Visual selection and response selection without effector selection in tasks with circular arrays.
Charles Eriksen and colleagues conducted influential visual-search experiments with circular arrays for which the responses were either vocal naming or unimanual left-right switch movements. These methods have the advantages of the stimuli being equidistant from a centered fixation point and allowing study of visual selection and response selection when effector selection is not required, as in the more typical case in which responses are key presses of distinct fingers. Other researchers have used similar spatial arrangements, but with aimed movements of the limb or of a mouse-controlled cursor to study effects of stimulus identification, visual search, spatial stimulus-response compatibility, response-effect compatibility, and practice/transfer in isolation and jointly. We systematically review studies in these areas that include visual selection and response selection and execution, and examine implications of their results for the role of effector selection. Also, we illustrate that as one moves from simpler to more complex tasks, the results are consistent with a basic information-processing framework in which stimulus identification and selection of a target response location are distinct from selecting, planning, and moving an effector to the targeted location.


Immunohistopathologic findings in proliferative diabetic retinopathy.
We performed immunopathologic studies on pars plana specimens obtained by biopsy in patients with diabetes mellitus type I or II and by autopsy in diabetic patients and normal subjects. Frozen sections were treated with several antisera, including anti-IgG, complement components, and major histocompatibility complex antigens, as well as anti-factor VIII to detect vascular structures. The results showed IgG in a linear pattern at the basal pole of pigment epithelial cells and complement deposits of C3c, C3d, and C4 at the same location and in the stroma. HLA-DR expression was found at the level of the pigmented cells. These data suggest that some autoimmune processes may be involved in proliferative diabetic retinopathy at the level of the pigment epithelium, but it is unknown whether they are an epiphenomenon of neovascularization or if they play a role in its initiation.


Increased healthcare costs in COVID-19 patients with unhealthy habits: The case of smoking.
This study aims to analyze the differences in the prognosis and cost of COVID-19 patients in terms of mortality and occurrence of complications due to tobacco use. This study was conducted using a unique Spanish electronic database built by health professionals during the first wave of the pandemic on the admission and evolution of a patient infected by the SARS-CoV-2 virus. Data were collected on all patients admitted to La Paz hospital (Madrid) from the pandemic's beginning until 15 July 2020. Demographic factors and the incidence of complications in smoker and non-smoker patients were compared using the Mann-Whitney U-test or chi-squared test. Survival analysis was performed using the Kaplan-Meier estimator and Cox regression. Finally, the costs between the two groups were estimated using a Generalized Linear Model. A total of 3521 patients were included in the analysis, with median age of 62 years (IQR: 47-78), 51.09% were women, and 16.42% were smokers. Patients who smoked had a higher incidence of complications during their hospital stay, especially complications related to the respiratory and cardiac systems. They were also associated with a worse prognosis in terms of the need for ICU admission and mortality, leading to an increase in the management cost of the smoking COVID-19 patients by 14.72%. Healthcare in Spain is mainly financed by the national tax system, so introducing an additional financing system for pathologies related to the consumption of addictive substances and associated diseases and complications would decrease the burden on the economy in terms of healthcare.


Engineering a twist in 9,10-diethynylanthracenes by steric interactions.
A series of 9,10-bis(phenylethynyl)anthracenes decorated with sterically demanding tert-butyl substituents have been prepared and spectroscopically characterised. We demonstrate that the introduction of two bulky substituents in the ortho position of the phenyl rings effectively locks the ground state into a conformation in which the three rings are orthogonal. Fluorescence spectroscopy reveals evidence for partial planarisation of this compound in the excited state at ambient temperature, but this is prevented in low temperature solvent glasses.


Development of arteriosclerosis after i.v. administered fat in rabbits.
In our previous studies performed in rats sclerotic alterations were induced in the vessel wall by Lipofundin-S 20% (Braun, Melsungen) administration for 8 or 16 days, respectively. In order to elucidate eventual species dependency, studies on rabbits were also performed. Alterations were examined in rabbits after Lipofundin treatment for 8 days. Electron microscopic studies revealed the accumulation of granular basement membrane-like material in the subendothelial space due to the activity of immigrated smooth muscle cells. The formation of elastic granules and elastic laminae was also observed among others due to the coalescence of microfibrillary structures formed from glycoseaminoglycans. The lesion developed was similar to that observed in the rat, the only difference was that in the rabbit the accumulation of the collagen fibres was more prominent. Results of our studies obtained in rabbits appear to support our previous findings: Lipofundin-S treatment induces aortic smooth muscle cell mobilization, fibrillogenesis and enlargement of the subendothelial space, namely sclerotic manifestations are induced.


Single-step, homogeneous and sensitive detection for microRNAs with dual-recognition steps based on luminescence resonance energy transfer (LRET) using upconversion nanoparticles.
A single-step, homogeneous and sensitive LRET assay is presented for the detection of miRNAs. The amplification-free assay provides a unique combination of high specificity with dual-recognition approach of different hybridization and ligation steps and preventing background auto-fluorescence in biological samples using upconversion nanoparticles (UCNPs) as signal-producing nanoprobes. The assay probe is composed of signal-producing unit (a pair of homogeneous upconversion luminescence resonance energy transfer (UC-LRET)-based oligonucleotides) and recognition unit (two adaptor oligonucleotides). In the presence of target miRNAs, the probe and target miRNAs leads to the formation of stable double-strands and semi-stable adaptor-miRNAs complexes with an adaptor nick. Ligation of the nick using ligase cause the formation of stable double-strands, resulting in UCNPs-to-dye UC-LRET for detection of the miRNAs with near-infrared radiation (980nm). Sensitive detection of miRNA-21 at concentrations of 200pM to 1.4nM and detection limits of 0.095nM with good precision of 3.9% (RSD) for seven repeated measurements of 500pM miRNAs demonstrate the feasibility of both high throughput and point-of-care clinical diagnostics. The homogeneous UC-LRET assay without any washing can be extended to the application in other important types of nucleic acid analysis.


Rab 23 is expressed in the glomerulus and plays a role in the development of focal segmental glomerulosclerosis.
Rab23, a member of the Rab family of small GTPase, has a function in antagonizing sonic hedgehog signal transduction. Both Rab-family and hedgehog-related proteins are involved in sclerosis and fibrosis in certain pathological states, but their roles in focal segmental glomerulosclerosis (FSGS) remain unclear. The FSGS model was established in Balb/c mice by a single injection of adriamycin. Serum, urine and mice kidneys were collected on Days 0, 7, 15 and 20. Western blot analysis was performed to detect the levels of Rab23 in the samples. Immunohistochemistry was used to examine the expressional profiles of Rab23 in kidneys. The expressions of transcripts of Rab23, extracellular matrix (ECM) proteins, and various hedgehog signalling pathway genes in kidneys or mesangial cells were evaluated by real-time RT-PCR. The effect of Rab23 on ECM protein expressions was evaluated by the knockdown or overexpression of Rab23 in mesangial cells. Our results show that elevations of Rab23 were observed in the urine, but not in the serum, of the FSGS mice. Rab23 and hedgehog signalling pathway genes were constitutively expressed in normal kidneys and were significantly up-regulated in the kidneys of FSGS mice. The basal expression of Rab23 was identified in glomeruli, and mesangial cells displayed obvious elevation of Rab23 in the FSGS state. The knockdown or overexpression of Rab23 affected the collagen expression in cultured mesangial cells. An autocrine loop of hedgehog signalling could be activated in mesangial cells in the FSGS state, and Rab23 may be elevated to suppress hedgehog signalling and/or influence collagen synthesis. Importantly, Rab23 could serve as a biomarker that indicates the severity of FSGS.


Survivin and molecular pathogenesis of colorectal cancer.
Colorectal cancer is thought to originate in the expansion of colonic crypt cells as a result of aberrant gene expression caused by transcription factors of the T-cell factor (TCF)/beta-catenin family. Survivin is a bifunctional regulator of cell death and cell proliferation expressed during embryonic development but undetectable in healthy adult tissues and re-expressed in many cancers, including colorectal cancer. We investigated gene expression by promoter analysis, mutagenesis, and electrophoretic mobility shift assay in colorectal cancer cells. Survivin expression in human and mouse embryonic intestine was determined by in-situ hybridisation and immunohistochemistry. Changes in apoptosis were monitored in cell lines engineered to express stabilising mutations in beta catenin. TCF/beta catenin stimulated a six-fold to 12-fold increased expression of the survivin gene in colorectal cancer cells. Three TCF-binding elements (TBE) in the survivin promoter were occupied by nuclear factors in colorectal cancer cells, and mutagenesis of the two proximal TBE sites abolished survivin gene expression by 75-79%. Strongly expressed at the bottom of human and mouse embryonic intestinal crypts, expression of survivin was lost in TCF-4 knockout animals, and a TCF-4 dominant negative mutant blocked survivin gene transcription in colorectal cancer cells. Expression of non-destructible beta catenin mutants increased survivin expression and protected against ultraviolet-B-induced apoptosis. Stimulation of survivin expression by TCF/beta catenin might impose a stem cell-like phenotype to colonic crypt epithelium coupling enhanced cell proliferation with resistance to apoptosis, and contribute to the molecular pathogenesis of colorectal cancer.


Azithromycin induces neurotoxicity in zebrafish by interfering with the VEGF/Notch signaling pathway.
Azithromycin (AZM) is a widely used antibiotic in both human and veterinary medicine, and its use has significantly increased during the COVID-19 pandemic. However, potential adverse effects of AZM on aquatic organisms have not been well studied. In this study, we explored the neurotoxicity of AZM in zebrafish and delved into its underlying mechanisms. Our results showed that AZM exposure resulted in a spectrum of detrimental effects in zebrafish, encompassing abnormal behaviors, damaged neuronal development, aberrant lateral line nervous system development, vascular malformations and perturbed expression of genes related to neural development. Moreover, we observed a concentration-dependent exacerbation of these neurotoxic manifestations with increasing AZM concentrations. Notably, AZM induced excessive cell apoptosis and oxidative stress damage. In addition, alterations in the expression levels of the genes involved in the VEGF/Notch signaling pathway were evident in AZM-exposed zebrafish. Consequently, we hypothesize that AZM may induce neurotoxicity by influencing the VEGF/Notch signaling pathway. To validate this hypothesis, we introduced a VEGF signaling inhibitor, axitinib, and a Notch signaling agonist, valproic acid, alongside AZM exposure. Remarkably, the administration of these rescue compounds significantly mitigated the neurotoxic effects induced by AZM. This dual verification provides compelling evidence that AZM indeed induces neurotoxicity during the early developmental stages of zebrafish, primarily through its interference with the VEGF/Notch pathway. Innovatively, our study reveals the molecular mechanism of AZM-induced neurotoxicity from the perspective of the close connection between blood vessels and nervous system. These findings provide new insights into the potential mechanisms underlying the neurotoxic effect of antibiotics and highlight the need for further investigation into the ecotoxicological effects of antibiotics on aquatic organisms and the potential risks to human health.


