Soluble intercellular adhesion molecule, vascular cell adhesion molecule, and impaired microvascular reactivity are early markers of vasculopathy in type 2 diabetic individuals without microalbuminuria.
Using von Willebrand Factor (vWF) as a marker of endothelial function, previous studies have shown that the development of microalbuminuria is associated with the onset of endothelial dysfunction in individuals with type 2 diabetes. We tested the hypothesis that endothelial dysfunction is already evident in normoalbuminuric individuals with type 2 diabetes. We used laser Doppler imaging scanning to measure vasodilation in the forearm skin in response to iontophoresis of 1% acetylcholine (endothelium-dependent) and 1% sodium nitroprusside (endothelium-independent). Multiple indicators of endothelial function--soluble intercellular adhesion molecule (sICAM), soluble vascular cell adhesion molecule (sVCAM), vWF, and microvascular reactivity--were measured in 20 healthy control subjects, 45 normoalbuminuric (urinary albumin/creatinine ratio < 30 micrograms/mg) individuals with type 2 diabetes, and 14 microalbuminuric (urinary albumin/creatinine ratio between 30 and 300 micrograms/mg) individuals with type 2 diabetes. Serum sICAM and sVCAM levels were elevated in the normoalbuminuric (305 +/- 120, 851 +/- 284 ng/ml) and microalbuminuric (300 +/- 89, 845 +/- 252 ng/ml) individuals with diabetes when compared with the healthy control subjects (213 +/- 58, 661 +/- 176 ng/ml) (P < 0.01). Furthermore, the microvascular endothelium-dependent and -independent vasodilation was reduced in the normoalbuminuric (76 +/- 44, 70 +/- 33) (percent increase in perfusion over baseline) and microalbuminuric (74 +/- 41, 73 +/- 28) individuals with diabetes compared with healthy control subjects (126 +/- 67, 120 +/- 47) (P < 0.05). In contrast, plasma vWF was elevated only in the microalbuminuric individuals with diabetes (129 +/- 35%) compared with the normoalbuminuric individuals with diabetes (110 +/- 34) and healthy control subjects (111.3 +/- 39) (P < 0.05). On stepwise multivariate analysis, fasting blood glucose was the most important contributing factor to the variation in microvascular reactivity and sVCAM, whereas insulin resistance (by homeostasis model assessment) was the most important contributing factor to the variation in sICAM. Addition of all clinical and biochemical measures explained only 15-22% of the variation in sICAM, sVCAM, and microvascular reactivity. Multiple markers of endothelial dysfunction were evident in normoalbuminuric individuals with type 2 diabetes. The pathogenic process of vasculopathy in type 2 diabetes occurs early and may be operative before the development of microalbuminuria.


Interleukin-1B (IL-1B) and interleukin-1 receptor antagonist (IL-1RN) gene polymorphisms are not associated with tubal pathology and Chlamydia trachomatis-related tubal factor subfertility.
Chlamydia trachomatis infections have been associated with tubal pathology. However, not all C.trachomatis-infected women actually develop tubal pathology. Recently, host genetic factors such as the interleukin-1 gene cluster have been linked to inflammatory and infectious diseases. Dutch Caucasian women were investigated for (i) the role of interleukin-1B (IL-1B) and interleukin-1 receptor antagonist (IL-1RN) gene polymorphisms in tubal pathology (group 1); and (ii) the presence of these gene polymorphisms in C.trachomatis IgG-positive women with and without tubal pathology (group 2). Group 1 consisted of women with (n = 40) or without (n = 95) tubal pathology, respectively, and group 2 of C.trachomatis IgG-positive women of whom 28 had tubal pathology at laparoscopy and 47 did not. IL-1B-511 and IL-1B+3954 gene polymorphisms were assessed by PCR-restriction fragment length polymorphism (RFLP), and the variable number of tandem repeats (VNTR) of the IL-1RN gene were assessed by a PCR-based assay. Neither IL-1B-511, IL-1B+3954 nor IL-1RN genotypes, allele or carrier frequencies showed significant association with tubal pathology or C.trachomatis post-infection-based tubal pathology. The data obtained suggest that specific IL-1 gene polymorphisms are not associated with the tubal pathology risk or to the development of C.trachomatis-based post-infectious severe sequelae.


Efficient generation of mutations mediated by CRISPR/Cas9 in the hairy root transformation system of Brassica carinata.
A protocol for the induction of site-directed deletions and insertions in the genome of Brassica carinata with CRISPR is described. The construct containing the Cas9 nuclease and the guide RNA (gRNA) was delivered by the hairy root transformation technique, and a successful transformation was monitored by GFP fluorescence. PAGE analysis of an amplified region, presumably containing the deletions and insertions, demonstrated up to seven different indels in one transgenic root and in all analyzed roots a wildtype allele of the modified gene was not detectable. Interestingly, many of these mutations consisted of relatively large indels with up to 112 bp. The exact size of the deletions was determined to allow an estimation whether the targeted gene was not functional due to a considerable deletion or a frame shift within the open reading frame. This allowed a direct phenotypic assessment of the previously characterized roots and, in fact, deletions in FASCICLIN-LIKE ARABINOGALACTAN PROTEIN 1 (BcFLA1)-a gene with an expression pattern consistent with a role in root hair architecture-resulted in shorter root hairs compared to control roots ectopically expressing an allele of the gene that cannot be targeted by the gRNA in parallel to the CRISPR construct. As an additional line of evidence, we monitored BcFLA1 expression with qPCR and detected a significant reduction of the transcript in roots with an active CRISPR construct compared to the control, although residual amounts of the transcript were detected, possibly due to inefficient nonsense-mediated mRNA decay. Additionally, the presence of deletions and insertions were verified by Sanger sequencing of the respective amplicons. In summary we demonstrate the successful application of CRISPR/Cas9 in hairy roots of B. carinata, the proof of its effectiveness and its effect on the root hair phenotype. This study paves the way for experimental strategies involving the phenotypic assessment of gene lesions by CRISPR which do not require germline transmission.


Transmission distortion and genetic incompatibilities between alleles in a multigenerational mouse advanced intercross line.
While direct additive and dominance effects on complex traits have been mapped repeatedly, additional genetic factors contributing to the heterogeneity of complex traits have been scarcely investigated. To assess genetic background effects, we investigated transmission ratio distortions (TRDs) of alleles from parent to offspring using an advanced intercross line (AIL) of an initial cross between the mouse inbred strains C57BL/6NCrl (B6N) and BFMI860-12 [Berlin Fat Mouse Inbred (BFMI)]. A total of 341 males of generation 28 and their respective 61 parents and 66 grandparents were genotyped using Mega Mouse Universal Genotyping Arrays. TRDs were investigated using allele transmission asymmetry tests, and pathway overrepresentation analysis was performed. Sequencing data were used to test for overrepresentation of nonsynonymous SNPs (nsSNPs) in TRD regions. Genetic incompatibilities were tested using the Bateson-Dobzhansky-Muller two-locus model. A total of 62 TRD regions were detected, many in close proximity to the telocentric centromere. TRD regions contained 44.5% more nsSNPs than randomly selected regions (182 vs 125.9 ± 17.0, P < 1 × 10-4). Testing for genetic incompatibilities between TRD regions identified 29 genome-wide significant incompatibilities between TRD regions [P(BF) < 0.05]. Pathway overrepresentation analysis of genes in TRD regions showed that DNA methylation, epigenetic regulation of RNA, and meiotic/meiosis regulation pathways were affected independent of the parental origin of the TRD. Paternal BFMI TRD regions showed overrepresentation in the small interfering RNA biogenesis and in the metabolism of lipids and lipoproteins. Maternal B6N TRD regions harbored genes involved in meiotic recombination, cell death, and apoptosis pathways. The analysis of genes in TRD regions suggests the potential distortion of protein-protein interactions influencing obesity and diabetic retinopathy as a result of disadvantageous combinations of allelic variants in Aass, Pgx6, and Nme8. Using an AIL significantly improves the resolution at which we can investigate TRD. Our analysis implicates distortion of protein-protein interactions as well as meiotic drive as the underlying mechanisms leading to the observed TRD in our AIL. Furthermore, genes with large amounts of nsSNPs located in TRD regions are more likely to be involved in pathways that are related to the phenotypic differences between the parental strains. Genes in these TRD regions provide new targets for investigating genetic adaptation, protein-protein interactions, and determinants of complex traits such as obesity.


[Ultrasonographic findings in a cow with extraskeletal chondroblastic osteosarcoma of the neck region].
This case report describes the clinical, ultrasonographic and pathological findings in a five-year-old Swiss Braunvieh cow with extraskeletal chondroblastic osteosarcoma of the neck region. The cow was referred because of a firm, non-painful swelling, approximately 25 cm in diameter, which was situated mainly on the lower left side of the neck but extended to the right. Ultrasonographic examination of the mass revealed a chambered structure containing echoic material that was separated by hyperechoic septa. Chondroblastic osteosarcoma was diagnosed based on histological evaluation of a biopsy sample, and the diagnosis was confirmed by postmortem examination.


Mediastinal lymphadenopathy detected with an integrated positron emission tomographic/computed tomographic scan: a differential diagnosis to consider before minimally invasive staging.
Integrated positron emission tomography combined with plain computed tomography is commonly employed as the diagnostic tool for patients presenting with lung opacities. This technology is non-invasive and does not put the patient at risk of contrast reaction. We report a case of a man who presented with fever, septicaemia, and a left apical lung opacity on chest radiography. His positron emission tomography and plain computed tomography scans showed increased uptake by the left apical lung opacity together with a huge anterior mediastinal mass, suggestive of lung cancer with mediastinal lymph node involvement, and a right upper lobe shadow. After an initial futile bronchoscopy, an endobronchial ultrasound-guided transbronchial needle aspiration of the mediastinal node was planned but a contrast computed tomographic scan of the thorax revealed no significantly enlarged mediastinal lymph nodes. The differential diagnoses of these findings, together with the limitations of positron emission tomography and plain computed tomography, are discussed.


Microbial production of a novel trihydroxy unsaturated fatty acid from linoleic acid.
A bacterium isolated from a dry soil sample collected from McCalla, AL, USA, converted linoleic acid to a novel compound, 12,13,17-trihydroxy-9 (Z)-octadecenoic acid (THOA). The organism is a Gram-positive, non-motile rod (0.5 microns x 2 microns). It was identified as a species of Clavibacter ALA2. The product was purified by high pressure liquid chromatography, and its structure was determined by 1H and 13C nuclear magnetic resonance and Fourier transform infrared spectroscopies, and by mass spectrometer. Maximum production of THOA with 25% conversion of the substrate was reached after 5-6 days of reaction. THOA was not further metabolized by strain ALA2. This is the first report of a 12,13,17-trihydroxy unsaturated fatty acid and its production by microbial transformation. Some dihydroxy intermediates were also detected. THOA has a structure similar to those of known plant self-defense substances.


