Brief report: Evaluation of working memory deficits in children with ADHD using the NIH list sorting working memory task.
Variability in working memory (WM) task selection likely contributes to heterogeneity in effect size estimates of deficiencies in youth with attention-deficit/hyperactivity disorder (ADHD). This has resulted in the development of brief, easy to administer assessments such as the NIH List Sorting Working Memory (LSWM) task from the NIH Cognitive Toolbox in hopes of standardizing measurement of this construct. Unfortunately, substantial questions persist regarding the specific constructs being evaluated by this task (e.g., visuospatial [VS] or phonological [PH] WM) as well as the ability of this task to detect WM deficits in previously identified impaired groups (e.g., ADHD). The current study examines the extent to which the LSWM task is associated with VS and PHWM performance as well as symptoms of ADHD. Additionally, we examined the magnitude of differences between ADHD and Typically Developing (TD) youth on this task relative to empirically derived WM tasks utilized in the past. Forty-six children (25 ADHD, 21 TD) completed multiple WM tasks. The LSWM task was moderately associated with PHWM and demonstrated relatively weaker associations with VSWM. Symptoms of inattention and hyperactivity/impulsivity were unrelated to the LSWM task; whereas tasks assessing PH and VSWM were moderately associated with inattention and weakly associated with hyperactivity (VSWM only). No significant between-group differences in performance emerged on the LSWM task; however, significant large-magnitude group differences were observed on both the PH and VSWM tasks. These findings suggest that the LSWM task may lack the ability to detect WM difficulties in youth with ADHD.


Display of native proteins on Bacillus subtilis spores.
In principle, protein display is enabled by fusing target proteins to naturally secreted, surface-anchored protein motifs. In this work, we developed a method of native protein display on the Bacillus spore surface that obviates the need to construct fusion proteins to display a motif. Spore coat proteins are expressed in the mother cell compartment and are subsequently assembled and deposited on the surface of spores. Therefore, target proteins overexpressed in the mother cell compartment during the late sporulation phase were expected to be targeted and displayed on the spore surface. As a proof of principle, we demonstrated the display of carboxymethylcellulase (CMCase) in its native form on the spore surface. The target protein, CMCase, was expressed under the control of the cry1Aa promoter, which is controlled by σ(E) and σ(K) and is expressed in the mother cell compartment. The correct display was confirmed using enzyme activity assays, flow cytometry, and immunogold electron microscopy. In addition, we demonstrated the display of a β-galactosidase tetramer and confirmed its correct display using enzyme activity assays and protein characterization. This native protein display system, combined with the robust nature of Bacillus spores, will broaden the range of displayable target proteins. Consequently, the applications of display technology will be expanded, including high-throughput screening, vaccines, biosensors, biocatalysis, bioremediation, and other innovative bioprocesses.


Corneal Nerve Ablation Abolishes Ocular Immune Privilege by Downregulating CD103 on T Regulatory Cells.
Severing corneal nerves during orthotopic corneal transplantation elicits the elaboration of the neuropeptide substance P (SP), which induces the generation of CD11c+ contrasuppressor (CS) cells. CS cells disable T regulatory cells (Tregs) that are induced when antigens enter the anterior chamber (AC), either by direct injection or by orthotopic corneal transplantation. This study examined the crucial cell surface molecules on Tregs that are adversely affected by CS cells that are generated by severing corneal nerves. CS cells were induced by producing shallow 2.0-mm circular incisions in the corneal epithelium in BALB/c mice. CD8+ Tregs were generated by injecting ovalbumin into the AC. The effects of CS cells and SP on the expression and function of two cell surface molecules (CD103 and the receptor of interferon-γ) that are crucial for the induction and function of CD8+ Tregs were analyzed. SP converted CD11c+, but not CD11c- , dendritic cells (DCs) to CS cells. Severing corneal nerves resulted in a 66% reduction in the expression of CD103 on CD8+ AC-associated immune deviation (ACAID) Tregs, and a 50% reduction in the interferon-γ receptor (IFN-γR). These effects could be mimicked in vitro by coculturing CS cells with CD8+ ACAID Tregs. The elaboration of SP in response to corneal nerve ablation converts CD11c+ DCs to CS cells. CS cells disable CD8+ ACAID Tregs by downregulating two crucial cell surface molecules, CD103 and IFN-γR, by an SP-dependent pathway. Blocking this pathway may provide a means of restoring ocular immune privilege in corneas subjected to corneal nerve injury.


An AfsK/AfsR system involved in the response of aerial mycelium formation to glucose in Streptomyces griseus.
In Streptomyces coelicolor A3(2), a protein serine/threonine kinase (AfsK) and its target protein (AfsR) control secondary metabolism. AfsK and AfsR homologues (AfsK-g and AfsR-g) from Streptomyces griseus showed high end-to-end similarity in amino acid sequence with the respective S. coelicolor A3(2) proteins, as determined by cloning and nucleotide sequencing. AfsK-g and a fusion protein between AfsK-g and thioredoxin (TRX-AfsK-g) produced in high yield as inclusion bodies in Escherichia coli were solubilized with urea, purified by column chromatography and then refolded to an active form by dialysis to gradually remove the urea. AfsR-g was also fused to glutathione S-transferase (GST-AfsR-g); the fusion product in the soluble fraction in E. coli was purified. Incubation of AfsK-g or TRX-AfsK-g in the presence of [gamma-32P]ATP yielded autophosphorylated products containing phosphoserine and phosphothreonine residues. In addition, TRX-AfsK-g phosphorylated serine and threonine residues of GST-AfsR-g in the presence of [gamma-32P]ATP. Disruption of chromosomal afsK-g had no effect on A-factor or streptomycin production, irrespective of the culture conditions. The afsK-g disruptants did not form aerial mycelium or spores on media containing glucose at concentrations higher than 1%, but did form spores on mannitol- and glycerol-containing media; this suggests that afsK-g is essential for morphogenesis in the presence of glucose. Introduction of afsK-g restored aerial mycelium formation in the disruptants. The phenotype of afsR-g disruptants was similar to that of afsK-g disruptants; introduction of afsR-g restored the defect in aerial mycelium formation on glucose-containing medium. Thus the AfsK/AfsR system in S. griseus is conditionally needed for morphological differentiation, whereas in S. coelicolor A3(2) it is conditionally involved in secondary metabolism.


Overexpressing lncRNA SNHG16 inhibited HCC proliferation and chemoresistance by functionally sponging hsa-miR-93.
Long noncoding RNAs (lncRNAs) have been identified as prognostic biomarkers and functional regulators in human cancers. The present study aimed to determine the expressions and functions of an lncRNA, Small Nucleolar RNA Host Gene 16 (SNHG16), in hepatocellular carcinoma (HCC). SNHG16 expressions were tested by quantitative real-time PCR (qRT-PCR) in HCC cell lines, as well as 43 pairs of HCC tissues and pair-matched healthy hepatic tissues. It was overexpressed in Hep3B and HuH7 cells. The effects of SNHG16 overexpression in HCC in vitro proliferation, 5-fluorouracil (5-FU) chemoresistance, and in vivo tumor growth were tested. A potential microRNA (miRNA) sponge target of SNHG16, hsa-miR-93, was tested by luciferase reporter assay and qRT-PCR. In addition, hsa-miR-93 was upregulated in SNHG16-overexpressed HCC cells to examine its effect on SNHG16-mediated cancer cell functional regulation in HCC. SNHG16 levels were markedly downregulated in both HCC cell lines and HCC tissues. Lentivirus-mediated SNHG16 overexpression inhibited HCC cell proliferation, 5-FU chemoresistance, and in vivo tumor growth. Hsa-miR-93 was confirmed to be directly sponging on SNHG16. Its upregulation in HCC cells reversed SNHG16 overexpression and induced tumor-suppressing effects in HCC cells. Our data demonstrate that SNHG16 plays a critical role in HCC development via functionally sponging hsa-miR-93.


Longitudinal mathematics development of students with learning disabilities and students without disabilities: a comparison of linear, quadratic, and piecewise linear mixed effects models.
Effective instructional planning and intervening rely heavily on accurate understanding of students' growth, but relatively few researchers have examined mathematics achievement trajectories, particularly for students with special needs. We applied linear, quadratic, and piecewise linear mixed-effects models to identify the best-fitting model for mathematics development over elementary and middle school and to ascertain differences in growth trajectories of children with learning disabilities relative to their typically developing peers. The analytic sample of 2150 students was drawn from the Early Childhood Longitudinal Study - Kindergarten Cohort, a nationally representative sample of United States children who entered kindergarten in 1998. We first modeled students' mathematics growth via multiple mixed-effects models to determine the best fitting model of 9-year growth and then compared the trajectories of students with and without learning disabilities. Results indicate that the piecewise linear mixed-effects model captured best the functional form of students' mathematics trajectories. In addition, there were substantial achievement gaps between students with learning disabilities and students with no disabilities, and their trajectories differed such that students without disabilities progressed at a higher rate than their peers who had learning disabilities. The results underscore the need for further research to understand how to appropriately model students' mathematics trajectories and the need for attention to mathematics achievement gaps in policy.


Neuroimaging studies of word reading.
This review discusses how neuroimaging can contribute to our understanding of a fundamental aspect of skilled reading: the ability to pronounce a visually presented word. One contribution of neuroimaging is that it provides a tool for localizing brain regions that are active during word reading. To assess the extent to which similar results are obtained across studies, a quantitative review of nine neuroimaging investigations of word reading was conducted. Across these studies, the results converge to reveal a set of areas active during word reading, including left-lateralized regions in occipital and occipitotemporal cortex, the left frontal operculum, bilateral regions within the cerebellum, primary motor cortex, and the superior and middle temporal cortex, and medial regions in the supplementary motor area and anterior cingulate. Beyond localization, the challenge is to use neuroimaging as a tool for understanding how reading is accomplished. Central to this challenge will be the integration of neuroimaging results with information from other methodologies. To illustrate this point, this review will highlight the importance of spelling-to-sound consistency in the transformation from orthographic (word form) to phonological (word sound) representations, and then explore results from three neuroimaging studies in which the spelling-to-sound consistency of the stimuli was deliberately varied. Emphasis is placed on the pattern of activation observed within the left frontal cortex, because the results provide an example of the issues and benefits involved in relating neuroimaging results to behavioral results in normal and brain damaged subjects, and to theoretical models of reading.