Small birth weight and later body composition and fat distribution in adolescents: the Avena study.
To assess the association between birth weight and body composition and fat distribution in adolescents, and to test the possible sex-specific effect in these relationships. A total of 1,223 adolescents 13-18.5 years old (553 male adolescents and 670 female adolescents) born at >35 weeks, were selected from a cross-sectional multicenter study conducted in five Spanish cities in 2000-2002. BMI was calculated from weight and height. Triceps and subscapular skinfold thickness (ST) were measured on the left side, and fat mass (FM) and fat-free mass (FFM) were estimated according to the equations of Slaughter et al. Subscapular skinfold adjusted by tricipital (ST) and waist circumference were used as markers of central adiposity. Birth weight Z-score was positively associated with FFM in female adolescents (P<0.001), but not in male adolescents, after controlling for age, pubertal stage, gestational age, socioeconomic status, physical activity, and current height (P<0.001 for interaction between adjusted birth weight Z-score and sex). Adjusted birth weight Z-score was inversely associated with central adiposity in male and female adolescents as measured by ST (P=0.026). These results provide further evidence that gender has an important influence on the programming effect of birth weight on later FFM in adolescents because the effect was only observed in female adolescents. Our results suggest that small size for gestational age at birth could program more central subcutaneous fat deposition in adolescents of both sexes, but further research is needed on this issue.


Pptidase activities in group N streptococci.
Several peptidase activites in the 3 species of Group N streptococci were partly separated by gel filtration on Sephadex G-200. The peptidases identified were a general aminopeptidase of wide specificity, a tripeptidase, a proline iminopeptidase (prolyl-beta-napthylamidase), a proline iminodipeptidase and an aminopeptidase-P. The effects of temperature and pH on the stability of the enzyme activities, and the influence of the type of N source used in the growth medium on the elution pattern of the enzymes were examined.


Antithrombotic activity of a symmetrical triglyceride with eicosapentaenoic acid and gamma-linolenic acid in guinea pig mesenteric microvasculature.
The antithrombotic effect of a synthetic symmetrical triglyceride having eicosapentaenoic acid (EPA) at positions 1 and 3, and gamma-linolenic acid (GLA) at position 2 was investigated. Administration of the triglyceride significantly increased thrombus formation time and thrombotic occlusion time induced by light irradiation and a fluorescent dye in guinea pigs after 14 days administration compared to that of soybean oil. The antithrombotic effect of the triglyceride was similar to that of EPA ethyl ester. Administration of the triglyceride increased GLA, dihomo-gamma-linolenic acid (DGLA) and EPA contents in plasma and the liver, and the ratio of DGLA to arachidonic acid. These results might be responsible for this antithrombotic effect.


Stress echocardiography for risk stratification of asymptomatic patients with type 2 diabetes mellitus.
The purpose of this study was to assess stressechocardiography (SE) for risk stratification of asymptomatic type 2 diabetic patients (DM2) without known coronary artery disease CAD. A total of 211 consecutive, asymptomatic DM2 patients underwent exercise (n=177) or dobutamine (n=34) SE and were followed up for 11+/-2 months. Primary endpoint was a major cardiac or vascular event (MACCE; all-cause-death, non-fatal myocardial infarction, coronary revascularization procedures, cerebrovascular event, acute limb-ischemia). During follow-up 39 of these patients suffered a MACCE. SE correctly identified 33 of these 39 patients by demonstrating silent ischemia in advance. In a multivariate logistic regression analysis a positive SE turned out to be an independent predictor for the occurrence of a MACCE during 11+/-2 months. SE represents an effective tool for risk stratification of asymptomatic DM2 patients.


Theoretical quantification of the effects of plastic wall thickness on phantom measurements in electromagnetic hyperthermia.
Phantom experiments are a staple of research and development in electromagnetic hyperthermia. Phantom containers and compartments are typically constructed from plastics which are readily available in a wide variety of thicknesses and material compositions. The perturbation effects of these plastics on the electric fields to be measured may be important, especially if one is trying to obtain quantitative results such as when comparing with a numerical model. This communication presents a theoretical investigation into the effects of plastic wall thickness on the computed electric field. Design curves are reported which aid in the selecting of an acceptable wall thickness given a maximum degree of wall perturbation that can be tolerated. Many other materials such as rubbers and polystyrenes also have electrical properties within the ranges considered herein; hence, the results should apply to a variety of commonly used phantom construction materials.


Membrane-localized neoantigens predict the efficacy of cancer immunotherapy.
Immune checkpoint immunotherapy (ICI) can re-activate immune reactions against neoantigens, leading to remarkable remission in cancer patients. Nevertheless, only a minority of patients are responsive to ICI, and approaches for prediction of responsiveness are needed to improve the success of cancer treatments. While the tumor mutational burden (TMB) correlates positively with responsiveness and survival of patients undergoing ICI, the influence of the subcellular localizations of the neoantigens remains unclear. Here, we demonstrate in both a mouse melanoma model and human clinical datasets of 1,722 ICI-treated patients that a high proportion of membrane-localized neoantigens, particularly at the plasma membrane, correlate with responsiveness to ICI therapy and improved overall survival across multiple cancer types. We further show that combining membrane localization and TMB analyses can enhance the predictability of cancer patient response to ICI. Our results may have important implications for establishing future clinical guidelines to direct the choice of treatment toward ICI.


Synthesis of potent inhibitors of β-ketoacyl-acyl carrier protein synthase III as potential antimicrobial agents.
Mycobacterium tuberculosis FabH, an essential enzyme in the mycolic acid biosynthetic pathway, is an attractive target for novel anti-tubercolosis agents. Structure-based design and synthesis of 1-(4-carboxybutyl)-4-(4-(substituted benzyloxy)phenyl)-1H-pyrrole-2-carboxylic acid derivatives 7a-h, a subset of eight potential FabH inhibitors, is described in this paper. The Vilsmeier-Haack reaction was employed as a key step. The structures of all the newly synthesized compounds were identified by IR, ¹H-NMR, ¹³C-NMR, ESI-MS and HRMS. The alamarBlue™ microassay was employed to evaluate the compounds 7a-h against Mycobacterium tuberculosis H₃₇Rv. The results demonstrate that the compound 7d possesses good in vitro antimycobacterial activity against Mycobacterium tuberculosis H₃₇Rv (Minimum Inhibitory Concentration value [MIC], 12.5 µg/mL).These compounds may prove useful in the discovery and development of new anti-tuberculosis drugs.


A novel radiographic analysis system for subaxial cervical spine pedicle screw placement.
Precise pedicle screw placement of the subaxial cervical spine is difficult. Not every hospital is equipped with a guidance system that can provide effective help. Computed tomography (CT) scanning is almost a routine preoperative examination for cervical spine surgery in all hospitals. Appropriate measurement and analysis of the CT images could assist optimal cervical pedicle screw placement. The purpose of this study is to propose a new and universal method using computed tomography (CT) morphological parameters analysis to assist optimal cervical pedicle screw placement from C3 to C7. A localization system with six parameters was designed based on preoperative CT reconstruction to guide subaxial cervical spine pedicle screw placement. The six parameters were distance from the starting point to the midline [D1], distance from the starting point to the lower edge of the inferior articular process [D2], transverse section angle [TSA], sagittal section angle [SSA], pedicle width [PW], and pedicle height [PH]. The six parameters were analyzed in 53 participants. Combining D1 and D2 could localize the entrance of the pedicle screw, and we concluded that D1 and TSA and D2 and SSA could be a new standard for determination of the transverse and sagittal orientation of the pedicle screw. The six parameters were closely related to the patient's gender, height, and weight. PH and PW were linearly correlated and could guide selection of the appropriate pedicle screw. SSA was an independent parameter of the relevant vertebral body, and changes in SSA had nothing to do with the curvature or posture of the cervical spine. Understanding and applying the six-parameter localization system are essential for achieving accurate and optimal pedicle screw placement in subaxial cervical spine, regardless of cervical sagittal alignment.


[Allogeneic stem cell transplantation for acute myeloid leukemia and HIV infection--case 3/2012].
A 27-year-old male patient with a past medical history of HIV presented with acute myeloid leukemia for allogeneic hematopoietic stem cell transplantation (HSCT). Highly active anti-retroviral therapy suppressed the viral load below detection threshold. There were no contraindications for allogeneic HSCT. Myeloablative conditioning consisted of total body irradiation and cyclophosphamide. Anti-thymocyte globulin, tacrolimus and mycophenolate mofetil were used for immunosuppression. Combined anti-retroviral therapy (nucleoside and nucleotide analog reverse-transcriptase inhibitor, boostered protease inhibitor, maraviroc and raltegravir) was maintained for allogeneic HSCT and viral load remained below detection threshold. No graft-versus-host disease or serious infectious complications occurred. The patient showed good graft function with stable hematopoiesis. Localized Kaposi's sarcoma was diagnosed six months after allogeneic HSCT and treated successfully with surgical excision and reduction of immunosuppression. Almost one year after allogeneic HSCT, the CD4+ cell count is rising and viral load remains below detection threshold with combined anti-retroviral therapy. Allogeneic HSCT can be safely performed in HIV positive patients. Kaposi's sarcoma is a rare event after allogeneic HSCT and linked to strong immunosuppression.


[Influence of the irritant used to provoke the peritoneal exudate used in the macrophage migration inhibition test (author's transl)].
The effect of the irritant chosen to provoke the peritoneal exudate in donors of cells for the in vitro migration inhibition test was studied. Guinea-pigs with delayed hypersensitivity to tubercle bacilli were used. Irritants studied included starch, Bayol and thioglycollate medium. The exudates obtained with thioglycollate were found to show greater inhibition than those obtained with the other irritants in the in vitro response to PPD, Inhibition of migration was seen when 1 microgram/ml of PPD was added to the culture medium; with this dose the inhibition of migration was 53%. The state of activation of macrophages caused by the irritant thus appears to have some importance for their responsiveness to factors liberated by specifically sensitized lymphocytes.