Effect of ultrasonic treatment on enzymes: Decoupling the relation between the ultrasonic driven conformational change and enzyme activity.
Considering the growing interest in enzyme-based extraction technology as a safe and eco-friendly extraction technique, along with the relatively high cost associated with enzymatic applications, it became necessary to explore novel strategies aimed to improve enzyme activity. In this study, the impact of ultrasonic treatment on commercial cellulase and pectinase was investigated. As this effect may be influenced by various ultrasonic and enzyme-related parameters, changes in enzyme conformation were explored under optimal and non-optimal enzyme conditions. The intrinsic fluorescence spectrum was utilized as a tool for monitoring these changes. Additionally, the enzyme's catalytic potential was also assessed under the same conditions. Results indicated that the impact of ultrasonic treatment on enzyme conformation primarily depends on the total ultrasonic energy delivered to the system, rather than other ultrasonic parameters such as power, sample volume, treatment time, or duty cycle. The maximum relative decrease in intrinsic fluorescence intensity of Pectinex® Ultra Clear (PUC) and Pectinex® Ultra SPL (PUS) after ultrasonic treatment was approximately 51% and 55%, respectively, while the decrease induced by thermal denaturation was 25% and 30% respectively. Furthermore, a blue shift in the fluorescence spectrum of both pectinases was observed upon sonication for all process conditions indicating a change in enzyme conformation. However, ultrasonic treatment did not result in a significant change in enzyme activity, suggesting that these conformational adjustments may occur in regions other than the active sites. Moreover, ultrasonicated pectinases and cellulases did not exhibit any improvement in their catalytic potential under either optimal or non-optimal conditions.


ActivePapers: a platform for publishing and archiving computer-aided research.
The lack of replicability and reproducibility of scientific studies based on computational methods has lead to serious mistakes in published scientific findings, some of which have been discovered and publicized recently. Many strategies are currently pursued to improve the situation. This article reports the first conclusions from the ActivePapers project, whose goal is the development and application of a computational platform that allows the publication of computational research in a form that enables installation-free deployment, encourages reuse, and permits the full integration of datasets and software into the scientific record. The main finding is that these goals can be achieved with existing technology, but that there is no straightforward way to adapt legacy software to such a framework.


The crystal structure of human glycosylation-inhibiting factor is a trimeric barrel with three 6-stranded beta-sheets.
Glycosylation-inhibiting factor (GIF) is a cytokine that is involved in the regulation of IgE synthesis. The crystal structure of recombinant human GIF was determined by the multiple isomorphous replacement method. The structure was refined to an R factor of 0.168 at 1.9 angstrom resolution. The overall structure is seen to consist of three interconnected subunits forming a barrel with three 6-stranded beta-sheets on the inside and six alpha-helices on the outside. There is a 5-angstrom-diameter "hole" through the middle of the barrel. The barrel structure of GIF in part resembles other "trefoil" cytokines such as interleukin 1 and fibroblast growth factor. Each subunit has a new class of alpha + beta sandwich structure consisting of two beta-alpha-beta motifs. These beta-alpha-beta motifs are related by a pseudo-twofold axis and resemble both interleukin 8 and the peptide binding domain of major histocompatibility complex protein, although the topology of the polypeptide chain is quite different.


Systemic FasL neutralization increases eosinophilic inflammation in a mouse model of asthma.
Eosinophils and lymphocytes are pathogenically important in allergic inflammation and sensitive to Fas-mediated apoptosis. Fas ligand (FasL) activity therefore should play a role in regulating the allergic immune response. We aimed to characterize the role of FasL expression in airway eosinophilia in Aspergillus fumigatus (Af)-induced sensitization and to determine whether FasL neutralization alters the inflammatory response. Sensitized Balb/c mice were killed before (day 0) and 1, 7 and 10 days after a single intranasal challenge with Af. Animals received either neutralizing antibody to FasL (clone MFL4) or irrelevant hamster IgG via intraperitoneal injection on days -1 and 5. FasL expression, BAL and tissue inflammatory cell and cytokine profile, and apoptosis were assessed. Postchallenge FasL gene expression in BAL cells and TUNEL positivity in the airways coincided with the height of inflammatory cell influx on day 1, while soluble FasL protein was released on day 7, preceding resolution of the inflammatory changes. Although eosinophil numbers showed a negative correlation with soluble FasL levels in the airways, MBP(+) eosinophils remained TUNEL negative in the submucosal tissue, throughout the 10-day period after Af challenge. Systemic FasL neutralization significantly enhanced BAL and tissue eosinophil counts. This effect was associated with increased activation of T cells and release of IL-5, IL-9, and GM-CSF in the BAL fluid of mice, indicating an involvement of pro-eosinophilic survival pathways. FasL activity may play an active role in resolving eosinophilic inflammation through regulating T cells and pro-eosinophilic cytokine release during the allergic airway response.


Mathematical and Ex Vivo Thermal Modeling for Renal Tumor Radiofrequency Ablation with Pyeloperfusion.
Radiofrequency ablation (RFA) is an effective technique for the treatment of patients with small renal tumors, although it is often limited to tumors at least 2 cm from the renal pelvis or ureter. Retrograde pyeloperfusion (PPF) of the pelvis with cold saline during RFA may protect the pelvis and ureter. We designed a mathematical and ex vivo model of RFA to investigate the effects of PPF. Our theoretical model uses heat transfer principles simplifying the RFA probe to a heat-emitting cylinder within a material. In the ex vivo model, an RFA probe was placed 18 mm from the pelvis in porcine kidneys and with temperature probes on either side of the RFA probe. Control trials with no PPF were compared with either cold saline (2°C), warm saline (38°C), or antifreeze (-20°C) pumped into the renal calix at a rate of 60 mL/min. Ablated volumes were measured and confirmed histologically. The average steady state temperatures at each probe were highest with no PPF, followed by warm saline, cold saline, then antifreeze. Compared with no PPF, temperatures were significantly (P<0.05) colder with warm saline (-8.4°C), cold saline (-18°C), and significantly colder at the calix (warm -14°C, cold -27°C). While RFA output a constant voltage, significantly lower resistances in warm (171Ω) and cold (124Ω) PPF vs no PPF (363Ω) translated to significantly greater power outputs in warm (40 W) and cold (42 W) vs no PPF (14 W). The ablated volumes were significantly higher in warm saline (2.3 cm(3)) vs cold saline (0.84 cm(3)) and no PPF (1.1 cm(3)). Mathematical modeling produced a predictive temperature curve with R2=0.44. PPF lowers temperatures throughout the entire kidney during RFA, most notably near the collecting system and is dependent on the temperature of the liquid used. In addition, PPF may cause less charring of the tissue around the probe resulting in lower resistance and higher power outputs.


The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium.
The psychrophilic cellulase, Cel5G, from the Antarctic bacterium Pseudoalteromonas haloplanktis is composed of a catalytic module (CM) joined to a carbohydrate-binding module (CBM) by an unusually long, extended and flexible linker region (LR) containing three loops closed by three disulfide bridges. To evaluate the possible role of this region in cold adaptation, the LR was sequentially shortened by protein engineering, successively deleting one and two loops of this module, whereas the last disulfide bridge was also suppressed by replacing the last two cysteine residue by two alanine residues. The kinetic and thermodynamic properties of the mutants were compared with those of the full-length enzyme, and also with those of the cold-adapted CM alone and with those of the homologous mesophilic enzyme, Cel5A, from Erwinia chrysanthemi. The thermostability of the mutated enzymes as well as their relative flexibility were evaluated by differential scanning calorimetry and fluorescence quenching respectively. The topology of the structure of the shortest mutant was determined by SAXS (small-angle X-ray scattering). The data indicate that the sequential shortening of the LR induces a regular decrease of the specific activity towards macromolecular substrates, reduces the relative flexibility and concomitantly increases the thermostability of the shortened enzymes. This demonstrates that the long LR of the full-length enzyme favours the catalytic efficiency at low and moderate temperatures by rendering the structure not only less compact, but also less stable, and plays a crucial role in the adaptation to cold of this cellulolytic enzyme.


Differential subcellular localisation of the tumour suppressor protein LIMD1 in breast cancer correlates with patient survival.
The tumour suppressor gene (TSG) LIM domain containing protein 1 (LIMD1) has been associated with transformation of epithelial cells of the lung and its expression is downregulated in all lung tumour samples tested compared to normal lung matched controls. In the first study of its kind we used an anti-LIMD1 specific monoclonal antibody to investigate expression/localisation of the LIMD1 protein in a well-characterised tissue microarray of breast cancers and normal adjacent epithelia. Comparison of tumour with adjacent normal and distant normal tissue demonstrated that LIMD1 expression is moderate to high compared to tumour. There was also a significant correlation with histological grade (p = 0.0001), tumour size (p = 0.013) and tumour type (p = 0.004) indicating an association with aggressive disease. Cytoplasmic LIMD1 expression was seen in 99.3% of cases, with 43.1% showing both nuclear and cytoplasmic localisation. Absence/loss of nuclear staining showed a strong correlation with patient survival and was indicative of poor prognosis (p = 0.033). There was no association with lymph node status and other clinicopathological parameters. Nuclear staining was more pronounced in better prognosis tumours and normal tissue. This study demonstrates that LIMD1 represents a novel prognostic marker for breast cancer. Combined with the fact that LIMD1 expression is downregulated in lung cancers this clearly indicates that LIMD1 may represent a critical TSG, the function of which is deregulated via overall loss of expression and/or relocalisation within the cell during tumour development. The possible functions of LIMD1 localisation within the nucleus and cytoplasm and its relationship to tumour prognosis are discussed.


NMDA receptors are essential for the acquisition, but not expression, of conditional fear and associative spike firing in the lateral amygdala.
We examined the contribution of N-methyl-D-aspartate (NMDA) receptors (NMDARs) to the acquisition and expression of amygdaloid plasticity and Pavlovian fear conditioning using single-unit recording techniques in behaving rats. We demonstrate that NMDARs are essential for the acquisition of both behavioral and neuronal correlates of conditional fear, but play a comparatively limited role in their expression. Administration of the competitive NMDAR antagonist +/--3-(2-carboxypiperazin-4-yl) propyl-1-phosphonic acid (CPP) prior to auditory fear conditioning completely abolished the acquisition of conditional freezing and conditional single-unit activity in the lateral amygdala (LA). In contrast, CPP given prior to extinction testing did not affect the expression of conditional single-unit activity in LA, despite producing deficits in conditional freezing. Administration of CPP also blocked the induction of long-term potentiation in the amygdala. Together, these data suggest that NMDARs are essential for the acquisition of conditioning-related plasticity in the amygdala, and that NMDARs are more critical for regulating synaptic plasticity and learning than routine synaptic transmission in the circuitry supporting fear conditioning.


