Identification of proximal sequence element nucleotides contributing to the differential expression of variant U4 small nuclear RNA genes.
The two U4 genes in the chicken genome code for distinct sequence variants of U4 small nuclear RNA that are differentially expressed during development. Whereas U4B RNA is constitutively expressed, U4X RNA is specifically down-regulated relative to U4B in a tissue-specific manner during development. To investigate mechanisms controlling the differential expression of the U4B and U4X genes, chimeric U4 genes were constructed and their transcriptional activities assayed by injection into Xenopus oocytes or by transfection of CV-1 cells. The proximal regulatory region of the U4B gene and the enhancers of both the U4B and U4X genes functioned efficiently in each expression system. However, the proximal region of the U4X gene was inactive. To localize and identify the responsible nucleotides, reciprocal point mutations were introduced into the U4X and U4B proximal regulatory regions. The results indicate that the U4X gene contains a suboptimal proximal sequence element, and that this results primarily from the identities of the nucleotides at positions -61 and -57 relative to the transcription start site.


Atezolizumab Monotherapy for Non-small Cell Lung Cancer Patients: An Observational Study in Ibaraki Group (ATTENTION-IBARAKI).
Atezolizumab is a monoclonal antibody that targets programmed death-ligand 1 (PD-L1) expressed on cancer cells derived from various organs and antigen-presenting cells and is currently commonly used in combination with chemotherapy. We conducted a study to clarify the current status of response to atezolizumab monotherapy in clinical practice and clarify the factors that contribute to long-term response and survival. We conducted a retrospective review of patients with advanced non-small cell lung cancer (NSCLC) treated with atezolizumab monotherapy from April 2018 to March 2023 at 11 Hospitals. The 147 patients evaluated had a progression-free survival (PFS) of 3.0 months and an overall survival of 7.0 months. Immune-related adverse events of any grade were observed in 13 patients (8.8%), grade 3 or higher in nine patients (6.1%), and grade 5 with pulmonary toxicity in one patient (0.7%). Favorable factors related to PFS were 'types of NSCLC other than adenocarcinoma'. Favorable factors for overall survival were 'performance status 0-1' and 'treatment lines up to 3'. There were 16 patients (10.9%) with PFS >1 year. No characteristic clinical findings were found in these 16 patients compared to the remaining 131 patients. Efficacy and immune-related adverse events of NSCLC patients associated with atezolizumab monotherapy were comparable to those of previous clinical trial results. Knowledge of characteristics of patients who are most likely to benefit from atezolizumab monotherapy is a crucial step towards implementing appropriate prescribing.


Higher-order Multiples.
Higher-order multiple gestations have increased since the advent of advanced reproductive technologies. These pregnancies present unique risks to both mothers and fetuses. It is imperative that early diagnosis of chronicity be determined and that proper counseling is performed, so patients understand the risks, evaluation, and management needed.


Hemorheological Alteration in Patients Clinically Diagnosed with Chronic Liver Diseases.
Since liver function is changed by chronic liver diseases, chronic liver disease can lead to different hemorheological alterations during the course of the progression. This study aims to compare alterations in whole blood viscosity in patients with chronic liver disease, focusing on the gender effect. Chronic liver diseases were classified into three categories by patient's history, serologic markers, and radiologic findings: nonalcoholic fatty liver disease (NAFLD) (n = 63), chronic viral hepatitis B and C (n = 50), and liver cirrhosis (LC) (n = 35). Whole blood viscosity was measured by automated scanning capillary tube viscometer, while liver stiffness was measured by transient elastography using FibroScan®. Both systolic and diastolic whole blood viscosities were significantly lower in patients with LC than NAFLD and chronic viral hepatitis (P < 0.001) in male patients, but not in female patients. In correlation analysis, there were inverse relationships between both systolic and diastolic whole blood viscosity and liver stiffness (systolic: r = -0.25, diastolic: r = -0.22). Whole blood viscosity was significantly lower in male patients with LC than NAFLD or chronic viral hepatitis. Our data suggest that whole blood viscosity test can become a useful tool for classifying chronic liver disease and determining the prognosis for different types of chronic liver diseases.


Application of a colorimetric chain-termination assay for characterization of reverse transcriptase from 3'-azido-2',3'-deoxythymidine-resistant HIV isolates.
Two different enzyme assays, both based on the interaction of native reverse transcriptase (RT) and 3'-azido-2',3'-deoxythymidine triphosphate (AZT-TP), were used to characterize the enzymes from 18 HIV-1 isolates with decreased sensitivity to AZT in cell culture. The first assay, which measures the balance between incorporation and excision of AZT monophosphate in the presence of dNTP substrate (in terms of IC(50)), gave an approx. 9-fold variation in sensitivity to AZT-TP. There was a correlation between the IC(50) values and the sensitivity of the corresponding virus to AZT in cell culture (r=0.60, P<0.01). The second assay, which was designed specifically for measurement of chain termination in the absence of dNTP substrate (as the concentration of AZT-TP giving 50% residual primer function, or CT(50)), revealed a more than 600-fold difference between the different isolate RTs. For the majority of enzymes there was a strict correlation between the results from the two assays; however, four isolates exhibited significantly higher CT(50)/IC(50) ratios than the other isolates. These differences were not related to sensitivity of the corresponding viruses to AZT but to the occurrence of certain mutations in their pol gene. The four deviating isolates contained either a minimum of four AZT-specific substitutions, including Thr-215-->Tyr (isolates 134 and 143), or some of the known specific substitutions combined with Thr-39-->Ala (isolates 80 and 157). The Thr-39-->Ala substitution has previously been recorded in connection with AZT/Foscarnet combination therapy.


Palmar filiform hyperkeratosis (FH) associated with underlying pathology?
We describe two patients with palmar filiform hyperkeratosis, characterized by multiple thin spiny keratotic projections on the palms. The condition has been associated with an underlying malignancy in some cases. One patient has myelofibrosis, an association not previously described. In addition we describe a further patient with filiform hyperkeratoses of both palms and soles with no associated underlying disorder. In view of the relatively high risk of underlying malignancy occurring in patients with filiform hyperkeratosis, these patients should always be investigated fully.


Trastuzumab for treatment of refractory/relapsed HER2-positive adult B-ALL: results of a phase 2 GRAALL study.
The aim of this phase 2 study was to evaluate the efficacy and safety of trastuzumab, a humanized monoclonal antibody targeted against the human epidermal growth factor receptor 2 (HER2), for adult patients with relapsed/refractory HER2-positive B-ALL. Fifteen patients, with a median age of 62 years, received trastuzumab according to the schedule approved for breast cancer patients (ie, 4 mg/kg intravenous loading dose followed by 2 mg/kg weekly). The overall response rate was 13% with 2 patients achieving partial response and partial remission cytolytic response, respectively. Two other patients were documented with blast clearance. Only 1 reversible grade 3 cardiac toxic event occurred. This phase 2 study showed that trastuzumab monotherapy can allow for some responses in a very high-risk refractory/relapsed HER2-positive adult B-ALL population. Combination of trastuzumab with chemotherapy or other therapeutic monoclonal antibodies should be tested in the future.


Ultrasound and photochemical procedures for nanocatalysts preparation: application in photocatalytic biomass valorization.
Nano-photocatalysis is becoming increasingly important due to its multiple applications and multidisciplinary aspects. Applications such as water/air purification, solar energy storage, chemicals production and optoelectronics are some of the most promising. In recent years, the development of novel environmental friendly and cost efficient methods for materials preparation that could replace the old ones is on demand. Unconventional and "soft" techniques such as sonication and photochemistry offer huge possibilities for the synthesis of a broad spectrum of nanostructured materials (e.g., nano-photocatalysts). In the present study, I focus on ultrasound and photochemical procedures for the preparation of nanostructured photocatalysts (e.g., supported metals, metal oxides) and their application in food organic wastes valorization.


[Organizational climate, trust and burnout in a rehabilitation center].
Today human resources are considered of fundamental importance and necessity for the success of the organizations that provide health services. The aims of the present study were: 1) to investigate the perception that rehabilitation medical staff employees have of their hospital organization, 2) to quantify their evaluation concerning organizational trust, 3) to identify eventual burnout symptoms and their relationship with perceived organizational climate and trust. The sample consisted of 131 employees, subdivided into 6 professional categories. Three questionnaires were administered to the employees. The results evidenced significant differences between the various professional groups regarding the climate perceived and trust in the organization. Administrative personnel, nurses and medical graduates revealed a greater satisfaction, responsibility and work integration than the other employee groups (p = 0.023). All the scales which measured organizational climate correlated (inversely) with those measuring burnout (p" 0.05), indicating the existence of a close relationship between a work climate perceived as collaborative (r = -0.33) and characterized by a continuous exchange of information about the hospital organization (r = -0.50), and the psychological well-being experienced by the employees. This study confirms the importance of promoting organizational strategies aimed at mutual reinforcement and support characterized by regular and constructive feedback, wherein there is a reciprocal recognition of each employee's role through a clear, open communication.


[Digital radiography of the thorax with selenium detector. Clinical results and optimization potential of the technique].
We compared the image quality of the chest radiograph obtained with a digital selenium detector and with a conventional system and investigated the possible improvements in the digital technique resulting from kilovoltage (kV) lowering, antiscattering grid addition and image format reduction. 150 subjects in the first series were submitted to posteroanterior chest radiography with both the selenium and the conventional systems. Image quality was compared by giving a score to the depiction of anatominal and pathologic findings. Thirty-two and 31 subjects from two other series were submitted to two digital chest radiographs each: the former at high (150) and low (90) kV, and the latter at 150 kV with the antiscattering grid and at 90 kV without the grid. Comparisons were made by choosing the better of the two images of each subjects. A score was given to the depiction quality of several difficult-to-detect findings in full-size and small-size format digital images obtained in another series of 27 subjects. As for anatomical detailing, digital selenium images were of much better quality than conventional images: the mean scores given by 3 observers to digital images (5.32; 5.55; 6.68) are higher than those given to the corresponding conventional images (4.49; 5.02; 5.81) and the difference is statistically significant (p < 0.001 in all cases). The advantage of digital over conventional images is also significant with reference to diagnostic confidence in the identification of pathologic findings (p < 0.001; p < 0.005; p < 0.01), but to a lessere extent (mean scores: 3.98; 4.22; 3.60 for the digital system, versus 3.43; 3.69; 3.38 for the conventional system). The digital images acquired at lower kV (90 kV) were much more frequently chosen by the two observes (87.5% and 96.8% of cases) than the images acquired at 150 kV; the entry dose at lower energies (91 muGy using an anthropomorphic phantom) is not significantly higher than the dose given at 150 kV (85 muGy). No significant difference was found in the two observers' choice between the digital images taken at 90 kV without antiscattering grid and those taken at 150 kV with the grid, the former being preferred in 38.7% and 58% of cases. The level of diagnostic confidence in the detailing of difficult-to-detect findings was slightly higher in full-size digital images (mean scores: 5.33 and 6.77) than in small-size ones (4.88 and 5.96). Digital selenium images always exibit better quality than conventional images: the difference is very marked relative to anatomical detailing and not so striking, though still significant, in showing pathologic findings. Digital selenium image quality can be improved relative to the manufacturer's guidelines (150 kV exposure with no grid), by lowering the kV and adding the antiscattering grid, without increasing patient exposure too much. Digital image format reduction allows cost containment without affecting diagnostic reliability.