Restructuring reward: A pilot study to enhance natural reward response in adults with cannabis use disorder.
Hedonic dysregulation is a core mechanism of addiction. There is a dearth of research on hedonic dysregulation in cannabis use disorder (CUD). The current study tested whether personalized scripted imagery may be an efficacious approach to remediate reward functioning in adults with CUD. Adults with CUD (n=10) and non-CUD controls (n=12) completed a single session personalized scripted imagery procedure. Non-drug (i.e. natural) reward and neutral scripts were transcribed and participants listened to the scripts in counterbalanced order. Primary outcomes included positive affect (PA), galvanic skin response (GSR), and cortisol and were assessed at four timepoints. Mixed effects models were used to compare between and within subject effects. Mixed effects models revealed a Condition (reward vs. neutral) X Group (CUD vs. control) interaction (p=0.01) on PA response, indicating blunted PA response to the neutral script relative to the reward script in CUD participants. Likewise, GSR response in CUD participants was decreased in response to the neutral script relative to the reward script (p=0.034; interaction n.s.). An interaction effect of Group X PA on cortisol response was found (p=.036) indicating that cortisol was positively correlated with PA in healthy control participants, but not CUD participants. Adults with CUD may demonstrate acute deficits in hedonic tone under neutral conditions relative to healthy controls. Personalized scripted imagery may be an efficacious tool to remediate hedonic dysregulation in CUD. Cortisol may play a role in healthy positive affect regulation warranting further investigation.


Liver injury associated with the beta-interferons for MS: a comparison between the three products.
A population-based retrospective chart review of the biochemical liver tests of 844 patients with multiple sclerosis prescribed a beta-interferon (IFNbeta) product in British Columbia, Canada was performed between 1995 and 2001. Overall, 36.9% (243/659) of patients developed new elevations of alanine aminotransferase. All the IFNbetas caused elevated aminotransferase levels compared with pretreatment levels (p < 0.005) and were higher than reported in clinical trials. Their relative effect on aminotransferases can be approximated as IFNbeta-1b(subcutaneous [SC]) = IFNbeta-1a(SC) > IFNbeta-1a(IM).


An energy optimization study of the electric arc furnace from the steelmaking process with hot metal charging.
A study of energy optimization derived from simulation software has been done for the high-intensity energy of an Electric Arc Furnace (EAF), hot metal charging within the cooler duct dedusting system. The present work aims to develop a dynamic model of the EAF operation based on mass and energy balances integrated with simulation of the dedusting system with hot metal charging system using MATLAB® and Computational Fluid Dynamics (CFD). The effect of various percentages of hot metal charging on EAF performance and the dedusting system was simulated and validated from the real EAF data plant from one of the steel manufacturer companies in Indonesia. Three cases for the EAF with various hot metal (HM), sponge iron, and scrap iron charging compositions and four cases for the EAF with post-combustion CO have been developed. Careful observation shows that the electric arc power consumption can be reduced down to 72.9 MWh from 87.4 MWh (ca. 16% more efficient) while at the same time increasing the HM charging temperature at the endpoint of the duct dedusting system up to 900 °C from 540 °C (app. 65% higher). Additionally, advanced simulation of an EAF with post-combustion CO shows that power consumption can be decreased to 59.9 MWh (ca. 30% more efficient).


The radiologic approach to malignant tumors of the head and neck, with emphasis on computed tomography.
Computed tomography is the single most effective tool for the investigation and presurgical mapping of the extent of malignant tumors of the upper aerodigestive tract, paranasal sinuses, orbit, skull base, larynx, and neck. Close cooperation between the primary physician and the radiologist is essential for planning an efficient and cost-effective diagnostic plan. The radiologist may also assist the surgeon by obliterating tumor hypervascularity with preoperative fluoroscopically controlled arterial embolization and occlusion.


Interferon-alfa corrects thrombocytosis in patients with myeloproliferative disorders.
During previous therapeutic trials with interferon, decreased levels of peripheral platelet counts have been observed. Taking advantage of this effect, we investigated the efficacy of recombinant interferon (rec-IFN) in the treatment of thrombocytosis in myeloproliferative diseases. A total of 15 patients with polycythemia vera, essential thrombocytosis, or chronic myeloid leukemia received rec-IFN-alfa at initial doses of 25-70 x 10(6) units/week; maintenance therapy following week 8 of treatment consisted of 20-35 x 10(6) units/week rec-IFN. Observation periods ranged from 24 to 48 weeks. Significant reductions in the number of platelets were noted in all cases; 12/15 patients achieved platelet counts below 440 x 10(9)/l and maintained those normal values for at least 4 weeks. The number of bone marrow megakaryocytes, which had been increased prior to treatment, diminished during rec-IFN therapy, while the previously shortened platelet half-life further decreased with rec-IFN treatment. During rec-IFN-induced remission, the plasma levels of platelet factors, the activity of natural killer cells, and platelet aggregation showed changes between slight improvement and normal values. Severe side effects were only observed with the highest rec-IFN doses; dosage adjustments were effective in improving or eliminating all treatment-related symptoms. Rec-IFN may prove to be a valuable therapeutic alternative to cytostatic treatment of thrombocytosis in myeloproliferative disorders.


Effects of birth weight and growth on childhood wheezing disorders: findings from the Born in Bradford Cohort.
To examine the effects of birth weight and childhood growth on childhood wheezing disorders. We hypothesised that low birth weight and fast growth during early age would increase the risk of wheezing disorders. Observational secondary analysis of data from the Born in Bradford cohort. All children who were born at the Bradford Royal Infirmary hospital between March 2007 and December 2010 were eligible for the study. A total of 13,734 and 1598 children participated in the analyses of the effects of birth weight and growth on wheezing disorders, respectively. Wheezing disorders diagnosis (diagnosed as asthma or had wheezing symptom) during the ages of 0-7 years were the primary outcome measures. Diagnosis of asthma and occurrence of wheezing during the same period were secondary outcome measures. Birth weight was classified as normal (2.5-4.0 kg), low (<2.5 kg) and high (>4.0 kg). Growth mixture models were used to drive growth pattern outcomes which were classified as 'normal', 'fast' and 'slow' growth based on their velocities between birth and 36 months. The adjusted relative risks (RRs) of wheezing disorders diagnosis for the low and high birthweight children were 1.29 (95% CI 1.12 to 1.50; p=0.001) and 0.91 (95% CI 0.79 to 1.04; p=0.17), respectively. The adjusted RRs of wheezing disorders diagnosis were 1.30 (95% CI 0.56 to 3.06; p=0.54) and 0.60 (95% CI 0.16 to 2.18; p=0.44), respectively, for the 'fast' and 'slow' growth as compared with the 'normal' growth. Low birth weight is associated with an increased risk of wheezing disorders; however, there is a weak evidence that suggests high birthweight children have a reduced risk in this birth cohort. Low birth weight coupled with a slower growth until 3 months and a sharp growth between 3 and 12 months has an increased risk of wheezing disorders diagnosis.


Is conservative organ-sparing treatment of penile carcinoma justified?
The results of different conservative organ-sparing methods - radiotherapy, chemotherapy and radiochemotherapy - in the treatment of penile carcinoma were studied. Conservatively treated 223 patients with penile carcinoma seen in three hospitals between 1959 and 1996 years were studied retrospectively. Among them 155 received radiotherapy, 33 chemotherapy, and 35 radiochemotherapy. The local control was achieved in 135 (60.5%) of 223 patients. The difference in local control rate among these three groups of patients has not reached statistical significance. The efficacy of conservative treatment was highly associated with three factors: tumor size, grade and patient's age. Overall, 24 of 135 patients (17.7%) had local recurrence. Ten patients (4.5%) developed regional metastases. The recurrence rate did not correlate with tumor size and grade and was similar for all treatment modalities. Long-term results of each method were approximatley equal and 5-year survival varied from 78 to 88%. Conservative organ-sparing treatment of early-stage penile carcinomas is justified. Failure should be corrected by surgery without compromising survival.


Dexamethasone reduces bilirubin-induced toxicity and IL-8 and MCP-1 release in human NT2-N neurons.
The mechanisms of neurotoxicity induced by unconjugated bilirubin (UCB) in newborns are incompletely understood. UCB may cause both necrotic and apoptotic neuronal death. We explored UCB toxicity and release of cytokines in human NT2-N neurons and the effect of dexamethasone on these processes. Cultured NT2-N neurons were exposed to UCB, and neuronal damage was evaluated by LDH release and MTT cleavage. After 96 hours, 2 μM UCB significantly increased release of IL-8 and MCP-1, but not IL-13, IP-10, PDGF, or VEGF. Dexamethasone significantly lowered the UCB-induced increase in MCP-1 release, and attenuated UCB-induced neuronal damage assessed with MTT cleavage and LDH release. For comparison, the effects of hydrogen peroxide on cytokine formation and neuronal damage were tested. Hydrogen peroxide increased MCP-1, IP-10, and VEGF, but not IL-8, IL-13, or PDGF. Dexamethasone inhibited the hydrogen peroxide-induced increase in MCP-1 and IP-10. We conclude that UCB causes release of IL-8 and MCP-1 in cultured human NT2-N neurons. Dexamethasone reduces UCB-induced cytokine release and protects against UCB-induced toxicity.


Steering single-element lead zirconate titanate ultrasound transducers using biaxial driving.
Previous work has shown that biaxial driving using two phase-offset orthogonal electric fields (propagation and lateral) improves the efficiency of ferroelectric materials by reducing coercivity and, hence, energy dissipation. In the current investigation, we demonstrated the capability of the biaxial method to steer ultrasound waves in single-element piezoceramic transducers made of prismatic lead zirconate titanate (PZT). We conducted finite element analysis simulations for 133 kHz (model 1) and 470 kHz biaxial (model 2) transducers models. We performed experimental validation with biaxially driven single-element transducers (n = 3) operating at an average frequency of 131 kHz with the same characteristics as model 1. For both models, we found non-symmetric steering that was a function of both the phase and power of the second electric field. At a constant electrical power (1 W) on the propagation electrodes, simulations for the 133 kHz model predicted maximal steering of 10.3°, 22.6°, and 30.9° for lateral electrode powers of 0.1 W, 0.5 W, and 1.0 W, respectively. Experimentally, for model 1, the maximal steering was 11.7° ± 1.9°, 23.5° ± 3.5°, and 30.2° ± 4.4° for the lateral electrode powers of 0.1 W, 0.5 W, and 1.0 W, respectively. Simulations for the 470 kHz model predicted maximal steering of 8.8°, 16.1°, and 27° for lateral electrode powers of 0.1 W, 0.5 W, and 1.0 W, respectively. Simulations showed that the cause of the steering asymmetry was a non-uniform shear deformation associated with the slightly off-resonance lateral electric field driving frequency. This is the first demonstration of ultrasound steering using a single-element transducer, which can have important applications for ultrasound focusing with phased arrays.