Cardiac Dysfunction Rather Than Aortic Valve Stenosis Severity Drives Exercise Intolerance and Adverse Hemodynamics.
To study the impact of heart failure with preserved ejection fraction (HFpEF) versus aortic stenosis (AS) lesion severity on left ventricular (LV) hypertrophy, diastolic dysfunction, left atrial (LA) dysfunction, hemodynamics, and exercise capacity. Patients (n = 206) with at least moderate AS (aortic valve area ≤0.85 cm/m2) and discordant symptoms underwent cardiopulmonary exercise testing with simultaneous echocardiography. The population was stratified according to the probability of underlying HFpEF by the H2FPEF score [0-5 (AS/HFpEF-) vs. 6-9 points (AS/HFpEF+)] and AS severity (Moderate vs. Severe). Mean age was 73 ± 10 years with 40% women. Twenty-eight patients had Severe AS/HFpEF + (14%), 111 Severe AS/HFpEF- (54%), 13 Moderate AS/HFpEF + (6%), and 54 Moderate AS/HFpEF- (26%). AS/HFpEF + versus AS/HFpEF- patients, irrespective of AS severity, had a lower LV global longitudinal strain, impaired diastolic function, reduced LV compliance, and more pronounced LA dysfunction. The pulmonary arterial pressure-cardiac output slope was significantly higher in AS/HFpEF + versus AS/HFpEF- (5.4 ± 3.1 vs. 3.9 ± 2.2 mmHg/L/min, respectively; p = 0.003), mainly driven by impaired cardiac output and chronotropic reserve, with signs of right ventricular-pulmonary arterial uncoupling. AS/HFpEF + versus AS/HFpEF- was associated with a lower peak aerobic capacity (11.5 ± 3.7 vs. 15.9 ± 5.9 mL/min/kg, respectively; p < 0.0001), but did not differ between Moderate and Severe AS (14.7 ± 5.5 vs. 15.2 ± 5.9 mL/min/kg, respectively; p = 0.6). A high H2FPEF score is associated with a reduced exercise capacity and adverse hemodynamics in patients with moderate to severe AS. Both exercise performance and hemodynamics correspond better with intrinsic cardiac dysfunction than AS severity.


Delayed wound healing in the absence of intercellular adhesion molecule-1 or L-selectin expression.
Inflammatory cells play a crucial role in wound healing, but the role of adhesion molecules including L-selectin and intercellular adhesion molecule-1 (ICAM-1) is not known in this process. We examined skin wound repair of excisional wounds in mice lacking L-selectin, ICAM-1, or both. The loss of ICAM-1 inhibited wound healing, keratinocyte migration from the edges of the wound toward the center, and granulation tissue formation. By contrast, L-selectin deficiency alone did not affect any of these parameters. However, the loss of both L-selectin and ICAM-1 resulted in inhibition of keratinocyte migration and granulation tissue formation beyond those caused by loss of ICAM-1 alone. Treatment of platelet-derived growth factor to the wounds normalized delayed wound healing in ICAM-1(-/-) mice, but not in L-selectin/ICAM-1(-/-) mice. Therefore, although ICAM-1 contributes to wound repair to a greater extent than L-selectin, a role for L-selectin was revealed in the absence of ICAM-1. The impaired wound repair was associated with reduced infiltration of neutrophils and macrophages in ICAM-1(-/-) and L-selectin/ICAM-1(-/-) mice. These results demonstrate a distinct role of ICAM-1 and L-selectin in wound healing and that the delayed wound healing in the absence of these molecules is likely because of decreased leukocyte accumulation into the wound site.


Female gender hormones regulate mRNA levels and function of the rat lung epithelial Na channel.
The epithelial Na channel (ENaC) plays a critical role in the active reabsorption of alveolar fluid at the time of birth or during pulmonary edema. Although rat (r) ENaC is regulated by glucocorticoids during fetal development, there are no data regarding the influence of gender hormones on ENaC expression or function. We report higher levels of mRNAs encoding the alpha-rENaC subunit or the cystic fibrosis transmembrane conductance regulator (CFTR) in the lungs of nonpregnant adult female relative to adult male Wistar rats. Combined, but not separate, administration of progesterone and 17 beta-estradiol increased mRNA levels encoding alpha-rENaC, gamma-rENaC, and CFTR within 24 h. We also found a dose-dependent increase in rENaC functional activity (as assessed by the amiloride-sensitive short-circuit current across primary monolayer cultures of alveolar epithelial cells mounted in Ussing chambers) after a 5-day incubation of cells in medium containing progesterone and 17 beta-estradiol. These findings suggest a gender-dependent influence on the lung's ability to recover from pulmonary edema and on the degree of airway fluid hydration in cystic fibrosis.


Internal Carotid Artery Stenosis and Collateral Recruitment in Stroke Patients.
Leptomeningeal collaterals improve outcome in stroke patients. There is great individual variability in their extent. Internal carotid artery (ICA) stenosis may lead to more extensive recruitment of leptomeningeal collaterals. The purpose of this study was to evaluate the association of pre-existing ICA stenosis with leptomeningeal collateral filling visualized with computed tomography perfusion (CTP). From a prospective acute ischemic stroke cohort, patients were included with an M1 middle cerebral artery (MCA) occlusion and absent ipsilateral, extracranial ICA occlusion. ICA stenosis was determined on admission CT angiography (CTA). Leptomeningeal collaterals were graded as good (>50%) or poor (≤50%) collateral filling in the affected MCA territory on CTP-derived vessel images of the admission scan. The association between ipsilateral ICA stenosis ≥70% and extent of collateral filling was analyzed using logistic regression. In a multivariable analysis the odds ratio (OR) of ICA stenosis ≥70% was adjusted for complete circle of Willis, gender and age. We included 188 patients in our analyses, 50 (26.6%) patients were classified as having poor collateral filling and 138 (73.4%) as good. Of the patients 4 with poor collateral filling had an ICA stenosis ≥70% and 14 with good collateral filling. Unadjusted and adjusted ORs of ICA stenosis ≥70% for good collateral filling were 1.30 (0.41-4.15) and 2.67 (0.81-8.77), respectively. Patients with poor collateral filling had a significantly worse outcome (90-day modified Rankin scale 3-6; 80% versus 52%, p = 0.001). No association was found between pre-existing ICA stenosis and extent of CTP derived collateral filling in patients with an M1 occlusion.


Omalizumab Drug Survival in Chronic Urticaria: A Retrospective Multicentric French Study.
Omalizumab (OMA) dramatically improves disease control and quality of life in patients with chronic urticaria (CU). We aimed to evaluate the discontinuation patterns of OMA and their determinants in a cohort of French patients with CU. We conducted a retrospective multicenter study in 9 French tertiary referral hospitals. All patients diagnosed with either spontaneous (CSU) and/or inducible (CIndU) CU who received at least 1 injection of OMA between 2009 and 2021 were included. We analyzed OMA drug survival and investigated possible determinants using Kaplan-Meier curves and log-rank tests. A total of 878 patients were included in this study; 48.8% had CSU, 10.1% CIndU, and 41.1% a combination of both. OMA was discontinued in 408 patients, but the drug was later reintroduced in 50% of them. The main reason for discontinuing treatment was the achievement of a well-controlled disease in 50% of patients. Half of the patients were still being treated with OMA 2.4 years after the initiation of treatment. Drug survival was shorter in patients with CIndU and in those with an autoimmune background. In atopic patients, OMA was discontinued earlier in patients achieving a well-controlled disease. A longer OMA drug survival was observed in patients with a longer disease duration at initiation. In French patients with CU, the drug survival of OMA appears to be longer than that observed in previous studies conducted elsewhere, highlighting discrepancies in prescription and reimbursement possibilities. Further studies are warranted to develop customized OMA treatment schemes based on individual patterns.


Animal models for medications development targeting alcohol abuse using selectively bred rat lines: neurobiological and pharmacological validity.
The purpose of this review paper is to present evidence that rat animal models of alcoholism provide an ideal platform for developing and screening medications that target alcohol abuse and dependence. The focus is on the 5 oldest international rat lines that have been selectively bred for a high alcohol-consumption phenotype. The behavioral and neurochemical phenotypes of these rat lines are reviewed and placed in the context of the clinical literature. The paper presents behavioral models for assessing the efficacy of pharmaceuticals for the treatment of alcohol abuse and dependence in rodents, with particular emphasis on rats. Drugs that have been tested for their effectiveness in reducing alcohol/ethanol consumption and/or self-administration by these rat lines and their putative site of action are summarized. The paper also presents some current and future directions for developing pharmacological treatments targeting alcohol abuse and dependence.


Analysing the impact of private health insurance on inequities in health care utilization: a longitudinal study from China.
Since the early 2000s, Chinese government has sought to encourage the growth of private health insurance (PHI) while simultaneously expanding the breadth of coverage in its social health insurance (SHI) system. This paper examines how the prevalence of PHI has changed during this period and the extent to which PHI contributed to the growth of horizontal and geographical inequities with a focus on healthcare utilization. National data from China Health and Nutrition Survey between 2000 and 2015 were analysed using a multilevel modelling approach. The analysis investigated the impact of SHI membership as related to PHI uptake, PHI enrolees' utilization of health services and out-of-pocket (OOP) expenses. This study found being covered by an SHI scheme reduced the uptake of PHI between 2004 and 2015. Having PHI caused an increase in utilizing outpatient care but did not affect OOP expenses. Coverage prevalence of PHI in a residential community was positively associated with the average level of healthcare utilization. Coverage prevalence of PHI and its effects on healthcare utilization varied geographically. The findings suggest that expanding the role of PHI was not effective without clear support from government policy. Furthermore, the expansion of PHI may cause an increase in horizontal and geographical inequities in healthcare utilization.