Cephalometric Evaluation of Anterior Cranial Base Slope in Patients with Skeletal Class I Malocclusion with Low or High SNA and SNB Angles.
In the cephalometric analyses, it is observed that both SNA and SNB angles are higher or lower than normal for some skeletal Class I patients. The aim of this study was to assess the correlation between low or high SNA, SNB angles, and anterior cranial base (ACB) slope. One hundred and seventeen skeletal Class I patients (45 males with a mean age of 14.5 years, 72 females with a mean age of 14.4 years) were evaluated in three groups. Group 1(n=40): Control group, individuals with normal SNA(82°±2°), and SNB(80°±2°) values. Group 2 (n=37): Patients with SNA>84° and SNB >82°, Group 3 (n=40): Patients with both SNA and SNB values lower than 78°. On the cephalometric radiographs, three angulars (SN/FH; anterior cranial base, Ba-S/FH; posterior cranial base, SN-Ba; total cranial base) and seven linear (S-FH, N-FH, Δ, Ba-S, Ba-N, Ba-A, Ba-B) measurements were performed to analyze the vertical and horizontal positions of the S and N points and thereby the ACB slope. One-way ANOVA and Kruskal Wallis tests were used for statistical analysis. The ACB slope was observed to be relatively flatter in Group 2, and steeper in Group 3 (p<0.05). The location of the S and N points in the sagittal plane did not significantly affect the SNA and SNB. However, the vertical position of the S and N points was a factor determining the inclination of the ACB, therefore the SNA and SNB. ACB slope directly affected SNA and SNB measurements. ACB might lead to misleading results when used as a reference plane.


[Measles in the Italian regions: estimate of infection parameters].
Key epidemiological parameters (forces of infection, contact matrices, reproduction ratios) from the basic SEIR age structured model for childhood infectious diseases are estimated for all Italian regions from pre-vaccination case-notification data. Such parameters allow to summarise the pre-vaccination epidemiology of measles in the Italian regions, particularly the amount of effort needed for the eradication of the disease, consistently with the WHO targets. Despite the limited reliability paid to Italian case notifications data, the results show i) that the estimated eradication coverages are nor distant from the levels estimated from Northern-Europe; ii) that regions seemingly demanding the largest eradication effort seem also to be those characterised, up to now, by the lowest coverages; iii) the importance of achieving high coverages without delays in the age of administration of vaccination.


Effects of dietary glucose and fructose upon cannabinoid CB1 receptor functionality in the rat brain: a pilot study.
A high consumption of fructose leads not only to peripheral changes in insulin sensitivity and vascular function, but also to central changes in several brain regions. Given the role of the endogenous cannabinoid system in the control of energy intake, we undertook a pilot study to determine whether a high fructose diet produced changes in brain CB1 receptor functionality. Male rats given access ad libitum to normal chow were given either water, glucose or fructose solutions to drink. CB1 receptor functionality was measured autoradiographically as the increase in [(35)S]GTPγS binding produced by the agonist CP55,940. Seven regions were investigated: the prefrontal cortex, caudate-putamen, hippocampal CA1-CA3, dentate gyrus, amygdala, and dorsomedial and ventromedial hypothalami. Two-way robust Wilcoxon analyses for each brain region indicated that the dietary treatment did not produce significant main effects upon agonist-stimulated [(35)S]GTPγS binding in any of the regions, in contrast to a significant main effect upon both leptin and adiponectin levels in the blood. However, a MANCOVA of the data controlling for leptin and adiponectin as co-variables identified a significant effect of glucose and fructose treatment for five weeks upon the [(35)S]GTPγS response in the ventromedial hypothalamus, a region of importance for regulation of appetite. It is concluded from this pilot study that palatable solutions do not produce overt changes in brain CB1 receptor functionality, although subtle changes in discrete brain regions may occur.


Viscous food matrix influences absorption and excretion but not metabolism of blackcurrant anthocyanins in rats.
The aim of the present study was to investigate the effect of a simultaneous intake of food and anthocyanins (ACNs) on ACN absorption, metabolism, and excretion. Blackcurrant ACNs (BcACNs) were dissolved in water with or without the addition of oatmeal and orally administered to rats, providing approximately 250 mg total ACNs per kilogram BW. Blood, urine, digesta, and tissue samples of the stomach, jejunum, and colon were subsequently collected at 0.25, 0.5, 1, 2, 3, 7, and 24 h. Identification and quantification of ACNs were carried out by Reversed phase-high-performance liquid chromatography (RP-HPLC) and liquid chromatography-mass spectrometry (LC-MS). Four major ACNs were present in the blackcurrant extract: delphinidin 3-O-glucoside, delphinidin 3-O-rutinoside, cyanidin 3-O-glucoside, and cyanidin 3-O-rutinoside. In plasma, the 4 ACNs of blackcurrant were identified and quantified. The time to reach maximal total ACN plasma concentration (C(max) BcACN/water = 0.37 +/- 0.07 micromol/L; C(max) BcACN/oatmeal = 0.20 +/- 0.05 micromol/L) occurred faster after BcACN/water (t(max)= 0.25 h), than after BcACN/oatmeal administration (t(max)= 1.0 h). In digesta and tissue samples, the 4 original blackcurrant ACNs were detected. The relative concentration of rutinosides in the digesta increased during their passage through the gastrointestinal tract, while the glucosides decreased. Maximum ACN excretion in urine occurred later after BcACN/oatmeal than after BcACN/water administration (3 compared with 2 h). The 4 original ACNs of blackcurrant in their unchanged form, as well as several metabolites, were identified in the urine samples of both groups. The simultaneous intake of food affects ACN absorption and excretion in the urine, but not metabolism.


A novel model of highly lethal uncontrolled torso hemorrhage in swine.
A reproducible, lethal noncompressible torso hemorrhage model is important to civilian and military trauma research. Current large animal models balancing clinical applicability with standardization and internal validity. As such, large animal models of trauma vary widely in the surgical literature, limiting comparisons. Our aim was to create and validate a porcine model of uncontrolled hemorrhage that maximizes reproducibility and standardization. Seven Yorkshire-cross swine were anesthetized, instrumented, and splenectomized. A simple liver tourniquet was applied before injury to prevent unregulated hemorrhage while creating a traumatic amputation of 30% of the liver. Release of the tourniquet and rapid abdominal closure following injury provided a standardized reference point for the onset and duration of uncontrolled hemorrhage. At the moment of death, the liver tourniquet was quickly reapplied to provide accurate quantification of intra-abdominal blood loss. Weight and volume of the resected and residual liver segments were measured. Hemodynamic parameters were recorded continuously throughout each experiment. This liver injury was rapidly and universally lethal (11.2 ± 4.9 min). The volume of hemorrhage (35.8% ± 6% of total blood volume) and severity of uncontrolled hemorrhage (100% of animals deteriorated to a sustained mean arterial pressure <35 mmHg for 5 min) were consistent across all animals. Use of the tourniquet effectively halted preprocedure and postprocedure blood loss allowing for accurate quantification of amount of hemorrhage over a defined period. In addition, the tourniquet facilitated the creation of a consistent liver resection weight (0.0043 ± 0.0003 liver resection weight: body weight) and as a percentage of total liver resection weight (27% ± 2.2%). This novel tourniquet-assisted noncompressible torso hemorrhage model creates a standardized, reproducible, highly lethal, and clinically applicable injury in swine. Use of the tourniquet allowed for consistent liver injury and precise control over hemorrhage. Recorded blood loss was similar across all animals. Improving reproducibility and standardization has the potential to offer improvements in large animal translational models of hemorrhage. Level I.


Binder syndrome.
Patients with chondrodysplasia punctata (CDP) usually present with Binder-type features, and often CDP is misdiagnosed as Binder syndrome. This study reviewed the management and outcome of patients with Binder syndrome and CDP in a multidisciplinary setting. The notes and radiographs of the patients managed at the Australian Craniofacial Unit with a multidisciplinary setting since 1976 were reviewed, and data were collected on patient demographics, associated medical and surgical problems, subsequent management, and complications. Seventy-seven patients were treated over the 30-year period (5 patients were lost to follow-up); of the remaining 72 patients, 60 (83%) had Binder syndrome, and 12 (17%) were patients with CDP. Forty were males, and 32 were females, with an age range of 6 months to 47 years. Thirteen patients (18%) had a strong family history, and 65 patients (90%) have so far undergone surgical correction, and of those, 35 (54%) have completed their treatment, the longest follow-up time being 18 years. The mean number of surgical procedures was 2.4, and 18 patients (28%) had postoperative complications, which included partial necrosis of the maxilla, osteomyelitis of the mandible, facial nerve and inferior alveolar nerve neuropraxia, nasal bone graft exposure, and cellulitis. Because of the phenotypic characteristics shared by both Binder syndrome and CDP, it is most likely that Binder syndrome is not a syndrome, nor is it an entity, but most likely to be an "association." We would advocate that these patients should be managed in a multidisciplinary setting.


Semi-Supervised Generative Adversarial Nets with Multiple Generators for SAR Image Recognition.
As an important model of deep learning, semi-supervised learning models are based on Generative Adversarial Nets (GANs) and have achieved a competitive performance on standard optical images. However, the training of GANs becomes unstable when they are applied to SAR images, which reduces the feature extraction capability of the discriminator in GANs. This paper presents a new semi-supervised GANs with Multiple generators and a classifier (MCGAN). This model improves the stability of training for SAR images by employing multiple generators. A multi-classifier is introduced to the new GANs to utilize the labeled images during the training of the GANs, which shares the low level layers with the discriminator. Then, the layers of the trained discriminator and the classifier construct the recognition network for SAR images after having been finely tuned using a small number of the labeled images. Experiments on the Moving and Stationary Target Acquisition and Recognition (MSTAR) databases show that the proposed recognition network achieves a better and more stable recognition performance than several traditional semi-supervised methods as well as other GANs-based semi-supervised methods.


Localization of a pyrroloquinoline quinone biosynthesis gene near the methanol dehydrogenase structural gene in Methylobacterium organophilum DSM 760.
A partial Sau3AI genomic bank of Methylobacterium organophilum DSM 760 was constructed in the cosmid pSUP106 and moxF, the structural gene for methanol dehydrogenase, was isolated. In M. organophilum, pSUP106 behaves as a suicide plasmid. This property was used to insert Tn5 into the bacterial chromosome, in the vicinity of moxF, by marker exchange. Mobilization of the Tn5-labelled chromosomal region by a broad-host-range plasmid, pJB3J1 (an R68-45 derivative), allowed the selection of several large R' hybrid plasmids in Escherichia coli HB101. Most of them were able to complement both mutants of the moxF region and mutant MTM1, the first mutant of the pyrroloquinoline quinone (PQQ) biosynthesis pathway in M. organophilum. The gene involved, pqqA, was subcloned and localized.