Early life arsenic exposure and acute and long-term responses to influenza A infection in mice.
Arsenic is a significant global environmental health problem. Exposure to arsenic in early life has been shown to increase the rate of respiratory infections during infancy, reduce childhood lung function, and increase the rates of bronchiectasis in early adulthood. We aimed to determine if early life exposure to arsenic exacerbates the response to early life influenza infection in mice. C57BL/6 mice were exposed to arsenic in utero and throughout postnatal life. At 1 week of age, a subgroup of mice were infected with influenza A. We then assessed the acute and long-term effects of arsenic exposure on viral clearance, inflammation, lung structure, and lung function. Early life arsenic exposure reduced the clearance of and exacerbated the inflammatory response to influenza A, and resulted in acute and long-term changes in lung mechanics and airway structure. Increased susceptibility to respiratory infections combined with exaggerated inflammatory responses throughout early life may contribute to the development of bronchiectasis in arsenic-exposed populations.


Alagille Syndrome and Chronic Arthritis: An International Case Series.
We describe 10 children with Alagille syndrome and inflammatory arthritis. In our centers, the prevalence of chronic arthritis in patients with Alagille syndrome is approximately 50 times higher compared with the general population. Arthritis was refractory to most treatment. Patients with Alagille syndrome should routinely be screened for musculoskeletal symptoms.


Complementary water quality observations from high and medium resolution Sentinel sensors by aligning chlorophyll-a and turbidity algorithms.
High resolution imaging spectrometers are prerequisite to address significant data gaps in inland optical water quality monitoring. In this work, we provide a data-driven alignment of chlorophyll-a and turbidity derived from the Sentinel-2 MultiSpectral Imager (MSI) with corresponding Sentinel-3 Ocean and Land Colour Instrument (OLCI) products. For chlorophyll-a retrieval, empirical 'ocean colour' blue-green band ratios and a near infra-red (NIR) band ratio algorithm, as well as a semi-analytical three-band NIR-red ratio algorithm, were included in the analysis. Six million co-registrations with MSI and OLCI spanning 24 lakes across five continents were analysed. Following atmospheric correction with POLYMER, the reflectance distributions of the red and NIR bands showed close similarity between the two sensors, whereas the distribution for blue and green bands was positively skewed in the MSI results compared to OLCI. Whilst it is not possible from this analysis to determine the accuracy of reflectance retrieved with either MSI or OLCI results, optimizing water quality algorithms for MSI against those previously derived for the Envisat Medium Resolution Imaging Spectrometer (MERIS) and its follow-on OLCI, supports the wider use of MSI for aquatic applications. Chlorophyll-a algorithms were thus tuned for MSI against concurrent OLCI observations, resulting in significant improvements against the original algorithm coefficients. The mean absolute difference (MAD) for the blue-green band ratio algorithm decreased from 1.95 mg m-3 to 1.11 mg m-3, whilst the correlation coefficient increased from 0.61 to 0.80. For the NIR-red band ratio algorithms improvements were modest, with the MAD decreasing from 4.68 to 4.64 mg m-3 for the empirical red band ratio algorithm, and 3.73 to 3.67 for the semi-analytical 3-band algorithm. Three implementations of the turbidity algorithm showed improvement after tuning with the resulting distributions having reduced bias. The MAD reduced from 0.85 to 0.72, 1.22 to 1.10 and 1.93 to 1.55 FNU for the 665, 708 and 778 nm implementations respectively. However, several sources of uncertainty remain: adjacent land showed high divergence between the sensors, suggesting that high product uncertainty near land continues to be an issue for small water bodies, while it cannot be stated at this point whether MSI or OLCI results are differentially affected. The effect of spectrally wider bands of the MSI on algorithm sensitivity to chlorophyll-a and turbidity cannot be fully established without further availability of in situ optical measurements.


Vitamin D genetic risk scores in multiple sclerosis.
Low serum 25(OH)D3 (vD) is an environmental risk factor for multiple sclerosis (MS). Lower vD levels during early disease may be associated with long-term disability. Determinants of serum vD levels in healthy individuals include supplementation behaviour and genetic factors. These determinants have been less well studied in people with MS (pwMS). We developed a vD-weighted genetic risk score (GRS) and validated this in 373,357 UK Biobank participants without MS. We measured serum 25(OH)D3 and genotyped six vD-associated SNPs (rs12785878, rs10741657, rs17216707, rs10745742, rs8018720, rs2282679) in a cohort of pwMS (n = 315) with age and geographically matched controls (n = 232). We then assessed predictors of serum vD concentration in this cohort. The GRS was strongly associated with vD status in the Biobank cohort (p < 2 × 10-16). vD supplementation, having MS, lower BMI, increased age and supplementation dose were associated with higher vD levels (false discovery rate, FDR < 5%). In multivariable models adjusting for supplementation, BMI, age, sex, and MS status, the GRS was strongly associated with vD level (p = 0.004), but not in those who supplemented (p = 0.47). Our findings suggest that vD supplementation is the major determinant of vD level in pwMS, with genetic determinants playing a far smaller role.


IntaRNA: efficient prediction of bacterial sRNA targets incorporating target site accessibility and seed regions.
During the last few years, several new small regulatory RNAs (sRNAs) have been discovered in bacteria. Most of them act as post-transcriptional regulators by base pairing to a target mRNA, causing translational repression or activation, or mRNA degradation. Numerous sRNAs have already been identified, but the number of experimentally verified targets is considerably lower. Consequently, computational target prediction is in great demand. Many existing target prediction programs neglect the accessibility of target sites and the existence of a seed, while other approaches are either specialized to certain types of RNAs or too slow for genome-wide searches. We introduce INTARNA, a new general and fast approach to the prediction of RNA-RNA interactions incorporating accessibility of target sites as well as the existence of a user-definable seed. We successfully applied INTARNA to the prediction of bacterial sRNA targets and determined the exact locations of the interactions with a higher accuracy than competing programs. http://www.bioinf.uni-freiburg.de/Software/


Standard operative temperatures and cost of thermoregulation in the arctic ground squirrel, Spermophilus undulatus.
Body temperatures (T b) and daily activity patterns of free-living arctic ground squirrells (Spermophilus undulatus) were determined via telemetry at a field site in northern Alaska. Simultaneous measurements were made of ambient temperature (T a), wind speed (V), and incident solar radiation. The operative environmental temperature (T e) for ground squirrels was obtained from fur-covered, thin metal taxidermic models of the animals. Standard operative temperature (T es), a comparative index of heat flow, was calculated from T e, V, and laboratory measurements of thermal conductivity.During the period of the study (August), S. undulatus were active for about 14 h per day (06.00 to 20.00 h). T b was high throughout the daily cycle, averaging 38-39°C. Circadian variations in T b were slight; average T b values dropped <1°C at night. Daytime T b fluctuations were not closely correlated to activity or to changes in environmental conditions. Air temperatures during the study were low, usually between 10 and 15°C during the day. However, T es in exposed areas was normally higher, even though skies were generally overcast. During periods of sunshine, T es may be as high as 34°C. The absence of nocturnal activity may result from increased costs of thermoregulation at night, which sharply reduces foraging efficiency. The high and stable body temperatures of S. undulatus probably result from thermoneutral daytime T es, low activity levels, and the use of well-insulated nests.


LiZn2V3O8: a new geometrically frustrated cluster spin-glass.
We have investigated the structural and magnetic properties of a new cubic spinel LiZn2V3O8 (LZVO) through x-ray diffraction, dc and ac susceptibility, magnetic relaxation, aging, memory effect, heat capacity and 7Li nuclear magnetic resonance (NMR) measurements. A Curie-Weiss fit of the dc susceptibility [Formula: see text]([Formula: see text]) yields a Curie-Weiss temperature [Formula: see text] K. This suggests strong antiferromagnetic (AFM) interactions among the magnetic vanadium ions. The dc and ac susceptibility data indicate the spin-glass behavior below a freezing temperature T f [Formula: see text] 3 K. The frequency dependence of the T f is characterized by the Vogel-Fulcher law and critical dynamic scaling behavior or power law. From both fitting, we obtained the value of the characteristic angular frequency [Formula: see text] [Formula: see text] 3.56 [Formula: see text] 106 Hz, the dynamic exponent [Formula: see text] [Formula: see text] 2.65, and the critical time constant [Formula: see text] [Formula: see text] 1.82 [Formula: see text] 10-6 s, which falls in the conventional range for typical cluster spin-glass (CSG) systems. The value of relative shift in freezing temperature [Formula: see text] [Formula: see text] 0.039 supports a CSG ground states. We also found aging phenomena and memory effects in LZVO. The asymmetric response of the magnetic relaxation below T f supports the hierarchical model. Heat capacity data show no long-range or short-range ordering down to 2 K. Only about 25% magnetic entropy change [Formula: see text] signifies the presence of strong frustration in the system. The 7Li NMR spectra show a shift and broadening with decreasing temperature. The spin-lattice and spin-spin relaxation rates show anomalies due to spin freezing around 3 K as the bulk magnetization.


Newcastle disease virus evolution. I. Multiple lineages defined by sequence variability of the hemagglutinin-neuraminidase gene.
We compared the hemagglutinin-neuraminidase gene sequence among 13 strains of Newcastle disease virus (NDV) isolated over the last 50 years. Although overall homology was remarkably high, the sequence variability demonstrated the existence of at least three distinct lineages, which must have co-circulated for considerable periods. The sequence variability also appears to reflect some accumulation of mutations over time. Strictly correlating with the lineages, the translation products could be classified into three size classes. One class lacked the interchain disulfide bond, and another represented unusual precursor protein of biologically inactive form. The lineages correlated to some extent with virulence and place of isolation of the strains. However, antigenic variations, which were neither cumulative nor progressive, did not correlate with the lineages. These analyses showing multiple lineages were greatly facilitated by a precise calculation of synonymous substitutions, which had been largely free from selective pressures and had occurred frequently and evenly throughout the coding region.