What makes online substance-use interventions engaging? A systematic review and narrative synthesis.
Online substance-use interventions are effective in producing reductions in harmful-use. However, low user engagement rates with online interventions reduces overall effectiveness of interventions. Identifying optimal strategies with which to engage users with online substance-use interventions may improve usage rates and subsequent effectiveness. (1) To identify the most prevalent engagement promoting strategies utilised to increase use of online substance-use interventions. (2) To determine whether the identified engagement promoting strategies increased said use of online substance-use interventions. The reviewed followed Cochrane methodology. Databases were searched for online substance-use interventions and engagement promoting strategies limited by study type (randomised controlled trial). Due to heterogeneity between engagement promoting strategies and engagement outcomes, meta-analytic techniques were not possible. Narrative synthesis methods were used. Fifteen studies were included. Five different engagement promoting strategies were identified: (1) tailoring; (2) delivery strategies; (3) incentives; (4) reminders; (5) social support. The most frequently reported engagement promoting strategies was tailoring (47% of studies), followed by reminders and social support (40% of studies) and delivery strategies (33% of studies). The narrative synthesis demonstrated that tailoring, multimedia delivery of content and reminders are potential techniques for promoting engagement. The evidence for social support was inconclusive and negative for incentives. This review was the first to examine engagement promoting strategies in solely online substance-use interventions. Three strategies were identified that may be integral in promoting engagement with online substance-use interventions. However, the small number of eligible extracted studies, inconsistent reporting of engagement outcomes and diversity of engagement features prevent firmer conclusions. More high-quality trials examining engagement are required.


Elucidating the anticancer activities of guanidinium-functionalized amphiphilic random copolymers by varying the structure and composition in the hydrophobic monomer.
Rationale: Use of traditional anticancer chemotherapeutics has been hindered by the multifactorial nature of multi-drug resistance (MDR) development and metastasis. Recently, cationic polycarbonates were reported as novel unconventional anticancer agents that mitigated MDR and inhibited metastasis. The aim of this study is to explore structure-anticancer activity relationship. Specifically, a series of cationic guanidinium-based random copolymers of varying hydrophobicity was synthesized with a narrow polydispersity (Ð = 1.12-1.27) via organocatalytic ring-opening polymerization (OROP) of functional cyclic carbonate monomers, and evaluated for anticancer activity, killing kinetics, degradability and functional mechanism. Methods: Linear, branched and aromatic hydrophobic side chain units, such as ethyl, benzyl, butyl, isobutyl and hexyl moieties were explored as comonomer units for modulating anticancer activity. As hydrophobicity/hydrophilicity balance of the polymers determines their anticancer efficacy, the feed ratio between the two monomers was varied to tune their hydrophobicity. Results: Notably, incorporating the hexyl moiety greatly enhanced anticancer efficiency and killing kinetics on cancer cells. Degradation studies showed that the polymers degraded completely within 4-6 days. Flow cytometry and lactate dehydrogenase (LDH) release analyses demonstrated that anticancer mechanism of the copolymers containing a hydrophobic co-monomer was concentration dependent, apoptosis at IC50, and both apoptosis and necrosis at 2 × IC50. In contrast, the homopolymer without a hydrophobic comonomer killed cancer cells predominantly via apoptotic mechanism. Conclusion: The hydrophobicity of the polymers played an important role in anticancer efficacy, killing kinetics and anticancer mechanism. This study provides valuable insights into designing novel anticancer agents utilizing polymers.


Primary pulmonary extranodal natural killer/T-cell lymphoma (ENKTL), nasal type: Two case reports and literature review.
Extranodal natural killer/T-cell lymphoma (ENKTL) - nasal type is an aggressive form of malignant non-Hodgkin lymphoma with a very poor prognosis. Especially primary pulmonary ENKTL is a relatively rare form of non-Hodgkin lymphoma. Until now, the prevalence of primary pulmonary ENKTL is unknown. Since 2001, only 18 cases of primary pulmonary ENKTL have been published, in addition to the 2 cases reported here. We describe 2 cases of primary pulmonary ENKTL. Both patients were male non-smokers, aged 61 and 49 years. Their main clinical symptoms included cold-like symptoms and intermittent fever (39.3°C and 38.8°C) for some days (40 days and 3 weeks). Both patients had no relevant personal or family medical history. The patients were initially misdiagnosed with community-acquired pneumonia. Primary pulmonary ENKTL was confirmed by immunohistochemical staining of computed tomography-guided transthoracic needle biopsy specimens. Both cases were positive for CD56, CD3, and in situ hybridization for Epstein-Barr virus-encoded small RNA, but negative for CD20. Initially, both patients were treated inadequately with intravenous moxifloxacin administration (unknown dosage and 400 mg q.d) in their local hospitals. Once diagnosed with primary pulmonary ENKTL in our hospital, they received 3 cycles of chemotherapy with combined regimens of dexamethasone, methotrexate, ifosfamide, L-asparaginase, and etoposide (SMILE), and in the second patient, bone marrow transplantation was performed following the third chemotherapy cycle. Clinical follow-up after the chemotherapy showed that the condition of the first patient progressively deteriorated. He died 2 months following the initial diagnosis. However, the presence of the hemophagocytic lymphohistocytosis gradually improved in the second patient during chemotherapy. Ultimately, the second patient died of acute transplant rejection 6 months after the initial diagnosis. The diagnosis of ENKTL should be considered when patients present with fever and expansile consolidation of the lung not responding to antibiotics. The diagnosis depends on histopathology and immunophenotyping. Percutaneous transthoracic needle biopsy is a safe and effective biopsy method. Chemotherapy may improve the prognosis, but this should be confirmed by prospective multicenter studies.


Tuning bio-inspired skin-core structure of nascent fiber via interplay of polymer phase transitions.
The properties of polymer fibers are determined by their inner structures. We performed dynamic Monte Carlo simulations of early-stage solidification in the fluid filaments of stretched polymer solutions after extrusion into a coagulation bath upon fiber spinning. We observed that the radial temperature gradient dominates polymer crystallization to form an oriented crystalline skin (from single to multiple layers), while the radial non-solvent influx dominates phase separation to form a concentrated but less oriented core. The skin-core structure offers fibers a balanced performance between strength and toughness similar to plant stems, which can be tuned by the interplay of phase transitions. Our molecular-level observations facilitate a systematic understanding of the microscopic mechanism of fiber-spinning, which will pave a way towards making excellent polymer fibers.


Further characterization of Cys-type and Ser-type anaerobic sulfatase maturating enzymes suggests a commonality in the mechanism of catalysis.
The anaerobic sulfatase-maturating enzyme from Clostridium perfringens (anSMEcpe) catalyzes the two-electron oxidation of a cysteinyl residue on a cognate protein to a formylglycyl residue (FGly) using a mechanism that involves organic radicals. The FGly residue plays a unique role as a cofactor in a class of enzymes termed arylsulfatases, which catalyze the hydrolysis of various organosulfate monoesters. anSMEcpe has been shown to be a member of the radical S-adenosylmethionine (SAM) family of enzymes, [4Fe-4S] cluster-requiring proteins that use a 5'-deoxyadenosyl 5'-radical (5'-dA(•)) generated from a reductive cleavage of SAM to initiate radical-based catalysis. Herein, we show that anSMEcpe contains in addition to the [4Fe-4S] cluster harbored by all radical SAM (RS) enzymes, two additional [4Fe-4S] clusters, similar to the radical SAM protein AtsB, which catalyzes the two-electron oxidation of a seryl residue to a FGly residue. We show by size-exclusion chromatography that both AtsB and anSMEcpe are monomeric proteins, and site-directed mutagenesis studies of AtsB reveal that individual Cys → Ala substitutions at seven conserved positions result in an insoluble protein, consistent with those residues acting as ligands to the two additional [4Fe-4S] clusters. Ala substitutions at an additional conserved Cys residue (C291 in AtsB and C276 in anSMEcpe) afford proteins that display intermediate behavior. These proteins exhibit reduced solubility and drastically reduced activity, behavior that is conspicuously similar to that of a critical Cys residue in BtrN, another radical SAM dehydrogenase [Grove, T. L., et al. (2010) Biochemistry 49, 3783-3785]. We also show that wild-type anSMEcpe acts on peptides containing other oxidizable amino acids at the target position. Moreover, we show that the enzyme will convert threonyl peptides to the corresponding ketone product, and also allo-threonyl peptides, but with a significantly reduced efficiency, suggesting that the pro-S hydrogen atom of the normal cysteinyl substrate is stereoselectively removed during turnover. Lastly, we show that the electron generated during catalysis by AtsB and anSMEcpe can be utilized for multiple turnovers, albeit through a reduced flavodoxin-mediated pathway.


Desmoid-type fibromatosis: toward a holistic management.
Desmoid-type fibromatosis, a rare locally aggressive fibroblastic proliferation, is a treatment challenge. This review aimed to explore recent data about the management of desmoid-type fibromatosis. New data underline the role of kinases and ɣ-secretase in stimulating cell proliferation and invasiveness in desmoid-type fibromatosis. This explains the proven activity of multikinase inhibitors (sorafenib or pazopanib) in the management of desmoid-type fibromatosis or the emerging role of a ɣ-secretase inhibitor. An international guideline for management was recently published, and this guideline take into account patient point of view. Lastly, recent studies highlight the multidimensional burden of desmoid-type fibromatosis, particularly health-related quality of life (HRQoL). Active surveillance with planned MRI is the first-line management in desmoid-type fibromatosis. A site-specific and stepwise approach should be considered for progressive desmoid-type fibromatosis. Further, a risk-benefit analysis that considers the side effects and long-term sequelae should be conducted before deciding to start any treatment. A less aggressive approach should be considered. Multikinase inhibitors are effective, but their tolerability and side effects should be discussed with the patients. The symptoms and HRQoL should be integrated in decision-making. Desmoid-type fibromatosis patients should be offered support to address their needs supportive care.


[The combined chemotherapy with vincristine, prospidin, bleomycin and cyclophosphane of epidermoid lung cancer].
The effectiveness of combination chemotherapy with vincristine, prospidin, bleomycin, cyclophosphamide and prednisolone (VPBCP regimen) was studied in 71 patients with morphologically verified locally advanced and inoperable epidermoid lung cancer. A more than 50% regression of tumor was registered in 32.4% with 4.2% showing complete response. Tumor shrank by 25-50% in 15.5% but failed to respond in 29.6%. Toxicity proved low. A direct correlation--the more apparent regression, the longer remission and, correspondingly, longer survival--was established between prognosis and response.