Complementary Dual-Channel Gas Sensor Devices Based on a Role-Allocated ZnO/Graphene Hybrid Heterostructure.
Here, we present a new approach to dual-channel gas sensors on the basis of a role-allocated graphene/ZnO heterostructure, formed by the complementary hybridization of graphene and a ZnO thin film. The method enables cyclic and reproducible gas response as well as high gas response. The role allocation of graphene and ZnO was verified by studying the electrical transport properties of the heterostructure. The results indicated that the ZnO top layer and graphene bottom layer act as a gas adsorption layer and a carrier conducting layer, respectively. The charge interactions of the heterostructures were systematically explored by monitoring changes in transfer characteristics at room temperature and elevated temperature ( T = 250 °C) after introducing 20 ppm NO2. These results can be understood in terms of the dual-channel effect of the graphene/ZnO heterostructures. Remarkably, an abrupt and reliable gas response under periodic NO2 gas injection was unambiguously achieved by the heterostructure-based gas sensors and as ∼30 times higher than those of a graphene-based gas sensor. These proposed heterostructures represent a fundamental building block of a complementary hybrid gas sensor with highly sensitive and reproducible gas response.


Changing Social Contexts to Foster Equity in College Science Courses: An Ecological-Belonging Intervention.
In diverse classrooms, stereotypes are often "in the air," which can interfere with learning and performance among stigmatized students. Two studies designed to foster equity in college science classrooms (Ns = 1,215 and 607) tested an intervention to establish social norms that make stereotypes irrelevant in the classroom. At the beginning of the term, classrooms assigned to an ecological-belonging intervention engaged in discussion with peers around the message that social and academic adversity is normative and that students generally overcome such adversity. Compared with business-as-usual controls, intervention students had higher attendance, course grades, and 1-year college persistence. The intervention was especially impactful among historically underperforming students, as it improved course grades for ethnic minorities in introductory biology and for women in introductory physics. Regardless of demographics, attendance in the intervention classroom predicted higher cumulative grade point averages 2 to 4 years later. The results illustrate the viability of an ecological approach to fostering equity and unlocking student potential.


Synthesis of Ac-Asp-Gly-Ser and Ac-Asp-Pro-Leu-Gly-NH2.
The syntheses of Ac-Asp-Gly-Ser and Ac-Asp-Pro-Leu-Gly-NH2 are described. The two peptides were prepared in solution by stepwise elongation using the DCC method with additives (HOSu or HOBt). The push-pull method was also used in the synthesis of Ac-Asp-Gly-Ser. A racemization test of the amino acids in this peptide has been performed.


Long Distance Translocation of Sucrose, Serine, Leucine, Lysine, and Carbon Dioxide Assimilates: II. Oats.
To establish whether several amino acids were equally able to enter the phloem of oat (Avena sativa L.) plants and be transported, several (14)C-labeled amino acids were applied individually to an abraded spot on a fully expanded source leaf. The base of an immature sink leaf was monitored with a GM tube for time and rate of arrival of radioactivity. Transport of (14)C-sucrose and (14)CO(2) assimilates was measured for a comparison. The applied l-serine, l-lysine, and l-leucine, as well as sucrose, entered the phloem and were transported to the sink leaf at rates between 1.16 and 1.83 cm/min. Transport velocity for CO(2) assimilates was 1.57 cm/min. A heat girdle near the top of the source leaf sheath blocked most transport, which indicated that transport was primarily through the phloem. Mass transfer rates for amino acids were only 3% as great as that for sucrose, suggesting different mechanisms of entry for sucrose than for amino acids into the phloem. The higher percentage of CO(2) assimilates mobilized to the sink leaf was attributed to the greater surface area of minor veins accessible to loading, as compared to those compounds supplied via an abraded spot. Serine was extensively metabolized in the source leaf, and radioactive products in the sink leaf mirrored those in the source leaf. Most radioactivity of lysine and leucine remained within these compounds in the source, path, and sink tissues. We concluded that there was no barrier to entry of amino acids into the phloem and transport therein. Data do not suggest a specific mechanism for entry of amino acids into the phloem.


Body Mass Index and the Risk of Persistent Proteinuria in Middle-Aged Men: The Kansai Healthcare Study.
Proteinuria is a risk factor for end-stage renal failure. However, it is not known whether body mass index (BMI) is prospectively associated with the risk of future developing proteinuria, taking into account transient proteinuria. We enrolled 9,320 nondiabetic Japanese middle-aged men who had no proteinuria, an estimated glomerular filtration rate ≥60 mL/min/1.73 m2, no history of cancer, and no use of antihypertensive medications at baseline. "Any proteinuria" was defined as proteinuria detected for the first time during the follow-up period regardless of its frequency. "Persistent proteinuria" was defined as proteinuria that was detected at least twice consecutively at annual examinations and did not return to negative until the end of the follow-up. During the 11-year follow-up period, 1,972 cases of any proteinuria and 151 cases of persistent proteinuria were confirmed. Both lower and higher BMI were associated with the risk of any proteinuria. As for persistent proteinuria, in those with a BMI ≥20 kg/m2, higher BMI was associated with a higher risk of future persistent proteinuria. The association between BMI and the risk of persistent proteinuria was stronger than that between BMI and any proteinuria. In multiple-adjusted model, hazard ratios of persistent proteinuria for BMI <18.0, 18.0-19.9, 20.0-21.9, 22.0-23.9, 24.0-25.9, 26.0-27.9, and ≥28.0 kg/m2 were 1.52 (95% confidence interval 0.51-4.49), 1.07 (0.49-2.29), 1.00 (reference), 1.14 (0.64-2.01), 1.89 (1.09-3.27), 2.12 (1.15-3.93), and 3.85 (2.03-7.30), respectively. In those with a BMI ≥20 kg/m2, higher BMI was associated with a higher risk of future persistent proteinuria and any proteinuria. This relationship was stronger for persistent proteinuria than for any proteinuria.


The Eng1 β-Glucanase Enhances Histoplasma Virulence by Reducing β-Glucan Exposure.
The fungal pathogen Histoplasma capsulatum parasitizes host phagocytes. To avoid antimicrobial immune responses, Histoplasma yeasts must minimize their detection by host receptors while simultaneously interacting with the phagocyte. Pathogenic Histoplasma yeast cells, but not avirulent mycelial cells, secrete the Eng1 protein, which is a member of the glycosylhydrolase 81 (GH81) family. We show that Histoplasma Eng1 is a glucanase that hydrolyzes β-(1,3)-glycosyl linkages but is not required for Histoplasma growth in vitro or for cell separation. However, Histoplasma yeasts lacking Eng1 function have attenuated virulence in vivo, particularly during the cell-mediated immunity stage. Histoplasma yeasts deficient for Eng1 show increased exposure of cell wall β-glucans, which results in enhanced binding to the Dectin-1 β-glucan receptor. Consistent with this, Eng1-deficient yeasts trigger increased tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) cytokine production from macrophages and dendritic cells. While not responsible for large-scale cell wall structure and function, the secreted Eng1 reduces levels of exposed β-glucans at the yeast cell wall, thereby diminishing potential recognition by Dectin-1 and proinflammatory cytokine production by phagocytes. In α-glucan-producing Histoplasma strains, Eng1 acts in concert with α-glucan to minimize β-glucan exposure: α-glucan provides a masking function by covering the β-glucan-rich cell wall, while Eng1 removes any remaining exposed β-glucans. Thus, Histoplasma Eng1 has evolved a specialized pathogenesis function to remove exposed β-glucans, thereby enhancing the ability of yeasts to escape detection by host phagocytes. The success of Histoplasma capsulatum as an intracellular pathogen results, in part, from an ability to minimize its detection by receptors on phagocytic cells of the immune system. In this study, we showed that Histoplasma pathogenic yeast cells, but not avirulent mycelia, secrete a β-glucanase, Eng1, which reduces recognition of fungal cell wall β-glucans. We demonstrated that the Eng1 β-glucanase promotes Histoplasma virulence by reducing levels of surface-exposed β-glucans on yeast cells, thereby enabling Histoplasma yeasts to escape detection by the host β-glucan receptor, Dectin-1. As a consequence, phagocyte recognition of Histoplasma yeasts is reduced, leading to less proinflammatory cytokine production by phagocytes and less control of Histoplasma infection in vivo Thus, Histoplasma yeasts express two mechanisms to avoid phagocyte detection: masking of cell wall β-glucans by α-glucan and enzymatic removal of exposed β-glucans by the Eng1 β-glucanase.


Lanthanide cofactors accelerate DNA-catalyzed synthesis of branched RNA.
Most deoxyribozymes (DNA catalysts) require metal ions as cofactors for catalytic activity, with Mg(2+), Mn(2+), and Zn(2+) being the most represented activators. Trivalent transition-metal ions have been less frequently considered. Rare earth ions offer attractive properties for studying metal ion binding by biochemical and spectroscopic methods. Here we report the effect of lanthanide cofactors, in particular terbium (Tb(3+)), for DNA-catalyzed synthesis of 2',5'-branched RNA. We found up to 10(4)-fold increased ligation rates for the 9F7 deoxribozyme using 100 μM Tb(3+) and 7 mM Mg(2+), compared to performing the reaction with 7 mM Mg(2+) alone. Combinatorial mutation interference analysis (CoMA) was used to identify nucleotides in the catalytic region of 9F7 that are essential for ligation activity with different metal ion combinations. A minimized version of the DNA enzyme sustained high levels of Tb(3+)-assisted activity. Sensitized luminescence of Tb(3+) bound to DNA in combination with DMS probing and DNase I footprinting results supported the CoMA data. The accelerating effect of Tb(3+) was confirmed for related RNA-ligating deoxyribozymes, pointing toward favorable activation of internal 2'-OH nucleophiles. The results of this study offer fundamental insights into nucleotide requirements for DNA-catalyzed RNA ligation and will be beneficial for practical applications that utilize 2',5'-branched RNA.