Instability restricts signaling of multiple fibroblast growth factors.
Fibroblast growth factors (FGFs) deliver extracellular signals that govern many developmental and regenerative processes, but the mechanisms regulating FGF signaling remain incompletely understood. Here, we explored the relationship between intrinsic stability of FGF proteins and their biological activity for all 18 members of the FGF family. We report that FGF1, FGF3, FGF4, FGF6, FGF8, FGF9, FGF10, FGF16, FGF17, FGF18, FGF20, and FGF22 exist as unstable proteins, which are rapidly degraded in cell cultivation media. Biological activity of FGF1, FGF3, FGF4, FGF6, FGF8, FGF10, FGF16, FGF17, and FGF20 is limited by their instability, manifesting as failure to activate FGF receptor signal transduction over long periods of time, and influence specific cell behavior in vitro and in vivo. Stabilization via exogenous heparin binding, introduction of stabilizing mutations or lowering the cell cultivation temperature rescues signaling of unstable FGFs. Thus, the intrinsic ligand instability is an important elementary level of regulation in the FGF signaling system.


Hair cell regeneration: the identities of progenitor cells, potential triggers and instructive cues.
Hair cells are produced and accumulate in the ears of fish and amphibians as they grow during postembryonic life; hair cell regeneration occurs in lateral line organs in those groups and in the cochlea in birds. Continuous time-lapse microscopy has directly demonstrated that supporting cells divide to give rise to hair cells during regeneration in lateral line neuromasts. Supporting cells also appear to give rise to hair cells during regeneration in the avian ear, but additional cell types have been proposed as hair cell progenitors. Alternative interpretations of current evidence are discussed in relation to the possibility that supporting cells may be the common progenitor in all cases of hair cell regeneration. The regenerative proliferation of hair cells in birds occurs in populations of cells that are mitotically quiescent in undamaged ears. Evidence suggests that the extrusion of damaged hair cells and the breaking of intercellular junctional adhesions may be a trigger for regenerative proliferation. The potential triggering influence of phagocytes is also discussed. The differentiation of replacement cells during regeneration in the cochlea may be regulated by surface interactions between cells. A model that could account for the reconstitution of the mosaic pattern of hair cells and supporting cells is proposed.


Primary non-Hodgkin's lymphoma of the liver (case report).
Primary non-Hodgkin's lymphoma of the liver is very uncommon, and fewer than 100 cases have been reported in the literature. Most reports describe either solitary or multiple mass lesions in the liver. A diffuse lesion without nodule formation is a relatively rare form of the disease. The histologic feature of the disease is a predominantly large cell lymphoma of B-cell lineage. We report a case of primary B-cell non-Hodgkin's lymphoma which had diffusely infiltrated the liver without nodule formation.


Nucleoside exchange catalysed by the cytoplasmic 5'-nucleotidase.
The inhibition of the cytoplasmic 5'-nucleotidase (EC 188.8.131.52) by its product, inosine, was studied with a partially purified preparation of the enzyme from rat liver. Inhibition of Pi production was found to be due to exchange of the inosine moiety between inosine and IMP. Exchange was not catalysed by reversal of the hydrolytic reaction, suggesting, instead, the mediation of an enzyme-phosphate intermediate. Two models for the catalytic mechanism are proposed and rate equations for the dependence of Pi production on inosine concentration are derived. The experimentally determined dependence was consistent with a mechanism in which hydrolysis of the enzyme-phosphate intermediate occurred only when it was unoccupied by inosine. This conclusion suggests that inosine analogues that cannot participate in exchange should inhibit the enzyme. Such inhibitors might be useful in defining the enzyme's physiological role or as pharmacological agents to decrease breakdown of purine nucleotides. The possibility that nucleoside exchange provides an alternative route for the phosphorylation of mutagenic or cytotoxic nucleoside analogues should also be considered.


Long-term renal outcome in patients with malignant hypertension: a retrospective cohort study.
Malignant hypertension is frequently complicated by renal insufficiency. Although the survival of this hypertensive emergency has improved, recent data on renal outcome and its predictors are lacking. We assessed renal outcome and its predictors in patients with malignant hypertension. Retrospective analysis of patients admitted with malignant hypertension in Amsterdam, the Netherlands between August 1992-January 2010. Follow-up data on vital status, renal function and blood pressure (BP) were obtained from the outpatient department and from general practitioners. The primary composite endpoint was end-stage renal disease (ESRD) defined as the start of kidney replacement therapy (KRT) or ≥ 50% decline of estimated glomerular filtration rate (eGFR). The secondary endpoint was all cause mortality. A total of 120 patients admitted with malignant hypertension were included. After a median follow-up period of 67 months (IQR 28 to 108 months) the primary endpoint was reached by 37 (31%) patients, whereas 18 patients (15%) reached the secondary endpoint. Twenty-nine (24%) patients started KRT and 8 patients (7%) had an eGFR decline ≥ 50%. After the acute phase (> 3 months after admission), initial serum creatinine and follow-up BP were the main predictors of future ESRD with hazard ratios of 6.1 (95% CI, 2.2-17) for patients with initial serum creatinine ≥ 175 μmol /L and 4.3 (95% CI, 1.4-14) for patients with uncontrolled hypertension. Progressive renal function decline leading to ESRD remains a major threat to patients with malignant hypertension. BP control during follow-up was an important modifiable predictor of renal outcome.


Recent progress in clinical quantitative cytology.
The last several years have seen changes in quantitative cytologic procedures that have significantly expanded the clinical research and diagnostic applications of quantitative cytologic analysis. Intact nuclei can be isolated from formalin-fixed paraffin-embedded tissue, and the DNA distribution of these nuclei can be analyzed either with flow cytometry or with slide-based image-processing techniques. In addition, more informative DNA analysis has been achieved using antibodies to bromodeoxyuridine-modified DNA, proliferation-associated nuclear antigens, or tissue-type-specific cytoplasmic antigens as second stains for two-parameter analysis. Improvements in both staining and instrumentation have also increased the usefulness of flow cytometry in analysis of "rare events," ie, cell types comprising less than 0.1% to 0.5% of a total cell suspension. Commercially available slide-based image-processing systems have been developed that provide sophisticated cell and tissue analysis functions using inexpensive personal-computer-based systems.


Safety, tolerability, and efficacy of orally administered atogepant for the prevention of episodic migraine in adults: a double-blind, randomised phase 2b/3 trial.
Atogepant is an orally administered, small-molecule, calcitonin gene-related peptide (CGRP) receptor antagonist under investigation for treatment of migraine. We aimed to examine a range of oral doses for safety, tolerability, and efficacy for the preventive treatment of migraine. In this double-blind, phase 2b/3 trial, adults (aged 18-75 years), with a history (≥1 year) of migraine and 4-14 migraine days per month, were randomly assigned 2:1:2:2:1:1 (by means of a sequence generated by the statistical programming department of the sponsor, and operationalised through an automated interactive web-based response system) to receive placebo or atogepant 10 mg once daily, 30 mg once daily, 60 mg once daily, 30 mg twice daily, or 60 mg twice daily, in matching capsules. Participants, site personnel, and all study sponsor personnel were masked to treatment allocations. The study was done in 78 academic and private practice settings in the USA. The primary outcome was change from baseline in monthly migraine days across 12 weeks of treatment using a modified intention-to-treat approach. The overall type I error rate for multiple comparisons across active treatment doses was controlled at the 0·05 level by means of a graphic approach. The main outcomes to assess safety and tolerability were adverse event recordings. The trial is registered with ClinicalTrials.gov, NCT02848326 and is completed. Between Sept 6, 2016, and April 23, 2018, of 1772 individuals screened, 834 were randomly assigned and 825 received one dose or more of study medication: 186 received placebo, 93 atogepant 10 mg once daily, 183 atogepant 30 mg once daily, 186 atogepant 60 mg once daily, 86 atogepant 30 mg twice daily, and 91 atogepant 60 mg twice daily. Overall, 714 (87%) of 825 participants were female, 628 (76%) were white, median migraine duration was 17·5 years (IQR 10·0-28·0), and 232 (28%) had previously used preventive treatment. The primary efficacy analysis included 795 patients: 178 received placebo, 92 atogepant 10 mg once daily, 182 atogepant 30 mg once daily, 177 atogepant 60 mg once daily, 79 atogepant 30 mg twice daily, and 87 atogepant 60 mg twice daily. Across the 12-week treatment period, all five atogepant groups showed significant least-squares mean (SE) change from baseline in mean monthly migraine days versus placebo: atogepant 10 mg once daily -4·0 (0·3; p=0·024), 30 mg once daily -3·8 (0·2; p=0·039), 60 mg once daily -3·6 (0·2; p=0·039), 30 mg twice daily -4·2 (0·4; p=0·0034), and 60 mg twice daily -4·1 (0·3; p=0·0031); placebo -2·9 (0·2). The most common treatment-emergent adverse events (TEAEs) across all groups were nausea (range 5% [5/93] for 10 mg once daily to 12% [22/186] for 60 mg once daily vs 5% [9/186] for placebo) and fatigue (1% [1/93] for 10 mg once daily to 10% [9/91] for 60 mg twice daily vs 3% [6/186] for placebo). Treatment-related TEAE frequency ranged from 18% (17/93) for 10 mg once daily to 26% (24/91) for 60 mg twice daily, versus 16% (30/186) for placebo. Seven participants reported a total of eight serious TEAEs (two participants each in the placebo, 30 mg once-daily, and 60 mg once-daily groups, and one participant in the 10 mg once-daily group). TEAEs leading to discontinuation were reported in 33 (5%) of 639 atogepant participants and 5 (3%) of 186 of those randomised to placebo. All serious TEAEs were unrelated to treatment. All doses of oral atogepant were associated with a significant decrease in monthly migraine days over 12 weeks compared with placebo. Atogepant was safe and well tolerated over 12 weeks, supporting its phase 3 development for the preventive treatment of migraine. Allergan (before its acquisition by AbbVie).


