Sensitivity and specificity of a shortened elimination diet protocol for the diagnosis of food-induced atopic dermatitis (FIAD).
The gold standard to diagnose food allergy in dogs is an eight week elimination diet trial (EDT) followed by a re-challenge. A recent study demonstrated that a shorter EDT is possible if prednisolone is administered initially. The goal was to evaluate the sensitivity and specificity of the EDT based on the number of relapses after prednisolone discontinuation. In addition, the aim was to determine whether the initial treatment length or the replacement of prednisolone with oclacitinib would influence the outcome. Eighty-seven dogs with atopic dermatitis. Dogs were fed an elimination diet and treated with either prednisolone or oclacitinib for two to three weeks. Relapsing dogs were treated a second time. In the absence of a relapse after two weeks off medication, dogs then were challenged. Dogs never achieving two weeks off treatment without relapse received the regular EDT. Fifty-eight of 87 dogs completed the study. Thirty-nine of 58 dogs received prednisolone; 21 of these were diagnosed with FIAD and had no relapse (n = 14), one relapse (n = 6) or two relapses (n = 1). Nineteen of 58 dogs received oclacitinib; 11 of these were deemed food-allergic and had no relapse (n = 7) or two relapses (n = 4). The initial treatment duration did not influence the outcome. The threshold of one relapse or less for the diagnosis of FIAD was associated with a sensitivity of 95% for prednisolone and 63% for oclacitinib. The specificity was 100% for both drugs. Initial prednisolone or oclacitinib use in EDT shortens the time to diagnosis of FIAD.


Detection of Δ9-tetrahydrocannabinol in exhaled breath collected from cannabis users.
Exhaled breath has recently been proposed as a new possible matrix for drugs of abuse testing. A key drug is cannabis, and the present study was aimed at investigating the possibility of detecting tetrahydrocannabinol and tetrahydrocannabinol carboxylic acid in exhaled breath after cannabis smoking. Exhaled breath was sampled from 10 regular cannabis users and 8 controls by directing the exhaled breath by suction through an Empore C(18) disk. The disk was extracted with hexane/ethyl acetate, and the resulting extract was evaporated to dryness and redissolved in 100 μL hexane/ethyl acetate. A 3-μL aliquot was injected onto the LC-MS-MS system and analyzed using positive electrospray ionization and selected reaction monitoring. In samples collected 1-12 h after cannabis smoking, tetrahydrocannabinol was detected in all 10 subjects. The rate of excretion was between 9.0 and 77.3 pg/min. Identification of tetrahydrocannabinol was based on correct retention time relative to tetrahydrocannabinol-d(3) and correct product ion ratio. In three samples, peaks were observed for tetrahydrocannabinol carboxylic acid, but these did not fulfill identification criteria. Neither tetrahydrocannabinol or tetrahydrocannabinol carboxylic acid was detected in the controls. These results confirm older reports that tetrahydrocannabinol is present in exhaled breath following cannabis smoking and extend the detection time from minutes to hours. The results further support the idea that exhaled breath is a promising matrix for drugs-of-abuse testing.


Smooth muscle cell expression of type I cyclic GMP-dependent protein kinase is suppressed by continuous exposure to nitrovasodilators, theophylline, cyclic GMP, and cyclic AMP.
A key component of the nitric oxide-cyclic guanosine monophosphate (cGMP) pathway in smooth muscle cells (SMC) is the type I GMP-dependent protein kinase (PK-G I). Activation of PK-G I mediates the reduction of cytoplasmic calcium concentrations and vasorelaxation. In this manuscript, we demonstrate that continuous exposure of SMC in culture to the nitrovasodilators S-nitroso-N-acetylpenicillamine (SNAP) or sodium nitroprusside (SNP) results in approximately 75% suppression of PK-G I mRNA by 48 h. PK-G I mRNA and protein were also suppressed by continuous exposure to cGMP analogues 8-bromo- and 8-(4-chlorophenylthio) guanosine-3,5-monophosphate or the cAMP analogue dibutyryl cAMP. These results suggest that activation of one or both of the cyclic nucleotide-dependent protein kinases mediates PK-G I mRNA suppression. Using isoform-specific cDNA probes, only the PK-G I alpha was detected in SMC, either at baseline or after suppression, while PK-G I beta was not detected, indicating that isoform switch was not contributing to the gene regulation. Using the transcription inhibitor actinomycin D, the PK-G I mRNA half-life in bovine SMC was observed to be 5 h. The half-life was not affected by the addition of SNAP to actinomycin D, indicating no effect on PK-G I mRNA stability. Nuclear runoff studies indicated a suppression of PK-G I gene transcription by SNAP. PK-G I suppression was also observed in vivo in rats given isosorbide dinitrate in the drinking water, with a dose-dependent suppression of PK-G I protein in the aorta. PK-G I antigen in whole rat lung extract was also suppressed by administration of isosorbide or theophylline in the drinking water. These data may contribute to our understanding of nitrovasodilator resistance, a phenomenon resulting from continuous exposure to nitroglycerin or other nitrovasodilators.


Prevention of firearm injuries with gun safety devices and safe storage: An Eastern Association for the Surgery of Trauma Systematic Review.
Over 50,000 homicides and suicides occur each year. An estimated half of all US households are believed to have a firearm present, making them one of the most ubiquitous consumer products. Our goal was to determine if the manner of storage of a firearm in a home could potentially make a difference in the outcomes of intentional and unintentional injuries involving a firearm; specifically addressing the use of gun safes and devices that block/disable firearm function (trigger locks, cable locks, etc.). A comprehensive review of the literature was performed. We used Grading of Recommendations Assessment, Development, and Evaluation methodology to assess the breadth and quality of the data specific to our Population, Intervention, Comparator, Outcomes (PICO) questions. A total of 176 studies were initially identified, then, 120 more added after a subsequent literature review, with 97 removed as duplicates. One hundred ninety-one case reports, case series, and reviews were removed because they did not focus on prevention or did not address our comparators of interest. This left a total of two studies which merited inclusion for PICO 1, should gun locks be used to prevent firearm injuries and six studies which merited inclusion for PICO 2, should safe storage for guns be used to prevent firearm injuries. PICO 1: We conditionally recommend that gun locks be used to prevent unintentional firearm injury. PICO 2: Because of the large effect size and the reasonable quality of available evidence with safe storage of firearms, we recommend safe storage prevent firearm-related injuries. Systematic review, level III.


The conformational requirements for the mechanical precipitation of hemoglobin S and other mutants.
The mechanical stability of human hemoglobin mutants was studied for the specific effects of single and double amino acid substitutions, the ligand state of each chain, and the effect of hybrids between oxy and cyanmet partners on precipitability. It was found that the beta6Glu leads to Val and the beta73 Asp leads to Asn mutations increased the degree of mechanical precipitation in the liganded but not in the deoxy form. When these mutations occurred on the same chain, the effects were approximately additive. Heat labile mutants such as Hb Gun Hill and Hb Leiden exhibited mechanical instability, but probably through a different mechanism, as very little dependence on ligand state was apparent. Studies with valency hybrids of HbS(alpha2 betas2-and-alpha2 betas2 where = cyanmet) revealed that instability was primarily determined by the state of the betas chain, which must be liganded to confer instability on the tetramer. A good agreement between surface activity and mechanical precipitability of these mutants has been found.