Emerging clinical imaging techniques for cerebral cavernous malformations: a systematic review.
Cerebral cavernous malformations (CCMs) are divided into sporadic and familial forms. For clinical imaging, T2-weighted gradient-echo sequences have been shown to be more sensitive than conventional sequences. Recently more advanced imaging techniques such as high-field and susceptibility-weighted MR imaging have been employed for the evaluation of CCMs. Furthermore, diffusion tensor imaging and functional MR imaging have been applied to the preoperative and intraoperative management of these lesions. In this paper, the authors attempt to provide a concise review of the emerging imaging methods used in the clinical diagnosis and treatment of CCMs.


Hyperbilirubinemia due to minor blood group (anti-E) incompatibility in a newborn: a case report.
In addition to Rh and ABO incompatibilities subgroup incompatibilities may rarely play a role among the causes of hemolytic anemia and indirect hyperbilirubinemia in newborns. The most common minor blood group antigens that cause blood incompatibility between the mother and baby are C, c, E, e, Kell, Duffy, Diego, Kidd and MNSs antigens. In this article, a newborn in whom hyperbilirubinemia due to anti-E minor blood group incompatibility developed and was treated with phototherapy succesfully is presented and minor blood group incompatibilities due to anti-E are reviewed.


Molecular diagnosis of cystic fibrosis.
A review of the current molecular diagnosis of cystic fibrosis including an introduction to cystic fibrosis, the gene function, the phenotypic variation, who should be screened for which mutation, newborn and couple screening, quality assurance, phenotype-genotype correlation, methods and method limitations, options, statements, recommendations, useful Websites and treatments.


Aberrant splicing and drug resistance in AML.
The advent of next-generation sequencing technologies has unveiled a new window into the heterogeneity of acute myeloid leukemia (AML). In particular, recurrent mutations in spliceosome machinery and genome-wide aberrant splicing events have been recognized as a prominent component of this disease. This review will focus on how these factors influence drug resistance through altered splicing of tumor suppressor and oncogenes and dysregulation of the apoptotic signaling network. A better understanding of these factors in disease progression is necessary to design appropriate therapeutic strategies recognizing specific alternatively spliced or mutated oncogenic targets.


Body composition, glucose metabolism markers and serum androgens - association in women with polycystic ovary syndrome.
To evaluate the association between abdominal and gynoid fat, glucose and lipid metabolism markers, and serum androgens in women with polycystic ovary syndrome (PCOS). Anthropometric measurements were performed in 40 women with PCOS aged 19-49 years with body mass index (BMI) 18.7-53.8 kg/m2. Fasting serum glucose, lipids, insulin, leptin, LH, FSH, oestradiol, androgens, SHGB and TSH were estimated. Body composition was measured by DEXA scan. Four women (10%) were overweight, and 23 (57.5%) were obese. All subjects were hyperandrogenic (in 33 serum androgen levels were increased), and 16 of them were insulin resistant. All of the obese subjects had the abdominal type of obesity. Body weight, BMI, fat mass, fat mass of the trunk, abdominal and gynoidal fat mass correlated with serum triglyceride, glucose and insulin levels, and with HOMA index and blood pressure. Free androgen index (FAI) correlated with body weight (r = 0.43, p 〈 0.01), and BMI (r = 0.46, p 〈 0.05). Using the DEXA method, we demonstrated abdominal type of obesity in all our obese subjects. There were positive significant correlations between fatness, lipids and glucose metabolism indices and blood pressure. Direct positive correlations between free androgen index, body weight and BMI were found.


Sideburn reconstruction for postrhytidectomy deformity.
A posteriorly based random-pattern scalp flap is described for the correction of sideburn and temporal alopecia. The procedure can be readily incorporated into a coronal lift operation.


Null mutation of AtCUL1 causes arrest in early embryogenesis in Arabidopsis.
The SCF (for SKP1, Cullin/CDC53, F-box protein) ubiquitin ligase targets a number of cell cycle regulators, transcription factors, and other proteins for degradation in yeast and mammalian cells. Recent genetic studies demonstrate that plant F-box proteins are involved in auxin responses, jasmonate signaling, flower morphogenesis, photocontrol of circadian clocks, and leaf senescence, implying a large spectrum of functions for the SCF pathway in plant development. Here, we present a molecular and functional characterization of plant cullins. The Arabidopsis genome contains 11 cullin-related genes. Complementation assays revealed that AtCUL1 but not AtCUL4 can functionally complement the yeast cdc53 mutant. Arabidopsis mutants containing transfer DNA (T-DNA) insertions in the AtCUL1 gene were shown to display an arrest in early embryogenesis. Consistently, both the transcript and the protein of the AtCUL1 gene were found to accumulate in embryos. The AtCUL1 protein localized mainly in the nucleus but also weakly in the cytoplasm during interphase and colocalized with the mitotic spindle in metaphase. Our results demonstrate a critical role for the SCF ubiquitin ligase in Arabidopsis embryogenesis.


Synthesis of nestlike ZnO hierarchically porous structures and analysis of their gas sensing properties.
Nestlike 3D ZnO porous structures with size of 1.0-3.0 μm have been synthesized through annealing the zinc hydroxide carbonate precursor, which was obtained by a one-pot hydrothermal process with the assistance of glycine, Na(2)SO(4), and polyvinyl pyrrolidone (PVP). The nestlike 3D ZnO structures are built of 2D nanoflakes with the thickness of ca. 20 nm, which exhibit the nanoporous wormhole-like characteristic. The measured surface area is 36.4 m(2)g(-1) and the pore size is ca. 3-40 nm. The unique nestlike 3D ZnO porous structures provided large contacting surface area for electrons, oxygen and target gas molecules, and abundant channels for gas diffusion and mass transport. Gas sensing tests showed that the nestlike 3D ZnO porous structures exhibit excellent gas sensing performances such as high sensitivity and fast response and recovery speed, suggesting the potential applications as advanced gas sensing materials.


The circle of Willis in healthy older persons.
The current indication for carotid surgery is based upon the percentage of stenosis of the internal carotid artery. This approach shows many limitations, one of which is to presume the anatomical completeness of the circle of Willis in all patients who are evaluated. On the contrary, there is increasing evidence of a great natural variability in the configuration of the circle of Willis. The aim of the present paper was to investigate, by magnetic resonance angiography, the variability of the circle of Willis in a cohort of 118 healthy older persons. The circle of Willis showed an entirely complete configuration in 47% of the subjects, a complete configuration of its anterior part in 90% of the subjects, and a complete configuration of its posterior part in 48.5% of the subjects. These findings confirm the great variability of the circle of Willis even in healthy older persons and suggest that, in indicating carotid surgery, the configuration of the circle of Willis should also be taken into account.


A national study of the efficiency of hospitals in urban markets.
Using a sample of 3,000 urban hospitals, this article examines the contributions of selected hospital characteristics to variations in hospital technical efficiencies, while it accounts for multiple products and inputs, and controls for local environmental variations. Four hospital characteristics are examined: hospital size, membership in a multihospital system, ownership, and payer mix (managed care contracts, percent Medicare, and percent Medicaid). Ownership and percent Medicare are consistently found to be related significantly to hospital efficiency. Within the ownership variable, government hospitals tend to be more efficient and for-profit hospitals less efficient than other hospitals. Higher percentages of Medicare payment are negatively related to efficiency. While not consistently significant across all five of the MSA size categories in which the analyses are conducted, possession of managed care contracts, membership in a multihospital system, and size all are consistently related positively to hospital technical efficiency. These variables are also all significant when the hospitals are examined in a combined analysis. Percent Medicaid was not significant in any of the analyses. Implications for policy and the need for methodological work are discussed.


Intraoperative Management of Hypercapnia With an Extracorporeal Carbon Dioxide Removal Device During Giant Bullectomy.
Extracorporeal CO2-removal devices have been introduced in clinical practice to provide protective and ultraprotective ventilation strategies in different settings to avoid retention of carbon dioxide. The need to facilitate lung-protective ventilation is required not only for the treatment of acute respiratory distress syndrome but also in thoracic surgery during complex operations, especially in respiratory compromised patients. This report describes a case of giant bullectomy for vanishing lung syndrome in which intraoperative hypercapnia secondary to protective ventilation was managed with a CO2-removal device (Decap-Hemodec s.r.l., Salerno, Italy). To the best of our knowledge, this is the first report in the literature of the intraoperative use of the Decap system for giant bullectomy.


Clinical utilization of blinatumomab and inotuzumab immunotherapy in children with relapsed or refractory B-acute lymphoblastic leukemia.
The treatment paradigm for patients with relapsed/refractory B-cell acute lymphoblastic leukemia (rrALL) has been revolutionized given recent clinical trials demonstrating remarkable success of immunotherapies and leading to drug approvals by United States and European agencies. We report experience with commercial blinatumomab and inotuzumab use at two North American pediatric oncology centers in children and adolescents/young adults with B-ALL. Patients 0-25 years old treated with the CD19 × CD3 bispecific T cell-engaging antibody blinatumomab and/or the CD22 antibody-drug conjugate inotuzumab from January 1, 2010, to June 1, 2018, were eligible. Disease status included relapsed B-ALL in second or greater relapse, primary chemotherapy-refractory B-ALL, or B-ALL complicated by severe infection precluding delivery of conventional chemotherapy. We identified 27 patients who received blinatumomab and/or inotuzumab outside of clinical trials during the study period. Four of the 13 patients (31%) with relapsed disease achieved minimal residual disease (MRD)-negative remission, and five patients (39%) underwent hematopoietic stem cell transplant (HSCT). In the 12 patients with primary chemorefractory B-ALL treated with immunotherapy, 11 (92%) achieved MRD-negative remission as assessed by flow cytometry; 10 patients (83%) underwent subsequent HSCT. Two patients with B-ALL in MRD-negative remission received blinatumomab due to severe infection and remained in remission after chemotherapy continuation. Blinatumomab and inotuzumab can induce deep remissions in patients with rrALL and facilitate subsequent HSCT or other cellular therapies. Blinatumomab can also serve as an effective bridging therapy during severe infection. The optimal timing, choice of immunotherapeutic agent(s), and duration of responses require further investigation via larger-scale clinical trials.