[Osteonecrosis of the mandible induced by intravenous bisphosphonates in oncological patients].
Bisphosphonates are synthetic analogs of inorganic pyrophosphate, which are not only potent inhibitors of osteoclast-mediated bone resorption, but also present an important inhibitory activity of angiogenesis. Bisphosphonates become, since 1996, the standard of care in the management of patients with osteoporoses as well as bone metabolism alterations associated with neoplasias. In 2003 the first case-reports of bisphosphonate induced osteonecrosis of the jaws were published. The authors report 2 new cases of patients with osteonecrosis induced by bisphosphonates. Case 1: Seventy one years-old male patient, undergoing standard antineoplastic therapy and bisphosphonates due to multiple myeloma 4 years ago was referred to an oral surgeon because of bone exposure, after dental extraction. Case 2: Seventy years-old female patient undergoing treatment with bisphosphonates for adenocarcinoma of the breast was referred to an appointment of oral surgery due to bony exposure. The increasing incidence of this new entity, its physiopathological mechanisms and the nonexistence of consensual and evidence-based treatments, calls for a special attention on prevention, with special emphasis on a careful clinical examination of the oral cavity previously to the establishment of a therapeutic approach with intravenous bisphosphonates in oncological patients.


The gut microbiome and its potential role in paradoxical anaerobism in pupfishes of the Mojave Desert.
Pupfishes frequently enter paradoxical anaerobism in response to endogenously produced or exogenously supplied ethanol in a dose-dependent manner. To decipher the role of the gut microbiota in ethanol-associated paradoxical anaerobism, gut microbial communities were depleted using a cocktail of antibiotics and profiled using 16S rRNA gene sequencing. Compared to the control group (n = 12), microbiota-depleted fish (n = 12) spent more time in paradoxical anaerobism. Our analysis indicated that the bacterial phyla Proteobacteria, Fusobacteria, Bacteroidetes, Firmicutes, Actinobacteria, Patescibacteria, and Dependentiae dominated the pupfish gut, which is consistent with other fish gut microbiota. Although the gut microbial communities with and without antibiotic treatment were similarly diverse, they were distinct and the greatest contribution to the dissimilarity (27.38%) was the common fish commensal Cetobacterium. This study reports the first characterization of gut microbial communities of pupfish and suggests the microbiome may play a critical role in regulating metabolic strategies that are critical for survival in extremes of temperature and oxygen concentration. We speculate that Cetobacterium, a primary fermenter, also consumes ethanol through secondary fermentation via an alcohol dehydrogenase and therefore regulates the transition from paradoxical anaerobism to aerobic respiration in fish. Given the wide distribution and abundance of Cetobacterium in warm-water fishes, this process may be of broad importance, and suggests that the microbiome be carefully considered for both conservation and aquaculture.


A qualitative analysis of patients' experiences with an emergency department diagnosis of gastrointestinal cancer.
Optimally, cancer is diagnosed through periodic screening or detection of early symptoms in primary care settings. However, an estimated 23%-52% of gastrointestinal (GI) cancers are diagnosed in the emergency department (ED). Cancer diagnosed in the ED has been associated with worse clinical and patient-reported outcomes even after adjustment for cancer stage. We sought to explore patients' accounts of patient and health care system factors related to their diagnosis in the ED and their lived experience of receiving a diagnosis in this setting. Patients with an ED visit during or within 30 days of their GI cancer diagnosis at an urban academic hospital serving a largely disadvantaged population were recruited. Interviews were coded in NVivo 12 and analyzed using a thematic analysis approach. Patient-reported factors associated with their experiences included denial and avoidance of symptoms, mistrust of the health system, and lack of cancer screening knowledge. Health care system factors included misdiagnosis and delayed access to specialty care or tests. Experiences receiving a cancer diagnosis in the ED were overwhelmingly negative. This study highlights the unmet needs in identifying and diagnosing patients who ultimately present to the ED for evaluation and eventual diagnosis of cancer. Our results shed light on several modifiable factors, including the need for increased public awareness of the asymptomatic nature of cancer and the importance of cancer screening. Additionally, health care systems modifications beyond the ED are needed to improve access to timely care when symptoms arise.


An overview cost-utility analysis of prostate cancer screening.
The value of prostate cancer screening remains controversial because of the high prevalence of the disease and the fact that many tumors detected through screening are not destined to lead to morbidity or mortality, rendering treatment unnecessary. An ongoing NCI-sponsored screening trial may eventually put an end to the controversy. However, in the meantime, cost-utility estimates suggest that the cost per crude and quality-adjusted life year gained from prostate cancer screening and treatment ranges from $8,400 to $23,100 (with an estimated 1 to 2.68 QALYs gained from screening and treatment), and that these estimates are actually lower than, or well within the range of, the costs of many commonly accepted medical interventions, including screening mammography in women under age 50 ($232,000) and treatment of hypertension with captopril ($82,600) or hydrochlorothiazide ($23,500). Thus, we conclude that prostate cancer screening may indeed be cost effective and should be offered to men in the at-risk age range.


Observational and genetic plasma YKL-40 and cancer in 96,099 individuals from the general population.
Plasma YKL-40 is high in patients with cancer and in individuals who later develop cancer. Whether YKL-40 is only a marker or indeed a cause of cancer is presently unknown. We tested the hypothesis that observationally and genetically, high plasma YKL-40 is associated with high risk of cancer. For this purpose, we performed cohort and Mendelian randomization studies in 96,099 individuals from the Danish general population. Plasma levels of YKL-40 were measured in 21,643 and CHI3L1 rs4950928 was genotyped in 94,568 individuals. From 1943 through 2011, 2,291 individuals developed gastrointestinal cancer, 913 developed lung cancer, 2,863 women developed breast cancer, 1,557 men developed prostate cancer and 5,146 individuals developed other cancer. Follow-up was 100% complete. Multifactorially and CRP adjusted hazard ratio (HR) for gastrointestinal cancer was 1.82 (95%CI, 1.16-2.86) for 96-100% versus 0-33% YKL-40 percentile category. Corresponding HR were 1.71 (0.95-3.07) for lung cancer, but insignificant for breast cancer, prostate cancer and other cancers. CHI3L1 rs4950928 genotype was associated with plasmaYKL-40 levels, but not with risk of any cancer category. For gastrointestinal cancer, a doubling in YKL-40 was associated with a multifactorially and CRP adjusted observational HR of 1.14(1.05-1.23) for gastrointestinal cancer, but a corresponding genetic odds ratio of 1.06(0.94-1.18). For lung cancer, corresponding risk estimates were 1.11(1.00-1.22) observationally and 1.01(0.84-1.20) genetically. For other cancer categories, observational and genetic findings were insignificant. This study shows that high plasma YKL-40 levels were associated with high risk of gastrointestinal and likely of lung cancer, but genetic high levels were not.


High-dose cyclophosphamide, carmustine, and etoposide followed by autologous peripheral stem cell transplantation for patients with relapsed Hodgkin's disease.
Between February 1986 and March 1990, 56 patients with relapsed Hodgkin's disease treated with high-dose cyclophosphamide, carmustine, and etoposide (CBV) received an autologous peripheral stem cell transplantation (PSCT) rather than an autologous bone marrow transplantation (ABMT) because each patient had a marrow abnormality, either hypocellularity or tumor involvement. At least 6.5 x 10(8) [corrected] mononuclear cells/kg patient weight were collected from the peripheral blood of each patient, cyropreserved, and returned intravenously following CBV administration. Three patients had an early death 2, 22, and 25 days after PSCT. The actuarial event-free survival for these 56 patients at 3 years was 37% and was at least as good as that reported for relapsed Hodgkin's disease patients treated with CBV and ABMT. The 30 patients who had no marrow metastases at the time of PSC harvesting had an actuarial event-free survival of 47%, while those 26 patients with marrow metastases had a significantly different actuarial event-free survival of 27% (P = .02). CBV and PSCT for patients with relapsed Hodgkin's diseases who have marrow hypocellularity in traditional harvest sites or histopathologic evidence of BM metastases can result in long-term event-free survival.


KChIP-like auxiliary subunits of Kv4 channels regulate excitability of muscle cells and control male turning behavior during mating in Caenorhabditis elegans.
Voltage-gated Kv4 channels control the excitability of neurons and cardiac myocytes by conducting rapidly activating-inactivating currents. The function of Kv4 channels is profoundly modulated by K(+) channel interacting protein (KChIP) soluble auxiliary subunits. However, the in vivo mechanism of the modulation is not fully understood. Here, we identified three C. elegans KChIP-like (ceKChIP) proteins, NCS-4, NCS-5, and NCS-7. All three ceKChIPs alter electrical characteristics of SHL-1, a C. elegans Kv4 channel ortholog, currents by slowing down inactivation kinetics and shifting voltage dependence of activation to more hyperpolarizing potentials. Native SHL-1 current is completely abolished in cultured myocytes of Triple KO worms in which all three ceKChIP genes are deleted. Reexpression of NCS-4 partially restored expression of functional SHL-1 channels, whereas NCS-4(efm), a NCS-4 mutant with impaired Ca(2+)-binding ability, only enhanced expression of SHL-1 proteins, but failed to transport them from the Golgi apparatus to the cell membrane in body wall muscles of Triple KO worms. Moreover, translational reporter revealed that NCS-4 assembles with SHL-1 K(+) channels in male diagonal muscles. Deletion of either ncs-4 or shl-1 significantly impairs male turning, a behavior controlled by diagonal muscles during mating. The phenotype of the ncs-4 null mutant could be rescued by reexpression of NCS-4, but not NCS-4(efm), further emphasizing the importance of Ca(2+) binding to ceKChIPs in regulating native SHL-1 channel function. Together, these data reveal an evolutionarily conserved mechanism underlying the regulation of Kv4 channels by KChIPs and unravel critical roles of ceKChIPs in regulating muscle cell excitability and animal behavior in C. elegans.


Parental and health care provider understanding of childhood fever: a Canadian perspective.
Fever is common in children and causes misconceptions among parents. Many investigators have called for improved parental education to dispel "fever phobia." Our objectives were to assess parental and health care provider understanding of fever, its treatment, and beliefs about its consequences, as well as to identify parental sources of information about fever. Self-administered surveys were distributed to 3 parent groups and 4 health care provider groups. Parent groups included parents of children with fever presenting to the emergency department (ED) (fever group, n = 209), parents of children with an injury presenting to ED (injury group, n = 160), and parents of healthy school children (school group, n = 141). Provider groups included pediatric ED physicians (n = 16), pediatric ED nurses (n = 39), general pediatricians (n = 26) and family physicians (n = 79). Parent groups considered a temperature of 37.9 degrees C to be a fever, 39.1 degrees C to be a high fever, and 39.9 degrees C to be a dangerous fever. Parents were most concerned about discomfort, seizures and dehydration, and parents in the "fever group" worried more about dehydration (p = 0.01) and brain damage (p = 0.03) than other parents. Most physicians were concerned about dehydration and seizures, but family physicians were most likely to express concerns about brain damage (40.5%) and death (34.1%). Fever phobia exists among parents and health care providers and is most likely in parents of febrile children and family physicians. Health care providers varied in their knowledge of fever and its treatment. Greater education of health care workers is required in order to provide families with appropriate information.