A female infant with Frasier syndrome showing splice site mutation in Wilms' tumor gene (WT1) intron 9.
Wilms' tumor gene (WT1) abnormality leads to various disorders of differentiation as well as renal and urinary system abnormalities. Here we present a case of WT1 abnormality and steroid-resistant nephrotic syndrome in a female infant. The 3-year-old patient was initially diagnosed with proteinuria at an annual mass screening program for children aged three years and was referred to our hospital. She met the diagnostic criteria for nephrotic syndrome and showed normal renal function. The patient was treated with corticosteroids; however her condition showed resistance to corticosteroids. On renal biopsy, she was diagnosed with focal segmental glomerulosclerosis (FSGS). Because of the possibility of WT1 abnormality, an exon array analysis was conducted, which ruled out Denys-Drash Syndrome (DDS). The patient was then diagnosed with Frasier Syndrome (FS) on the basis of donor site mutation (IVS9+5G > A) of the splice site in the intron 9. Reports of female infants with FS are extremely rare. FS is one of the pre-mRNA splicing diseases, in which the occurrence of symptoms is associated with a decrease in the ratio of the lysine-threonine-serine (+/- KTS) isoform of the WT1 protein. A typical case exhibits 46 XY male karyotype and is characterized by male pseudohermaphroditism with cord-like gonadal structures as well as progressive nephropathy caused by FSGS. However, in female infants without such extrarenal signs, it is necessary to consider the analysis for WT1 intron 9 for conclusive diagnosis of FS, because the presence of nephropathy is the only symptom for possible detection.


Mutational analysis of bacteriophage lambda lysis gene S.
A plasmid carrying the bacteriophage lambda lysis genes under lac control was subjected to hydroxylamine mutagenesis, and mutations eliminating the host lethality of the S gene were selected. DNA sequence analysis revealed 48 single-base mutations which resulted in alterations within the coding sequence of the S gene. Thirty-three different missense alleles were generated. Most of the missense changes clustered in the first two-thirds of the molecule from the N terminus. A simple model for the disposition of the S protein within the inner membrane can be derived from inspection of the primary sequence. In the first 60 residues, there are two distinct stretches of predominantly hydrophobic amino acids, each region having a net neutral charge and extending for at least 20 residues. These regions resemble canonical membrane-spanning domains. In the model, the two domains span the bilayer as a pair of net neutral charge helices, and the N-terminal 10 to 12 residues extend into the periplasm. The mutational pattern is largely consistent with the model. Charge changes within the putative imbedded regions render the protein nonfunctional. Loss of glycine residues at crucial reverse-turn domains which would be required to reorient the molecule to reenter the membrane also inactivate the molecule. Finally, a number of neutral and rather subtle mutations such as Ala to Val and Met to Ile are found, mostly within the putative spanning regions. Although no obvious explanation exists for this subtle and heterogeneous class of mutations, it is noted that all of the changes result in a loss of alpha-helical character as predicted by Chou-Fasman theoretical analysis. Alternative explanations for some of these changes are also possible, including a reduction in net translation rate due to substitution of a rare codon for a common one. The model and the pattern of mutations have implications for the probable oligomerization of the S protein at the time of endolysin release at the end of the vegetative growth period.


Fibrin-hyaluronic acid hydrogel-based delivery of antisense oligonucleotides for ADAMTS5 inhibition in co-delivered and resident joint cells in osteoarthritis.
To date no disease-modifying drugs for osteoarthritis (OA) are available, with treatment limited to the use of pain killers and prosthetic replacement. The ADAMTS (A Disintegrin and Metallo Proteinase with Thrombospondin Motifs) enzyme family is thought to be instrumental in the loss of proteoglycans during cartilage degeneration in OA, and their inhibition was shown to reverse osteoarthritic cartilage degeneration. Locked Nucleic Acid (LNA)-modified antisense oligonucleotides (gapmers) released from biomaterial scaffolds for specific and prolonged ADAMTS inhibition in co-delivered and resident chondrocytes, is an attractive therapeutic strategy. Here, a gapmer sequence identified from a gapmer screen showed 90% ADAMTS5 silencing in a monolayer culture of human OA chondrocytes. Incorporation of the gapmer in a fibrin-hyaluronic acid hydrogel exhibited a sustained release profile up to 14 days. Gapmers loaded in hydrogels were able to transfect both co-embedded chondrocytes and chondrocytes in a neighboring gapmer-free hydrogel, as demonstrated by flow cytometry and confocal microscopy. Efficient knockdown of ADAMTS5 was shown up to 14 days in both cell populations, i.e. the gapmer-loaded and gapmer-free hydrogel. This work demonstrates the use applicability of a hydrogel as a platform for combined local delivery of chondrocytes and an ADAMTS-targeting gapmer for catabolic gene modulation in OA.


Crop bromide concentrations following methyl bromide fumigation for pale cyst nematode in southeastern Idaho.
Methyl bromide (MeBr) is a sterilizing fumigant used to control quarantine pests that is restricted due to its detrimental atmospheric effects. The degradation of injected MeBr produces crop-available Br- . Up to five applications of MeBr were used in southeastern Idaho fields to combat the pale cyst nematode (Globodera pallida). Data regarding the uptake and partitioning of Br- in crops following MeBr application in the region were unavailable. Research determined background concentrations of Br- in alfalfa (Medicago sativa L.), barley (Hordeum vulgare L.), corn (Zea mays L.), potato (Solanum tuberosum L.), and wheat (Triticum aestivum L.) compared to MeBr-treated fields. Background Br- concentrations ranged from nondetectable (ND) to 33.0 mg Br- kg-1 ; vegetative tissue concentrations were greater than reproductive, except corn where there was no difference. Nearly all crops grown in MeBr-treated fields had greater Br- concentrations than background. Background-baled-alfalfa tissue Br- concentration was 33.0 mg kg-1 compared to 117.8 mg Br- kg-1 from a MeBr-treated field. Br- concentration in green alfalfa decreased from 79.8 to 36.5 mg Br- kg-1 at the final cutting in a MeBr-treated field, where time after application decreased crop Br- concentrations. Small grains had low Br- concentrations in reproductive tissue (1.7 mg Br- kg-1 ) compared to vegetative tissue (106.5 mg Br- kg-1 ). Corn stover concentration (12.7 mg Br- kg-1 ) was low relative to small-grain straw, but corn ear (5.8 mg Br- kg-1 ) was greater than small-grain reproductive tissue in the MeBr-treated field. Crop selection following MeBr applications should consider the likelihood of elevated Br- concentration for the plant fractions intended end use.


Design, implementation and evaluation of a new core learning objectives curriculum for a urology clerkship.
Until recently, medical students at the University of Wisconsin School of Medicine and Public Health participated in a traditional 2-week urology clerkship. We hypothesized that a new curriculum with core learning objectives and student oriented didactic sessions would increase learning and satisfaction compared to a traditional clerkship. Between July 2008 and June 2009, 55 medical students completed the urology clerkship following the traditional curriculum. Between July 2009 and June 2010, 51 students followed the core learning objectives curriculum. We compared the curriculum outcomes using objective and subjective measures. Overall student participation was 90%, with 95 of 106 students completing both assessment tools. The objective scores of the students following the core learning objectives were higher than those of the students following the traditional curriculum. The t test to evaluate the difference between the 2 curricula was statistically significant (t = 2.845, df = 93, p <0.05). Subjective scores for the core learning objectives group were lower in all but 1 category. Student perception of knowledge attainment for the core learning objectives cohort was higher than that of the traditional cohort, but none of the subjective scores was statistically significant. This study demonstrated that a core learning objectives curriculum was associated with higher objective test scores compared to a traditional model, suggesting that the core learning objectives curriculum increased student learning compared to the traditional curriculum. However, the core learning objectives cohort did not show greater satisfaction than students following the traditional curriculum.


Rationally Designed PI3Kα Mutants to Mimic ATR and Their Use to Understand Binding Specificity of ATR Inhibitors.
ATR, a protein kinase in the PIKK family, plays a critical role in the cell DNA-damage response and is an attractive anticancer drug target. Several potent and selective inhibitors of ATR have been reported showing significant antitumor efficacy, with most advanced ones entering clinical trials. However, due to the absence of an experimental ATR structure, the determinants contributing to ATR inhibitors' potency and specificity are not well understood. Here we present the mutations in the ATP-binding site of PI3Kα to progressively transform the pocket to mimic that of ATR. The generated PI3Kα mutants exhibit significantly improved affinity for selective ATR inhibitors in multiple chemical classes. Furthermore, we obtained the X-ray structures of the PI3Kα mutants in complex with the ATR inhibitors. The crystal structures together with the analysis on the inhibitor affinity profile elucidate the roles of individual amino acid residues in the binding of ATR inhibitors, offering key insights for the binding mechanism and revealing the structure features important for the specificity of ATR inhibitors. The ability to obtain structural and binding data for these PI3Kα mutants, together with their ATR-like inhibitor binding profiles, makes these chimeric PI3Kα proteins valuable model systems for structure-based inhibitor design.


Dual-locus DNA metabarcoding reveals southern hairy-nosed wombats (Lasiorhinus latifrons Owen) have a summer diet dominated by toxic invasive plants.
Habitat degradation and summer droughts severely restrict feeding options for the endangered southern hairy-nosed wombat (SHNW; Lasiorhinus latifrons). We reconstructed SHNW summer diets by DNA metabarcoding from feces. We initially validated rbcL and ndhJ diet reconstructions using autopsied and captive animals. Subsequent diet reconstructions of wild wombats broadly reflected vegetative ground cover, implying local rather than long-range foraging. Diets were all dominated by alien invasives. Chemical analysis of alien food revealed Carrichtera annua contains high levels of glucosinolates. Clinical examination (7 animals) and autopsy (12 animals) revealed that the most degraded site also contained most individuals showing signs of glucosinolate poisoning. We infer that dietary poisoning through the ingestion of alien invasives may have contributed to the recent population crashes in the region. In floristically diverse sites, individuals appear to be able to manage glucosinolate intake by avoidance or episodic feeding but this strategy is less tractable in the most degraded sites. We conclude that recovery of the most affected populations may require effective Carrichtera management and interim supplementary feeding. More generally, we argue that protection against population decline by poisoning in territorial herbivores requires knowledge of their diet and of those food plants containing toxic principles.