Multimodality therapy of localized unresectable pancreatic adenocarcinoma.
Eighty-eight patients with localized unresectable carcinoma of the pancreas were treated at Thomas Jefferson University Hospital between 1974 and 1981. Four treatment regimens were used which were sequential modifications of the technique based on the experience in the preceding group of patients. Each treatment changed the course of the disease, and as patterns of failure were identified, the treatment was altered to deal with them. Initially, all patients were treated with external beam radiation. Subsequently, Iodine-125 implantation was added to improve local control; low-dose preoperative radiotherapy to reduce the risk of peritoneal seeding; and adjuvant chemotherapy to reduce the risks of distant metastases. The addition of 125I implantation increased the local control from 22% to 81%, but did not increase the median survival, which was unchanged from 7 months. The addition of adjuvant chemotherapy increased the median survival from 7 months to 14 months, but had no impact on the control of the pancreatic tumor. Adjunctive chemotherapy and low-dose preoperative radiotherapy appear synergistic in reducing the risk of peritoneal seeding. The combination of 125I implantation, external beam radiation, and adjunctive chemotherapy is safe and effective. This regimen produces excellent local control with acceptable morbidity. This regimen produced a 30% survival at 18 months. The patterns of failure among these patients suggest future modifications of the technique.


Ascaris lumbricoides infestation as a cause of intestinal obstruction in children: experience with 87 cases.
The purpose of this study is to describe the occurrence and management of bowel obstruction caused by Ascaris lumbricoides, a common parasite in warm climates that affects children with limited socioeconomic means. Eighty-seven patients with intestinal infestation owing to Ascaris lumbricoides were treated in the past 10 years (1984-1994). There were 48 (55.2%) girls and 39 (44.8%) boys. The mean age was 4.6 years, with peak occurrence at 2 years of age. Half the patients had a history of passing worms by mouth or anus. The majority of patients, 64 (73.5%), presented with a subacute clinical course; 23 (26.5%) had acute presentation, with severe abdominal pain, fever, dehydratation, vomiting, and abdominal distension and required vigorous fluid resuscitation and emergency surgical intervention. Diagnosis was achieved with plain abdominal roentgenograms, which showed a "whirlpool" pattern of intraluminal worms in most cases. Six patients had been incorrectly diagnosed as having appendicitis; two cases had appendicitis owing to Ascaris in the cecum and distal ileum. The majority of cases with a subacute presentation respond to medical (anthelmintic) management using oral administration of racine oil and piperazine. Of the 23 patients taken to the operating room, 11 required external "milking" of the obstructing bolus of worms from the ileum into the colon, six required intestinal resection and end-to-end anastomosis, six had an appendectomy, and three needed an enterotomy to manually extract the worms. In one case, initial management consisted of an ileostomy because of intraoperative instability owing to sepsis. Subsequently, after stabilization and treatment with anthelmintic agents, closure of the stoma with an end-to-end ileocolostomy was performed. There was no significant postoperative morbidity or mortality. These observations suggest that physicians should have a high index of suspicion for parasitic infestation in warm climates where economically deprived children present with symptoms of intestinal obstruction. Ascaris lumbricoides may be the cause of these events in endemic areas. Oral piperazine and racine oil can successfully resolve most subacute cases; however, aggressive resuscitation and prompt surgical intervention in patients with intestinal obstruction result in a satisfactory outcome.


Effects of moxibustion on dynorphin and endomorphin in rats with chronic visceral hyperalgesia.
To observe the analgesic effects of moxibustion in rats with chronic visceral hyperalgesia and its influence on the concentration of dynorphin (Dyn) and endomorphin (EM) in spinal cord. The rat model of chronic visceral hyperalgesia was established by colorectal distention (CRD). In moxibustion (MX) group, moxibustion was applied once daily for 7 d; in sham moxibustion (SM) group, moxibustion was given to the same acupoints but with the non-smoldered end of the moxa stick. Model control (MC) group and normal control group were also studied. The scoring system of abdominal withdrawal reflex was used to evaluate visceral pain for behavioral assessment. Enzyme linked immunosorbent assay was performed to determine the concentrations of Dyn and EM in spinal cord. Moxibustion significantly decreased visceral pain to CRD in this rat model, and no significant difference was detected between the SM group and the MC group. In MX group, moxibustion also increased the concentrations of Dyn and EM in spinal cord, and no significant difference was found between the SM group and the MC group. Moxibustion therapy can significantly enhance the pain threshold of rats with chronic visceral hyperalgesia, and the effect may be closely related to the increased concentration of Dyn and EM in spinal cord.


Mouse embryonic stem cells established in physiological-glucose media express the high KM Glut2 glucose transporter expressed by normal embryos.
Glut2 is one of the facilitative glucose transporters expressed by preimplantation and early postimplantation embryos. Glut2 is important for survival before embryonic day 10.5. The Glut2 KM (∼16 mmol/liter) is significantly higher than physiologic glucose concentrations (∼5.5 mmol/liter), suggesting that Glut2 normally performs some essential function other than glucose transport. Nevertheless, Glut2 efficiently transports glucose when extracellular glucose concentrations are above the Glut2 KM. Media containing 25 mmol/liter glucose are widely used to establish and propagate embryonic stem cells (ESCs). Glut2-mediated glucose uptake by embryos induces oxidative stress and can cause embryo cell death. Here we tested the hypothesis that low-glucose embryonic stem cells (LG-ESCs) isolated in physiological-glucose (5.5 mmol/liter) media express a functional Glut2 glucose transporter. LG-ESCs were compared with conventional D3 ESCs that had been cultured only in high-glucose media. LG-ESCs expressed Glut2 mRNA and protein at much higher levels than D3 ESCs, and 2-deoxyglucose transport by LG-ESCs, but not D3 ESCs, exhibited high Michaelis-Menten kinetics. Glucose at 25 mmol/liter induced oxidative stress in LG-ESCs and inhibited expression of Pax3, an embryo gene that is inhibited by hyperglycemia, in neuronal precursors derived from LG-ESCs. These effects were not observed in D3 ESCs. These findings demonstrate that ESCs isolated in physiological-glucose media retain a functional Glut2 transporter that is expressed by embryos. These cells are better suited to the study of metabolic regulation characteristic of the early embryo and may be advantageous for therapeutic applications.


Metabolic surgery for type 2 diabetes: appraisal of clinical evidence and review of randomized controlled clinical trials comparing surgery with medical therapy.
Bariatric surgical procedures were originally developed to treat morbid obesity where their benefits certainly outweigh their potential side effects. Although they are very beneficial in improving metabolic control in type 2 diabetes, there are many medical treatments that are also effective. The role of bariatric surgery as primary therapy for type 2 diabetes depends on whether the benefit exceeds the surgical and nutritional complications, which are significant. The ultimate role for bariatric surgery in treating type 2 diabetes can only be determined by large, long-term randomized clinical trials which compare clinical outcomes of bariatric surgery with those of current intensive medical treatment. The four reported small, mostly 1-year trials have shown superior glycemic control by surgery as compared with medical treatment, but at the expense of significant surgical complications and unknown nutritional liability. They show that future trials will have to be much larger and last for at least 5-10 years.


Screening examination and treatment of Trichophyton tonsurans infection in judo athletes affiliated with the University Judo Federation of Tokyo.
In Japan, Trichophyton tonsurans infection has become an increasing problem among combat sports participants. We investigated the prevalence of T. tonsurans infection in athletes affiliated to judo clubs in the 21 First Division universities that were registered with the University Judo Federation of Tokyo in 2008. Study procedures performed by the subjects included (i) completion of a questionnaire concerning lifestyle, risk factors for tinea corporis and medical history; (ii) scrubbing the scalp with a circular hairbrush to obtain samples for fungal culture; (iii) anti-fungal treatment as recommended by a dermatologist, based on the number of fungal colonies isolated from the hairbrush; and (iv) repeat testing using the hairbrush method 3 months after treatment recommendations were received. Of 902 study subjects, 102 (11.3%) yielded positive hairbrush culture results. Of these, 14 individuals (13.7%) had tinea corporis; the remainder were asymptomatic. Conversion to negative fungal culture was observed in 85 of 96 culture-positive individuals who performed the second hairbrush culture test following treatment. Control of T. tonsurans infection among judo athletes could be achieved by educating athletes, trainers and coaches in judo clubs concerning detection, prevention, and treatment of T. tonsurans infection.


Decreased Bioenergetic Health Index in monocytes isolated from the pericardial fluid and blood of post-operative cardiac surgery patients.
Monitoring the bioenergetics of leucocytes is now emerging as an important approach in translational research to detect mitochondrial dysfunction in blood or other patient samples. Using the mitochondrial stress test, which involves the sequential addition of mitochondrial inhibitors to adherent leucocytes, we have calculated a single value, the Bioenergetic Health Index (BHI), which represents the mitochondrial function in cells isolated from patients. In the present report, we assess the BHI of monocytes isolated from the post-operative blood and post-operative pericardial fluid (PO-PCF) from patients undergoing cardiac surgery. Analysis of the bioenergetics of monocytes isolated from patients' PO-PCF revealed a profound decrease in mitochondrial function compared with monocytes isolated from their blood or from healthy controls. Further, patient blood monocytes showed no significant difference in the individual energetic parameters from the mitochondrial stress test but, when integrated into the BHI evaluation, there was a significant decrease in BHI compared with healthy control monocytes. These data support the utility of BHI measurements in integrating the individual parameters from the mitochondrial stress test into a single value. Supporting our previous finding that the PO-PCF is pro-oxidant, we found that exposure of rat cardiomyocytes to PO-PCF caused a significant loss of mitochondrial membrane potential and increased reactive oxygen species (ROS). These findings support the hypothesis that integrated measures of bioenergetic health could have prognostic and diagnostic value in translational bioenergetics.


Update in medical therapy of ulcerative colitis: a practical approach.
The bewildering array of medications in the therapy of inflammatory bowel disease (IBD) often confounds the clinician in the choice of specific agents regarding the balance between safety and efficacy. This review surveys and evaluates currently available IBD therapies as well as those used in clinical trials of ulcerative colitis. The primary purpose is to provide the busy clinician with a practical guide to the use of established and newly emerging medical therapies of IBD.