Prominence of beta 2-microglobulin, class I heavy chain conformation, and tapasin in the interactions of class I heavy chain with calreticulin and the transporter associated with antigen processing.
Newly synthesized class I heavy (H) chain/beta 2m heterodimers awaiting peptides in the endoplasmic reticulum are associated with the transporter associated with Ag processing (TAP). We present evidence here that calreticulin, but not calnexin, displays steady state association with class I/TAP complexes. To separate the ability of beta 2m to bind with TAP and calreticulin from that of H chain, we studied human cell lines that lack expression of beta 2m or H chain. Little if any H chain was detected in association with TAP and calreticulin in the beta 2m- cell line Daudi. By contrast, high levels of beta 2m are found with TAP and calreticulin in the H chain-deficient cell line LCL 721.221, even after preclearance of the trace amount of class IB protein expressed by this cell line. Thus, beta 2m appears to bind TAP in the absence of H chain, providing an elegant mechanism to retain beta 2m in the endoplasmic reticulum at the site of peptide loading. To investigate whether other molecules participate in the binding of beta 2m and H chain to TAP and calreticulin, we analyzed the deletion mutant cell line LCL 721.220, which lacks tapasin. In 721.220, TAP and calreticulin are not associated with each other. Also, in these cells, H chain/beta 2m are not associated with TAP, but H chain and a low level of beta 2m are associated with calreticulin. These results suggest that tapasin is an obligatory mediator of the assemblage of calreticulin/H chain/beta 2m with TAP.


Validation of the EQ-5D in Taiwan using item response theory.
Our study aims to provide validity evidence for the EuroQol five dimensions questionnaire (EQ-5D) in the National Health Interview Survey of Taiwan in the 2013 wave and further interpret the EQ-5D scores for patients with chronic diseases. Another goal of the study was to use item response theory (IRT) to identify items that are informative for assessing quality of life using EQ-5D. There were 17,260 participants, aged 12-64, who completed the interviews in our study. Psychometric methods, including factor analysis and the IRT model known as the Graded Response Model (GRM), were used to assess the unidimensionality of EQ-5D and its item properties. Correlation analysis was used to assess whether EQ-5D scores are associated with scores from the 36-Item Short Form Survey (SF-36). The EQ-5D scores have moderate internal consistency (Cronbach's alpha: 0.60) and a scree plot suggests that the EQ-5D measure is unidimensional. The item information function analysis from the IRT model demonstrates that the first 3 items, "mobility," "self-care," and "usual activities" are the most informative items for patients who have chronic diseases and health-related quality of life below the 10th percentile. The EQ-5D scores have a moderate correlation (r: 0.61) with SF-36 scores. The EQ-5D scale shows promise for use in the general population. The IRT model informs our interpretation of the EQ-5D scores. Given the time constraints in clinical settings, we suggest using the first three items in EQ-5D to measure the health-related quality of life for patients with chronic diseases.


Solutol and cremophor products as new additives in suppository formulation.
Our research has a double purpose. On the one hand, doctors have expressed the need to formulate a rectal suppository dosage form from diuretic ethacrynic acid, which would add to the choice of treatment methods and thereby increase the possibilities of individual cure. On the other hand, the liberation and thereby the bioavailability of poorly-soluble ethacrynic acid needs to be enhanced, and for this purpose solubility-increasing additives new to rectal therapy were used. Solutol HS 15, Cremophor RH 40, and Cremophor RH 60 were used as additives in concentrations of 1, 3, 5, and 10%. The quantity of drug released changed as a function of additive concentration. Depending on the acceptor phase, the best results were achieved with an additive concentration of 1-3%, which is related to the optimal additive quantity accumulated on the boundary surface.


Using discrete choice experiments to understand preferences for quality of life. Variance-scale heterogeneity matters.
Health services researchers are increasingly using discrete choice experiments (DCEs) to model a latent variable, be it health, health-related quality of life or utility. Unfortunately it is not widely recognised that failure to model variance heterogeneity correctly leads to bias in the point estimates. This paper compares variance heterogeneity latent class models with traditional multinomial logistic (MNL) regression models. Using the ICECAP-O quality of life instrument which was designed to provide a set of preference-based general quality of life tariffs for the UK population aged 65+, it demonstrates that there is both mean and variance heterogeneity in preferences for quality of life, which covariate-adjusted MNL is incapable of separating. Two policy-relevant mean groups were found: one group that particularly disliked impairments to independence was dominated by females living alone (typically widows). Males who live alone (often widowers) did not display a preference for independence, but instead showed a strong aversion to social isolation, as did older people (of either sex) who lived with a spouse. Approximately 6-10% of respondents can be classified into a third group that often misunderstood the task. Having a qualification of any type and higher quality of life was associated with smaller random component variances. This illustrates how better understanding of random utility theory enables richer inferences to be drawn from discrete choice experiments. The methods have relevance for all health studies using discrete choice tasks to make inferences about a latent scale, particular QALY valuation exercises that use DCEs, best-worst scaling and ranking tasks.


Predictive biomarkers with potential of converting conventional chemotherapy to targeted therapy in patients with metastatic colorectal cancer.
The availability of systemic chemotherapy regimens for the treatment of patients with metastatic colorectal cancer (mCRC) is based on the results from large prospective, randomized studies. The main chemotherapeutic drugs used in treatment of mCRC are the fluoropyrimidines (5-fluorouracil (5-FU); capecitabine) in combination with either oxaliplatin (FOLFOX) or irinotecan (FOLFIRI). The objective response rate to either combination is approximately 50%, where no significant differences with regard to progression free survival or overall survival have been observed. Interestingly, a number of preclinical and clinical studies have indicated lack of full cross resistance between oxaliplatin based and irinotecan based treatment. Therefore, it is possible that certain mCRC patient subpopulations would benefit more from one drug combination rather than the other. To address this clinical problem there has been much focus on development and validation of predictive biomarkers for these three drugs. Here, we present a thorough review on the current status of predictive biomarkers for 5-FU, oxaliplatin and irinotecan treatment of mCRC patients. The overall conclusions were as follows: Several promising biomarker candidates were identified, notably thymidylate synthase for 5-FU, topoisomerase I for irinotecan and ERCC1 for oxaliplatin. However, these candidates warrant further analysis, where assay performance and clinical trial design should be in focus.


Avascular necrosis of the talus after McKay clubfoot release for idiopathic congenital clubfoot.
Avascular necrosis of the talus is a serious potential complication of clubfoot surgery. In the few cases described in the literature, the necrosis has involved the entire talus and resulted in progressive fragmentation and collapse. Serial postoperative radiographs of 96 idiopathic clubfeet in 70 patients are reviewed here to determine the incidence of avascular necrosis after McKay soft tissue release. Based on criteria in the literature for making the diagnosis, no cases of avascular necrosis were seen. Growth lines were observed in the cuboids and calcanei of all the feet during the follow-up period. Eight feet failed to develop growth lines in the talus during follow-up. Five of these feet showed flattening of the dome of the talus and three hypoplasia of the talar head and neck at the most recent follow-up. Absence of normal growth lines in the talus after operation seems to predict talar abnormalities.


Expressed sequence tag analysis of the response of apple (Malus x domestica'Royal Gala') to low temperature and water deficit.
Leaf, bark, xylem and root tissues were used to make nine cDNA libraries from non-stressed (control) 'Royal Gala' apple trees, and from 'Royal Gala' trees exposed to either low temperature (5 degrees C for 24 h) or water deficit (45% of saturated pot mass for 2 weeks). Over 22 600 clones from the nine libraries were subjected to 5' single-pass sequencing, clustered and annotated using blastx. The number of clusters in the libraries ranged from 170 to 1430. Regarding annotation of the sequences, blastx analysis indicated that within the libraries 65-72% of the clones had a high similarity to known function genes, 6-15% had no functional assignment and 15-26% were completely novel. The expressed sequence tags were combined into three classes (control, low-temperature and water deficit) and the annotated genes in each class were placed into 1 of 10 different functional categories. The percentage of genes falling into each category was then calculated. This analysis indicated a distinct downregulation of genes involved in general metabolism and photosynthesis, while a significant increase in defense/stress-related genes, protein metabolism and energy was observed. In particular, there was a three-fold increase in the number of stress genes observed in the water deficit libraries indicating a major shift in gene expression in response to a chronic stress. The number of stress genes in response to low temperature, although elevated, was much less than the water deficit libraries perhaps reflecting the shorter (24 h) exposure to stress. Genes with greater than five clones in any specific library were identified and, based on the number of clones obtained, the fold increase or decrease in expression in the libraries was calculated and verified by semiquantitative polymerase chain reaction. Genes, of particular note, that code for the following proteins were overexpressed in the low-temperature libraries: dehydrin and metallothionein-like proteins, ubiquitin proteins, a dormancy-associated protein, a plasma membrane intrinsic protein and an RNA-binding protein. Genes that were upregulated in the water deficit libraries fell mainly into the functional categories of stress (heat shock proteins, dehydrins) and photosynthesis. With few exceptions, the overall differences in downregulated genes were nominal compared with differences in upregulated genes. The results of this apple study are similar to other global studies of plant response to stress but offer a more detailed analysis of specific tissue response (bark vs xylem vs leaf vs root) and a comparison between an acute stress (24-h exposure to low temperature) and a chronic stress (2 weeks of water deficit).


Genetic interactions between naturally occurring alleles at quantitative trait loci and mutant alleles at candidate loci affecting bristle number in Drosophila melanogaster.
Previously, we mapped quantitative trait loci (QTL) affecting response to short-term selection for abdominal bristle number to seven suggestive regions that contain loci involved in bristle development and/or that have adult bristle number mutant phenotypes, and are thus candidates for bristle number QTL in natural populations. To test the hypothesis that the factors contributing to selection response genetically interact with these candidate loci, high and low chromosomes from selection lines were crossed to chromosomes containing wild-type or mutant alleles at the candidate loci, and the numbers of bristles were recorded in trans heterozygotes. Quantitative failure to complement, detected as a significant selection line*cross effect by analysis of variance, can be interpreted as evidence for allelism or epistasis between the factors on selected chromosomes and the candidate loci. Mutations at some candidate loci (bb, emc, h, Dl, Hairless) showed strong interactions with selected chromosomes, whereas others interacted weakly (ASC, abd, Scr) or not at all (N, mab, E(spl)). These results support the hypothesis that some candidate loci, initially identified through mutations of large effect on bristle number, either harbor or are close members in the same genetic pathway as variants that contribute to standing variation in bristle number.