A Mixed-Methods Study of Early Childhood Education and Care in South Korea: Policies and Practices During COVID-19.
This study of South Korea's response to COVID-19 has three purposes. First, it uses document analysis to examine policies, strategies, and resources offered by the South Korean government and public organizations to support young children and families during the first 6 months of the pandemic. Next, it uses open-ended surveys with 30 directors of early childhood institutions to explore institutional-level supports and needs during the pandemic. Finally, it looks at the discrepancies between stated policies outlining the South Korea's response to COVID-19 and the lived experiences of early childhood educators as a route to arriving at recommendations for education policymakers and other stakeholders. To that end, we reviewed government documents (n = 84) containing early childhood education-related responses to Covid-19 established by the Ministry of Education, the Ministry of Health and Welfare, and other relevant government sectors. An online survey with 17 kindergarten and 13 child care center directors was also analyzed. Using content analysis, the findings revealed that the government's policies and guidance for Early Childhood Education and Care (ECEC) as well as the institutional supports for children and families were overall comprehensive in its scope. The analysis, based on the five tenets of the Whole Child approach, also indicated that the government's policy responses and services for ECEC focused mainly on the 'Safe' and 'Supported' tenets, while 'Challenged' was given the least amount of consideration. The survey responses demonstrated different measures taken by kindergartens and child care centers highlighting the separate nature of 'education' and 'care' in South Korea, while also indicating limited resources for supporting children's psychological well-being and for children and families in need. This overview provides a foundation for further discussion and research on the impact of Covid-19 on ECEC in South Korea and beyond.


Can proteomics lead to the discovery of real biomarkers for HCC?
The development of proteomics technologies has lead to a great deal of effort being focused on the identification of biomarkers for cancers. Although many papers have reported candidate biomarkers for hepatocellular carcinomas (HCCs) in particular, so far none of these candidate biomarkers have been used either for diagnosis or therapy intreating patients. The question remains: Can proteomics identify real biomarkers for HCCs?


Molecular Detection of Alpha Thalassemia: A Review of Prevalent Techniques.
Alpha thalassemia (α-thalassemia) is an autosomal recessive disorder due to the reduction or absence of α globin chain production. Laboratory diagnosis of α-thalassemia requires molecular analysis for the confirmatory diagnosis. A screening test, comprising complete blood count, blood smear and hemoglobin quantification by high performance liquid chromatography and capillary electrophoresis, may not possibly detect all the thalassemia diseases. This review focused on the molecular techniques used to detect α-thalassemia, and the advantages and disadvantages of each technique were highlighted. Multiplex gap-polymerase chain reaction, single-tube multiplex polymerase chain reaction, multiplex ligation-dependent probe amplification, and loop-mediated isothermal amplification were used to detect common deletion of α-thalassemia. Furthermore, the reverse dot blot analysis and a single tube multiplex polymerase chain reaction could detect non-deletion mutation of the α-globin gene. Sanger sequencing is widely used to detect non-deletion mutations of α-thalassemia. Recently, next-generation sequencing was introduced in the diagnosis of both deletion and point mutations of α-thalassemia. Despite the advantages and disadvantages of different techniques, the routine method employed in the laboratory should be based on the facility, expertise, available equipment, and economic conditions.


Endovascular repair of direct carotid-cavernous fistula in Ehlers-Danlos type IV.
Ehlers-Danlos syndrome (EDS) type IV is a collagen vascular disease with an autosomal dominant inheritance caused by COL3A1 mutation. Patients with EDS type IV can present with organ rupture, spontaneous arterial dissections and ruptured aneurysms. Because of their propensity to form arterial dissections, aneurysms and rupture, they can develop carotid-cavernous fistula (CCF) after minor trauma or spontaneously. In EDS, it has been reported that even conventional catheter diagnostic angiography may result in large artery dissections and vessel rupture. In addition, the treatment of CCF in EDS type IV can result in up to 59% mortality after initial treatment, of which 23% is attributed to direct complications of treatment. We present the case of a patient with EDS type IV who previously had spontaneous dissection and multiple pseudoaneurysms of both the iliac and femoral arteries and the distal abdominal aorta. Several years later the patient developed a direct type A CCF which was successfully treated with endovascular embolization using a transvenous approach with detachable coils. The literature pertaining to CCF in EDS type IV and its treatment is reviewed.


Use of thermonuclease testing to identify Staphylococcus aureus by direct examination of blood cultures.
Blood cultures submitted to the Clinical Microbiology Laboratory, Queen Alia Military Hospital, Amman during 1999-2001 were examined to evaluate thermonuclease testing for identifying Staphylococcus aureus in blood culture broths growing gram-positive cocci. Of 170 cultures studied, 129 yielded gram-positive staphylococci and 41 yielded other gram-positive cocci. Toluidine blue-deoxynucleic acid agar plates were used to test for thermonuclease activity. Standard tube coagulase tests were performed on the isolates. Direct detection of thermonuclease activity in 76 blood culture broths containing gram-positive staphylococci showed 100% correlation with subsequent tube coagulase tests. The thermonuclease test provides a fast, specific and reliable confirmation of S. aureus bacteraemia by direct examination of blood culture broths that contain gram-positive cocci. This allows for timely, optimal antibiotic therapy.


Pleiotropic skeletal and ocular phenotypes of the mouse mutation congenital hydrocephalus (ch/Mf1) arise from a winged helix/forkhead transcriptionfactor gene.
Congenital hydrocephalus is an etiologically diverse, poorly understood, but relatively common birth defect. Most human cases are sporadic with familial forms showing considerable phenotypic and etiologic heterogeneity. We have studied the autosomal recessive mouse mutation congenital hydrocephalus ( ch ) to identify candidate human hydrocephalus genes and their modifiers. ch mice have a congenital, lethal hydrocephalus in association with multiple developmental defects, notably skeletal defects, in tissues derived from the cephalic neural crest. We utilized positional cloning methods to map ch in the vicinity of D13Mit294 and confirm that the ch phenotype is caused by homozygosity for a nonsense mutation in a gene encoding a winged helix/forkhead transcription factor ( Mf1 ). Based on linked genetic markers, we performed detailed phenotypic characterization of mutant homozygotes and heterozygotes to demonstrate the pleiotropic effects of the mutant gene. Surprisingly, ch heterozygotes have the glaucoma-related distinct phenotype of multiple anterior segment defects resembling Axenfeld-Rieger anomaly. We also localized a second member of this gene family ( Hfh1 ), a candidate for other developmental defects, approximately 470 kb proximal to Mf1.


Photoreception of Paramecium cilia: localization of photosensitivity and binding with anti-frog-rhodopsin IgG.
Paramecium bursaria is photosensitive and accumulates in a lighted area. The cells can be deciliated by a brief suspension in dilute ethanol. Both intact and deciliated cells showed depolarization in response to light stimulation by a step-increase from dark to above 0.7 mW cm-2 (550 nm). On the other hand, after a step-increase to below 0.4 mW cm-1, intact cells showed hyperpolarization, while the deciliated cells showed no change in membrane potential. This difference in membrane potential response between ciliated and deciliated cells suggests that both somatic and ciliary structures are photosensitive. In our search for the photoreceptive molecules, a polyclonal antibody induced in rabbits against frog rhodopsin was found to cross-react with a 63x10(3) Mr protein of P. bursaria, by immunoelectrophoresis. Immunocytochemical studies showed that the antibody labeling was localized on both the ciliary and the somatic membranes. These results raise the possibility that P. bursaria may contain a rhodopsin-like protein as a photoreceptor molecule.


Methanesulfinic acid reaction with OH: mechanism, rate constants, and atmospheric implications.
The mechanism for the atmospheric oxidation of methanesulfinic acid (MSIA) has been studied. This is the first theoretical study of the reaction between MSIA and the OH radical. All the possible channels in this reaction have been studied theoretically, and their corresponding rate constants have been evaluated under the variational transition-state theory (VTST) formalism. Two different products can be formed: the CH3S(O)2 radical (which had been experimentally proposed as the only one), and sulfurous acid (H2SO3). The CH3S(O)2 radical can be formed directly or can form via an intermediate adduct, which yields to the radical through the elimination of a water molecule. For the first time, it is theoretically demonstrated that SO2 is formed in the addition channel of the DMS + OH reaction. The consequences of this result in the interpretation of the T-dependence of the SO4(2-)/MSA (methanesulfonic acid) quocient are analyzed. The competition between the unimolecular dissociation of the CH3S(O)2 radical and OH-addition to yield MSA is proposed as one of the possible multiple branching points (along the DMS + OH degradation scheme) influencing the T-dependence of the SO4(2-)/MSA relation.


Insulin stimulates the halting, tethering, and fusion of mobile GLUT4 vesicles in rat adipose cells.
Glucose transport in adipose cells is regulated by changing the distribution of glucose transporter 4 (GLUT4) between the cell interior and the plasma membrane (PM). Insulin shifts this distribution by augmenting the rate of exocytosis of specialized GLUT4 vesicles. We applied time-lapse total internal reflection fluorescence microscopy to dissect intermediates of this GLUT4 translocation in rat adipose cells in primary culture. Without insulin, GLUT4 vesicles rapidly moved along a microtubule network covering the entire PM, periodically stopping, most often just briefly, by loosely tethering to the PM. Insulin halted this traffic by tightly tethering vesicles to the PM where they formed clusters and slowly fused to the PM. This slow release of GLUT4 determined the overall increase of the PM GLUT4. Thus, insulin initially recruits GLUT4 sequestered in mobile vesicles near the PM. It is likely that the primary mechanism of insulin action in GLUT4 translocation is to stimulate tethering and fusion of trafficking vesicles to specific fusion sites in the PM.


The effects of electronic prescribing by community-based providers on ambulatory medication safety.
US federal policies are incentivizing use of electronic prescribing (e-prescribing) to improve safety. However, little is known about e-prescribing's actual impact on medication safety over time. A study was conducted to assess the effect of implementing a commercial electronic health record (EHR) with e-prescribing on rates and types of prescribing errors. Understanding safety effects from e-prescribing will be important as providers increasingly e-prescribe. Prescriptions written by 20 community-based primary care providers in the Hudson Valley region of New York from November 2008 to November 2009 were retrospectively studied. All providers adopted a commercial EHR with robust clinical decision support and extensive technical support to aid in prescribing. Errors were identified by standardized prescription and chart review. Some 1,629 prescriptions were analyzed at three months postimplementation, and 1,738 prescriptions were analyzed at one year postimplementation. Use of e-prescribing resulted in relatively low error rates (6.0 errors per 100 prescriptions). These rates were sustained over time but without further improvement (6.0 versus 4.5 errors per 100 prescriptions, p = .15). Antibiotics were the class of medications most frequently involved (12.7% of overall errors), and direction errors were most common (24% of errors). This study is the first, as far as known, to quantitatively evaluate prescribing errors early after EHR implementation and after sustained use among community-based primary care providers. Relatively low rates of errors with e-prescribing were found early and after prolonged use. Extensive support for providers before, during, and after implementation may mitigate potential safety threats from implementation of an EHR system and result in sustained safety benefits over the long-term.