Divergent membrane properties of mouse cochlear glial cells around hearing onset.
Spiral ganglion neurons (SGNs) are the primary afferent neurons of the auditory system, and together with their attendant glia, form the auditory nerve. Within the cochlea, satellite glial cells (SGCs) encapsulate the cell body of SGNs, whereas Schwann cells (SCs) wrap their peripherally- and centrally-directed neurites. Despite their likely importance in auditory nerve function and homeostasis, the physiological properties of auditory glial cells have evaded description. Here, we characterized the voltage-activated membrane currents of glial cells from the mouse cochlea. We identified a prominent weak inwardly rectifying current in SGCs within cochlear slice preparations (postnatal day P5-P6), which was also present in presumptive SGCs within dissociated cultures prepared from the cochleae of hearing mice (P14-P15). Pharmacological block by Ba2+ and desipramine suggested that channels belonging to the Kir4 family mediated the weak inwardly rectifying current, and post hoc immunofluorescence implicated the involvement of Kir4.1 subunits. Additional electrophysiological profiles were identified for glial cells within dissociated cultures, suggesting that glial subtypes may have specific membrane properties to support distinct physiological roles. Immunofluorescence using fixed cochlear sections revealed that although Kir4.1 is restricted to SGCs after the onset of hearing, these channels are more widely distributed within the glial population earlier in postnatal development (i.e., within both SGCs and SCs). The decrease in Kir4.1 immunofluorescence during SC maturation was coincident with a reduction of Sox2 expression and advancing neurite myelination. The data suggest a diversification of glial properties occurs in preparation for sound-driven activity in the auditory nerve.


Early clinical and radiological results of unilateral posterior pedicle instrumentation through a Wiltse approach with lateral lumbar interbody fusion.
To assess the clinical outcomes of 44 patients who underwent single-level lateral lumbar interbody fusion (LLIF) with unilateral pedicle screw instrumentation through a paramedian Wiltse approach. Demographic, comorbidity, clinical assessment, peri-operative, and complication data were assessed. Visual analog scale (VAS), Oswestry disability index (ODI), and short form-12 (SF-12) were used to assess clinical outcomes. Post-operative plain radiographs were assessed for subsidence, cage migration, and fusion. Average age of patients at surgery was 60.8±10.6 years (range, 32-79 years), with 15 males and 29 females. Recombinant human bone morphogenic protein-2 (rhBMP-2) was used in 32 cases (73%) and 13 posterolateral fusions (30%). Average duration of surgery was 195.2±36.9 minutes (range: 111-295 minutes), with an estimated blood loss of 159.3±90.8 cc (range, 50-500 cc). There were no intra-operative complications. Average length of hospital stay was 4.2±2.5 days (range, 2-13 days). Four patients (9%) experienced neurological deficit, 2 of which resolved by 3-month follow-up and 2 of which improved but did not resolve by final follow-up at 11 and 16 months. There was significant improvement in VAS (P<0.001), ODI (P<0.001), and SF-12 physical component (P<0.001), but not for SF-12 mental component (P=0.053). Patients with minimum 6 months radiographic follow-up demonstrated successful fusion in 90% of cases (35/39), with 2 cases of grade 1 (5%) subsidence of the adjacent cranial vertebra, and no cases higher than grade 0 subsidence of the adjacent caudal vertebra. Unilateral pedicle screw instrumentation following LLIF was associated with significant improvement in clinical outcomes and favorable radiographic outcomes.


Comparison of 2 Immunosuppression Minimization Strategies in Kidney Transplantation: The ALLEGRO Trial.
Evidence on the optimal maintenance of immunosuppressive regimen in kidney transplantation recipients is limited. The Amsterdam, LEiden, GROningen trial is a randomized, multicenter, investigator-driven, noninferiority, open-label trial in de novo kidney transplant recipients, in which 2 immunosuppression minimization strategies were compared with standard immunosuppression with basiliximab, corticosteroids, tacrolimus, and mycophenolic acid. In the minimization groups, either steroids were withdrawn from day 3, or tacrolimus exposure was reduced from 6 mo after transplantation. The primary endpoint was kidney transplant function at 24 mo. A total of 295 participants were included in the intention-to-treat analysis. Noninferiority was shown for the primary endpoint; estimated glomerular filtration rate at 24 mo was 45.3 mL/min/1.73 m2 in the early steroid withdrawal group, 49.0 mL/min/1.73 m2 in the standard immunosuppression group, and 44.7 mL/min/1.73 m2 in the tacrolimus minimization group. Participants in the early steroid withdrawal group were significantly more often treated for rejection (P = 0.04). However, in this group, the number of participants with diabetes mellitus during follow-up and total cholesterol at 24 mo were significantly lower. Tacrolimus minimization can be considered in kidney transplant recipients who do not have an increased immunological risk. Before withdrawing steroids the risk of rejection should be weighed against the potential metabolic advantages.


Aluminum-induced mitogenesis in MC3T3-E1 osteoblasts: potential mechanism underlying neoosteogenesis.
We recently reported that aluminum administration to beagles stimulates uncoupled bone formation in the marrow cavity which increases trabecular bone volume and generates new osseous networks within the axial skeleton. To investigate whether this osteogenic process results from direct stimulation of bone cell replication, we examined the mitogenic effects of aluminum on undifferentiated osteoblasts derived from the MC3T3-E1 clonal cell line. Addition of AlCl3 (1-50 microM) to serum-free culture medium of quiescent osteoblasts resulted in a dose-dependent increase in [3H]thymidine incorporation into DNA and a concordant increase in cell number to 48% of the density achieved at the maximum replicative rate induced by fetal bovine serum (FBS). The time course of aluminum-induced mitogenesis was similar to that of FBS, with onset of DNA synthesis detectable by 12 h and progressive increases in replicative rates observed over a 48-h study period. Moreover, maximal stimulation of DNA synthesis by AlCl3 and that by FBS were not additive, whereas aluminum exerted additional effects on cell replication when combined with low FBS concentrations. Analysis of cell cycle kinetics indicated that aluminum, analogous to FBS, influences the osteoblast replicative activity by inducing transition from the G0 to the S phase of the cell cycle. In addition, exposure of cells to aluminum resulted in rapid accumulation of c-fos mRNA by 30 min, indicating that aluminum, like fetal bovine serum, induced expression of growth-regulating genes. Deferoxamine mesylate, a chelator of aluminum, blocked the replicative actions of aluminum in a dose-dependent fashion. In contrast, pertussis toxin, a specific inhibitor of certain G-proteins, had no effect on the mitogenic effects of aluminum, indicating that aluminum-induced mitogenesis occurs by a pertussis toxin-insensitive pathway. Though the particular cellular pathway remains to be defined, these data provide initial evidence that aluminum-induced neoosteogenesis may depend upon direct stimulation of osteoblast replication.


Ameloblastoma diagnosis by fine-needle aspiration cytology supplemented by cell block samples.
Ameloblastomas are rarely encountered in clinical practice, accounting for only 1% of tumors and cysts of the jaw although they are one of the most common odontogenic neoplasms. The cytological features are described in a few case reports only. The aim of this study was to describe the morphological features of ameloblastoma in fine-needle aspiration cytology (FNAC) and highlight the contribution of cell blocks in their diagnosis. Three cases of ameloblastoma diagnosed on FNAC (FNAC) with cell block samples were retrieved and evaluated in detail. Radiological correlation was performed in three cases. Follow-up histopathology of the resected specimen was available in two cases. Cytology smears showed clusters of basaloid cells with high nucleocytoplasmic(N/C) ratio and dense chromatin. Focal squamoid differentiation was present in one case and cystic change predominated in one case. The characteristic morphology of the tumor was better appreciated on cell block section with cribriform and trabecular arrangement of tumor cells with peripheral nuclear palisading with foci of squamoid differentiation and cystic change. Cell blocks from aspirates act as mini-biopsies and contribute to accurate diagnoses of ameloblastomas of the mandible, thereby emphasizing their contribution to the proper management of these uncommon neoplasms.


A cross-species spatiotemporal proteomic analysis identifies UBE3A-dependent signaling pathways and targets.
Angelman syndrome (AS) is a severe neurodevelopmental disorder caused by the loss of neuronal E3 ligase UBE3A. Restoring UBE3A levels is a potential disease-modifying therapy for AS and has recently entered clinical trials. There is paucity of data regarding the molecular changes downstream of UBE3A hampering elucidation of disease therapeutics and biomarkers. Notably, UBE3A plays an important role in the nucleus but its targets have yet to be elucidated. Using proteomics, we assessed changes during postnatal cortical development in an AS mouse model. Pathway analysis revealed dysregulation of proteasomal and tRNA synthetase pathways at all postnatal brain developmental stages, while synaptic proteins were altered in adults. We confirmed pathway alterations in an adult AS rat model across multiple brain regions and highlighted region-specific differences. UBE3A reinstatement in AS model mice resulted in near complete and partial rescue of the proteome alterations in adolescence and adults, respectively, supporting the notion that restoration of UBE3A expression provides a promising therapeutic option. We show that the nuclear enriched transketolase (TKT), one of the most abundantly altered proteins, is a novel direct UBE3A substrate and is elevated in the neuronal nucleus of rat brains and human iPSC-derived neurons. Taken together, our study provides a comprehensive map of UBE3A-driven proteome remodeling in AS across development and species, and corroborates an early UBE3A reinstatement as a viable therapeutic option. To support future disease and biomarker research, we present an accessible large-scale multi-species proteomic resource for the AS community ( https://www.angelman-proteome-project.org/ ).


[The identification of ephedrine and its analogues in urine by gas chromatography].
A sensitive and reliable method of analysis for ephedrine and its analogues was established by using GC and GC-MS. The columns used for both GC and GC-MS were HP-5, and the carrier gas was helium. NPD was used as the detector for GC. The urine sample was extracted with ether and submitted to GC. If positive, it was then submitted to GC-MS to identify its structure. The detection limits for ephedrine and its analogues are lower than 200 ng/ml urine. The recoveries are above 80%.


Clinical validation study of percutaneous cochlear access using patient-customized microstereotactic frames.
Percutaneous cochlear implant (PCI) surgery consists of drilling a single trough from the lateral cranium to the cochlea avoiding vital anatomy. To accomplish PCI, we use a patient-customized microstereotactic frame, which we call a "microtable" because it consists of a small tabletop sitting on legs. The orientation of the legs controls the alignment of the tabletop such that it is perpendicular to a specified trajectory. Prospective. Tertiary referral center. Thirteen patients (18 ears) undergoing traditional cochlear implant surgery. With institutional review board approval, each patient had 3 fiducial markers implanted in bone surrounding the ear. Temporal bone computed tomographic scans were obtained, and the markers were localized, as was vital anatomy. A linear trajectory from the lateral cranium through the facial recess to the cochlea was planned. A microtable was fabricated to follow the specified trajectory. After mastoidectomy and posterior tympanotomy, accuracy of trajectories was validated by mounting the microtables on the bone-implanted markers and then passing sham drill bits across the facial recess to the cochlea. The distance from the drill to vital anatomy was measured. Microtables were constructed on a computer-numeric-control milling machine in less than 5 minutes each. Successful access across the facial recess to the cochlea was achieved in all 18 cases. The mean +/- SD distance was 1.20 +/- 0.36 mm from midportion of the drill to the facial nerve and 1.25 +/- 0.33 mm from the chorda tympani. These results demonstrate the feasibility of PCI access using customized microstereotactic frames.