Ethanol production from food waste at high solids content with vacuum recovery technology.
Ethanol production from food wastes does not only solve environmental issues but also provides renewable biofuels. This study investigated the feasibility of producing ethanol from food wastes at high solids content (35%, w/w). A vacuum recovery system was developed and applied to remove ethanol from fermentation broth to reduce yeast ethanol inhibition. A high concentration of ethanol (144 g/L) was produced by the conventional fermentation of food waste without a vacuum recovery system. When the vacuum recovery is applied to the fermentation process, the ethanol concentration in the fermentation broth was controlled below 100 g/L, thus reducing yeast ethanol inhibition. At the end of the conventional fermentation, the residual glucose in the fermentation broth was 5.7 g/L, indicating incomplete utilization of glucose, while the vacuum fermentation allowed for complete utilization of glucose. The ethanol yield for the vacuum fermentation was found to be 358 g/kg of food waste (dry basis), higher than that for the conventional fermentation at 327 g/kg of food waste (dry basis).


Segmental duplications in the silkworm genome.
Segmental duplications (SDs) or low-copy repeats play important roles in both gene and genome evolution. SDs have been extensively investigated in many organisms, however, there is no information about SDs in the silkworm, Bombyx mori. In this study, we identified and annotated the SDs in the silkworm genome. Our results suggested that SDs constitute ~1.4% of the silkworm genome sequence (≥1 kb in length and ≥90% in the identity of sequence); the number is similar to that in Drosophila melanogaster but smaller than mammalian organisms. Almost half (42%) of the SD sequences are not assigned to chromosomes, indicating that the SDs are challenges for the assembling of genome sequences. We also provided experimental validation of large duplications using qPCR. The analysis of SD content indicated that the genes related to immunity, detoxification, reproduction, and environmental signal recognition are significantly enriched in the silkworm SDs. Our results suggested that segmental duplications have been problematic for sequencing and assembling of the silkworm genome. SDs may have important biological significances in immunity, detoxification, reproduction, and environmental signal recognition in the silkworm. This study provides insight into the evolution of the silkworm genome and an invaluable resource for insect genomics research.


Sex Differences, Sleep Disturbance and Risk of Persistent Pain Associated With Groin Hernia Surgery: A Nationwide Register-Based Cohort Study.
Persistent pain after groin hernia repair is a major health problem. Sleep disturbance is associated with heightened pain sensitivity. The main objective of this study was to examine the role of sleep disturbance in the development and long-term maintenance of chronic postherniorrhaphy inguinal pain (CPIP), with exploration of sex differences. From 2012 to 2017, a national cohort of patients with prior groin hernia repair (n = 2084;45.8% females) were assessed for the development of CPIP 12 months after surgery. Patients then underwent long-term (median 5.0 years) follow-up to evaluate the contribution of sex and sleep disturbance on the maintenance of CPIP. Associations between pre- and postoperative sleep problems (assessed at long-term follow-up) and CPIP were tested using logistic regression. Females had higher rates of CPIP with negative impact on daily activities 12 months after surgery as compared to males (14.6 vs 9.2%, P < .0005), and were more likely to have moderate-severe CPIP in the long-term (3.1 vs 1.2%, P = .003). Preoperative sleep problems predicted development of CPIP 12 months after surgery (adjusted odds ratio [aOR] 1.76 [95%CI 1.26-2.46], P = .001) and CPIP in the long-term (aOR 2.20 [1.61-3.00] , P < .0001). CPIP was associated with insomnia and depression. Sleep disturbance may increase the risk for CPIP, and contribute to maintenance of postsurgical pain. PERSPECTIVE: Females are at heightened risk for CPIP as compared to males. Increased severity of pain symptoms are linked to poorer sleep and psychiatric morbidity. Given the robust associations between sleep disturbance and CPIP, interventions which consolidate and promote sleep, especially in females, may improve long-term pain control.


NEDD4-2 as a potential candidate susceptibility gene for epileptic photosensitivity.
Photosensitive seizures occur most commonly in childhood and adolescence, usually as a manifestation of complex idiopathic generalized epilepsies (IGEs). Molecular mechanisms underlying this condition are yet to be determined because no susceptibility genes have been identified. The NEDD4-2 (Neuronally Expressed Developmentally Downregulated 4) gene encodes a ubiquitin protein ligase proposed to regulate cell surface levels of several ion channels, receptors and transporters involved in regulating neuronal excitability, including voltage-gated sodium channels (VGSCs), the most clinically relevant of the epilepsy genes. The regulation of NEDD4-2 in vivo involves complex interactions with accessory proteins in a cell type specific manner. We screened NEDD4-2 for mutations in a cohort of 253 families with IGEs. We identified three NEDD4-2 missense changes in highly conserved residues; S233L, E271A and H515P in families with photosensitive generalized epilepsy. The NEDD4-2 variants were as effective as wild-type NEDD4-2 in downregulating the VGSC subtype Na(v)1.2 when assessed in the Xenopus oocyte heterologous expression system showing that the direct interaction with the ion channel was not altered by these variants. These data raise the possibility that photosensitive epilepsy may arise from defective interaction of NEDD4-2 with as yet unidentified accessory or target proteins.


A Bayesian transformation model for wavelet shrinkage.
Wavelet shrinkage estimators, in general, make the additive normal noise assumption and disregard the nonlinear nature of contamination. We develop Bayesian wavelet shrinkage estimators (based on the power transformations in the linear model) to accommodate a broad class of noise models in image processing applications. We intend to admit, under one roof, the widespread additive model, the product models common in imaging (such as in synthetic aperture radar (SAR) imagery), as well as noise that may exist amid these two extremes. Tactful prior elicitation in this model, such as the simultaneous assignment of mixture priors for wavelet coefficients and the transformation, imparts flexibility and ample insight into the underlying structure. The model permits estimation with unknown noise structure for reasonably unimodal and well-behaved (on the tails) distributions, wherein it can outperform common shrinkage estimators. Extensions with multiple transformations and Markov random field priors are also considered for adaptation to local variations in contamination. Modern Markov chain Monte Carlo (MCMC) Bayesian computation has been used for simulations and several examples are reported in the paper.


[Achalasia, alacrima without adrenal insufficiency with peripheral and autonomic neurological dysfunction (Allgrove's syndrome)].
To present a patient with Allgrove's syndrome and peripheral and autonomic neurological dysfunction. This pathological entity has not been previously reported in national medical literature. Allgrove's syndrome is comprised of achalasia, alacrima and adrenal insufficiency. It is a rare disease. Some isolated cases have been informed in the literature. The most extensive series was published by Grant et al in 1993. In this study, 20 patients from various european countries were analyzed. We describe a patient with diagnosis suggestive of Allgrove's syndrome and expose the results of a review of the medical literature concerning this syndrome. We emphasize in neurological dysfunction of these patients. Such abnormalities are not actually well defined in this syndrome. A twelve-year-old male patient was admitted to the hospital for dysphagia. A diagnosis of achalasia was performed with endoscopy and esophagogram and confirmed with esophageal manometry. Physical examination showed alacrima and neurological dysfunction. Stimulation of adrenal function with ACTH (Cortosin) was normal. Neurologic examination revealed: peripheral neuropathy, autonomic dysfunction; corticospinal, psychomotor and intellectual impairment. Allgrove's syndrome must be in mind in physicians who see children with achalasia. Nevertheless adrenal insufficiency is not always present in cases when diagnosis is performed. It may appear several years afterwards. Neurological dysfunction seems to be the most prominent feature.


The use of magnetic resonance to evaluate tissue oxygenation in renal artery stenosis.
Vascular occlusive disease poses a threat to kidney viability, but whether the events leading to injury and eventual fibrosis actually entail reduced oxygenation and regional tissue ischemia is unknown. Answering this question has been difficult because of the lack of an adequate method to assess tissue oxygenation in humans. BOLD (blood oxygen-level-dependent) magnetic resonance imaging detects changes in tissue deoxyhemoglobin during maneuvers that affect oxygen consumption, therefore this technique was used to image and analyze cortical and medullary segments of 50 kidneys in 25 subjects undergoing magnetic resonance (MR) angiography to diagnose renal artery stenosis (RAS). Magnetic rate of relaxation (R2*) positively correlates with deoxyhemoglobin levels and was therefore used as a surrogate measure of tissue oxygenation. Furosemide was administered to examine the effect of inhibiting energy-dependent electrolyte transport on tissue oxygenation in subjects with renovascular disease. In 21 kidneys with normal nephrograms, administration of furosemide led to a 20% decrease in medullary R2* (P < 0.01) and an 11.2% decrease in cortical R2*. In normal-size kidneys downstream of high-grade renal arterial stenoses, R2* was elevated at baseline, but fell after furosemide. In contrast, atrophic kidneys beyond totally occluded renal arteries demonstrated low levels of R2* that did not change after furosemide. In kidneys with multiple arteries, localized renal artery stenoses produced focal elevations of R2*, suggesting areas of deoxyhemoglobin accumulation. These results suggest that BOLD MR coupled with a method to suppress tubular oxygen consumption can be used to evaluate regional tissue oxygenation in the human kidney affected by vascular occlusive disease.


[Prevention of hemodynamic disorders during anesthesia in middle-aged and elderly patients with essential hypertension. Components of anesthesia. II].
Hemodynamic parameters were evaluated during various stages of anesthesia in two groups of patients aged 60-85 years with mild essential hypertension. In group 1, routine preoperative treatment with hypotensive drugs was carried out and in group 2 differentiated preoperative treatment with calcium antagonists was carried out with consideration for a hemodynamic type. Group I patients were operated on under traditional neuroleptic analgesia (NLA), group 2 under NLA with 40% lower drug doses than in group 1 and with addition of calcium antagonists. To patients with hypo- and eukinetic hemodynamics, nifedipine was injected (bolus injection) in a dose of 14.2 micrograms/kg before induction and then was infused in a dose of 6.0-14.2 micrograms/kg/min. Patients with hyperkinetic hemodynamics were injected verapamil in a dose of 70-140 micrograms/kg (bolus injection) and then the same dose every hour of the operation. Differentiated use of calcium antagonists during anesthesia helped transform the hyper- and hypokinetic types of hemodynamics into a more rational eukinetic type, which was associated with a decrease of MVO2. Calcium antagonists, exerting analgesic, automatic stabilizing, and antihypoxic effects, prevented the hyperdynamic reactions complicating NLA in 30% cases and precluded complications of concomitant ischemic diseases, which were observed in 40% of group 1 patients.