A survey on the state of nanosafety research in the European Union and the United States.
Researchers in the field of environmental safety and health of nanomaterials (nanosafety) were surveyed between September 2017 and January 2018 to obtain a snapshot on the state of investigations in this field. The data received from 84 respondents give information on exposure pathways, methods, biologic and toxic effects, dose metrics and range, criteria for selecting benchmark materials, and on problems that are considered urgent in the field. The results can help guide research strategies and funding schemes related to nanosafety of nanoparticles. It is intended to repeat this survey every few years, to follow developments of the field.


Intracellular energetics and critical PO2 in resting ischemic human skeletal muscle in vivo.
During ischemia and some types of muscular contractions, oxygen tension (Po(2)) declines to the point that mitochondrial ATP synthesis becomes limited by oxygen availability. Although this critical Po(2) has been determined in animal tissue in vitro and in situ, there remains controversy concerning potential disparities between values measured in vivo and ex vivo. To address this issue, we used concurrent heteronuclear magnetic resonance spectroscopy (MRS) to determine the critical intracellular Po(2) in resting human skeletal muscle in vivo. We interleaved measurements of deoxymyoglobin using (1)H-MRS with measures of high-energy phosphates and pH using (31)P-MRS, during 15 min of ischemia in the tibialis anterior muscles of 6 young men. ATP production and intramyocellular Po(2) were quantified throughout ischemia. Critical Po(2), determined as the Po(2) corresponding to the point where PCr begins to decline (PCr(ip)) in resting muscle during ischemia, was 0.35 ± 0.20 Torr, means ± SD. This in vivo value is consistent with reported values ex vivo and does not support the notion that critical Po(2) in resting muscle is higher when measured in vivo. Furthermore, we observed a 4.5-fold range of critical Po(2) values among the individuals studied. Regression analyses revealed that time to PCr(ip) was associated with critical Po(2) and the rate of myoglobin desaturation (r = 0.83, P = 0.04) but not the rate of ATP consumption during ischemia. The apparent dissociation between ATP demand and myoglobin deoxygenation during ischemia suggests that some degree of uncoupling between intracellular energetics and oxygenation is a potentially important factor that influences critical Po(2) in vivo.


Genomic DNA functions as a universal external standard in quantitative real-time PCR.
Real-time quantitative PCR (qPCR) is a powerful tool for quantifying specific DNA target sequences. Although determination of relative quantity is widely accepted as a reliable means of measuring differences between samples, there are advantages to being able to determine the absolute copy numbers of a given target. One approach to absolute quantification relies on construction of an accurate standard curve using appropriate external standards of known concentration. We have validated the use of tissue genomic DNA as a universal external standard to facilitate quantification of any target sequence contained in the genome of a given species, addressing several key technical issues regarding its use. This approach was applied to validate mRNA expression of gene candidates identified from microarray data and to determine gene copies in transgenic mice. A simple method that can assist achieving absolute quantification of gene expression would broadly enhance the uses of real-time qPCR and in particular, augment the evaluation of global gene expression studies.


Systemic sclerosis and cryoglobulinemia: our experience with overlapping syndrome of scleroderma and severe cryoglobulinemic vasculitis and review of the literature.
Systemic sclerosis (SSc) is an immune-mediated disorder characterized by multiple organ fibrotic alterations and diffuse microangiopathy. The SSc can be associated with other connective tissue diseases and less frequently with systemic vasculitides, including cryoglobulinemic vasculitis (CV). The aim of the present study was to investigate the prevalence of CV in a large series of SSc patients. The presence of serum cryoglobulins was detected in 246 SSc patients (24 M and 222 F, age 61±13.5 SD years, disease duration 9.3±6.7 SD years); the observed clinico-serological findings, in particular the presence of SSc-CV overlapping syndrome, were carefully analyzed and compared with previous data reported in the literature. The presence of circulating cryoglobulins was found in 7/246 (2.8%) of SSc patients; namely, 2 subjects only trace amounts of cryoglobulins, while 5 (2%) showed mixed cryoglobulinemia (type II, IgG-IgMk), low C4, rheumatoid factor seropositivity, and hepatitis C virus infection. Among SSc patients with serum mixed cryoglobulins, 4 (1.6%) developed a clinically overt CV, while the other one was totally asymptomatic with regard to typical vasculitic manifestations. Patients with SSc-CV overlapping syndrome had limited cutaneous SSc with serum anticentromere antibodies, pulmonary hypertension, clinico-serological features of HCV-related CV, and non-healing skin ulcers of the lower limbs. In all cases, the diagnosis of SSc preceded the clinical onset of CV, from 3 to 17years. The treatment with rituximab was useful on skin ulcers of lower limb in 2/3 patients; however, the overall clinical outcome of the four SSc-CV patients was unusually severe: one with very severe skin ulcers complicated by gangrene required bilateral through-the knee amputation, the other three subjects died because of severe heart failure, and in two cases because of untreatable pulmonary hypertension. In the literature, the prevalence of mixed cryoglobulinemia in scleroderma patients is quite rare (range 0.3-2%); while, the association of SSc with clinically overt CV is only anecdotally described, always in the absence of HCV infection. The SSc-CV overlapping syndrome described here is characterized by markedly severe vascular manifestations responsible for very poor prognosis; these peculiar clinical manifestations suggest a synergic activity of typical scleroderma microangiopathy and cryoglobulinemic vasculitis.


Excitation of surface plasmons by finite width beams.
The field properties of surface plasmons produced by realistically bounded beams incident in various attenuated total reflection (ATR) geometries are examined. Analytical results are first derived for the general case dealing with a beam field incident in a multilayered configuration. We show that, at the phase matching condition, the reflected field can be severely distorted in comparison with the incident beam shape. We also find that the power intensity inside the metal medium can be much smaller than that expected under the assumption of plane wave incidence. However, when the beamwidth is larger than the proportion range of the excited plasmon, the power intensity profiles and find that they exhibit distinguishing characteristics. In particular, for an incident Gaussian beam, the location of maximum power density on the metal surface shifts with respect to the center of the incident beam by a distance of the order of the plasmon propagation length. For a rectangular beam incident at the phase matching condition, on the other hand, the propagation range of the coupled surface plasmon can be found directly from the profile of the reflected field. We also show the overall process of beam wave coupling in the ATR geometry can be simulated by a spatial operating system having the response of either a differentiator (for the reflected field) or an integrator (for the transmitted field).


High-speed gallop locomotion in the Thoroughbred racehorse. II. The effect of incline on centre of mass movement and mechanical energy fluctuation.
During locomotion on an incline, mechanical work is performed to move an animal up the slope and increase the potential energy (PE) of the trunk and hence the centre of mass (CoM). Thus, at a given speed the total net mechanical work increases with the PE of the animal. In this study we investigate the mechanical energy (ME) fluctuations and the mechanical cost of transport (MCT) in six horses galloping up a range of gradients. We captured trunk movement with a six degrees-of-freedom inertial sensor mounted over the dorsal spinous process of the fourth to sixth thoracic vertebrae of the horse. Footfall timings were measured using a previously validated system of limb-mounted accelerometers. Speed was measured using a Global Positioning System (GPS) data logger. A track survey provided detailed incline information for the track. Linear (craniocaudal, mediolateral and dorsoventral) and rotational (roll, pitch and heading) kinematic parameters (displacement, velocity and acceleration) were calculated at speeds ranging from 9.0 to 12.0 m s(-1) during routine training over a range of inclines. Estimates of ME fluctuations and the MCT were made. Results showed the effect of incline on trunk motion during galloping was small. Increases in linear mechanical work and MCT were primarily explained by an increase in the work required to move the animal up the slope (and increase the PE of the CoM). Within the stride the majority of the work was performed during hindlimb stance. Our results have provided new insights into how horses power uphill locomotion.


Impact of saccharides on the drying kinetics of agarose gels measured by in-situ interferometry.
Agarose gels are viscoelastic soft solids that display a porous microstructure filled with water at 90% w/w or more. Despite an extensive use in food industry and microbiology, little is known about the drying kinetics of such squishy solids, which suffers from a lack of time-resolved local measurements. Moreover, only scattered empirical observations are available on the role of the gel composition on the drying kinetics. Here we study by in-situ interferometry the drying of agarose gels of various compositions cast in Petri dishes. The gel thinning is associated with the displacement of interference fringes that are analyzed using an efficient spatiotemporal filtering method, which allows us to assess local thinning rates as low as 10 nm/s with high accuracy. The gel thinning rate measured at the center of the dish appears as a robust observable to quantify the role of additives on the gel drying kinetics and compare the drying speed of agarose gels loaded with various non-gelling saccharides of increasing molecular weights. Our work shows that saccharides systematically decrease the agarose gel thinning rate up to a factor two, and exemplifies interferometry as a powerful tool to quantify the impact of additives on the drying kinetics of polymer gels.


Adrenocortical tumorigenesis, luteinizing hormone receptor and transcription factors GATA-4 and GATA-6.
Luteinizing hormone (LH/hCG) responsiveness of normal and pathological human adrenal glands as well as the possibility of constitutive expressions of luteinizing hormone receptor (LHR) in adrenal cortex has been reported. Some recent studies showed a correlation between the LHR and abundant GATA-4 expression in both metastasizing and non-metastasizing human adrenocortical tumors, but not in normal adrenals, implicating the putative relevance of LHR and GATA-4 for adrenocortical pathophysiology. However, the physio- and pathophysiological significance of LHR and GATA-4 in the mechanism of adrenocortical tumorigenesis remains unclear. The paucity of suitable models for adrenal tumorigenesis makes the establishment of proper animal models highly important. LHR expression in the murine adrenal gland is an exception and not found in wild-type (WT) animal. We have previously shown that ectopic LHR expression in the murine adrenal gland can be induced by chronically elevated LH levels. We have generated a gonadotropin-responsive adrenal tumor model in gonadectomized transgenic (TG) mice expressing the inhibin alpha promoter/Simian Virus 40 T antigen transgene (inhalpha/Tag). Given the induction of expression and regulation of GATA-4 and GATA-6 zinc finger transcription factors in the gonads by gonadotropins, this review will explore their relationship to LHR expression and their role in adrenocortical tumorigenesis. A functional link between LHR and GATA-4 actions in the adrenal pathophysiology is proposed.


