, impact of schistosomiasis control programs in Schistosoma mansoni-endemic areas is monitored primarily by assessment of parasitologic indicators only. Our study was conducted to evaluate the use of antibody responses as a way to measure the impact of schistosomiasis control programs. A total of 3,612 serum samples collected at three time points from children 1-5 years of age were tested for antibody responses to two schistosome antigens (soluble egg antigen [SEA] and Sm25) by multiplex bead assay. The overall prevalence of antibody responses to SEA was high at baseline (50.0%). After one round of mass drug administration (MDA), there was minimal change in odds of SEA positivity (odds ratio [OR] = 1.02, confidence interval [CI] = 0.79-1.32, P = 0.89). However, after two rounds of treatment, there was a slight decrease in odds of SEA positivity (OR = 0.80, CI = 0.63-1.02, P = 0.08). In contrast to the SEA results, prevalence of antibody responses to Sm25 was lowest at baseline (14.1%) and higher in years 2 (19.8%) and 3 (18.4%). After one round of MDA, odds of Sm25 positivity increased significantly (OR = 1.51, CI = 1.14-2.02, P = 0.005) and remained significantly higher than baseline after two rounds of MDA (OR = 1.37, CI = 1.07-1.76, P = 0.01). There was a significant decrease in the proportion of 1-year-olds with positive SEA responses from 33.1% in year 1 to 13.2% in year 3 and a corresponding decrease in the odds (OR = 3.25, CI = 1.75-6.08, P < 0.001). These results provide preliminary evidence that schistosomiasis program impact can be monitored using serologic responses.


Sex commonalities and differences in the relationship between resilient personality and the intrinsic connectivity of the salience and default mode networks.
Increased resilience is associated with better health outcomes and reduced morbidity in response to injury and homeostatic perturbations. Proper functioning of the salience network (SN) and modulation of the default mode network (DMN) by SN may play a role in adaptively responding to stress. Here, we demonstrate that resilient personality in healthy subjects is associated with SN and DMN connectivity patterns and that these patterns are influenced by sex. While connectivity of SN with several brain regions including right anterior insula was significantly associated with resilient personality in both men and women, results suggest that increased functional integration of anterior DMN preferentially benefits women while increased functional integration of posterior DMN preferentially benefits men in terms of resilience. These findings may relate to previous demonstrations that men and women engage different information processing and behavioral strategies to achieve resilience and highlight the importance of considering sex in resilience research.


[Evaluation of in vitro antibacterial potencies of oral antibiotics against sputum isolates].
Susceptibility of 162 sputum isolates to oral antibiotics was measured by an agar dilution method. The sputum isolates included S. pneumoniae 25 strains, S. aureus 30 strains, H. influenzae 37 strains, K. pneumoniae 51 strains and E. coli 19 strains. Minimal inhibitory concentration (MIC) values of cefaclor (CCL), cephalexin (CEX), ampicillin (ABPC) and minocycline (MINO) were measured for each strains. Eighty percent of S. pneumoniae strains were inhibited at 0.024 to 0.05 micrograms/ml of ABPC, 0.39 to 0.78 micrograms/ml of CCL, and 1.56 to 3.13 micrograms/ml of CEX and MINO. ABPC, CCL and CEX were considered to be effective clinically when they were used with the usual dosage. However, about 30% of strains were resistant to the usual dosage of orally administrated MINO. Eighty percent of S. aureus strains were inhibited at 0.20 to 0.39 microgram/ml of MINO and 3.13 to 6.25 micrograms/ml of the other 3 drugs. MINO is the most effective with the usual dosage. Twenty to 40% of strains showed resistance to CCL, CEX and ABPC. Eighty percent of H. influenzae strains were inhibited at 0.39 micrograms/ml of ABPC, 0.78 to 1.56 micrograms/ml of MINO, 3.13 micrograms/ml of CCL and 12.5 to 25 micrograms/ml of CEX. ABPC should be selected as the first choice antibiotic. However, there were 2 ABPC-resistant strains that were highly susceptible to CCL. Eighty percent of K. pneumoniae strains were inhibited at 0.39 to 0.78 micrograms/ml of CCL, 3.13 to 6.25 micrograms/ml of MINO and CEX, and 12.5 to 25 micrograms/ml of ABPC. CCL seemed to be only effective oral antibiotic for K. pneumoniae infection.(ABSTRACT TRUNCATED AT 250 WORDS)