Regenerative process after experimental injury of hypoglossal nerves in guinea pigs.
The nerve regenerative process has been investigated by many studies. However, the quantification of the degree of crush of peripheral nerves has not yet been performed. The aim of this study was to determine and standardize the ligature intensity and crush level of the hypoglossal nerve of guinea pigs. The compound action potentials evoked by electric stimulation were used as an index of the degree of nerve crush. To demonstrate nerve regeneration after ligating and crushing of the right hypoglossal nerve, fluorescein isothiocynate conjugated cholera toxin-B subunit (CTb-FITC) was injected into the intact fiber of the left hypoglossal nerve, and the central side from the crushed region of the right hypoglossal nerve fiber. The total cross sectional area of the retrograde-labeled hypoglossal motoneurons was investigated under a confocal laser scanning microscope. The results of the evaluation using CTb-FITC indicated that the nerve regeneration occurred from two weeks after crush and recovered in six weeks.


Combined Schwannoma and Kappa-Restricted Plasma Cell Neoplasm: A Case Report and Review of the Literature.
The patient is a 78-year-old woman with a popliteal soft tissue mass that was tender to palpation with shooting pain on physical examination. A schwannoma was seen on biopsy with subsequent excision demonstrating a concomitant kappa-restricted plasma cell neoplasm. Follow-up did not show evidence of a systemic plasma cell neoplasm. MRI studies showed no evidence of focal lesions, although PET-CT revealed presence of multiple lytic lesions. The patient is currently being monitored every six months. This case is the first kappa-restricted plasma cell neoplasm reported in association with a schwannoma and the first reported in the extremities.


Iron Supplementation Effects on Redox Status following Aseptic Skeletal Muscle Trauma in Adults and Children.
Exercise-induced skeletal muscle microtrauma is characterized by loss of muscle cell integrity, marked aseptic inflammatory response, and oxidative stress. We examined if iron supplementation would alter redox status after eccentric exercise. In a randomized, double blind crossover study, that was conducted in two cycles, healthy adults (n = 14) and children (n = 11) received daily either 37 mg of elemental iron or placebo for 3 weeks prior to and up to 72 h after an acute eccentric exercise bout. Blood was drawn at baseline, before exercise, and 72 h after exercise for the assessment of iron status, creatine kinase activity (CK), and redox status. Iron supplementation at rest increased iron concentration and transferrin saturation (p < 0.01). In adults, CK activity increased at 72 h after exercise, while no changes occurred in children. Iron supplementation increased TBARS at 72 h after exercise in both adults and children; no changes occurred under placebo condition. Eccentric exercise decreased bilirubin concentration at 72 h in all groups. Iron supplementation can alter redox responses after muscle-damaging exercise in both adults and children. This could be of great importance not only for healthy exercising individuals, but also in clinical conditions which are characterized by skeletal muscle injury and inflammation, yet iron supplementation is crucial for maintaining iron homeostasis. This study was registered at Clinicaltrials.gov Identifier: NCT02374619.


Detection of herpes simplex virus in pseudoexfoliation syndrome and exfoliation glaucoma.
The pathogenesis of pseudoexfoliation syndrome (PEX) remains unknown. An infection, possibly viral, is one of the proposed pathogenetic mechanisms. This study examines the presence of herpes simplex virus (HSV) and varicella-zoster virus (VZV) in iris and anterior capsule specimens of PEX and non-PEX patients. Iris and anterior capsule specimens were obtained from 64 patients with PEX (study group, SG) and 61 patients without PEX (control group, CG). The presence of HSV and VZV DNA was evaluated with a polymerase chain reaction (PCR). Herpes simplex virus type I was detected significantly more often in iris specimens from the SG (13.79%), compared to those from the CG (1.75%). Varicella-zoster virus DNA was not detected in any of the examined specimens. Results imply a possible relationship between HSV type I and PEX, although no aetiological role of HSV infection in PEX pathogenesis can be established. Results also advocate against any association between VZV and PEX.


Transition metal alkynyl complexes by transmetallation from Au(C[triple bond]CAr)(PPh3) (Ar = C6H5 or C6H4Me-4).
Facile acetylide transfer reactions take place between gold(I) complexes Au(C[triple bond]CAr)(PPh3) (Ar = C6H5 or C6H4Me-4) and a variety of representative inorganic and organometallic complexes MXL, (M = metal, X = halide, L, = supporting ligands) featuring metals from groups 8-11, to afford the corresponding metal-alkynyl complexes M(C[triple bond]CR)Ln in modest to good yield. Reaction products have been characterised by spectroscopic methods, and molecular structure determinations are reported for Fe(C[triple bond]CC6H4Me-4)(dppe)Cp, Ru(C[triple bond]CC6H4Me-4)(dppe)Cp*, Ru(C=CCsF,)(l2-O2)(PPh3)Cp*, Ir(C-CC6H4Me-4)(eta2-O2)(CO)(PPh3), Ni(C[triple bond]CC6H4Me-4)(PPh3)Cp and trans-Pt(C[triple bond]CAr)2L2 (Ar = C6H5, L = PPh3; Ar = C6H4Me-4, L = PPh3, PMe3).


Deciding about patients' requests for extraction: ethical and legal guidelines.
and Overview. Dentists frequently are faced with patients' requests for an extraction, sometimes of the entire dentition. In this article, the authors offer guidelines to help dentists and oral surgeons make decisions regarding such requests for extraction. In most cases of patients' requesting extractions, the ethical principle of nonmaleficence will play a decisive role in the dentist's decision making. In cases in which the request appears influenced by a specific mental condition such as a phobia of dental treatment, extraction rarely is justifiable. Practice Implications. Dental professionals should keep in mind that they cannot be forced to carry out treatment that is at odds with the ethical principle of nonmaleficence or that is outside of the bounds of accepted treatment. To aid dentists in making treatment decisions in such cases, the authors present a flowchart that integrates possible considerations.


Imaging of Hepatocellular Carcinoma Response After 90Y Radioembolization.
The purpose of this study is to discuss the imaging modalities and response criteria used for assessing hepatocellular carcinoma (HCC) response to 90Y radioembolization, as well as the imaging appearances of treated tumors. An understanding of the appearance of HCC after 90Y radioembolization is crucial for accurate evaluation of treatment response. Residual tumor necrosis and enhancement are essential for assessing response. Multiparametric MRI, including DWI and perfusion imaging, plays an emerging role in response assessment and outcome prediction.


Increasing cholesterol synthesis in 7-dehydrosterol reductase (DHCR7) deficient mouse models through gene transfer.
Smith-Lemli-Opitz syndrome (SLOS) is caused by deficiency in the terminal step of cholesterol biosynthesis: the conversion of 7-dehydrocholesterol (7DHC) to cholesterol (C), catalyzed by 7-dehydrocholesterol reductase (DHCR7). This disorder exhibits several phenotypic traits including dysmorphia and mental retardation with a broad range of severity. There are few proven treatment options. That most commonly used is a high cholesterol diet that seems to enhance the quality of life and improve behavioral characteristics of patients, although these positive effects are controversial. The goal of our study was to investigate the possibility of restoring DHCR7 activity by gene transfer. We constructed an adeno-associated virus (AAV) vector containing the DHCR7 gene. After we infused this vector into affected mice, the introduced DHCR7 gene could be identified in liver, mRNA was expressed and a functional enzyme was produced. Evidence of functionality came from the ability to partially normalize the serum ratio of 7DHC/C in treated animals, apparently by increasing cholesterol production with concomitant decrease in 7DHC precursor. By 5 weeks after treatment the mean ratio (for 7 animals) had fallen to 0.05 while the ratio for untreated littermate controls had risen to 0.14. This provides proof of principle that gene transfer can ameliorate the genetic defect causing SLOS and provides a new experimental tool for studying the pathogenesis of this disease. If effective in humans, it might also offer a possible alternative to exogenous cholesterol therapy. However, it would not offer a complete cure for the disorder as many of the negative implications of defective synthesis are already established during prenatal development.


Simultaneous lack of catalase and beta-toxin in Staphylococcus aureus leads to increased intracellular survival in macrophages and epithelial cells and to attenuated virulence in murine and ovine models.
Staphylococcus aureus produces a variety of virulence factors that allow it to cause a wide range of infections in humans and animals. In the latter, S. aureus is a leading cause of intramammary infections. The contribution of catalase (KatA), an enzyme implicated in oxidative stress resistance, and beta-toxin (Hlb), a haemolysin, to the pathogenesis of S. aureus is poorly characterized. To investigate the role of these enzymes as potential virulence factors in S. aureus, we examined the intracellular survival of DeltakatA, Deltahlb and DeltakatA Deltahlb mutants in murine macrophages (J774A.1) and bovine mammary epithelial cells (MAC-T), and their virulence in different murine and ovine models. Catalase was not required for the survival of S. aureus within either J774A.1 or MAC-T cells. However, it was necessary for the intracellular proliferation of the bacterium after invasion of MAC-T cells. In addition, catalase was not needed for the full virulence of S. aureus in mice. Deletion of the hlb gene had no effect on the intracellular survival of S. aureus in J774A.1 cells but did cause a slight increase in survival in MAC-T cells. Furthermore, like catalase, beta-toxin was not required for complete virulence of S. aureus in murine models. Unexpectedly, the DeltakatA Deltahlb mutant showed a notably increased persistence in both cell lines, and was significantly less virulent for mice than were the wild-type strain and single mutants. Most interestingly, it was also markedly attenuated in intramammary and subcutaneous infections in ewes and lambs.


Effects of work in cold stores on man.
A work physiological study was conducted during three different types of work in cold stores, i.e., at dairies and in the cutting and packaging of meat. Peripheral cooling, especially of the fingers and hands, was noted in all the subjects. The work load was relatively light for the packers but varied considerably in the dairy work. In both dairy and packaging work, clothing was unsatisfactory in terms of thermal insulation and flexibility. The butchers were subjected to less cooling, as a result of a relatively intense but even work rate and clothing adapted to the work. The subjects were very capable of perceiving temperature changes in the peripheral parts of their bodies. Individual variations were considerable, especially with respect to cooling and the perception of temperature and discomfort.