Biopsy and Noninvasive Methods to Assess Progression of Nonalcoholic Fatty Liver Disease.
Nonalcoholic fatty liver disease (NAFLD) comprises a spectrum of histopathologic features, ranging from isolated hepatic steatosis, to steatohepatitis with evidence of hepatocellular injury and fibrosis, to cirrhosis. The diagnosis and determination of NAFLD prognosis requires clinical and histopathologic assessments. Liver biopsy still is regarded as the reference for differentiating steatosis (NAFL) from nonalcoholic steatohepatitis, for staging hepatic fibrosis, and for identifying NAFLD in patients with other chronic liver disease. Standardized grading and staging histologic scoring systems, such as the NAFLD activity score and the steatosis, activity, and fibrosis score, can help guide clinical decisions and assess outcomes of clinical trials. Improved understanding of the pathophysiology of NAFLD and technologic advances have led to algorithms that can be used to assess serum biomarkers and imaging methods that are noninvasive alternatives to biopsy collection and analysis. We review the advantages and limitations of biopsy analysis and noninvasive tests as diagnostic and prognostic tools for patients with NAFLD. We also discuss techniques to improve dynamic histopathology assessment, and emerging blood and imaging biomarkers of fibrogenesis.


In-vivo measurement of swine myocardial resistivity.
We used a four-terminal plunge probe to measure myocardial resistivity in two directions at three sites from the epicardial surface of eight open-chest pigs in-vivo at eight frequencies ranging from 1 Hz to 1 MHz. We calibrated the plunge probe to minimize the error due to stray capacitance between the measured subject and ground. We calibrated the probe in saline solutions contained in a metal cup situated near the heart that had an electrical connection to the pig's heart. The mean of the measured myocardial resistivity was 319 ohm x cm at 1 Hz down to 166 ohm x cm at 1 MHz. Statistical analysis showed the measured myocardial resistivity of two out of eight pigs was significantly different from that of other pigs. The myocardial resistivity measured with the resistivity probe oriented along and across the epicardial fiber direction was significantly different at only one out of the eight frequencies. There was no significant difference in the myocardial resistivity measured at different sites.


The mineralocorticoid receptor (MR) regulates ENaC but not NCC in mice with random MR deletion.
Aldosterone binds to the mineralocorticoid receptor (MR) and increases renal Na(+) reabsorption via up-regulation of the epithelial Na(+) channel (ENaC) and the Na(+)-K(+)-ATPase in the collecting system (CS) and possibly also via the NaCl cotransporter (NCC) in the distal convoluted tubule (DCT). However, whether aldosterone directly regulates NCC via MR or indirectly through systemic alterations remains controversial. We used mice with deletion of MR in ∼20 % of renal tubule cells (MR/X mice), in which MR-positive (MR(wt)) and -negative (MR(ko)) cells can be studied side-by-side in the same physiological context. Adult MR/X mice showed similar mRNA and protein levels of renal ion transport proteins to control mice. In MR/X mice, no differences in NCC abundance and phosphorylation was seen between MR(wt) and MR(ko) cells and dietary Na(+) restriction up-regulated NCC to similar extent in both groups of cells. In contrast, MR(ko) cells in the CS did not show any detectable alpha-ENaC abundance or apical targeting of ENaC neither on control diet nor in response to dietary Na(+) restriction. Furthermore, Na(+)-K(+)-ATPase expression was unaffected in MR(ko) cells of the DCT, while it was lost in MR(ko) cells of the CS. In conclusion, MR is crucial for ENaC and Na(+)-K(+)-ATPase regulation in the CS, but is dispensable for NCC and Na(+)-K(+)-ATPase regulation in the DCT.


Coupled neural systems underlie the production and comprehension of naturalistic narrative speech.
Neuroimaging studies of language have typically focused on either production or comprehension of single speech utterances such as syllables, words, or sentences. In this study we used a new approach to functional MRI acquisition and analysis to characterize the neural responses during production and comprehension of complex real-life speech. First, using a time-warp based intrasubject correlation method, we identified all areas that are reliably activated in the brains of speakers telling a 15-min-long narrative. Next, we identified areas that are reliably activated in the brains of listeners as they comprehended that same narrative. This allowed us to identify networks of brain regions specific to production and comprehension, as well as those that are shared between the two processes. The results indicate that production of a real-life narrative is not localized to the left hemisphere but recruits an extensive bilateral network, which overlaps extensively with the comprehension system. Moreover, by directly comparing the neural activity time courses during production and comprehension of the same narrative we were able to identify not only the spatial overlap of activity but also areas in which the neural activity is coupled across the speaker's and listener's brains during production and comprehension of the same narrative. We demonstrate widespread bilateral coupling between production- and comprehension-related processing within both linguistic and nonlinguistic areas, exposing the surprising extent of shared processes across the two systems.


The effect of relaxation exercises on quality of sleep among the elderly: holistic nursing practice review copy.
The aim of this study was to determine the effect of relaxation exercises on quality of sleep of the elderly people staying in a nursing home. Control group had 44 elderly persons and the intervention group had 20 elderly persons in a nursing home. Quality of sleep in the intervention group increased statistically (P = .000). The elderly people should be encouraged to perform relaxation exercises.


Leaf Gas Exchange and Chlorophyll a Fluorescence Imaging of Rice Leaves Infected with Monographella albescens.
This study was intended to analyze the photosynthetic performance of rice leaf blades infected with Monographella albescens by combining chlorophyll (Chl) a fluorescence images with gas exchange and photosynthetic pigment pools. The net CO2 assimilation rate, stomatal conductance, transpiration rate, total Chl and carotenoid pools, and Chl a/b ratio all decreased but the internal CO2 concentration increased in the inoculated plants compared with their noninoculated counterparts. The first detectable changes in the images of Chl a fluorescence from the leaves of inoculated plants were already evident at 24 h after inoculation (hai) and increased dramatically as the leaf scald lesions expanded. However, these changes were negligible for the photosystem II photochemical efficiency (Fv/Fm) at 24 hai, in contrast to other Chl fluorescence traits such as the photochemical quenching coefficient, yield of photochemistry, and yield for dissipation by downregulation; which, therefore, were much more sensitive than the Fv/Fm ratio in assessing the early stages of fungal infection. It was also demonstrated that M. albescens was able to impair the photosynthetic process in both symptomatic and asymptomatic leaf areas. Overall, it was proven that Chl a fluorescence imaging is an excellent tool to describe the loss of functionality of the photosynthetic apparatus occurring in rice leaves upon infection by M. albescens.