Influence of age and co-medication on the steady-state pharmacokinetics of valproic acid in Tunisian patients with epilepsy.
Valproic acid (VPA) is a widely prescribed broad-spectrum antiepileptic drug. However, the use of VPA is complicated in clinical practice by its remarkably wide variability of pharmacokinetics. The objective of this study was to investigate the effects of demographic factors and associated therapies on steady-state plasma VPA concentrations in patients with epilepsy. This retrospective cohort study was carried out using the routine therapeutic drug monitoring (TDM) database. Stepwise logistic regression analysis was used to compare serum VPA levels in 78 epilepsy patients treated with VPA in association with at least one other drug that could have interacted with CYP2C9, CYP2C19 or UGT enzymes. The frequency of subtherapeutic serum VPA levels was significantly increased with younger age (P<0.02), the number of co-medications (P<0.007) and use of enzyme-inducing co-medications (P<0.02). No significant correlations between VPA dose and trough plasma concentrations were found, as the latter did not increase in proportion to the dose. Routine monitoring of VPA serum levels would be extremely useful in epilepsy patients in the pediatric age group and in those who require associated enzyme-inducing medications.


The impact of polysubstance use patterns on engagement of substance use disorder treatment among emergency department patients at high risk of opioid overdose.
Substance use patterns are diverse, and multiple substances are often involved in fatal and nonfatal overdoses. Additionally, polysubstance use is associated with greater difficulty accessing and remaining in substance use disorder (SUD) treatment. The aim of this study was to identify substance use patterns and determine their association with SUD treatment engagement among emergency department (ED) patients at risk of opioid overdose. This was a sub-analysis of a randomized controlled trial comparing two behavioral interventions for individuals at two EDs in Rhode Island from 2018 to 2021. Past six-month substance use frequency for eight substances plus injection drug use was self-reported at trial enrollment, and SUD treatment engagement within 90 days after enrollment was obtained using administrative data linkages. Latent class analysis identified substance use patterns and multivariable log-binomial models estimated the association with SUD treatment engagement. Among 607 participants, there were four substance use patterns: 1) low reported use (n = 295), 2) frequent injection and heroin use (n = 131), 3) high frequency broad polysubstance use (n = 62), and 4) low frequency broad polysubstance use (n = 119). Compared to participants with the low reported use pattern, those with the frequent injection and heroin pattern had a greater likelihood of SUD treatment engagement (adjusted risk ratio = 1.28; 95% confidence interval = 1.02-1.61). Distinct and meaningful polysubstance use patterns showed differential SUD treatment engagement after ED discharge. Nuanced relationships between substance use patterns and treatment highlight the necessity for tailored harm reduction, treatment, and recovery services.


Purification and properties of a fructose-1,6-diphosphate activated L-lactate dehydrogenase from Staphylococcus epidermidis.
L-(+)-lactate dehydrogenase (LDH) from Staphylococcus epidermidis ATCC 14990 was purified by affinity chromatography. The purified enzyme was specifically activated by fructose-1,6-diphosphate (FDP). The concentration of FDP required for 50% maximal activity was about 0.15 mM. The enzyme activity was inhibited by adenosine diphosphate (ADP) and oxamate. The inhibition by ADP appeared to be competitive with respect to reduced nicotinamide adenine dinucleotide (NADH). The catalytic activity of the LDH for pyruvate reduction exhibited an optimum at pH 5.6. The enzyme is composed of four, probably identical, subunits. Sephadex gel filtration and sedimentation velocity at pH 5.6 Yielded molecular weights of about 130 000 and 126 000, respectively. The molecular weight at pH 6.5 and 7.0 was found to be only about 68 000. Polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate and sedimentation velocity at pH 2.0 or 8.5 revealed monomeric subunits with an approximate molecular weight of 36000. The thermostability of the heat labile enzyme was increased in the presence of FDP, NADH and pyruvate. The purified LDH exhibited an anomalous type of kinetic behavior. Plots of initial velocity vs. different concentrations of pyruvate, NADH or FDP led to saturation curves with intermediary plateau regions. As a consequence of these plateau regions the Hill coefficient alternated between lower and higher n-values. Some distinguishing properties of the S. epidermidis LDH and other LDHs activated by FDP are discussed.


Superconducting Phases in Lithium Decorated Graphene LiC6.
A study of possible superconducting phases of graphene has been constructed in detail. A realistic tight binding model, fit to ab initio calculations, accounts for the Li-decoration of graphene with broken lattice symmetry, and includes s and d symmetry Bloch character that influences the gap symmetries that can arise. The resulting seven hybridized Li-C orbitals that support nine possible bond pairing amplitudes. The gap equation is solved for all possible gap symmetries. One band is weakly dispersive near the Fermi energy along Γ → M where its Bloch wave function has linear combination of [Formula: see text] and dxy character, and is responsible for [Formula: see text] and dxy pairing with lowest pairing energy in our model. These symmetries almost preserve properties from a two band model of pristine graphene. Another part of this band, along K → Γ, is nearly degenerate with upper s band that favors extended s wave pairing which is not found in two band model. Upon electron doping to a critical chemical potential μ1 = 0.22 eV the pairing potential decreases, then increases until a second critical value μ2 = 1.3 eV at which a phase transition to a distorted s-wave occurs. The distortion of d- or s-wave phases are a consequence of decoration which is not appear in two band pristine model. In the pristine graphene these phases convert to usual d-wave or extended s-wave pairing.


Caspase 2 and p53 Reunited in Tumor Control.
Recent findings (Tsabar et al.) demonstrate that human cancer cells that evade the cell cycle blockage normally imposed by DNA damage experience sustained p53 signaling upon MDM2 degradation by caspase 2. Such a response may be connected to the delivery of immunostimulatory signals to ensure the elimination of genetically instable cancer cells.


Complement C3 gene polymorphism in renal transplantation (an Iranian experience).
The C3 component of complement has different roles in kidney disease and its local production in donor kidney may affect allograft function and rejection after organ transplantation. A single base substitution in c3 gene (rs2230199), defines two common allelic variants with different mobility on gel electrophoresis: S (Slow) and F (Fast). In order to evaluate the effect of this polymorphism on acute renal allograft rejection, one hundred samples of donor and recipients were collected and genotyping was done by PCR-RFLP method. The allelic frequencies were: C3S=0.791, C3F=0.209. There was no significant association between recipient's genotype and acute rejection (p value<0.05). No correlation between donor genotype and acute rejection was also present. Patients were divided into four groups, according to the recipient and donor genotypes: SS recipients and FS or FF donor, SS recipient and donor, FF or FS recipient and SS donor and FS or FF recipient and donor. There was no significant difference in rate of acute rejection between groups. Although the results didn't find any association between C3 complement polymorphisms and acute allograft rejection, there was no acute rejection in FS or FF donors and SS recipient group.


Childhood diarrhoea in Danish day care centres could be associated with infant colic, low birthweight and antibiotics.
Diarrhoea is very common in children attending day care centres. The aim of this study was to examine certain predisposing risk factors for an association with diarrhoea, including foreign travel, treatment with antibiotics, having household pets, infant colic, bottle feeding, using a pacifier and low birthweight. A dynamic one-year follow-up cohort study comprising 179 children from 36 day care centres was conducted from September 2009 to July 2013 in Copenhagen, Denmark. Questionnaires were sent to the children's parents or legal guardians every two months for a year, requesting information on gastrointestinal symptoms and exposure. A logistic regression was performed to identify the odds ratios of different risk factors for diarrhoea. The odds ratios for diarrhoea were 1.97 (0.93-4.20) for children with a history of infant colic, 1.91 (0.90-4.04) for low birthweight children and 1.45 (0.74-2.82) for children who had used antibiotics. Having a pet in the household had a possible protective effect towards diarrhoeal events, with an odds ratio of 0.47 (0.20-1.09). A history of infant colic, low birthweight, and to a lesser extent antibiotic use, possibly increased the risk of diarrhoea in Danish children in day care centres.


The Dermatological Effects of Box Jellyfish Envenomation in Stinging Victims in Thailand: Underestimated Severity.
The dermatological effects of box jellyfish envenomation among stinging victims in Thailand are not well reported, particularly concerning chronic effects. For first aid, different recommendations indicate the necessity for the removal of tentacles in life-threatening situations. This study aimed to describe the dermatological effects of box jellyfish envenomation and propose recommendations regarding first aid for victims in urgent care or life-threatening situations. Surveillance systems and Toxic Jellyfish Networks were established to improve detection and investigation. The networks investigated all severe victims of jellyfish envenomation. A retrospective study was conducted, and victims of stinging by box jellyfish investigated from 1999 to 2021 were included. One hundred and twenty-four victims were recorded. The majority of victims were males (55%), tourists (69%), and Thai nationals (49%). Direct contact had more severe consequences than indirect contact. Dermatological effects included edema, erythematous caterpillar track-like rash, blistering, bullae, papular eruption, necrosis, digital gangrene, recurrent dermatitis, dermal hypersensitivity, numbness, lichenification, hyperpigmentation, keloids, and scarring. Suffering and healing continued from several weeks to many years. Victims with multiple-tentacle box jellyfish stings had papular eruptions and greater severity of skin issues. All fatally envenomed victims collapsed within a few minutes and received incorrect/no first aid. The proposed first aid for life-threatening box jellyfish stings is continuous irrigation of the wound with vinegar for at least 30 s and initiation of cardiopulmonary resuscitation if there is no respiration or heartbeat. Tentacles often detach spontaneously, and removal is not always necessary, thus saving time. The findings provide input for improving diagnosis and treatment guidelines.


Absence of the major dense line in myelin of the mutant mouse "shiverer".
The myelin of the central nervous system (CNS) of the mutant mouse Shiverer is characterized by the absence of the major dense line (MDL). The intraperiod line, as seen in conventional electron micrographs and in freeze-fractured replicas, appears normal. Peripheral myelin, as seen in ventral and dorsal roots of spinal cord, is unaffected by the mutation. During the period of active myelination, the cytoplasm of most oligodendrocytes (ODs) is packed with electron-lucent vacuoles in continuity with the Golgi apparatus and with bundles of microtubules. It is concluded that a metabolic pathway possibly involving the Golgi apparatus, and contributing to the formation of the MDL is selectively affected in this mutant.