Disparities in Rural Breast Cancer Care: Factors Affecting Choice of Breast Reconstruction in a West Virginia Tertiary Care Hospital.
The decision to pursue immediate postmastectomy breast reconstruction (IBR) is not uniform across all groups of women in the United States. We sought to investigate if the challenges of caring for a socioeconomically and geographically challenged rural population affected the incidence of IBR and to identify predictive factors of IBR in this population. Using our institution's cancer registry, we conducted a 10-year review of women with early-stage, unilateral breast cancer who received mastectomy. Demographics, insurance status, comorbidities, and surgeon graduation year were compared for patients receiving IBR versus no reconstruction. Variables with P < 0.05 on univariate analysis were included into a multivariate logistic regression model to determine independent predictors of IBR. From 2006 to 2015, 53/423 (12.5%) patients underwent IBR. On multivariate analysis, three factors independently predicted the decision to undergo IBR: age (P = 0.004), insurance type (P = 0.034), and use of contralateral prophylactic mastectomy (CPM, P < 0.001). Our data indicate that age, insurance type, and utilization of CPM influence the decision to pursue reconstruction. Additionally, the rate of IBR was found to be much lower in our West Virginia population than reported nationally, suggesting disparities in the care of women with early-stage, unilateral breast cancer in a rural population, even in a tertiary care environment.


[Knowledge and attitudes towards reproductive health: survey among moroccans living in rural areas].
The objective is to describe the knowledge, attitudes and practices (KAP) of men living in a landlocked area towards reproductive health. We conducted a mixed methods study (focus group and KAP survey ) in a mountainous region near Marrakech. This descriptive survey involved 74 men ; we used two sampling methods (souk visitors on a weekly basis and persons accompanying the expectant mothers to their antenatal visits as part of an antenatal clinic caravan campaign). Data collection (March - April 2014) was made using a questionnaire translated into respondents' dialect and administered by trained Amazigh-speaking investigators. Quantitative data analysis was descriptive and bivariate (statistical significance level at 5%). Focus group data were analyzed after recording and transcription. The median age was 38 years (19-80 years). Of the 64 men living together (86.5%), half (55.4%) opted for the pill. Two thirds recognized the importance of importance of pregnancy monitoring, while 37% of husbands were unaware of the frequency of follow-up visits. The non-use of services was 56.9% (N = 58) of which 42.4% without any reason. Educational attainment was associated with spouse's pregnancy screening (p = 0.015) and with a positive attitude toward the importance of pregnancy monitoring (p = 0.011). The lack of awareness regarding possible complications (35.1% of respondents) was a factor associated with lack of postnatal care among spouses (p = 0.021). The barriers identified in focus group analysis were cultural, geographic and socio-economic. Information, education, communication programmes should mostly aim to involve male participants in similar contexts.


Protein ligand interaction analysis against new CaMKK2 inhibitors by use of X-ray crystallography and the fragment molecular orbital (FMO) method.
CaMKK2 (calcium/calmodulin dependent protein kinase kinase 2) is a serine/threonine protein kinase that regulates phosphorylation of CaM kinases (CaMKs) such as CaMKI, CaMKIV, and AMP-activated protein kinase (AMPK). From a pathological perspective, CaMKK2 plays a role in obesity, diabetes, and prostate cancer. Therefore, CaMKK2 is an attractive target protein for drug design. Here, we tried to find new CaMKK2 inhibitors by using ligand-based and structure-based drug design approaches. From the in silico hit compounds, we identified new inhibitors by using a CaMKK2 kinase assay. We solved X-ray crystallography structures of the CaMKK2-inhibitor complexes and performed Fragment Molecular Orbital (FMO) calculations to analyze the protein-ligand interactions, identify the key residues in inhibitor binding, and quantitatively measure their contribution. We experimentally determined five CaMKK2-inhibitor structures and calculated the binding energies of the inhibitors by the FMO method plus MM-PBSA (Molecular Mechanics Poisson-Boltzmann Surface Area) approach. The results showed a high correlation (R = -0.89) between experimentally measured inhibitory activity (pIC50) and the predicted ligand binding energy. We then quantitatively evaluated the contribution of each binding site residue in CaMKK2 by the IFIE (Inter-fragment Interaction Energy)/PIEDA (Pair Interaction Energy Decomposition Analysis) method. The IFIE values indicated that Lys194 and Glu236, which formed hydrogen bonds with the carboxylate groups of the inhibitors, were key residues for ligand binding. PIEDA revealed that the dispersion interaction of inhibitors with hydrophobic residues, such as Ile171, Phe267, and Leu319, contributed highly to ligand binding; we considered that this was due to CH-π interactions with methoxy groups and/or aromatic rings contained in our CaMKK2 inhibitor. These results from the quantitative interaction analysis by the FMO method are useful not only for future CaMMK2 inhibitor development but for application of the FMO method to in silico drug design.