A new concept in denture design for patients with few remaining teeth: case reports.
This article presents an alternative to the conventional swing-lock de sign of removable partial denture construction. It incorporates the principles of sectional dentures as well as the swing-lock concept and overcomes some of the limitations of conventional swing-lock dentures. I-bar struts and stainless steel keepers form part of the first section of the denture. The second part of the denture consists of the denture base with the artificial teeth and magnetic retention units. This system has been used successfully in a number of patients whose situations were suitable for the conventional swing-lock design.


Effect of a Natural Supplement Containing Curcuma Longa, Guggul, and Chlorogenic Acid in Patients With Metabolic Syndrome.
The impact of a natural supplement (Kepar; Rikrea, Italy), containing several plant extracts such as curcuma longa, silymarin, guggul, chlorogenic acid, and inulin, was evaluated in 78 patients with metabolic syndrome (MetS; 45 men; age: 62 ± 9 years). Kepar at a dose of 2 pills/d was given for 4 months as add-on therapy to the ongoing treatment, maintained at fixed doses for the entire study. Anthropometric variables, plasma lipids, glucose parameters, and oxidative stress were measured at baseline and after 4 months. We found significant reductions in body weight (from 81.1 ± 13.5 to 79.4 ± 12.5 kg, P < .0001), body mass index (from 29.6 [23.7] to 29.3 [21.9] kg/m(2), P = .001), and waist circumference (from 105 ± 11 to 102 ± 10 cm, P = .0004) as well as in fasting glucose (from 6.5 [11.7] to 6.4 [7.6] mmol/L, P = .014) and total cholesterol (from 4.8 ± 1.4 to 4.5 ± 1.0 mmol/L, P = .03). No significant changes were found in the other appraised parameters, including oxidative stress. In conclusion, after few months of treatment Kepar seems to exert beneficial effects in patients with MetS. Larger studies with a longer follow-up period are needed to confirm these preliminary findings.


Insulin and leptin levels in patients with schizophrenia or related psychoses--a comparison between different antipsychotic agents.
Conventional as well as newer antipsychotics cause weight gain, and, in the regulation of body weight, both insulin and leptin are hormones involved. The aim of the present study was to compare these hormonal levels in patients on treatment with different antipsychotics. Nineteen patients receiving conventional antipsychotics, 14 patients receiving clozapine and 14 patients receiving olanzapine, were studied. Fasting blood samples for insulin, leptin, glucose, and drug serum concentrations were analyzed. In addition, body mass index (BMI) was calculated. The median insulin level was significantly higher in the patients receiving olanzapine than in those receiving conventional agents, whereas there was no significant difference in insulin between the clozapine and the other two groups. However, in the clozapine group, insulin levels were positively correlated to the drug serum concentration. BMI was elevated in about half of the patients, with no difference being found between the groups. The leptin level was significantly higher in the women than in the men in the conventional agent group, but not in the olanzapine or clozapine groups. The higher insulin level in the patients receiving olanzapine than in those receiving conventional antipsychotics, despite similar BMI, points to a probable influence of olanzapine on insulin secretion. The correlation between the insulin levels and the clozapine concentration indicates, in addition, an influence of clozapine on insulin secretion. The gender difference in leptin, i.e. females normally having higher leptin levels than males, was found in the conventional agent group, but not in the olanzapine or clozapine groups, suggesting that also leptin regulation is altered during olanzapine or clozapine treatments. Moreover, it was mainly due to an increase of leptin in the males that leptin levels were equalized between sexes in the olanzapine group. We conclude that the influence of olanzapine and clozapine on both insulin and leptin levels might be associated with their weight-gain-inducing ability, while other mechanisms may be involved in the weight gain caused by conventional antipsychotics.


Resveratrol in Rodent Models of Parkinson's Disease: A Systematic Review of Experimental Studies.
Parkinson's disease (PD) is a common neurodegenerative disease featured by progressive degeneration of nigrostriatal dopaminergic neurons (DA) accompanied with motor function impairment. Accumulating evidence has demonstrated that natural compounds from herbs have potent anti-PD efficacy in PD models. Among those compounds, resveratrol, a polyphenol found in many common plants and fruits, is more effective against PD. Resveratrol has displayed a potent neuroprotective efficacy in several PD animal models. However, there is still no systematic analysis of the quality of methodological design of these studies, nor of their results. In this review, we retrieved and analyzed 18 studies describing the therapeutic effect of resveratrol on PD animal models. There are 5 main kinds of PD rodent models involved in the 18 articles, including chemical-induced (MPTP, rotenone, 6-OHDA, paraquat, and maneb) and transgenic PD models. The neuroprotective mechanisms of resveratrol were mainly concentrated on the antioxidation, anti-inflammation, ameliorating mitochondrial dysfunction, and motor function. We discussed the disadvantages of different PD animal models, and we used meta-analysis approach to evaluate the results of the selected studies and used SYRCLE's risk of bias tool to evaluate the methodological quality. Our analytical approach minimized the bias of different studies. We have also summarized the pharmacological mechanisms of resveratrol on PD models as reported by the researchers. The results of this study support the notion that resveratrol has significant neuroprotective effects on different PD models quantified using qualitative and quantitative methods. The collective information in our review can guide researchers to further plan their future experiments without any hassle regarding preclinical and clinical studies. In addition, this collective assessment of animal studies can provide a qualitative analysis of different PD animal models, either to guide further testing of these models or to avoid unnecessary duplication in their future research.


Conceptualizing and measuring global interpersonal mistrust-trust.
Global interpersonal mistrust is conceptualized as a general mistrust of the motives of others in situations related to one's well-being: a general tendency to view others as mean, selfish, malevolent, or unreliable people who are, thus, not to be depended on to treat one well. The authors developed an 18-item unidimensional self-report inventory measuring interpersonal mistrust as a negative cognitive orientation toward others. The measure comprises items describing perceptions of specific hypothetical interpersonal situations rather than items asking respondents to describe their own general behavior. The measure was reliable and evidenced construct validity in a heterogeneous sample of Australians.


Are circular external fixators weakened by the use of hemispheric washers?
To compare the axial mechanical stability of 3 circular external fixators systems with and without hemispheric washers. Experimental study. The axial stiffness and load necessary to produce 0.5 and 1 mm of displacement of 10 circular external fixator constructs from 3 manufacturers were tested on a materials testing machine. The constructs tested included the Small Bone fixator (SBF; Hofmann S.a.S., Monza, Italy), the IMEX ring fixator (IMEX Inc., Longview, TX), and the Multiplanar C-Fix (MCF; PanVet Distribuzione, Seriate, Italy). Five configurations were tested for each construct: (1) conventional nut fixation, (2) hemispheric washer fixation with connecting rods offset by 0, (3) 1, and (4) 2 holes, and (5) with a ring placed at maximum angulation. The loads resisted at 0.5 and 1 mm of displacement did not differ when frame configurations were compared (P =.25733 and.33769, respectively). The linear stiffness of the following configurations were decreasingly stiff: standard constructs, hemispheric washers with connecting rods perpendicular to rings, hemispheric washers with connecting rods offset by 1 hole, hemispheric washers with connecting rods offset by 2 holes, and ring offset in relation to bone model. The SBF constructs tested were 34% and 41% more rigid than the IMEX and MCF constructs tested despite the larger diameter of the connecting rods for the IMEX frames (6 mm) compared with the SBF frames (5 mm). The IMEX constructs tested were 6% more rigid than the MCF constructs tested. Adding hemispheric washers and angling connecting rods in relation to rings did not influence the loads resisted at 0.5 and 1 mm displacement but decreased construct stiffness. The use of hemispheric washers had minor effects on the biomechanical performance of fixator frames tested in this study when used to angle a ring in relation to connecting rods for circular external fixators.