Measurements of nuclear data parameters of 201Tl by gamma-ray spectrometry.
201Tl is frequently used in radiopharmaceutical applications, and therefore the gamma-ray emission probabilities and half-life have been re-determined by means of gamma-ray spectroscopy. While the activity was calibrated using the sum-peak coincidence method, the half-life was obtained by the reference source method based on simultaneous counting of a reference source and the sample. Both the measurement techniques and assignment of uncertainties are presented and discussed, and the resulting data are shown to be in good agreement with previously published studies.


Localization of myosin II regulatory light chain in the cerebral vasculature.
The cytoskeleton of cerebral microvascular endothelial cells is a critical determinant of blood-brain barrier (BBB) function. Barrier integrity appears to be particularly sensitive to the phosphorylation state of specific residues within myosin regulatory light chain (RLC), one of two accessory light chains of the myosin II motor complex. Phosphorylation of myosin RLC by myosin light chain kinase (MLCK) has been implicated in BBB dysfunction associated with alcohol abuse and hypoxia, whereas dephosphorylation may enhance BBB integrity following exposure to lipid-lowering statin drugs. Using immunohistochemistry we provide evidence of widespread myosin II RLC distribution throughout the cerebral vasculature of the mouse. Light microscopy revealed immunolocalization of myosin II RLC protein in the endothelium of brain capillaries, the endothelial cell layer of arterioles and in association with venules. Immunolabeling of myosin RLC in non-muscle endothelial cells could be distinguished from myosin RLC immunoreactivity associated with the smooth muscle layer of the tunica media in larger muscular arterioles. These findings support an emerging role for myosin II RLC as a component of the actomyosin cytoskeleton of cerebral endothelial cells with the potential to contribute to the selective vulnerability of the brain in vivo.


[Determination of Hg in biological samples by inductively coupled plasma mass spectrometry].
To establish an inductively coupled plasma mass spectrometry (ICP-MS) method for determination of Hg in biological samples. The samples were digested with microwave digestion instrument. ICP-MS was applied to detect Hg in blood, urine and hair specimens by using 115In as an internal marker. The ability of gold to eliminate the memory effect of mercury was investigated with the gold amalgamate produced by gold and mercury. The limits of detection were in the 0.01 microg/L, and the accuracy of the method ranged from 97.0% to 107.1%. The concentration of gold was 10 microg/L and the memory effect of mercury was resolved. The method is accurate, rapid, sensitive and suitable for the cases of mercury poisoning and the clinical diagnosis and monitoring for patients with mercury poisoning.


Cloning and molecular characterization of the ORF5 gene from a PRRSV-SN strain from Southwest China.
To monitor the genetic variation of PRRSV, the ORF5 gene of the PRRSV-SN strain found in Suining City, Sichuan Province, was cloned and sequenced. The results showed that the PRRSV-SN strain was a highly pathogenic PRRSV (HP-PRRSV) variant strain with the North American (NA) genotype. Homology analysis showed that the ORF5 gene of the PRRSV-SN isolate shared 89.4% (86.5%) nucleotide (amino acid) sequence similarity with the North American strain VR-2332, 98.8% (96%) similarity with JXA1, and 63.8% (57.7%) similarity with the European type representative strain Lelystad virus. Phylogenetic analysis showed that PRRSV-SN belongs to the NA genotype and has the same subtype as other highly pathogenic PRRSV strains. Amino acid sequence analysis showed that compared with the VR2332 strain, PRRSV-SN has different degrees of variation in the signal peptide, transmembrane region (TM), primary neutralizing epitope (PNE), non-neutral epitopes and N-glycosylation sites. Antigenicity analysis showed that the PRRSV-SN ORF5 gene products and JXA1 have similar antigenic characteristics, and the antigenic epitopes are mainly located in aa30-39, aa50-60, aa128-141, aa146-155 and aa161-183 regions. In contrast, the antigenic characteristics of PRRSV-SN are quite different from those of the VR2332 strain. The main differences were that the PRRSV-SN strain was significantly narrower than the VR2332 strain in the aa30-39 and the aa50-60 regions but was significantly wider in the aa136-141 region. The results of this study showed that the epidemic strains that cause PRRSV outbreaks in the farm are still mainly JXA1 variants, but due to the more frequent use of live vaccine immunizations, the genes of the PRRSV epidemic strain still show constant variation. Vaccination with live PRRSV should be reduced, and surveillance of PRRSV strains should be enhanced.


Absence of simian virus 40 (SV40) DNA in lymphoma samples from Tasmania, Australia.
It has been postulated that the recent world-wide increase in the incidence of non-Hodgkin's lymphoma (NHL) may have been caused by human infection with simian virus 40 (SV40) (a lymphotropic monkey virus that was introduced to man from contaminated poliovirus vaccines between 1955 and 1963); therefore, we set out to determine the incidence of SV40 DNA positivity in lymphoma samples from patients in Tasmania, Australia. One hundred lymph node samples, 50 from patients with lymphomas and 50 from controls, were tested using PCR amplification of three SV40-specific primer pairs followed by dot-blot hybridisation. All of the samples tested contained amplifiable DNA, but none contained amplifiable SV40 sequences with any of the primer sets used. Our results demonstrate absence of SV40 in the lymphoid tissues of our study population in Tasmania, Australia. SV40 does not explain the increasing incidence of NHL in our population.


Effects of Boesenbergia rotunda (L.) Mansf. on sexual behaviour of male rats.
To study the effects of Boesenbergia rotunda (Krachai) on sexual behaviour in male albino rats. Thirty-two male Wistar rats were equally divided into four groups: experimental groups were gavaged with the ethanolic extract of the rhizome of B. rotunda at doses of 60, 120 and 240 mg/kg and a control group received distilled water, for 60 days. Sexual behaviour, reproductive organs, diameter of seminiferous tubule, epididymal sperm density, and androgenic hormones were evaluated. Within 30-min observation, there was no significant difference of courtship behaviour, mount frequency (MF), intromission frequency (IF), mount latency (ML), intromission latency (IL), copulatory efficiency or intercopulatory interval in male rats. In three 10-min intervals over a 30-min period, courtship behaviour and MF during the first 10-min were significantly higher than those in the second and third 10-min observation in all groups, whereas IF had no significant difference. All doses of B. rotunda extract significantly increased the relative testicular weight and the diameter of the seminiferous tubules. The dose of 60 mg/kg also significantly increased the relative weight of the seminal vesicle. Nevertheless, the sperm density, serum testosterone and androstenedione levels were not affected by the B. rotunda extract. B. rotunda does not affect sexual behaviour nor serum androgenic levels.


The effect of interactions between soil compaction and phenol contamination on plant growth characteristics: Implications for scaling bioremediation at industrial sites.
In phenol contaminated soils from an industrial site, soil compaction and soil chemical contaminants influenced germination, emergence, and early plant growth (roots and shoots) for Thrift (Ameria maratima) and Blackbutt oats (Avena sativa). The specific effects were of soil compaction (low, 100 psi and high, 800 psi) at two concentrations of phenol (biotreated, ∼200 mg kg-3; untreated, ∼1150 mg kg-3). Plant responses to compaction and phenol varies between the two species. Emergence of Thrift showed greater sensitivity to increasing soil compaction but only under high phenol concentrations. Low phenol inhibited Thrift root length, as is the case with Balckbutt oats, although high phenol had no significant impact on Thrift. While shoot length of Blackbutt oats is not significantly affected by increasing soil compaction, it is negatively impacted at the increased phenol concentrations in the untreated soils indicating an interaction effect with compaction. This is the first study that examines the impact of soil contamination on oats. The oat variety tested had known drought tolerance and hardiness indicating an ability to withstand compaction, and which may explain those characteristics. Thrift was most resistant to contamination which is consistent with its previously reported tolerance of pollutants. Implications of the study are that Blackbutt oats represent a potential candidate species for land rehabilitation under high compaction conditions such as at degraded soil sites at decommissioned mining operations, polluted industrial sites, or degraded (compacted) agricultural land.


Corrigendum: A two-year longitudinal study of retinal vascular impairment in patients with amnestic mild cognitive impairment.
[This corrects the article DOI: 10.3389/fnagi.2022.993621.].


Lessons from application of the UNRES force field to predictions of structures of CASP10 targets.
The performance of the physics-based protocol, whose main component is the United Residue (UNRES) physics-based coarse-grained force field, developed in our laboratory for the prediction of protein structure from amino acid sequence, is illustrated. Candidate models are selected, based on probabilities of the conformational families determined by multiplexed replica-exchange simulations, from the 10th Community Wide Experiment on the Critical Assessment of Techniques for Protein Structure Prediction (CASP10). For target T0663, classified as a new fold, which consists of two domains homologous to those of known proteins, UNRES predicted the correct symmetry of packing, in which the domains are rotated with respect to each other by 180° in the experimental structure. By contrast, models obtained by knowledge-based methods, in which each domain is modeled very accurately but not rotated, resulted in incorrect packing. Two UNRES models of this target were featured by the assessors. Correct domain packing was also predicted by UNRES for the homologous target T0644, which has a similar structure to that of T0663, except that the two domains are not rotated. Predictions for two other targets, T0668 and T0684_D2, are among the best ones by global distance test score. These results suggest that our physics-based method has substantial predictive power. In particular, it has the ability to predict domain-domain orientations, which is a significant advance in the state of the art.


Incorporation of atmospheric 15NO2-nitrogen into free amino acids by Norway spruce Picea abies (L.) Karst.
During spring and autumn 1991, potted 6-yearold spruce trees (Picea abies (L.) Karst.) were fumigated with 60 nl·1-115NO2 for 4 days under controlled conditions in constant light. Current and previous flush needles, the bark and the fine roots were analysed for total 15N content and incorporation of 15N into the α-amino nitrogen of free amino acids. In addition, in vitro nitrate reductase activity and stomatal conductance of the needles were measured. Nitrate reductase activity was significantly higher in the needles of fumigated trees compared to control trees exposed to filtered air. With an average of 9.1% 15N, free glutamate was the pool with the most label. Taking into account the time-course of the labelling of this pool, this figure can be taken as an estimate of the minimum contribution of NO2 to the N nutrition of the needles. 15N-labelled amino acids were also detected in the bark and the roots, indicating export from the needles.