Rural-Urban Differences in Workers' Access to Paid Sick Leave.
Paid sick leave (PSL) is associated with health care access and health outcomes. The COVID-19 pandemic highlighted the importance of PSL as a public health strategy, yet PSL is not guaranteed in the United States. Rural workers may have more limited PSL, but research on rural PSL has been limited. We estimated unadjusted and adjusted PSL prevalence among rural versus urban workers and identified characteristics of rural workers with lower PSL access using the 2014-2017 Medical Expenditure Panel Survey. We found rural workers had lower access to PSL than urban workers, even after adjusting for worker and employment characteristics. Paid sick leave access was lowest among rural workers who were Hispanic, lacked employer-sponsored insurance, and reported poorer health status. Lower rural access to PSL poses a threat to the health and health care access of rural workers and has implications for the COVID-19 public health emergency and beyond.


Morphological changes of the basilar artery, ventricles, and choroid plexus after experimental SAH.
Experimental subarachnoid hemorrhage (SAH) was induced in adult rabbits by three repeated cisterna magna injections of autologous nonheparinized blood. One week after the last blood injection the animals were sacrificed and examined for morphological change. No vasculopathy was noted in the basilar arteries and no periarterial thrombus was seen around the majority of these arteries. Gross and microscopic changes were observed in both the ventricles and choroid plexus. Ventricular changes included dilation of the lateral ventricles, destruction of ciliated ependymal cells, and deposition of small amounts of blood throughout the ventricular system. These ventricular changes are similar to the pathological sequelae of SAH in patients. Choroid plexus changes included electron-dense cytoplasmic inclusions and dilation of the lateral and subcellular spaces. The ventricular surface and the choroid plexus are both affected by the intracisternal injection of blood. The hydrocephalus that follows SAH may be potentiated by ependymal disruption and loss of ciliated activity in the ventricles. Changes in choroid plexus function may also occur.


Failure of intrapartum antibiotics to prevent culture-proved neonatal group B streptococcal sepsis.
Early-onset group B streptococci (GBS-EOS) sepsis may be prevented by intrapartum antibiotics administered for GBS maternal colonization, premature labor, or prolonged rupture of membranes. We sought to identify cases of neonatal GBS sepsis after apparent failure of intrapartum chemotherapy and to determine the factors associated with failure of intrapartum antibiotics in these cases. We identified 96 GBS blood culture-positive infants at five military medical centers from 1987 to 1990. Eighteen (18.7%) of these infants had mothers who had received intrapartum antibiotics; 16 of 18 cases were early-onset disease, 15 of which initially had symptoms at less than 1 hour of age. Two infants had late-onset disease develop at 3 weeks of age. At least one perinatal risk factor (prematurity, prolonged rupture of membranes > 12 hours, maternal fever) was present in each of the 16 cases. Indications for intrapartum antibiotics were suspected chorioamnionitis (13 cases), GBS colonization and prolonged rupture of membranes or prematurity (3), and GBS colonization alone (2). Maternal antibiotics included ampicillin (14 cases), cephadyl (1), vancomycin (1), clindamycin (1), and gentamicin alone (1). The median number of doses of ampicillin before delivery was 1 (range, 1 to 21), which was administered at a median of 4 hours (range, 1 to 84) before birth. The mean dose of ampicillin was 1.8 gm/dose (range, 1 to 2 gm/dose). Two of 16 (12.5%) infants with GBS-EOS died as a result of GBS sepsis. In our population of neonates with GBS-EOS, 18.4% (16 of 87) of the infants had positive blood cultures despite intrapartum antibiotics. Intrapartum antibiotics may fail to prevent GBS sepsis in a number of infants born to mothers colonized with GBS or to those with acute chorioamnionitis.


High Affinity Mannotetraose as an Alternative to Dextran in ConA Based Fluorescent Affinity Glucose Assay Due to Improved FRET Efficiency.
Diabetes mellitus affects millions of people worldwide and requires that individuals tightly self-regulate their blood glucose levels to minimize the associated secondary complications. Continuous monitoring devices potentially offer patients a long-term means to tightly monitor their glucose levels. In recent years, fluorescent affinity sensors based on lectins (e.g., Concanavalin A (ConA)) have been implemented in such devices. Traditionally, these sensors pair the lectin with a multivalent ligand, like dextran, in order to develop a competitive binding assay that changes its fluorescent properties in response to the surrounding glucose concentrations. This work introduces a new type of fluorescent ligand for FRET-based assays in an attempt to improve the sensitivity of such assays. This ligand is rationally designed to present a core trimannose structure and a donor fluorophore in close proximity to one another. This design decreases the distance between the FRET donor and the FRET acceptors on ConA to maximize the FRET efficiency upon binding of the ligand to ConA. This work specifically compares the FRET efficiency and sensitivity of this new competing ligand with a traditional dextran ligand, showing that the new ligand has improved characteristics. This work also tested the long-term thermal stability of the assay based on this new competing ligand and displayed a MARD of less than 10% across the physiological range of glucose after 30 days incubation at 37 °C. Ultimately, this new type of fluorescent ligand has the potential to significantly improve the accuracy of continuous glucose monitoring devices based on the competitive binding sensing approach.