[Correlation between clinical manifestation, prognosis and antibody in anti-muscle-specific tyrosine kinase antibody positive myasthenia gravis].
Objective: To determine the correlation between the titer of anti-muscle-specific tyrosine kinase (anti-MuSK) antibodies (Abs) and the severity and prognosis of Musk-associated myasthenia gravis (Musk-MG). Methods: A total of 33 MuSK-MG patients diagnosed at Department of Neurology, Peking Union Medical College Hospital from May 2018 to June 2020 were prospectively included. Patients were divided into different groups according to immune state, and the immune naive patients were further divided by the Myasthenia Gravis Foundation of America (MGFA) classification. There were 28 Musk-MG patients who completed the follow-up and subdivided into different groups according to post-intervention status (PIS). Twenty-five patients who received immunotherapy were divided into corticosteroid monotherapy group (n=17) and corticosteroid combined with immunosuppressant group (n=8). The comparison of Ab titers between different MGFA groups and PIS groups was determined by Kruskal-Wallis method, and the comparison of Ab titers between different time points was analyzed by Mann-Whitney U method. Results: There were 11 males and 22 females included in the study, with an onset age of 48 (18, 73) years, of which 16 cases were immune naive and 17 cases were treated with corticosteroids or immunosuppressant at least once. In immune naive population, a significant difference of Ab titers among different MGFA phenotypes was detected (P=0.04). Ab titers were reduced by immunosuppression therapy (the median value decreased from 1.20 to 0.87, P=0.01). Twenty-four (85.7%) MuSK-MG patients achieved a good prognosis (PIS-PR/MM), 1 (3.6%) case achieved improvement (PIS-I), and 3 (10.7%) patients' condition worsened (PIS-W), there was no significant difference of Ab titers among the three groups (P=0.21). Meanwhile, there was no significant difference of Ab titers between different treatment groups (P=0.95). Conclusions: In the immune naive state, the concentration of MuSK-Ab is consistent with the severity of the disease, and the Ab titers decrease after immunotherapy. Change of Ab titers is related to the daily dosage of corticosteroid and is not consistent with PIS grades.


Traumatic lingual ulcer in a child: Riga-Fede disease.
Riga-Fede disease RFD is an extremely rare, benign inflammatory disorder characterized by reactive, traumatic ulceration of the oral mucosa especially located on the tongue. It is most commonly associated with natal or neonatal teeth in newborns. Mucosal lesions are often caused by repetitive traumatic damage due to backward and forward motions of the tongue over the lower incisors. Failure to diagnose and treat these lesions properly may result in inadequate food intake, growth retardation and permanent lingual deformity. We report a 15-month-old healthy infant with tongue ulcer diagnosed as RFD based on history and clinical features.


Effect of linsidomine on the human radial artery.
The response of the human radial artery to a direct NO donor, linsidomine or 3-morpholino-sydnonimine (Sin 1), in the therapeutic management of peri-operative spasm may increase the patency rates of these grafts in the short, medium and long term. Evaluation of the effects of Sin 1 on the human radial artery is of even greater interest as it has not been published previously. Ninety-six human radial artery rings were studied with two protocols. Rings were mounted in an isolated organ bath between two stainless steel metallic rods connected to stress gauges. Protocol 1 studied the vasorelaxant effect of Sin 1 and nitroglycerin (TNT). Protocol 2 studied the reactivity of the radial artery to the vasoconstrictor agents arginine vasopressin (AVP) and angiotensin II (ANG II). The vasorelaxant effect of Sin 1 on the human radial artery was comparable with that of TNT, but with no tolerance effect. After Sin 1 pre-incubation, the vasoconstrictor effect of ANG II was abolished, whereas AVP induced maximum vasoconstriction similar to that of the control (not statistically significant), but with a shift in the EC50 to higher concentrations, EC50=15+/-20 nM. Sin 1 vasorelaxation of rings precontracted by ANG II was maximal, whereas after contraction by AVP, relaxation remained less than 70%; Sin 1 is a potent vasorelaxant on the human radial artery, which does not exhibit cross-tolerance with nitrates. This compound may be used pre- or post-operatively, and would undoubtedly be of benefit in the peri-operative preparation bath.


Spectrophotometric system based on a device created by 3D printing for the accommodation of a webcam chamber as a detection system.
The development of a simple and economical spectrophotometric system based on the use of a device created by 3D printing and the electronics necessary to control the intensity of the radiation source was described. The measurements are made with a low-cost digital webcam. The entire system is only powered through the USB outputs of a computer, which makes the portable and really practical system for the measurements in the field. This method was applied to determine iron (II) in waters using o-phenanthroline as chromogenic reagent giving a red complex, and also to hypochlorite determination using tetramethylbenzidine as the reagent providing a yellow color. The calibration curves were built using a mathematical algorithm making a RGB deconvolution. The intense of colors obtained from a webcam in each concentration of analyte had a relationship with the absorbance values. In order to confirm the accuracy and precision of this method, a traditional spectrophotometer was used for validation.


Emergency CT brain: preliminary interpretation with a tablet device: image quality and diagnostic performance of the Apple iPad.
Tablet devices have recently been used in radiological image interpretation because they have a display resolution comparable to desktop LCD monitors. We identified a need to examine tablet display performance prior to their use in preliminary interpretation of radiological images. We compared the spatial and contrast resolution of a commercially available tablet display with a diagnostic grade 2 megapixel monochrome LCD using a contrast detail phantom. We also recorded reporting discrepancies, using the ACR RADPEER system, between preliminary interpretation of 100 emergency CT brain examinations on the tablet display and formal review on a diagnostic LCD. The iPad display performed inferiorly to the diagnostic monochrome display without the ability to zoom. When the software zoom function was enabled on the tablet device, comparable contrast detail phantom scores of 163 vs 165 points were achieved. No reporting discrepancies were encountered during the interpretation of 43 normal examinations and five cases of acute intracranial hemorrhage. There were seven RADPEER2 (understandable) misses when using the iPad display and 12 with the diagnostic LCD. Use of software zoom in the tablet device improved its contrast detail phantom score. The tablet allowed satisfactory identification of acute CT brain findings, but additional research will be required to examine the cause of "understandable" reporting discrepancies that occur when using tablet devices.


"Cutting" on cancer: attitudes about cancer spread and surgery among primary care patients in the U.S.A.
Many underserved groups in the United States experience disparities in cancer survival. Part of the disparity may be due to differences in treatment or treatment uptake. Previous studies uncovered patient beliefs that surgery could cause cancer to spread and have suggested that this belief may affect uptake of cancer treatment. We explored patients' explanations about surgical treatment of cancer and cancer spread, as well as the perceived impact on decision-making among primary care patients from an underserved area. Focus groups and interviews were conducted with patients (n = 42) at a primary care federally qualified health center in 2006 and 2007. Focus groups/interviews were semi-structured and were audio-taped and transcribed. An inductive text analysis with multiple coders was used to analyze the data and extract themes. We found that nearly all respondents had heard that surgery ("cutting") and exposing cancer to the air would hasten cancer spread and result in worse outcomes. Most participants expressed agreement with this belief. Many participants said this concern would influence their decision about whether to have surgery and/or reported that a family member had refused surgery for this reason. A smaller group of respondents disagreed with this belief and offered experiential evidence to the contrary or hypotheses about its origination. The idea that "cutting" and "air" during surgery can cause cancer to spread may be more prevalent among patients than suspected, based on this sample of predominantly African American patients. While we were unable to disentangle the ideas about "cutting" from those about "exposure to air", this set of beliefs, when held strongly, can negatively influence patients' or family members' decisions to seek surgical care and, if it is more prevalent in underserved groups, may contribute to cancer disparities.


Nitrogen removal in vermifiltration: Mechanisms, influencing factors, and future research needs.
To meet global health and sanitation goals, there is a continued need for sustainable wastewater treatment alternatives that require minimal energy and investment. Vermifiltration, a technology gaining relevance in Africa and Asia, may be an alternative to traditional wastewater treatment systems due to its cost-effectiveness, ease of application and maintenance, and sustainability. However, nitrogen removal in vermifiltration is not well understood since most of the prior research focuses on organics removal. Thus, a state of the art review is necessary to separately focus on the mechanisms associated with nitrogen removal in vermifiltration, along with the factors affecting nitrogen removal. For the first time, this review attempts to present the types of vermifilter based on their flow pattern. The review further discusses the current status of the application of vermifiltration, along with the benefits and limitations associated with the adoption of this technology. It also explores possible strategies that could be adopted to maximize the nitrogen removal potential of vermifilters as optimizing nitrogen removal is critical for improving the performance of vermifiltration based treatment systems.


Sexual Dimorphism of a Genetic Risk Score for Obesity and Related Traits among Chinese Patients with Type 2 Diabetes.
Obesity is more prevalent in men than in women in China, especially within the middle-aged population. The present study aims to determine the contribution of sexual dimorphisms to obesity and related traits in terms of the mechanisms involving the obesity-related genetic variants among patients of Chinese Han ancestry with type 2 diabetes. In the Chinese National Diabetes and Metabolic Disorders Study, 2,555 out of 4,036 patients with type 2 diabetes were treatment naive, including 1,142 men and 1,413 women. Single-nucleotide polymorphisms (SNP) from 18 genomic loci previously found to be associated with obesity-related traits were successfully genotyped, and a genetic risk score (GRS) was constructed by summing the risk alleles for obesity. Single SNP analysis showed that genetic variants in SLC30A10, TMEM18, GNPDA2, PRL, TFAP2B, BDNF, MTCH2, FTO, and MC4R were nominally associated with waist circumference (WC), BMI, and risk for abdominal or general obesity in the untreated patients with type 2 diabetes, as well as in the total group of patients with type 2 diabetes (untreated and treated) (p < 0.05). Interactions between sex and SNP in PRL, MTCH2,and FTO were detected (p < 0.05). In the untreated patients with diabetes, the GRS was nominally associated with WC (β = 0.0032, SE = 0.0011; p = 0.003), BMI (β = 0.0030, SE = 0.0013; p = 0.027), and increased risk for abdominal (OR = 1.08; 95% CI 1.02-1.13; p = 0.004) or general obesity (OR = 1.07; 95% CI 1.02-1.13; p = 0.011) in men but not in women. GRS-sex interactions were detected in the determinant of WC (p = 0.019) and abdominal obesity (p = 0.016). Among patients aged 30-60 years, GRS was found to be significantly associated with WC (β = 0.0050, SE = 0.0016; p = 0.002) and abdominal obesity (OR = 1.10; 95% CI 1.04-1.17; p = 0.001) and nominally associated with BMI (β = 0.0057, SE = 0.0020; p = 0.005) and general obesity (OR = 1.07; 95% CI 1.01-1.14; p = 0.027) in men, whereas in women none of the associations were detected. GRS-sex interactions were present in the determinant of WC (p = 0.015), BMI (p = 0.032), and abdominal obesity (p = 0.012). Among patients aged 60 years or older, neither an association of GRS with obesity-related traits nor GRS-sex interactions were detected. Genetic factors contribute to obesity-related traits in a sex-dependent pattern among middle-aged Chinese, and men tend to be more susceptible to the genetic risk of obesity.