Clinician perceptions of pediatric growth chart use and electronic health records in Kentucky.
Growth chart recording is a key component of pediatric care. EHR systems could provide several growth charting functionalities compared to paper methods. To our knowledge, there has been no U.S. study exploring clinicians' perceptions and practices related to recording of growth parameters as they adapt to electronic methods. To explore clinician practices regarding recording growth parameters as they adapt to electronic health records (EHR) and to investigate clinician perceptions of electronic growth charting using EHR. An online survey of pediatricians and family practitioners in Kentucky inquiring about EHR usage, specifically use of growth charting with EHR, was conducted. Forty-six percent of respondents utilized EHRs, with pediatricians lagging family practitioners, and academic pediatricians lagging non-academicians. There was no consensus on EHR platforms being used. Almost a third of those who used EHR did not utilize electronic growth charting. Clinicians using EHR reported that electronic growth charts would improve clinician satisfaction and clinical efficiency as well as parent satisfaction and parent education. Only 12% of respondents provided copies of growth charts to parents at the end of their visit and discussed growth parameters with parents, with clinicians using EHR more likely to engage in these activities than non-EHR users. Although Kentucky clinicians continue to slowly adopt EHRs, clinician perceptions and practices reflect enduring barriers to widespread use of electronic growth charting in pediatric and family practice. However, our results suggest that electronic growth charting has important benefits for both clinicians and patients, and greater adoption is expected as EHRs become standard across health care systems.


Attitudes Toward Patient Safety among Medical Students in Malaysia.
The biggest challenge in moving toward a safer healthcare system is patient safety culture-that is, the prevention of harm to patients. Safe medical practices can prevent doing harm to the patients. For this, healthcare professionals must have good attitudes toward patient safety. Medical education plays an important role in promoting patient safety and patient safety attitudes. A study was conducted among medical students in Malaysia to assess their perceptions toward patient safety, using the 26-items Attitudes Toward Patient Safety Questionnaire (APSQ-III). In the analysis, the average percentage of positive responses (APPR) were computed for each domain, and APPR values of ≥75 were used as an indicator of positive perception. Out of the nine domains of APSQ, the students' attitude was positive in six-Safety Training (85.2%), Error Reporting (76.3%), Working Hours (89.5%), Error Inevitability (86.1%), Team Functioning (94.6%), and Patient Involvement (80.1%). The desired level of positive attitude was not met in Disclosure Responsibility (68.5%), Professional Incompetence (70.0%), and Safety Curriculum (71.1%). APRR for disclosure responsibility was high among the first-year students, but, generally, the effect wore off over the years of study. The results support the need to enhance perception on Disclosure Responsibility, Professional Incompetence, and Safety Curriculum among the medical students in Malaysia.


Predicting risk factors for radiation pneumonitis after stereotactic body radiation therapy for primary or metastatic lung tumours.
To investigate risk factors for radiation-induced pneumonitis (RP) after hypofractionated stereotactic body radiotherapy (SBRT) in patients with lung tumours. From May 2004 to January 2016, 66 patients with 71 primary or metastatic lung tumours were treated with SBRT; these 71 cases were retrospectively analyzed for RP. To explore the risk factors for RP, the following factors were investigated: age, sex, performance status, operability, number of treatments, respiratory gating, pulmonary emphysema, tumour location and subclinical interstitial lung disease (ILD). Irradiated underlying lung volumes of more than 5 Gy, 10 Gy, 20 Gy and 30 Gy (Lung V5, V10, V20 and V30), mean lung dose and volumes of gross tumour volume (in cubic centimetre) and planning target volume were calculated for possible risk factors of RP. The median follow-up period was 32 months. RP of Grade 2 or more, according to the Common Terminology Criteria for Adverse Events v. 4.0, was detected in 6 (8.4%) of the 71 cases. Grade 5 RP was identified in two cases. Of the risk factors of RP, subclinical ILD was the only factor significantly associated with the occurrence of RP of Grade 2 or more (p < 0.001). Both cases with Grade 5 RP had ILD with a honeycombing image. Subclinical ILD was the only significant factor for Grade 2-5 RP. In addition, the cases with honeycombing had a high potential for fatality related to severe RP. Patients with subclinical ILD should be carefully monitored for the occurrence of severe RP after SBRT. Advances in knowledge: Hypofractionated SBRT for primary or metastatic lung tumours provides a high local control rate and safe treatment.


p58IPK is an inhibitor of the eIF2α kinase GCN2 and its localization and expression underpin protein synthesis and ER processing capacity.
One of the key cellular responses to stress is the attenuation of mRNA translation and protein synthesis via the phosphorylation of eIF2α (eukaryotic translation initiation factor 2α). This is mediated by four eIF2α kinases and it has been suggested that each kinase is specific to the cellular stress imposed. In the present study, we show that both PERK (PKR-like endoplasmic reticulum kinase/eIF2α kinase 3) and GCN2 (general control non-derepressible 2/eIF2α kinase 4) are required for the stress responses associated with conditions encountered by cells overexpressing secreted recombinant protein. Importantly, whereas GCN2 is the kinase that is activated following cold-shock/hypothermic culturing of mammalian cells, PERK and GCN2 have overlapping functions since knockdown of one of these at the mRNA level is compensated for by the cell by up-regulating levels of the other. The protein p58IPK {also known as DnaJ3C [DnaJ heat-shock protein (hsp) 40 homologue, subfamily C, member 3]} is known to inhibit the eIF2α kinases PKR (dsRNA-dependent protein kinase/eIF2α kinase 2) and PERK and hence prevent or delay eIF2α phosphorylation and consequent inhibition of translation. However, we show that p58IPK is a general inhibitor of the eIF2α kinases in that it also interacts with GCN2. Thus forced overexpression of cytoplasmic p58 delays eIF2α phosphorylation, suppresses GCN2 phosphorylation and prolongs protein synthesis under endoplasmic reticulum (ER), hypothermic and prolonged culture stress conditions. Taken together, our data suggest that there is considerable cross talk between the eIF2α kinases to ensure that protein synthesis is tightly regulated. Their activation is controlled by p58 and the expression levels and localization of this protein are crucial in the capacity the cells to respond to cellular stress via control of protein synthesis rates and subsequent folding in the ER.


Sensory peptides as neuromodulators of wound healing in aged rats.
An intact nociceptor system of primary afferent sensory nerves is important for the initiation of the inflammatory process and successful tissue repair. Dysfunction of this system with age could be a contributing factor for delayed wound healing in the elderly. This study was designed to examine the role of sensory nerves (using capsaicin-pretreated rats) and the effect of aging on the healing of a thermal wound induced on the interscapular region (using a CO2 laser). The healing endpoint was the time when full wound contraction had occurred. The ability of the sensory peptides, substance P (SP) and calcitonin gene-related peptide (CGRP), in modulating wound healing in aged rats was examined by taking into account the modulatory interaction effects between these peptides. A blister model in the rat hind footpad combined with a laser Doppler flowmeter was used initially to establish these vascular interaction effects. The results showed a significant delay in thermal wound healing in both the capsaicin-treated and aged rats. Using the blister model, we demonstrated the ability of SP and CGRP to induce a limited and a prolonged vasodilator response, respectively. When tested together, SP attenuated the vascular effect of CGRP, an effect that was reversed using the tachykinin antagonist spantide II. When injected at the wound site, SP and CGRP were equally potent in promoting wound healing in aged rats. The beneficial effect of taking the interaction effects into account was most evident in a group of rats that received the tachykinin antagonist as part of their initial treatment with CGRP and 6 h later, an injection of SP; the time to complete wound closure was 11.3 +/- 0.6 days compared to 21.0 +/- 0.9 days in aged controls. We contend that would healing in aged rats can be accelerated by exogenous administration of sensory peptides. Furthermore, modulatory interaction effects between sensory peptides should be taken into consideration when designing any treatment protocol that purports to accelerate wound healing.


Diagnosis and management of trauma to the kidney.
Because trauma is an unplanned event, guidelines for managing patients sustaining upper urinary tract injury must already exist in the surgeon's mind before laying eyes on the injured patient. In order to simplify and standardize the evaluation and treatment of such patients, urologists have applied a system of staging paradigms and treatment algorithms, analogous to systems successfully used in the less chaotic arena of, for example, urologic oncology. Advancements in imaging modalities, improvements and renovations of surgical technique, and critical review of outcomes data have impacted on the management of genitourinary trauma and will be likely to influence the way we treat trauma patients in the future.


Digital social support for undergraduate students during COVID-19: Pivotal support for the digital transformation.
In a time of crisis, such as the COVID-19 pandemic, digital social support can prove helpful in buffering the stress and anxiety especially for students whose lives were being affected both socially and academically. The present study aims to explore the emotional and informational social support available to students during the online transformation of their education. It further investigates factors affecting anxiety and stress among students. An online survey was administered among 479 undergraduate students at a private university in Saudi Arabia from 2nd March till 17th June 2020. Descriptive analysis was used to report the findings of students' self-reported data. The results revealed that more than 70% of the students reported that they were satisfied with the informational and emotional support available to them via digital platforms. However, they felt less connected with the teachers and academic administration. Feeling sick (46%), being locked down at homes and isolation (43%) were major causes of stress and anxiety. 71% of the students reported their ability to keep themselves busy, however, only 55% of students mentioned that they did not feel lonely while confined to their homes. In these circumstances, the continuation of studies via online platforms helped students reduce stress and anxiety (52%). Although this online transformation was not planned, the university was able to provide adequate informational support to the students. However, students were dependent upon friends, family and relatives for emotional support. The results of the study point towards the need of training for teachers and academic administrative staff in order to better address students' needs for social support during the crisis. The results also contribute to the body of data reporting the various effects of the COVID-19 pandemic on students in a way to reach a sustainable framework for supporting students.


Static and Dynamical Properties of Antiferromagnetic Skyrmions in the Presence of Applied Current and Temperature.
Skyrmions are topologically protected entities in magnetic materials which have the potential to be used in spintronics for information storage and processing. However, Skyrmions in ferromagnets have some intrinsic difficulties which must be overcome to use them for spintronic applications, such as the inability to move straight along current. We show that Skyrmions can also be stabilized and manipulated in antiferromagnetic materials. An antiferromagnetic Skyrmion is a compound topological object with a similar but of opposite sign spin texture on each sublattice, which, e.g., results in a complete cancellation of the Magnus force. We find that the composite nature of antiferromagnetic Skyrmions gives rise to different dynamical behavior due to both an applied current and temperature effects.