Antifungal activity of steroidal glycosides from Yucca gloriosa L.
The antifungal activity of a crude steroidal glycoside extract from Yucca gloriosa flowers, named alexin, was investigated in vitro against a panel of human pathogenic fungi, yeasts as well as dermatophytes and filamentous species. The minimal inhibitory concentration (MIC) was determined by an agar dilution method. Alexin had a broad spectrum of antifungal activity, found to reside entirely in the spirostanoid fraction. The major tigogenyl glycosides, yuccaloeside B and yuccaloeside C, exhibited MICs between 0.39 and 6.25 microg[sol ]mL for all the tested yeast strains except for two (C. lusitaniae and C. kefyr). They were also active against several clinical Candida isolates known to be resistant to the usual antifungal agents. The MICs for the dermatophytes were between 0.78 and 12.5 microg[sol ]mL. The most sensitive filamentous species was A. fumigatus (MIC = 1.56 microg[sol ]mL). For most of the strains, the MICs of both glycosides were similar to those of the reference antifungal agent.


Prevalence and significance of anti-HCV antibodies.
The actual ELISA test for anti-HCV defects an antibody to a non-structural part of the virus. It is present in 0.3 to 1.5% of the blood donors and in about 60% of patients with chronic non-A, non-B hepatitis. Seroconversion occurs 4 to 6 months after the beginning of the disease. The risk of a sporadic transmission is very low but unknown. An ALT elevation occurred only in 7% of the sexual partners of positive chronic liver patients but nobody seroconverted. Vertical, mother to infant, transmission is rare. In all risk groups for HCV infection such as drug abuse, multiple transfusion, haemodialysis, the presence of the antibody is increased between 15 to 80%. There is also evidence that HCV may be an oncogenic factor for hepatocellular carcinoma. It is present in 30 to 60% of the cases.


Insect juvenile hormone resistance gene homology with the bHLH-PAS family of transcriptional regulators.
Juvenile hormone analog (JHA) insecticides are relatively nontoxic to vertebrates and offer effective control of certain insect pests. Recent reports of resistance in whiteflies and mosquitoes demonstrate the need to identify and understand genes for resistance to this class of insect growth regulators. Mutants of the Methoprene-tolerant (Met) gene in Drosophila melanogaster show resistance to both JHAs and JH, and previous biochemical studies have demonstrated a mechanism of resistance involving an intracellular JH binding-protein that has reduced ligand affinity in Met flies. We cloned the Met+ gene by transposable P-element tagging and found reduced transcript level in several mutant alleles, showing that underproduction of the normal gene product can lead to insecticide resistance. Transformation of Met flies with a Met+ cDNA resulted in susceptibility to methoprene, indicating that the cDNA encodes a functional Met+ protein. MET shows homology to the basic helix-loop-helix (bHLH)-PAS family of transcriptional regulators, implicating MET in the action of JH at the gene level in insects. This family also includes the vertebrate dioxin receptor, a transcriptional regulator known to bind a variety of environmental toxicants. Because JHAs include a diverse array of chemicals with JH activity, a mechanism whereby they can exert effects in insects through a common pathway is suggested.


Sinusoidal-wavelength-scanning interferometer with double feedback control for real-time distance measurement.
In addition to a conventional phase a the interference signal of a sinusoidal-wavelength-scanning interferometer has a phase-modulation amplitude Zb that is proportional to the optical path difference L and amplitude b of the wavelength scan. L and b are controlled by a double feedback system so that the phase alpha and the amplitude Zb are kept at 3pi/2 and pi, respectively. The voltage applied to a device that displaces a reference mirror to change the optical path difference becomes a ruler with scales smaller than a wavelength. Voltage applied to a device that determines the amplitude of the wavelength scan becomes a ruler marking every wavelength. These two rulers enable one to measure an absolute distance longer than a wavelength in real time.


Antimicrobial sensitivity pattern of organisms causing urinary tract infection in children with sickle cell anemia in Maiduguri, Nigeria.
Patients with sickle cell disease have increased tendency to develop frequent and severe infections, especially of bones and urinary tract. The knowledge of antimicrobial sensitivity pattern of common etiological agents will serve as a guide to empiric treatment while results of urine culture and sensitivity are being awaited. Antimicrobial sensitivity test was carried out on bacterial isolates from the urine of febrile children with sickle cell anemia (SCA) and children with HbAA in Maiduguri. Urine specimens were collected and cultured by standard methods. Sensitivity to 15 antimicrobials (based on availability of sensitivity disc) was tested using the disc-diffusion technique of stokes. Significant bacteriuria was obtained from 65 (26%) of the 250 children with SCA and 51 (20.4%) of the 250 controls. The isolates were E. coli, Klebsiella, Coliforms, Proteus, Staph aureus and Salmonella. Sensitivity was highest to 3 rd generation cephalosporins, followed by the quinolones: ciprofloxacin (86.2%), ofloxacin (83.1%), and peflacine (73.8%). Sensitivity of the organisms to some of the commonly used antibiotics like ampicillin, cotrimoxazole, and nalidixic acid was generally low. In general, the pattern of bacteriuria and their sensitivity in the SCA group was similar to the pattern in the control group. Etiological agents of childhood urinary tract infections (UTI) in this environment are resistant to most of the drugs commonly recommended for its treatment. Amoxicillin/clavulanic acid, cefuroxime, and gentamicin, are recommended as first-line drugs for treatment of UTI while awaiting results of culture and sensitivity. Ceftriaxone and ceftazidime should be reserved for case of non response to first-line drugs and severe infections.


The metacognitive beliefs account of hallucinatory experiences: a literature review and meta-analysis.
An influential model of hallucinations proposed by Morrison et al. (1995. Behavioural and Cognitive Psychotherapy, 23(3), 265-280) assumes that dysfunctional metacognitive beliefs lead to the misattribution of intrusive thoughts to external sources, therefore generating hallucinatory experiences. Following a comprehensive review of the literature, a series of meta-analyses was carried out to summarize the empirical findings on the association between hallucination-proneness and different metacognitive beliefs. The results of this research synthesis found little support for the existence of specific associations between hallucinations and dysfunctional metacognitive beliefs. While metacognitive beliefs are robustly associated with hallucination-proneness in non-clinical studies, they were only moderately associated with hallucinations in clinical samples. Additional analyses revealed that, after controlling for the effect of comorbid symptoms, hallucination-proneness was only weakly associated with metacognitive beliefs, suggesting that the large associations observed in previous research might stem from the failure to consider the covariation between different symptoms. These findings have important implications in relation to the role of metacognitive factors in psychopathological symptoms, as well as for the implementation of metacognitive-focused cognitive behavioural techniques for the treatment of psychosis.