Evidence for erythrocyte membrane glycoproteins being carriers of blood-group P1 determinants.
The contribution of different membrane constituents to the bloodgroup P1 activity of human erythrocytes was investigated. Pronase digestion of native red cell stroma or partition between butanol and water had no serologically detectable effect, whereas pronase-treatment of previously butanol-extracted membranes liberated virtually all blood-group P1 determinants from the ghosts. On Laemmli gels, all P1 activity was found in the band 4.5 region. Thus it is concluded that, in addition to the well-documented P1 glycolipid, also membrane glycoproteins are carriers of blood-group P1 determinants.


Minimizing the echo time in diffusion imaging using spiral readouts and a head gradient system.
Diffusion weighted imaging (DWI) is commonly limited by low signal-to-noise ratio (SNR) as well as motion artifacts. To address this limitation, a method that allows to maximize the achievable signal yield and increase the resolution in motion robust single-shot DWI is presented. DWI was performed on a 3T scanner equipped with a recently developed gradient insert (gradient strength: 200 mT/m, slew rate: 600 T/m/s). To further shorten the echo time, Stejskal-Tanner diffusion encoding with a single-shot spiral readout was implemented. To allow non-Cartesian image reconstruction using such strong and fast gradients, the characterization of eddy current and concomitant field effects was performed based on field-camera measurements. An echo time of only 19 ms was achieved for a b-factor of 1000 s/mm2 . An in-plane resolution of 0.68 mm was encoded by a single-shot spiral readout of 40.5 ms using 3-fold undersampling. The resulting images did not suffer from off-resonance artifacts and T 2 ∗ blurring that are common to single-shot images acquired with regular gradient systems. Spiral diffusion imaging using a head gradient system, together with an accurate characterization of the encoding process allows for a substantial reduction of the echo time, and improves the achievable resolution in motion-insensitive single-shot DWI.


Epidemiology and treatment of pediatric tibial fractures in Sweden: a nationwide population-based study on 5828 fractures from the Swedish Fracture Register.
Pediatric tibial fractures have been described internationally as mainly caused by fall during leisure activities and organized sports and showing a higher incidence in boys. Still, most studies are single center studies or have a small sample size. This study aimed to analyze sex and age distribution, seasonal variation, injury mechanisms and treatment of pediatric tibial fractures based on the nationwide Swedish Fracture Register (SFR). All tibial fractures in patients < 16 years at injury and registered in 2015-2019 were extracted from the SFR. We analyzed patient characteristics such as sex and age, injury mechanism, fracture location and treatment. The study cohort consisted of 5828 pediatric tibial fractures in 5719 patients. Median age of the patients was 7 years and 58% were boys. Shaft fractures were most common, followed by the distal and proximal tibia. The lowest incidence was observed during autumn. The most common cause of injury was fall mostly involving winter sports, stumbles and fall from play equipment. Play/free time and sports were the most common activities, common places of injury were sports facility and home. 1% were open fractures. 78% were treated non-surgically. Screw fixation was performed in 52% of surgically treated fractures, predominantly in the distal segment. Injury mechanism differs between age groups; play/free time injuries are common in younger children compared with sport activities in older children. Most patients are treated non-surgically. Open fractures are rare. Information on injury patterns is useful working preventively, for example safety work in playgrounds.


Biological Activities of Phenolics from the Fruits of Phyllanthus emblica L. (Euphorbiaceae).
Seven phenolic compounds, 1 - 7, including a new organic acid gallate, mucic acid 1-ethyl 6-methyl ester 2-O-gallate (7), were isolated from the MeOH extract of the fruits of Phyllanthus emblica L. (Euphorbiaceae). The structures were elucidated on the basis of extensive spectroscopic analysis and comparison with literature data. Upon evaluated for their antioxidant abilities by 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assays. The inhibitory activities against melanogenesis in B16 melanoma cells induced by α-MSH, as well as cytotoxic activities against four human cancer cell lines were also evaluated. All phenolic compounds, 1 - 7, exhibited potent antioxidant abilities (DPPH: IC50 5.6 - 12.9 μm; ABTS: 0.87 - 8.43 μm Trolox/μm; FRAP: 1.01 - 5.79 μm Fe2+ /μm, respectively). Besides, 5 - 7, also exhibited moderate inhibitory activities against melanogenesis (80.7 - 86.8% melanin content), even with no or low toxicity to the cells (93.5 - 101.6% cell viability) at a high concentration of 100 μm. Compounds 1 - 3 exhibited cytotoxic activity against one or more cell lines (IC50 13.9 - 68.4%), and compound 1 with high tumor selectivity for A549 (SI 3.2).


Regulation and the ponderostat.
Control theory is concerned mainly with the treatment of signals. This article takes into account that living beings not only treat information, but they are open systems traversed by flows of energy and mass. A block diagram of the regulation process is proposed, taking into account this fundamental difference between engineered and living systems. This new diagram is used to describe the stability of body weight. The ponderostat is hypothesized to work as a regulation of blood glucocorticoids with a hypothalamic set-point related to CRH concentration.


Acute liver failure associated with antiretroviral treatment for HIV: a report of six cases.
Severe hepatotoxicity is a rare but potentially fatal side effect of all antiretrovirals. We report a series of six human immunodeficiency virus (HIV)-infected patients admitted with acute liver failure (ALF) over a 25-month period, of whom five died. All had been treated with a range of antiretroviral therapy and only two had had acquired immune deficiency syndrome (AIDS) defining illnesses. Median duration of antiretroviral therapy was 12.5 months (range 1-23). Median time from the introduction of new antiretroviral therapy to the onset of ALF was 8 weeks (range 2-12). The development of ALF was unrelated to duration of HIV treatment or type of antiretroviral therapy, and was not predicted by close out-patient supervision and monitoring of liver function. Biochemical investigations were variable but revealed a predominantly hepatocellular pattern. Liver biopsy revealed typical features of mitochondrial toxicity in only one case, with confluent hepatocellular necrosis, inflammation and cholestasis seen in the others. There is a need to increase awareness of the potential hepatotoxicity of antiretroviral therapy and to develop means of predicting its development. With increasing usage of antiretroviral therapy, severe hepatotoxicity and ALF may arise more frequently, and the outcome is poor despite intensive supportive therapy.


Effect of immune serum and role of individual Fcgamma receptors on the intracellular distribution and survival of Salmonella enterica serovar Typhimurium in murine macrophages.
Immune serum has a protective role against Salmonella infections in mice, domestic animals and humans. In this study, the effect of antibody on the interaction between murine macrophages and S. enterica serovar Typhimurium was examined. Detailed analysis at the single-cell level demonstrated that opsonization of the bacteria with immune serum enhanced bacterial uptake and altered bacterial distribution within individual phagocytic cells. Using gene-targeted mice deficient in individual Fc gamma receptors it was shown that immune serum enhanced bacterial internalization by macrophages via the high-affinity immunoglobulin G (IgG) receptor, Fc gamma receptor I. Exposure of murine macrophages to S. enterica serovar Typhimurium opsonized with immune serum resulted in increased production of superoxide, leading to enhanced antibacterial functions of the infected cells. However, opsonization of bacteria with immune serum did not increase either nitric oxide production in response to S. enterica serovar Typhimurium or fusion of phagosomes with lysosomes.


From non-A, non-B hepatitis to hepatitis C virus cure.
The hepatitis C virus (HCV) was discovered in the late 1980s. Interferon (IFN)-α was proposed as an antiviral treatment for chronic hepatitis C at about the same time. Successive improvements in IFN-α-based therapy (dose finding, pegylation, addition of ribavirin) increased the rates of sustained virologic response, i.e. the rates of curing HCV infection. These rates were further improved by adding the first available direct-acting antiviral (DAA) drugs to the combination of pegylated IFN-α and ribavirin. An IFN-free era finally started in 2014, yielding rates of sustained virologic response over 90% in patients treated for 8 to 24 weeks with all-oral regimens. Major challenges however remain in implementation of these new treatment strategies, not only in low- to middle-income countries, but also in high-income countries where the price of these therapies is still prohibitive. Elimination of HCV infection through treatment in certain areas is possible but raises major public health issues.


After proper optimization of carvedilol dose, do different child classes of liver disease differ in terms of dose tolerance and response on a chronic basis?
Literature regarding safe doses of carvedilol is limited, and safe doses across different Child classes of chronic liver disease are not clear. A total of 102 consecutive cirrhotic patients with significant portal hypertension were included in this study. Hepatic venous pressure gradient was measured at baseline and 3 months after dose optimization. A total of 102 patients (63 males, 39 females) with a mean age of 58.3 ± 6.6 years were included. Among these patients, 42.2% had Child Class A, 31.9% had Class B, and 26.6% had Child Class C liver disease. The mean baseline hepatic venous pressure gradient was 16.75 ± 2.12 mmHg, and after dose optimization and reassessment of hepatic venous pressure gradient at 3 months, the mean reduction in the hepatic venous pressure gradient was 5.5 ± 1.7 mmHg and 2.8 ± 1.6 mmHg among responders and nonresponders respectively. The mean dose of carvedilol was higher in nonresponders (19.2 ± 5.7 mg) than responders (18.75 ± 5.1 mg). However, this difference was not statistically significant (P > 0.05). The univariate analysis determined that the absence of adverse events, the absence of ascites, and low baseline cardiac output were significantly associated with chronic response, whereas, the etiology, Child class, variceal size (large vs small), and gender were not. On multivariate analysis, the absence of any adverse event was determined to be an independent predictor of chronic response (OR 11.3, 95% CI; 1.9-67.8). The proper optimization of the dose of carvedilol, when administered chronically, may enable carvedilol treatment to achieve a greater response with minimum side effects among different Child classes of liver disease.


Ultrasound assisted design of sulfur/carbon cathodes with partially fluorinated ether electrolytes for highly efficient Li/S batteries.
A one-step, eco-friendly ultrasound-assisted process is established for the rapid synthesis of sulfur-carbon composite cathode materials, avoiding the widely used, energy inefficient "melt-down" process for Li-S batteries. It is demonstrated that, without inserting sulfur into pores of carbon, the coulombic efficiency of SC/Li cell in the new DOL/D2 electrolyte is greater than 96% for 100 cycles, which is far superior to the reported numerous electrolyte formulations.