Payment for performance (P4P): any future in Italy?
Pay for Performance (P4P) programs, based on provision of financial incentives for service quality, have been widely adopted to enhance quality of care and to promote a more efficient use of health care resources whilst improving patient outcomes. In Italy, as in other countries, the growing concern over the quality of health services provided and the scarcity of resources would make P4P programs a useful means of improving their performance. The aim of this paper is to evaluate whether it is possible to implement P4P programs in the Lombardy Region, in Italy, based on the existing data set. Thirteen quality measures were identified regarding four clinical conditions (acute myocardial infarction (AMI), heart failure (HF), ischemic stroke and hip and knee replacement) on the basis of an international literature review. Data was collected using the database of three institutions, which included hospital discharge records (Scheda di Dimissione ospedaliera-SDO-) and letters of discharge. The study population was identified using both the Principal ICD-9-CM diagnosis codes and the discharge date. A Statistical Analysis System (SAS) program was used for the text analysis. It was possible to calculate almost all the parameters pertaining to the three hospitals as all the data required was available with the exception of inpatient mortality in two hospitals and smoking cessation advice/counseling in one hospital. On the ground of this analysis, we believe that it is possible to implement a P4P program in the Lombardy Region. However, for this program to be initiated, all necessary data must be available in electronic format and uniformly collected. Moreover, several other factors must be assessed: which clinical conditions should be included, the threshold for each quality parameter, the amount of financial incentives offered and how they will be provided.


Facilitating mitigation of agricultural non-point source pollution and improving soil nutrient conditions: The role of low temperature co-pyrolysis biochar in nitrogen and phosphorus distribution.
The current study generated co-pyrolysis biochar by pyrolyzing rice straw and pig manure at 300°C and subsequently applying it in a field. Co-pyrolysis biochar demonstrated superior efficiency in mitigating agricultural non-point source pollution compared to biochar derived from individual sources. Furthermore, it displayed notable capabilities in retaining and releasing nutrients, resulting in increased soil levels of total nitrogen, total phosphorus, and organic matter during the maturation stage of rice. Moreover, co-pyrolysis biochar influences soil microbial communities, potentially impacting nutrient cycling. During the rice maturation stage, the soil treated with co-pyrolysis biochar exhibited significant increases in available nutrients and rice yield compared to the control (p<0.05). These findings emphasize the potential of co-pyrolysis biochar for in-situ nutrient retention and enhanced soil nutrient utilization. To summarize, the co-pyrolysis of agricultural waste materials presents a promising approach to waste management, contributing to controlling non-point source pollution, improving soil fertility, and promoting crop production.


The neuropathology of human immunodeficiency virus infection. The Dallas, Texas, experience.
Postmortem neuropathologic changes were evaluated in 141 consecutive patients dying with human immunodeficiency virus infection at publicly supported hospitals affiliated with the University of Texas Southwestern Medical Center in Dallas, between August 1984 and September 1990. Morphologic abnormalities were identified in 112 cases (79%). Cytomegalovirus was the most common opportunistic infection encountered, with characteristic viral inclusions identified in 23 patients, and presumptive evidence of infection in six additional patients. Progressive multifocal leukoencephalopathy was present in four patients. Gram-positive bacterial infections were identified in six patients, and mycobacterial infections in three patients. Opportunistic fungal infections included cryptococcosis (13 cases), histoplasmosis (two cases), and coccidioidomycosis (one case). Toxoplasmosis was uncommon, with active or quiescent lesions identified in five patients. Lymphoma was present in nine patients and was primary in the central nervous system in five patients. Multinucleate giant cell (human immunodeficiency virus) encephalitis was identified in 28 patients. In an additional 26 patients, microglial nodules and/or more generalized white-matter abnormalities were encountered in the absence of multinucleate giant cells, cytomegalovirus inclusions, or systemic cytomegalovirus infection. Vacuolar change was present in 21% of spinal cords, and was highly correlated with cytomegalovirus infection in the nervous system. Mixed infections and/or neoplasms were identified in 24 patients. This survey documents a high frequency of neuropathologic abnormalities in human immunodeficiency virus-infected individuals in a geographical region of the United States not represented in previous series. Variations noted in the frequencies of specific central nervous system disorders between this and other study populations reinforce the need for continuing documentation of geographical trends in human immunodeficiency virus-associated disorders.


ABC-VED Analysis of a Drug Store in the Department of Community Medicine of a Medical College in Delhi.
A matrix based on coupling of cost (always, better and control) analysis and criticality (vital, essential and desirable) analysis was employed for drug inventory containing 129 items of drug store in the Department of Community Medicine of a Medical College in Delhi. The annual drug expenditure incurred on 129 drug items for the year 2010-2011 was found to be Rs. 4,35,847.85. On always, better and control analysis, 18.6, 24.0 and 57.4% drugs were found to be always, better and control category items, respectively, amounting for 69.1, 20.8 and 10.1% of annual drug expenditure. About 13.2 (17), 38.8 (50) and 48.0% (62) items were found to be vital, essential and desirable category items, respectively, amounting for 18.7, 49.5 and 31.8% of annual drug expenditure. Based on always, better and control-vital, essential and desirable matrix analysis there were 37 (28.68%) items in category I, 53 (41.09%) items in category II and 39 (30.23%) items in category III, amounting for 73.0, 22.2 and 4.8% of annual drug expenditure, respectively. To conclude, scientific inventory management tools are needed to be applied in routine for efficient management of the pharmacy stores as it contributes to not only in improvement in patient care but also judicious use of resources as well.


Combined modality therapy for adults with small noncleaved cell lymphoma (Burkitt's and non-Burkitt's types).
Between June 1979 and June 1984 18 adult patients with small noncleaved cell lymphoma (SNCL) (diffuse undifferentiated lymphoma, Burkitt's and non-Burkitt's types of the Rappaport classification) were treated with high-dose cyclophosphamide, doxorubicin, vincristine, prednisone, midcycle high-dose methotrexate, and intrathecal methotrexate. Early in the course of treatment, hyperfractionated radiotherapy (125 cGy, every two days, for 1,500 to 2,250 centigray [cGy]) was administered to unresected masses greater than 10 cm in their greatest dimension. Chemotherapy was administered every 21 days for six to ten cycles. Treatment was generally well tolerated; however, one patient died of probable tumor lysis syndrome. With a median follow-up of 1.2 years, actuarial survival was 66.8% and relapse-free survival (RFS) was 71.3% for the entire group. All treatment failures and deaths occurred in patients with stage D disease. RFS projected at 2 years was 100% for stages A and AR and 60.6% for stage B, C, and D (P = .13 Gehan). Two-year RFS for patients with stage A, AR, B, or C disease was 100 v 41% for those with stage D disease. Patients with adverse prognostic features (n = 7)--unresected bulk measuring greater than 10 cm, pretreatment serum lactate dehydrogenase (LDH) 500 IU/L (normal, 200) or involvement of CNS or bone marrow--had a projected RFS of 28.6% compared with 100% for those patients without these features (P = .002 Gehan). Too few patients received induction radiotherapy to assert its role in therapy. By using aggressive multiagent therapy, cure can be expected in a high percentage of adults with SNCL. In the subset with adverse prognostic features, more effective therapy is necessary.


Clinical features, diagnostic and therapeutic approaches to haematogenous vertebral osteomyelitis.
This article review the clinical features and the diagnostic approach to haematogenous vertebral osteomyelitis in order to optimise treatment strategies and follow-up assessment. Haematogenous spread is considered to be the most important route: the lumbar spine is the most common site of involvement for pyogenic infection and the thoracic spine for tuberculosis infection. The risk factors for developing haematogenous vertebral osteomyelitis are different among old people, adults and children: the literature reports that the incidence seems to be increasing in older patients. The source of infection in the elderly has been related to the use of intravenous access devices and the asymptomatic urinary infections. In young patients the increase has been correlated with the growing number of intravenous drug abusers, with endocarditis and with immigrants from areas where tuberculosis is still endemic. The onset of symptoms is typically insidious with neck or back pain often underestimated by the patient. Fever is present in 10-45% of patients. Spinal infections may cause severe neurological compromise in few cases, but mild neurological deficit, limited to one or two nerve roots, was detected in 28-35% of patients. The diagnosis of haematogenous vertebral osteomyelitis may be very difficult, as the symptoms can be sometimes not specific, vague or almost absent. The usual delay in diagnosis has been reported to be two to four months, despite the use of imaging techniques: in the early diagnosis of vertebral ostemyelitis is important the role of bone scintigraphy. The general principles for the management of spine infections are non operative, consisting of external immobilization and intravenous antibiotics, followed by oral antibiotics. Indications for surgery should be given in case of absence of clinical improvement after 2-3 weeks of intravenous antibiotics, persistent back pain and systemic effects of chronic infection and with presence or progression of neurological deficit in elderly or in cervical infection. Chronic ostemyelitis may require surgery in case of a development of biomechanical instability and/or a vertebral collapse with progressive deformity.


Biomarkers for Evaluating the Inflammation Status in Patients with Cancer.
Inflammation can be a causative factor for carcinogenesis or can result from a consequence of cancer progression. Moreover, cancer therapeutic interventions can also induce an inflammatory response. Various inflammatory parameters are used to assess the inflammatory status during cancer treatment. It is important to select the most optimal biomarker among these parameters. Additionally, suitable biomarkers must be examined if there are no known parameters. We briefly reviewed the published literature for the use of inflammatory parameters in the treatment of patients with cancer. Most studies on inflammation evaluated the correlation between host characteristics, effect of interventions, and clinical outcomes. Additionally, the levels of C-reactive protein, albumin, lymphocytes, and platelets were the most commonly used laboratory parameters, either independently or in combination with other laboratory parameters and clinical characteristics. Furthermore, the immune parameters are classically examined using flow cytometry, immunohistochemical staining, and enzyme-linked immunosorbent assay techniques. However, gene expression profiling can aid in assessing the overall peri-interventional immune status. The checklists of guidelines, such as STAndards for Reporting of Diagnostic accuracy and REporting recommendations for tumor MARKer prognostic studies should be considered when designing studies to investigate the inflammatory parameters. Finally, the data should be interpreted after adjusting for clinically important variables, such as age and cancer stage.