Clinical applications of pharmacogenetics in psychiatry.
Pharmacogenetic research has identified response-related mutant variants in metabolic enzymes and drug-targeted receptors. Allelic variants of dopaminergic and serotonergic receptors have been associated with clinical outcome and adverse events such as movement disorders. Deficient metabolic enzymes have been related to drug accumulation and toxic events. This information will help to design safer and more efficient drugs. However, the field is moving rapidly towards a new goal: the application of pharmacogenetics as a clinical tool for the prediction of treatment outcome. The first studies in this direction have proved the feasibility of using genetic information for the prediction of response to antipsychotic drugs and to treatment of Alzheimer's disease. New strategies investigating genes related to specific symptoms and side-effects have produced encouraging results that can contribute to the improvement of the levels and accuracy of the predictions. This review tries to summarise recent advances and provides an overview of future clinical applications.


Bayesian identification of bacterial strains from sequencing data.
Rapidly assaying the diversity of a bacterial species present in a sample obtained from a hospital patient or an environmental source has become possible after recent technological advances in DNA sequencing. For several applications it is important to accurately identify the presence and estimate relative abundances of the target organisms from short sequence reads obtained from a sample. This task is particularly challenging when the set of interest includes very closely related organisms, such as different strains of pathogenic bacteria, which can vary considerably in terms of virulence, resistance and spread. Using advanced Bayesian statistical modelling and computation techniques we introduce a novel pipeline for bacterial identification that is shown to outperform the currently leading pipeline for this purpose. Our approach enables fast and accurate sequence-based identification of bacterial strains while using only modest computational resources. Hence it provides a useful tool for a wide spectrum of applications, including rapid clinical diagnostics to distinguish among closely related strains causing nosocomial infections. The software implementation is available at https://github.com/PROBIC/BIB.


A whole-heart motion-correction algorithm: Effects on CT image quality and diagnostic accuracy of mechanical valve prosthesis abnormalities.
We aimed to determine the effect of a whole-heart motion-correction algorithm (new-generation snapshot freeze, NG SSF) on the image quality of cardiac computed tomography (CT) images in patients with mechanical valve prostheses compared to standard images without motion correction and to compare the diagnostic accuracy of NG SSF and standard CT image sets for the detection of prosthetic valve abnormalities. A total of 20 patients with 32 mechanical valves who underwent wide-coverage detector cardiac CT with single-heartbeat acquisition were included. The CT image quality for subvalvular (below the prosthesis) and valvular regions (valve leaflets) of mechanical valves was assessed by two observers on a four-point scale (1 = poor, 2 = fair, 3 = good, and 4 = excellent). Paired t-tests or Wilcoxon signed rank tests were used to compare image quality scores and the number of diagnostic phases (image quality score≥3) between the standard image sets and NG SSF image sets. Diagnostic performance for detection of prosthetic valve abnormalities was compared between two image sets with the final diagnosis set by re-operation or clinical findings as the standard reference. NG SSF image sets had better image quality scores than standard image sets for both valvular and subvalvular regions (P < 0.05 for both). The number of phases that were of diagnostic image quality per patient was significantly greater in the NG SSF image set than standard image set for both valvular and subvalvular regions (P < 0.0001). Diagnostic performance of NG SSF image sets for the detection of prosthetic abnormalities (20 pannus and two paravalvular leaks) was greater than that of standard image sets (P < 0.05). Application of NG SSF can improve CT image quality and diagnostic accuracy in patients with mechanical valves compared to standard images.


Population genetic signatures of a climate change driven marine range extension.
Shifts in species distribution, or 'range shifts', are one of the most commonly documented responses to ocean warming, with important consequences for the function and structure of ecosystems, and for socio-economic activities. Understanding the genetic signatures of range shifts can help build our knowledge of the capacity of species to establish and persist in colonised areas. Here, seven microsatellite loci were used to examine the population connectivity, genetic structure and diversity of Octopus tetricus, which has extended its distribution several hundred kilometres polewards associated with the southwards extension of the warm East Australian Current along south-eastern Australia. The historical distribution and the range extension zones had significant genetic differences but levels of genetic diversity were comparable. The population in the range extension zone was sub-structured, contained relatively high levels of self-recruitment and was sourced by migrants from along the entire geographic distribution. Genetic bottlenecks and changes in population size were detected throughout the range extension axis. Persistent gene flow from throughout the historical zone and moderate genetic diversity may buffer the genetic bottlenecks and favour the range extension of O. tetricus. These characteristics may aid adaptation, establishment, and long-term persistence of the population in the range extension zone.


Cyclic AMP signaling reduces sirtuin 6 expression in non-small cell lung cancer cells by promoting ubiquitin-proteasomal degradation via inhibition of the Raf-MEK-ERK (Raf/mitogen-activated extracellular signal-regulated kinase/extracellular signal-regulated kinase) pathway.
The cAMP signaling system regulates various cellular functions, including metabolism, gene expression, and death. Sirtuin 6 (SIRT6) removes acetyl groups from histones and regulates genomic stability and cell viability. We hypothesized that cAMP modulates SIRT6 activity to regulate apoptosis. Therefore, we examined the effects of cAMP signaling on SIRT6 expression and radiation-induced apoptosis in lung cancer cells. cAMP signaling in H1299 and A549 human non-small cell lung cancer cells was activated via the expression of constitutively active Gαs plus treatment with prostaglandin E2 (PGE2), isoproterenol, or forskolin. The expression of sirtuins and signaling molecules were analyzed by Western blotting. Activation of cAMP signaling reduced SIRT6 protein expression in lung cancer cells. cAMP signaling increased the ubiquitination of SIRT6 protein and promoted its degradation. Treatment with MG132 and inhibiting PKA with H89 or with a dominant-negative PKA abolished the cAMP-mediated reduction in SIRT6 levels. Treatment with PGE2 inhibited c-Raf activation by increasing inhibitory phosphorylation at Ser-259 in a PKA-dependent manner, thereby inhibiting downstream MEK-ERK signaling. Inhibiting ERK with inhibitors or with dominant-negative ERKs reduced SIRT6 expression, whereas activation of ERK by constitutively active MEK abolished the SIRT6-depleting effects of PGE2. cAMP signaling also augmented radiation-induced apoptosis in lung cancer cells. This effect was abolished by exogenous expression of SIRT6. It is concluded that cAMP signaling reduces SIRT6 expression by promoting its ubiquitin-proteasome-dependent degradation, a process mediated by the PKA-dependent inhibition of the Raf-MEK-ERK pathway. Reduced SIRT6 expression mediates the augmentation of radiation-induced apoptosis by cAMP signaling in lung cancer cells.