Fluorescence Properties of Guard Cell Chloroplasts: EVIDENCE FOR LINEAR ELECTRON TRANSPORT AND LIGHT-HARVESTING PIGMENTS OF PHOTOSYSTEMS I AND II.
The presence of chloroplasts in guard cells from leaf epidermis, coleoptile, flowers, and albino portions of variegated leaves was established by incident fluorescence microscopy, thus confirming the notion that guard cell chloroplasts are remarkably conserved. Room temperature emission spectra from a few chloroplasts in a single guard cell of Vicia faba showed one major peak at around 683 nanometers. Low-temperature (77 K) emission spectra from peels of albino portions of Chlorophytum comosum leaves and from mesophyll chloroplasts of green parts of the same leaves showed major peaks at around 687 and 733 nanometers, peaks usually attributed to photosystem II and photosystem I pigment systems, respectively. Spectra of peels of V. faba leaves showed similar peaks. However, fluorescence microscopy revealed that the Vicia peels, as well as those from Allium cepa and Tulipa sp., were contaminated with non-guard cell chloroplasts which were practically undetectable under bright field illumination. These observations pose restrictions on the use of epidermal peels as a source of isolated guard cell chloroplasts. Studies on the 3-(3,4-dichlorophenyl)-1,1-dimethylurea-sensitive variable fluorescence kinetics of uncontaminated epidermal peels of C. comosum indicated that guard cell chloroplasts operate a normal, photosystem II-dependent, linear electron transport. The above properties in combination with their reported inability to fix CO(2) photosynthetically may render the guard cell chloroplasts optimally suited to supply the reducing and high-energy phosphate equivalents needed to sustain active ion transport during stomatal opening in daylight.


Bilingual effects of unilingual neuropsychological treatment of dyslexic adolescents: a pilot study.
Fourteen young adolescents with specific reading disabilities received short term neuropsychological treatment--specifically left hemisphere (LH) or right hemisphere (RH) stimulation--in a clinical pilot project. The effects on single-word and passage reading were evaluated when the language of treatment was either Dutch (mother tongue) or English (foreign language). Transfer effects across the two languages were also studied. In general, the results indicate that these short term interventions may prove successful for the treated language and with the additional transfer to a second (not treated) language.


COPD exacerbations by disease severity in England.
Exacerbations of chronic obstructive pulmonary disease (COPD) are associated with accelerated disease progression and are important drivers of health care resource utilization. The study aimed to quantify the rates of COPD exacerbations in England and assess health care resource utilization by severity categories according to the Global Initiative for Chronic Obstructive Lung Disease (GOLD) 2013. Data from the Clinical Practice Research Datalink linked to Hospital Episode Statistics were used to identify patients with a COPD diagnosis aged ≥40 years. Those with complete spirometric, modified Medical Research Council Dyspnea Scale information, and exacerbation history 12 months prior to January 1, 2011 (index date) were classified into GOLD severity groups. Study outcomes over follow-up (up to December 31, 2013) were exacerbation rates and resource utilization (general practitioner visits, hospital admissions). From the 44,201 patients in the study cohort, 83.5% were classified into severity levels GOLD A: 33.8%, GOLD B: 21.0%, GOLD C: 18.1%, and GOLD D: 27.0%. Mean age at diagnosis was 66 years and 52.0% were male. Annual exacerbation rates per person-year increased with severity, from 0.83 (95% confidence interval [CI]: 0.81-0.85) for GOLD A to 2.51 (95% CI: 2.47-2.55) for GOLD D. General practitioner visit rates per person-year also increased with severity, from 4.82 (95% CI: 4.74-4.93) for GOLD A to 7.44 (95% CI: 7.31-7.61) for GOLD D. COPD-related hospitalization rates per person-year increased from less symptoms (GOLD A: 0.28, GOLD C: 0.39) to more symptoms (GOLD B: 0.52, GOLD D: 0.84). Patients in the most severe category (GOLD D) experienced nearly three times the number of exacerbations and COPD-related hospitalizations as those in the least severe category (GOLD A), in addition to increased general practitioner visits. Better patient management to stabilize the disease progression could allow for an improvement in exacerbation frequency and a reduction in health care resource utilization.


Energetic determinants of tyrosine phosphorylation of focal adhesion proteins during hypoxia/reoxygenation of kidney proximal tubules.
Anaerobic mitochondrial metabolism of alpha-ketoglutarate and aspartate or alpha-ketoglutarate and malate can prevent and reverse severe mitochondrial dysfunction during reoxygenation after 60 minutes of hypoxia in kidney proximal tubules.(34) The present studies demonstrate that, during hypoxia, paxillin, focal adhesion kinase, and p130(cas) migrated faster by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, their phosphotyrosine (pY) content decreased to approximately 5% of that in oxygenated tubules without changes in total protein, and the normally basal immunostaining of beta1 and alpha6 integrin subunits, pY, and paxillin was lost or markedly decreased. During reoxygenation without supplemental substrates, recovery of pY and basal localization of the focal adhesion proteins was poor. alpha-Ketoglutarate and aspartate, which maintained slightly higher levels of ATP during hypoxia, also maintained 2.5-fold higher levels of pY during this period, and promoted full recovery of pY content and basal localization of focal adhesion proteins during subsequent reoxygenation. Similarly complete recovery was made possible by provision of alpha-ketoglutarate and aspartate or alpha-ketoglutarate and malate only during reoxygenation. These data emphasize the importance of very low energy thresholds for maintaining the integrity of key structural and biochemical components required for cellular survival and reaffirm the value of approaches aimed at conserving or generating energy in cells injured by hypoxia or ischemia.


An APC for every occasion: induction and expansion of human Ag-specific CD4 and CD8 T cells using cellular and non-cellular APC.
APC are used extensively to induce and expand Ag-specific T cells as well as to test their specificity and function. In the treatment of malignant and infectious diseases, APC are used to stimulate and expand Ag-specific T cells for adoptive transfer, or used directly in vivo to present Ag. The choice of APC to use depends on the particular application and on practical considerations, which include ease of production, availability, reproducibility and (for clinical use) established safety. The diversity of APC in use partly reflects the fact that no single technique of Ag presentation is ideal. For the clinician and laboratory worker alike the field can seem illogical and confusing. In this review we outline the functional requirements of APC for the induction of T cells, classify the APC in common use and describe their laboratory and clinical applications.


Quantitative comparison of pretreatment regimens used to sensitize in situ hybridization using oligonucleotide probes on paraffin-embedded brain tissue.
Paraffin embedding of tissue is generally perceived to dramatically reduce RNA detectability. As a consequence, in situ hybridization on paraffin-embedded tissue is largely confined to detection of high-copy RNA species (e.g., viral RNA) and/or to detection using typically more sensitive cDNA probes or riboprobes. In this study, several procedures for in situ hybridization on paraffin-embedded rat tissue using oligonucleotide probes complementary to cellular transcripts were developed and quantitatively compared. Certain pretreatments showed marked increases in sensitivity compared to untreated sections. Furthermore, through quantitative assessment using image analysis, sensitivity of optimal pretreatments was equal to that of routinely used fresh-frozen, postfixed tissue sections. The development of such techniques permitting in situ hybridization to be carried out on paraffin-embedded tissue allows a comparison of protein and mRNA distribution to be made in adjacent sections and provides the potential for double labeling by in situ hybridization and immunohistochemistry which may not be possible on post-fixed frozen sections.


Dickkopf-1 Promotes Angiogenesis and is a Biomarker for Hepatic Stem Cell-like Hepatocellular Carcinoma.
Cancer stemness evinces interest owing to the resulting malignancy and poor prognosis. We previously demonstrated that hepatic stem cell-like hepatocellular carcinoma (HpSC-HCC) is associated with high vascular invasion and poor prognosis. Dickkopf-1 (DKK-1), a Wnt signaling regulator, is highly expressed in HpSC-HCC. Here, we assessed the diagnostic and prognostic potential of serum DKK-1. Its levels were significantly higher in 391 patients with HCC compared with 205 patients with chronic liver disease. Receiver operating characteristic curve analysis revealed the optimal cutoff value of DKK-1 to diagnose HCC and predict the 3-year survival as 262.2 and 365.9 pg/mL, respectively. HCC patients with high-serum DKK-1 levels showed poor prognosis. We evaluated the effects of anti-DKK-1 antibody treatment on tumor growth in vivo and of recombinant DKK-1 on cell proliferation, invasion, and angiogenesis in vitro. DKK-1 knockdown decreased cancer cell proliferation, migration, and invasion. DKK-1 supplementation promoted angiogenesis in vitro; this effect was abolished by an anti-DKK-1 antibody. Co-injection of the anti-DKK-1 antibody with Huh7 cells inhibited their growth in NOD/SCID mice. Thus, DKK-1 promotes proliferation, migration, and invasion of HCC cells and activates angiogenesis in vascular endothelial cells. DKK-1 is a prognostic biomarker for HCC and a functional molecule for targeted therapy.


Magnetization transfer in human Achilles tendon assessed by a 3D ultrashort echo time sequence: quantitative examinations in healthy volunteers at 3T.
Magnetization transfer contrast (MTC) imaging provides insight into interactions between free and bounded water. Newly developed ultrashort echo time (UTE) sequences implemented on whole-body magnetic resonance (MR) scanners allow MTC imaging in tissues with extremely fast signal decay such as tendons. The aim of this study was to develop a technique for the quantification of the MT effect in healthy Achilles tendons in-vivo at 3 Tesla. 16 normal tendons of volunteers with no history of tendinopathy were examined using a 3D-UTE sequence with a rectangular on-resonant excitation pulse and a Fermi-shaped off-resonant MT preparation pulse. The frequency of the MT pulse was varied from 1 to 5 kHz. MT effects were calculated in terms of the MT ratio (MTR) between measurements without and with MT preparation. Direct saturation effects of MT preparation on the signal intensity were evaluated using numerical simulation of Bloch equations. One patient with tendinopathy was examined to exemplarily show changes of MTR under pathologic conditions. Calculation of MTR data was feasible in all examined tendons and showed a decrease from 0.53 ± 0.05 to 0.25 ± 0.03 (1 kHz to 5 kHz) for healthy volunteers. Evaluation of variation with gender and dominance of ankle revealed no significant differences (p > 0.05). In contrast, the patient with confirmed tendinopathy showed MTR values between 0.36 (1 kHz) and 0.19 (5 kHz). MT effects in human Achilles tendons can be reliably assessed in-vivo using a 3D UTE sequence at 3 T. All healthy tendons showed similar MTR values (coefficient of variation 10.0 ± 1.2 %). The examined patient showed a clearly different MT effect revealing a changed microstructure in the case of tendinopathy.