Cohen syndrome-associated protein, COH1, is a novel, giant Golgi matrix protein required for Golgi integrity.
Loss-of-function mutations in the gene COH1, also known as VPS13B, lead to autosomal recessive Cohen syndrome. However, the cellular distribution and function of the encoded protein COH1 (3997 amino acids), which lacks functional homologies to other mammalian proteins, have remained enigmatic. We show here that COH1 is a peripheral Golgi membrane protein that strongly co-localizes with the cis-Golgi matrix protein GM130. Consistent with its subcellular localization, COH1 depletion using RNAi causes fragmentation of the Golgi ribbon into ministacks. Disruption of Golgi organization observed in fibroblasts from Cohen syndrome patients suggests that Golgi dysfunction contributes to Cohen syndrome pathology. In conclusion, our findings establish COH1 as a Golgi-associated matrix protein required for Golgi integrity.


αII Spectrin Forms a Periodic Cytoskeleton at the Axon Initial Segment and Is Required for Nervous System Function.
Spectrins form a submembranous cytoskeleton proposed to confer strength and flexibility to neurons and to participate in ion channel clustering at axon initial segments (AIS) and nodes of Ranvier. Neuronal spectrin cytoskeletons consist of diverse β subunits and αII spectrin. Although αII spectrin is found in neurons in both axonal and somatodendritic domains, using proteomics, biochemistry, and superresolution microscopy, we show that αII and βIV spectrin interact and form a periodic AIS cytoskeleton. To determine the role of spectrins in the nervous system, we generated Sptan1f/f mice for deletion of CNS αII spectrin. We analyzed αII spectrin-deficient mice of both sexes and found that loss of αII spectrin causes profound reductions in all β spectrins. αII spectrin-deficient mice die before 1 month of age and have disrupted AIS and many other neurological impairments including seizures, disrupted cortical lamination, and widespread neurodegeneration. These results demonstrate the importance of the spectrin cytoskeleton both at the AIS and throughout the nervous system.SIGNIFICANCE STATEMENT Spectrin cytoskeletons play diverse roles in neurons, including assembly of excitable domains such as the axon initial segment (AIS) and nodes of Ranvier. However, the molecular composition and structure of these cytoskeletons remain poorly understood. Here, we show that αII spectrin partners with βIV spectrin to form a periodic cytoskeleton at the AIS. Using a new αII spectrin conditional knock-out mouse, we show that αII spectrin is required for AIS assembly, neuronal excitability, cortical lamination, and to protect against neurodegeneration. These results demonstrate the broad importance of spectrin cytoskeletons for nervous system function and development and have important implications for nervous system injuries and diseases because disruption of the spectrin cytoskeleton is a common molecular pathology.


Rapidly dissolving bilayer microneedle arrays - A minimally invasive transdermal drug delivery system for vitamin B12.
Vitamin B12 plays an essential role in one-carbon metabolism in the human body. A deficiency in this vitamin can lead to severe haematopoietic and neuropsychiatric disorders and is currently treated by oral or parenteral administration of exogenous vitamin. Unfortunately, the absorption of orally taken vitamin B12 is low and highly variable, while injections can cause pain and anxiety. Thus, an efficient alternative drug delivery system for overcoming these shortcomings is highly desirable. Novel polymeric microneedle (MN) arrays have the potential for minimally invasive transdermal treatment of vitamin B12 deficiency. Bilayer dissolving MN arrays (19 × 19 needles, 600 µm height) containing 135 µg vitamin B12 were cast using two different aqueous polymer blends. MN arrays showed sufficient mechanical strength for skin insertion, dissolved rapidly and delivered 72.92% of their drug load in vitro over 5 h. Ultimately, the potential of delivering a therapeutically relevant dose of vitamin B12 transdermally was demonstrated in vivo in Sprague-Dawley rats by comparison to subcutaneous injections. Maximum plasma levels of 0.37 µg/mL occurred 30 min post-MN application, highlighting the ability of fabricated MN arrays to rapidly deliver vitamin B12 transdermally.


Mouse A9 cells containing single human chromosomes for analysis of genomic imprinting.
To develop an systematic in vitro approach for the study of genomic imprinting, we generated a new library of human/mouse A9 monochromosomal hybrids. We used whole cell fusion and microcell-mediated chromosome transfer to generate A9 hybrids containing a single, intact, bsr-tagged human chromosome derived from primary fibroblasts. A9 hybrids were identified that contained either human chromosome 1, 2, 4, 5, 7, 8, 10, 11, 15, 18, 20, or X. The parental origin of these chromosomes was determined by polymorphic analysis using microsatellite markers, and matched hybrids containing maternal and paternal chromosomes were identified for chromosomes 5, 10, 11 and 15. The imprinted gene KVLQT1 on human chromosome 11p15.5 was expressed exclusively from the maternal chromosome in A9 hybrids, and the parental-origin-specific expression patterns of several other imprinted genes were also maintained. This library of human monochromosomal hybrids is a valuable resource for the mapping and cloning of human genes and is a novel in vitro system for the screening of imprinted genes and for their functional analysis.


Severe staphylococcal sepsis in adolescents.
During the three-year period beginning in early 1972, nine previously healthy children, ages 8 to 15 years, presented with manifestations of life-threatening septicemia due to coagulase-positive Staphylococcus aureus. In no patient was a predisposing cause for such infection, such as immunologic deficiency, drug abuse, or previous antibiotic or immunosuppressant therapy, discovered. Seven patients manifested evidence of bone and/or joint infection, while eight had pulmonary disease of varying degrees of severity, characterized initially by multiple small septic pulmonary emboli. One patient succumbed to the pulmonary complications of her disease. S. aureus was recovered from specimens of blood (seven patients), joint fluid (four), bone (four), pleural fluid (two), urine (one), sputum (one), and pericardial fluid (one). All organism isolated were methicillin-sensitive; isolates from only one patient were penicillin-sensitive. In several instances, lack of clinical response was related to delay in adequate surgical drainage of infected material. Review of hospital records for the preceding five-year period failed to reveal similar cases of severe systemic staphylococcal infection in healthy adolescents. Although no direct evidence for a causal relationship is available, the occurrence of these cases corresponds temporally with implementation of restrictions upon the use of hexachlorophene.


Different aspects of dysexecutive syndrome in patients with moyamoya disease and its clinical subtypes.
OBJECTIVE Dysexecutive syndrome is common in patients with moyamoya disease (MMD), a chronic cerebrovascular disease that is characterized by stenosis of the bilateral internal carotid arteries and progressive collateral revascularization, and MMD can be classified as ischemic or hemorrhagic according to the disease presentation and history. In this study, the authors aimed to determine which aspects of executive function are impaired in patients with MMD, in addition to the specific dysexecutive functions present among its clinical subtypes and the mechanisms underlying dysexecutive function in these patients. METHODS The authors administered 5 typical executive function tests (the Stroop test, the Hayling Sentence Completion Test [HSCT], the verbal fluency [VF] test, the N-back test, and the Sustained Attention to Response Task [SART]) to 49 patients with MMD and 47 IQ-, age-, education-, and social status-matched healthy controls. The dysexecutive questionnaire (DEX) was also used to assess participants' subjective feelings about their executive function. A total of 39 of the patients were evaluated by CT perfusion (CTP) before the assessments were performed, and the correlations among the performances of the patients on the above tests with the parameters of cerebral blood volume, cerebral blood flow (CBF), mean transit time (MTT), and time-to-peak (TTP) in the frontal lobes of these patients were also analyzed. RESULTS Many aspects of executive function in the patients with MMD were significantly poorer than those in the healthy controls, and the patients performed particularly poorer on the VF test, HSCT, N-back test, and SART. The patients with hemorrhagic MMD exhibited worse executive inhibition, executive processing, and semantic inhibition compared with those with ischemic MMD, but the latter group presented a worse working memory and poorer sustained attention. There were no significant differences in the DEX scores between the patients with MMD and healthy controls. The other findings were as follows: CBF was significantly positively correlated with the number correct on part B of the HSCT (r = 0.481, p = 0.01) and accuracy on the 0-back task of the N-back (r = 0.346, p = 0.031); MTT was significantly positively correlated with accuracy on the 2-back task of the N-back (r = 0.349, p = 0.034) and factor 5 of the DEX (r = 0.359, p = 0.032); and TTP was significantly positively correlated with the number correct on part B of the HSCT (r = 0.402, p = 0.034) and the 1-back reaction time of the N-back (r = 0.356, p = 0.026). CONCLUSIONS The patients with MMD exhibited impairments in semantic inhibition, executive processing, working memory, and sustained attention, but they were not aware of these deficits. Moreover, differences in dysexecutive function existed between the different subtypes of MMD. Hypoperfusion of the frontal lobe may be related to working memory and semantic inhibition impairments in patients with MMD.


Epidemiologic and clinical characteristics of multisystem inflammatory syndrome in adults: a rapid review.
Multisystem inflammatory disease in children (MIS-C) is one of the severe presentations of the coronavirus disease 2019 (COVID-19) that has been described in the literature since the beginning of the pandemic. Although MIS-C refers to children, cases with similar clinical characteristics have been recently described in adults. A description of the epidemiologic and clinical characteristics of multisystem inflammatory disease in adults (MIS-A) is a starting point for better knowledge and understanding of this emerging disease. We identified nine case reports of MIS-A in the literature, five from the United States, two from France and two from the United Kingdom. The case descriptions revealed similarities in clinical features, including occurrence during post-acute disease phase, fever, digestive symptoms, cardiac involvement and elevated inflammatory markers. All the patients were hospitalized, three required admission to the intensive care unit and one died. The most common treatments were intravenous immunoglobulin, prednisolone and aspirin. These findings suggest that MIS-A is a severe complication of COVID-19 disease that can lead to death. Further studies to improve our understanding of the pathogenesis of MIS-A, which will help improve treatment decisions and prevent sequelae or death.


Interaction of fine particles and nanoparticles with red blood cells visualized with advanced microscopic techniques.
So far, little is known about the interaction of nanoparticles with lung cells, the entering of nanoparticles, and their transport through the blood stream to other organs. The entering and localization of different nanoparticles consisting of differing materials and of different charges were studied in human red blood cells. As these cells do not have any phagocytic receptors on their surface, and no actinmyosin system, we chose them as a model for nonphagocytic cells to study how nanoparticles penetrate cell membranes. We combined different microscopic techniques to visualize fine and nanoparticles in red blood cells: (I) fluorescent particles were analyzed by laser scanning microscopy combined with digital image restoration, (II) gold particles were analyzed by conventional transmission electron microscopy and energy filtering transmission electron microscopy, and (III) titanium dioxide particles were analyzed by energy filtering transmission electron microscopy. By using these differing microscopic techniques we were able to visualize and detect particles < or = 0.2 microm and nanoparticles in red blood cells. We found that the surface charge and the material of the particles did not influence their entering. These results suggest that particles may penetrate the red blood cell membrane by a still unknown mechanism different from phagocytosis and endocytosis.