Retinal Microperimetry: A New Tool for Identifying Patients With Type 2 Diabetes at Risk for Developing Alzheimer Disease.
Type 2 diabetes is associated with a high risk of cognitive impairment and dementia. Therefore, strategies are needed to identify patients who are at risk for dementia. Given that the retina is a brain-derived tissue, it may provide a noninvasive way to examine brain pathology. The aims of this study were to evaluate whether retinal sensitivity 1) correlates with the specific parameters of brain imaging related to cognitive impairment and 2) discriminates patients with diabetes with mild cognitive impairment (MCI) from those with normal cognition and those with Alzheimer disease (AD). For this purpose, a prospective, nested case-control study was performed and included 35 patients with type 2 diabetes without cognitive impairment, 35 with MCI, and 35 with AD. Retinal sensitivity was assessed by Macular Integrity Assessment microperimetry, and a neuropsychological evaluation was performed. Brain neurodegeneration was assessed by MRI and fludeoxyglucose-18 positron emission tomography (18FDG-PET). A significant correlation was found between retinal sensitivity and the MRI and 18FDG-PET parameters related to brain neurodegeneration. Retinal sensitivity was related to cognitive status (normocognitive > MCI > AD; P < 0.0001). Our results suggest that retinal sensitivity assessed by microperimetry is related to brain neurodegeneration and could be a useful biomarker for identifying patients with type 2 diabetes who are at risk for developing AD.


Nebivolol, but not metoprolol, lowers blood pressure in nitric oxide-sensitive human hypertension.
Nebivolol, unlike other selective β1-receptor blockers, induces vasodilation attributable to increased NO bioavailability. The relative contribution of this mechanism to the blood pressure (BP)-lowering effects of nebivolol is unclear because it is normally masked by baroreflex buffering. Autonomic failure provides a unique model of hypertension devoid of autonomic modulation but sensitive to the hypotensive effects of NO potentiation. We tested the hypothesis that nebivolol would decrease BP in these patients through a mechanism independent of β-blockade. We randomized 20 autonomic failure patients with supine hypertension (14 men; 69±2 years) to receive a single oral dose of placebo, nebivolol 5 mg, metoprolol 50 mg (negative control), and sildenafil 25 mg (positive control) on separate nights in a double-blind, crossover study. Supine BP was monitored every 2 hours from 8:00 pm to 8:00 am. Compared with placebo, sildenafil and nebivolol decreased systolic BP during the night (P<0.001 and P=0.036, by mixed-effects model, maximal systolic BP reduction 8-hour postdrug of -20±6 and -24±9 mm Hg, respectively), whereas metoprolol had no effect. In a subanalysis, we divided patients into sildenafil responders (BP fall>20 mm Hg at 4:00 am) and nonresponders. Nebivolol significantly lowered systolic BP in sildenafil responders (-44±13 mm Hg) but not in nonresponders (1±11 mm Hg). Despite lowering nighttime BP, nebivolol did not worsen morning orthostatic tolerance compared with placebo. In conclusion, nebivolol effectively lowered supine hypertension in autonomic failure, independent of β1-blockade. These results are consistent with the hypothesis that NO potentiation contributes significantly to the antihypertensive effect of nebivolol.


Neutrophil heterogeneity and aging: implications for COVID-19 and wound healing.
Neutrophils play a critical role in the immune response to infection and tissue injury. However, recent studies have shown that neutrophils are a heterogeneous population with distinct subtypes that differ in their functional properties. Moreover, aging can alter neutrophil function and exacerbate immune dysregulation. In this review, we discuss the concept of neutrophil heterogeneity and how it may be affected by aging. We then examine the implications of neutrophil heterogeneity and aging for COVID-19 pathogenesis and wound healing. Specifically, we summarize the evidence for neutrophil involvement in COVID-19 and the potential mechanisms underlying neutrophil recruitment and activation in this disease. We also review the literature on the role of neutrophils in the wound healing process and how aging and neutrophil heterogeneity may impact wound healing outcomes. Finally, we discuss the potential for neutrophil-targeted therapies to improve clinical outcomes in COVID-19 and wound healing.


The Impact of Social Support Networks on Family Resilience in an Australian Intensive Care Unit: A Constructivist Grounded Theory.
This article discusses the findings of a grounded theory of family resilience in an Australian intensive care unit (ICU) with a specific focus on families' experiences of their interactions with other members within their own family, and the families of other patients in the ICU. A constructivist grounded theory methodology was adopted. Data were collected using in-depth interviews with 25 family members of 21 critically ill patients admitted to a tertiary-level ICU in Australia. The core category regaining control represents the families' journey toward resilience when in ICU. The major categories represent facilitators for, and barriers to, regaining control. One of the main facilitators is drawing strength, and it explains the manner with which families receive social support from their own and other family members to help them cope. This study offers a framework to improve patient- and family-centered care in the ICU by facilitating families' ability to manage their situation more effectively. Social support offered by family members facilitates the families' ability to regain control. An ICU family resilience theoretical framework, situated within the context of the Australian healthcare system, adds to what is currently known about the families' experiences in the ICU. The relationships that develop between families in the ICU may provide a source of social support; however, not all families welcome interactions with other ICU families, and it may cause further emotional distress. Further research is warranted to determine whether families suffer a secondary stress reaction from incidental interactions with other patients' families in the ICU. Furthermore, when family members pull together and offer social support to each other, they are better able to regain control. This process contributes to an ICU family resilience framework.


Dystrophin Dp71 in PC12 cell adhesion.
Previously, we reported that PC12 cells with decreased Dp71 expression (antisense-Dp71 cells) display deficient nerve-growth-factor-induced neurite outgrowth. In this study, we show that disturbed neurite outgrowth of antisense-Dp71 cells is accompanied by decreased adhesion activity on laminin, collagen and fibronectin. In wild-type cells, the immunostaining of Dp71 and beta1-integrin overlaps in the basal area contacting the substrate, but staining of both proteins decrease in the antisense-Dp71 cells. Morphology of antisense-Dp71 cells at the electron microscopic level is characterized by the lack of filopodia, cellular projections involved in adhesion. Our findings suggest that Dp71 is required for the efficient PC12 cell attachment to beta1-integrin-dependent substrata and that decreased adhesion activity of the antisense-Dp71 cells could determine their deficiency to extend neurites.