Rotational thromboelastometry in the diagnosis of coagulopathy in major pediatric surgical operations.
To examine the correlation between rotational thromboelastometry (ROTEM) and coagulopathy after major pediatric surgical operations. From November 2013 until April 2015, pediatric cases who underwent major noncardiac surgeries and met the coagulopathy-risk criteria were reviewed for postoperative clinically significant coagulopathy (CSC). Two ROTEM studies, EXTEM and INTEM, were performed at the immediately postoperatively without the results being taken into any clinical decision making. Seventy-seven operations on 73 patients were included in this analysis. CSC occurred following 24 operations (32%) with a significantly higher incidence when a patient had a higher coagulopathy risk. On univariate analysis, evidence of diffuse bleeding in the operative field and massive bleeding were the 2 parameters with the strongest association with CSC. INTEM and EXTEM had specificities in diagnosing CSC of 75.5% and 94.3%, respectively. When each individual EXTEM and INTEM item was analyzed against CSC using ROC analysis, clot forming time (CFT) gave the largest under the curve area. The cut-off CFTs that gave the highest sensitivity and specificity in this study were 120seconds for EXTEM and 100seconds for INTEM. Postoperative coagulopathy is a risk that should always be considered in pediatric surgical operations. Thromboelastometry can be a hemostatic test providing high predictive value for this condition.


New considerations for the cognitive locus of impairment in the irrelevant-sound effect.
The finding that serial recall performance for visually presented items is impaired by concurrently presented task-irrelevant speech or sounds is referred to as the irrelevant-speech/-sound effect (ISE). Substantial evidence has indicated that the impairment of serial rehearsal can result in an ISE, and this may be explained by several models. The present series of experiments has demonstrated an ISE in surprise nonserial recognition tasks in which participants were unaware of the need to maintain a large number of visual items for a later memory test, suggesting that neither the rehearsal nor maintenance of order information is necessary for observing the ISE. This effect was observed for both steady-state and changing-state irrelevant sounds, suggesting that the present results do not derive from a confusion of order information, but instead provide evidence that identity representations can also be impaired by irrelevant sound.


Changes in cellular prion protein expression, processing and localisation during differentiation of the neuronal cell line CAD 5.
Cellular prion protein (PrPC ) is infamous for its role in prion diseases. The physiological function of PrPC remains enigmatic, but several studies point to its involvement in cell differentiation processes. To test this possibility, we monitored PrPC changes during the differentiation of prion-susceptible CAD 5 cells, and then we analysed the effect of PrPC ablation on the differentiation process. Neuronal CAD 5 cells differentiate within 5 days of serum withdrawal, with the majority of the cells developing long neurites. This process is accompanied by an up to sixfold increase in PrPC expression and enhanced N-terminal β-cleavage of the protein, which suggests a role for the PrPC in the differentiation process. Moreover, the majority of PrPC in differentiated cells is inside the cell, and a large proportion of the protein does not associate with membrane lipid rafts. In contrast, PrPC in proliferating cells is found mostly on the cytoplasmic membrane and is predominantly associated with lipid rafts. To determine the importance of PrPC in cell differentiation, a CAD 5 PrP-/- cell line with ablated PrPC expression was created using the CRISPR/Cas9 system. We observed no considerable difference in morphology, proliferation rate or expression of molecular markers between CAD 5 and CAD 5 PrP-/- cells during the differentiation initiated by serum withdrawal. PrPC characteristics, such as cell localisation, level of expression and posttranslational modifications, change during CAD 5 cell differentiation, but PrPC ablation does not change the course of the differentiation process. Ablation of PrPC expression does not affect CAD 5 cell differentiation, although we observed many intriguing changes in PrPC features during the process. Our study does not support the concept that PrPC is important for neuronal cell differentiation, at least in simple in vitro conditions.


Measurement of carotid stenosis using duplex ultrasonography with a microconvex array transducer: a validation with cerebral angiography.
We aimed to evaluate the validity of duplex ultrasonography (DUS) using a microconvex array transducer (MAT) with enhanced flow imaging (EFI) for visualization of the distal, internal carotid artery (ICA) and the accurate assessment of ICA stenosis. Patients who underwent both DUS and digital subtraction angiography (DSA) were registered for this study. DUS was performed by using a linear array transducer (LAT) and an MAT with EFI. The visibility of the ICA was compared between the 2 transducers. ICA stenosis was evaluated by the North American Symptomatic Carotid Endarterectomy Trial (NASCET) method on DUS, and the peak systolic flow velocity (PSV) was evaluated by using an MAT. These results were compared with DSA. In 238 internal carotid arteries, the average length of visualized ICA was longer for DUS using an MAT than an LAT (38.7±11.7 mm versus 25.8±9.8 mm, P<.0001). In 68 stenotic, internal carotid arteries, the degree of ICA stenosis detected by the NASCET method on DUS was correlated to that on DSA (P<.0001, r=.969, and adjusted r2=.938). PSV also correlated to NASCET method on DSA (P<.0001, r=.804, and adjusted r2=.640). DUS using an MAT with EFI technology could reveal more extended distal views of the ICA and was strongly correlated with NASCET method on DSA.


[Clinical pathway "laparoscopic prostatectomy". Analysis of anesthesiological procedures in a randomized study].
In this study we investigated the anesthesiological module of a clinical pathway. We chose the pathway of "laparoscopic prostatectomy" as an example for time-consuming minimally invasive surgery and 40 patients were randomly assigned to 2 groups receiving either total intravenous anesthesia (TIVA) using propofol/ remifentanil or balanced minimal flow anesthesia using desflurane/ remifentanil. During this module the indicators of quality such as vigilance, pain, postoperative nausea and vomiting (PONV) and mobilization were measured. Costs were evaluated and analyzed by a bottom-up procedure. There were no anesthesia-related deviations from clinical pathways and both forms of anesthesia management were equally well tolerated by the patients. No significant difference was observed regarding hemodynamic measurements or PONV. The patients in the desflurane/ remifentanil group recovered more rapidly (p=0.037) and had more pain. The amount of analgesic agents given immediately following anesthesia was significantly higher than in the TIVA group (p=0.017). The median anesthesia costs per minute for laparoscopic prostatectomy in the propofol group were 2.79 EUR (minimum cost 2.41 EUR, maximum cost 3.21 EUR) and in the desflurane group 2.68 EUR (minimum cost 2.45 EUR, maximum cost 3.39 EUR). The total anesthesia costs for both groups were within the proceeds matrix range for diagnosis-related groups (DRG). However, the cost analysis for medication was slightly higher than the proceeds matrix range for DRGs. Both forms of anesthesia can be implemented for time-consuming surgical procedures and allow a cost-effective anesthesia management. Anesthesiological procedures must go hand-in-hand with the type of anesthesia selected. The prophylactic use of analgetics for desflurane/ remifentanil anesthesia should be given earlier and in higher doses than in propofol/ remifentanil anesthesia. The prophylactic use of antiemetics following laparoscopic procedures of long duration is indicated. Optimizing anesthesiological procedures could lead to a continuous improvement in the quality of therapeutic pathways.


Computer-assisted mutagenesis of ecotin to engineer its secondary binding site for urokinase inhibition.
Inhibitors of urokinase-type plasminogen activator (uPA) were selected in vitro from two ecotin phage-display libraries to study the effect on binding of amino acid substitutions at critical positions 108, 110, 112, and 113 within the 100s loop (RNKL, respectively, in wild type ecotin). The first, a focused library, was the result of a computation-assisted approach using the three-dimensional structure of the ecotin-trypsin complex to guide the modeling of amino acid substitutions predicted to increase affinity for uPA. The second, a complete library, allowed for all substitutions at the above identified positions. The consensus sequences selected from the focused, and complete libraries were RRWS and R(R/N)QL, respectively. Inhibition constant determinations showed ecotin variants containing these sequences to be similarly potent (K(i) = 1-2 nm). These substitutions were combined with previously identified substitutions in another critical region of ecotin. One of these combinations (D70R/M84R/RRQL) is the tightest (K(i) = 50 pm) ecotin variant inhibitor of uPA. The blending of combinatorial methods and computer algorithms designed to predict stronger binders has allowed us to obtain protein derivatives that exhibit greatly increased affinity for a predetermined target. This technology can be applied to select for enhanced binding interactions at protein-protein interfaces and accelerate the process of protease inhibitor development.


The Role of Formative Evaluation in Promoting Digitally-based Health Equity and Reducing Bias for Resilient Health Systems: The Case of Patient Portals.
Patient portals are increasingly implemented to improve patient involvement and engagement. We here seek to provide an overview of ways to mitigate existing concerns that these technologies increase inequity and bias and do not reach those who could benefit most from them. Based on the current literature, we review the limitations of existing evaluations of patient portals in relation to addressing health equity, literacy and bias; outline challenges evaluators face when conducting such evaluations; and suggest methodological approaches that may address existing shortcomings. Various stakeholder needs should be addressed before deploying patient portals, involving vulnerable groups in user-centred design, and studying unanticipated consequences and impacts of information systems in use over time. Formative approaches to evaluation can help to address existing shortcomings and facilitate the development and implementation of patient portals in an equitable way thereby promoting the creation of resilient health systems.


Effect of age on behavioral and enzymatic changes during thiamin deficiency.
Open field behavior and whole brain enzymatic activities were determined during thiamin deficiency in two strains of young, as well as in aged mice. In young CD-1 mice, thiamin deficiency reduced total distance traveled and vertical movements after 7 days and the decline was more than 50% by day 9. The behavioral deficit was highly correlated to decreases in 2-oxoglutarate dehydrogenase activity (KGDH). The open field behavior of Balb/c mice was about 40% less than in CD-1 mice and responded in a qualitatively different manner to thiamin deficiency. The activity of the Balb/c mice increased and then decreased with thiamin deficiency. The activity of 3 month old mice peaked on day 6 (126% of initial score), whereas 10 and 30 month mice showed a much greater increase (about 175% of initial scores), but on day 7. Although the activity of the thiamin dependent enzyme transketolase (TK) was affected similarly at all ages, the activity of KGDH in the aged brain was more sensitive to thiamin deficiency than in the young; KGDH activity declined 41%, 57% or 74% at 3, 10, or 30 months, respectively. Thus, the current mouse model is an attractive one to study the interaction of thiamin deficiency with aging.