Multiple Strategies for Light-Harvesting, Photoprotection, and Carbon Flow in High Latitude Microbial Mats.
Microbial mats are ubiquitous in polar freshwater ecosystems and sustain high concentrations of biomass despite the extreme seasonal variations in light and temperature. Here we aimed to resolve genomic adaptations for light-harvesting, bright-light protection, and carbon flow in mats that undergo seasonal freeze-up. To bracket a range of communities in shallow water habitats, we sampled cyanobacterial mats in the thawed littoral zone of two lakes situated at the northern and southern limits of the Canadian Arctic permafrost zone. We applied a multiphasic approach using pigment profiles from high performance liquid chromatography, Illumina MiSeq sequencing of the 16S and 18S rRNA genes, and metagenomic analysis. The mats shared a taxonomic and functional core microbiome, dominated by oxygenic cyanobacteria with light-harvesting and photoprotective pigments, bacteria with bacteriochlorophyll, and bacteria with light-driven Type I rhodopsins. Organisms able to use light for energy related processes represented up to 85% of the total microbial community, with 15-30% attributable to cyanobacteria and 55-70% attributable to other bacteria. The proportion of genes involved in anaplerotic CO2 fixation was greater than for genes associated with oxygenic photosynthesis. Diverse heterotrophic bacteria, eukaryotes (including metazoans and fungi) and viruses co-occurred in both communities. The results indicate a broad range of strategies for capturing sunlight and CO2, and for the subsequent flow of energy and carbon in these complex, light-driven microbial ecosystems.


Pregnancy Outcomes of Women Admitted to a Tertiary Care Centre with Short Cervix.
Our aim was to characterize the management and outcome of women admitted to a tertiary high-risk obstetrics unit with a short cervix (≤25 mm), measured on transvaginal ultrasound (TVS), and to determine the latency period from diagnosis to delivery. A retrospective chart review of women admitted to the High Risk Obstetrics Unit at Sunnybrook Health Sciences Centre between 2005-2011 with an ultrasound-identified short cervix was done. A total of 110 women admitted for a short cervix between 2005-2011 (N = 56 singletons; N = 54 twin gestations) met the inclusion criteria. The mean latency to delivery was 62.6 days; mean GA at delivery was 33.9 weeks. Following 7 and 14 days of admission, 5.5% and 11.8% of women had delivered, respectively. Preterm birth occurred in 70% of all women. Cervical length was a significant factor in predicting latency among singletons, whereas the length of stay in hospital for activity restriction was not. Although many women with a short cervix delivered preterm, only a small proportion delivered within 2 weeks of admission. Although it is not clear whether inpatient management improves the pregnancy outcome, these findings have implications for both patient care and health resources.


Live cell imaging of outward and inward vesiculation induced by the complement c5b-9 complex.
Cells resist death induced by the complement membrane attack complex (MAC, C5b-9) by removal of the MAC from their surface by an outward and/or inward vesiculation. To gain an insight into the route of MAC removal, human C9 was tagged with Alexa Fluor 488 and traced within live cells. Tagged C9-AF488 was active in lysis of erythrocytes and K562 cells. Upon treatment of K562 cells with antibody and human serum containing C9-AF488, C9-AF488 containing MAC bound to the cells. Within 5-10 min, the cells started shedding C5b-9-loaded vesicles (0.05-1 mum) by outward vesiculation. Concomitantly, C9-AF488 entered the cells and accumulated in a perinuclear, late recycling compartment, co-localized with endocytosed transferrin-Texas Red. Similar results were obtained with fixed cells in which the MAC was labeled with antibodies directed to a C5b-9 neoepitope. Inhibition of protein kinase C reduced endocytosis of C5b-9. Kinetic analysis demonstrated that peripheral, trypsin-sensitive C5b-9 was cleared from cells at a slower rate relative to fully inserted, trypsin-resistant C5b-9. MAC formation is controlled by CD59, a ubiquitously expressed membrane complement regulator. Analysis at a cell population level showed that the amount of C5b-9-AF488 bound to K562 cells after complement activation was highly heterogeneous and inversely correlated with the CD59 level of expression. Efficient C9-AF488 vesiculation was observed in cells expressing low CD59 levels, suggesting that the protective impact of MAC elimination by vesiculation increases as the level of expression of CD59 decreases.


US-guided biopsy of neck masses in postoperative management of patients with thyroid cancer.
High-frequency (10-MHz) sonography demonstrated a cervical mass or lymphadenopathy, or both, during postoperative follow-up of 52 patients who had undergone surgery for thyroid cancer. Percutaneous biopsy with ultrasonographic (US) guidance was performed in all 52 masses, 44 of which were nonpalpable. Malignant cells were obtained in 29 biopsies, and the results of 20 biopsies were negative, yielding benign lymphocytes only. Results in three biopsies were nondiagnostic due to hypocellular specimens. Therefore, 94% of biopsy results (49) of 52) were confidently assigned as either positive (56%) or negative (38%) for malignancy. There were no complications. High-frequency sonography can demonstrate clinically occult thyroid bed tumor recurrence and lymph node metastases. US-guided biopsy is an accurate and safe technique to confirm or exclude malignancy in patients at high risk of recurrence of thyroid cancer.


Evolution of dislocation substructures in metals via high-strain-rate nanoindentation.
Deformation at high strain rates often results in high stresses on many engineering materials, potentially leading to catastrophic failure without proper design. High-strain-rate mechanical testing is thus needed to improve the design of future structural materials for a wide range of applications. Although several high-strain-rate mechanical testing techniques have been developed to provide a fundamental understanding of material responses and microstructural evolution under high-strain-rate deformation conditions, these tests are often very time consuming and costly. In this work, we utilize a high-strain-rate nanoindentation testing technique and system in combination with transmission electron microscopy to reveal the deformation mechanisms and dislocation substructures that evolve in pure metals from low (10-2 s-1) to very high indentation strain rates (104 s-1), using face-centered cubic aluminum and body-centered cubic molybdenum as model materials. The results help to establish the conditions under which micro- and macro-scale tests can be compared with validity and also provide a promising pathway that could lead to accelerated high-strain-rate testing at substantially reduced costs.


Behavioral effects of flunitrazepam: reinforcing and discriminative stimulus effects in rhesus monkeys and prevention of withdrawal signs in pentobarbital-dependent rats.
Flunitrazepam was evaluated in several procedures that have been used extensively to study the behavioral effects and abuse potential of positive GABA(A) modulators. One group of monkeys (n=3) responded to receive injections of methohexital or saline (i.v.) while other groups (n=2-4/group) discriminated vehicle from either pentobarbital or triazolam. Other monkeys (n=2) received diazepam daily and discriminated flumazenil from vehicle. Finally, the ability of flunitrazepam to prevent the emergence of withdrawal signs in pentobarbital-treated rats was evaluated. Flunitrazepam maintained i.v. self-administration that was, on average, less than that maintained by methohexital and greater than that maintained by saline. In drug discrimination studies, flunitrazepam substituted for pentobarbital and for triazolam and failed to substitute for flumazenil. In rats (n=3-6/group), signs of withdrawal were not evident when flunitrazepam treatment replaced pentobarbital treatment; withdrawal signs emerged when either pentobarbital or flunitrazepam treatment was terminated. Taken together with data from previous studies, these data suggest that the abuse liability of flunitrazepam is comparable to that of other benzodiazepines.


Randomized Trial of Adding Parenteral Acetaminophen to Prochlorperazine and Diphenhydramine to Treat Headache in the Emergency Department.
Headaches represent over three million emergency department (ED) visits per year, comprising 2.4% of all ED visits. There are many proposed methods and clinical guidelines of treating acute headache presentations. However, data on intravenous acetaminophen usage in these settings are lacking. In this study, we sought to determine the efficacy of intravenous (IV) acetaminophen as an adjunct to a standard therapy for the treatment of patients who present to the ED with a chief complaint of "headache." We conducted a single site, randomized, double-blind, placebo-controlled trial investigating the clinical efficacy of IV acetaminophen as an adjunct to a standard therapy with prochlorperazine and diphenhydramine for the treatment of patients who present to the ED with a chief complaint of "headache" or variants thereof. (See below for variants). The primary outcome measure of the efficacy of parenteral acetaminophen as an adjunct treatment for headache in addition to a standard therapy was a threshold two-point reduction in visual analog scale (VAS) pain scores on a 1-10 level at 90 minutes. Secondary outcomes measures included assessment of decreased requirement of "rescue" pain medicines, defined as any analgesic medications outside of diphenhydramine, prochlorperazine and acetaminophen, with particular interest to potential opioid-sparing effects with parenteral acetaminophen. Additional secondary outcome measure included time to disposition from arrival in the ED. For the acetaminophen group the initial mean pain score was 8.67, for the placebo group 8.61. At 90 minutes pain score was 2.23 for the acetaminophen group and 3.99 for placebo (p<0.01, 95% confidence interval (CI) [0.8%-16%]. Of 45 patients in each group, we observed at least a threshold two-point decrease in pain score 36/45 (80%) with acetaminophen vs. 25/45 (55%) with placebo (p <0.01) 95% CI [5%-41%], number needed to treat (NNT) = 4). Secondary outcome measure did not demonstrate a difference in length of stay (161 minutes for acetaminophen arm and 159 minutes for placebo). However, 17/45 (38%) of patients who received IV acetaminophen required rescue analgesia, opposed to 24/45 (53%) of patients in the placebo group (p=0.13) 95% CI [-5%-34%]. IV acetaminophen when used with prochlorperazine and diphenhydramine to treat acute headaches in the ED resulted in statistically significant pain reduction compared with prochlorperazine and diphenhydramine alone as measured by both threshold of lowering VAS pain score by at least two points (NNT = 4) and overall decline in VAS pain score. Further study is required to validate these results.


Improved development of somatic cell cloned mouse embryos by vitamin C and latrunculin A.
Impaired development of embryos produced by somatic cell nuclear transfer (SCNT) is mostly associated with faulty reprogramming of the somatic nucleus to a totipotent state and can be improved by treatment with epigenetic modifiers. Here we report that addition of 100 μM vitamin C (VitC) to embryo culture medium for at least 16 h post-activation significantly increases mouse blastocyst formation and, when combined with the use of latrunculin A (LatA) during micromanipulation and activation procedures, also development to term. In spite of this, no significant effects on pluripotency (OCT4 and NANOG) or nuclear reprogramming markers (H3K14 acetylation, H3K9 methylation and DNA methylation and hydroxymethylation) could be detected. The use of LatA alone significantly improved in vitro development, but not full-term development. On the other hand, the simultaneous treatment of cloned embryos with VitC and the histone deacetylase inhibitor psammaplin A (PsA), in combination with the use of LatA, resulted in cloning efficiencies equivalent to those of VitC or PsA treatments alone, and the effects on pluripotency and nuclear reprogramming markers were less evident than when only the PsA treatment was applied. These results suggest that although both epigenetic modifiers improve cloning efficiencies, possibly through different mechanisms, they do not show an additive effect when combined. Improvement of SCNT efficiency is essential for its applications in reproductive and therapeutic cloning, and identification of molecules which increase this efficiency should facilitate studies on the mechanism of nuclear reprogramming and acquisition of totipotency.


Serological relations between twelve small RNA viruses of insects.
Serological tests were done to examine the relationships between twelve picorna-like viruses of insects. The results of the tests indicated that the majority of the viruses are unrelated. However, cricket paralysis virus, isolated from Australian wild field crickets, appeared to be identical to Drosophila C virus, independently isolated in France. Cricket paralysis virus was infective for adults of Drosophila melanogaster and its infectivity towards Galleria melonella was neutralised by Drosophila C virus antiserum. It is therefore concluded that cricket paralysis virus and Drosophila C virus are very closely related if not identical.


Micro-syringe chip-guided intratumoral administration of lipid nanoparticles for targeted anticancer therapy.
Nano-sized drug delivery system has been widely studied as a potential technique to promote tumor-specific delivery of anticancer drugs due to its passive targeting property, but resulting in very restricted improvements in its systemic administration so far. There is a requirement for a different approach that dramatically increases the targeting efficiency of therapeutic agents at targeted tumor tissues. To improve the tumor-specific accumulation of anticancer drugs and minimize their undesirable toxicity to normal tissues, a tumor-implantable micro-syringe chip (MSC) with a drug reservoir is fabricated. As a clinically established delivery system, six liposome nanoparticles (LNPs) with different compositions and surface chemistry are prepared and their physicochemical properties and cellular uptake are examined in vitro. Subsequently, MSC-guided intratumoral administration is studied to identify the most appropriate for the higher tumor targeting efficacy with a uniform intratumoral distribution. For efficient cancer treatment, pro-apoptotic anticancer prodrugs (SMAC-P-FRRG-DOX) are encapsulated to the optimal LNPs (SMAC-P-FRRG-DOX encapsulating LNPs; ApoLNPs), then the ApoLNPs are loaded into the 1 μL-volume drug reservoir of MSC to be delivered intratumorally for 9 h. The tumor accumulation and therapeutic effect of ApoLNPs administered via MSC guidance are evaluated and compared to those of intravenous and intratumoral administration of ApoLNP in 4T1 tumor-bearing mice. MSC is precisely fabricated to have a 0.5 × 4.5 mm needle and 1 μL-volume drug reservoir to achieve the uniform intratumoral distribution of LNPs in targeted tumor tissues. Six liposome nanoparticles with different compositions of 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine (PC), 1,2-dioleoyl-sn-glycero-3-phospho-L-serine (PS), 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy (polyethylene glycol)2000] (PEG2000-DSPE) are prepared with average sizes of 100-120 nm and loaded into the 1 μL-volume drug reservoir in MSC. Importantly negatively charged 10 mol% of PS-containing LNPs are very slowly infused into the tumor tissue through the micro-syringe of the MSC over 6 h. The intratumoral targeting efficiency of MSC guidance is 93.5%, effectively assisting the homogeneous diffusion of LNPs throughout the tumor tissue at 3.8- and 2.7-fold higher concentrations compared to the intravenous and intratumoral administrations of LNPs, respectively. Among the six LNP candidates 10 mol% of PS-containing LNPs are finally selected for preparing pro-apoptotic SMAC-P-FRRG-DOX anticancer prodrug-encapsulated LNPs (ApoLNPs) due to their moderate endocytosis rate high tumor accumulation and homogenous intratumoral distribution. The ApoLNPs show a high therapeutic effect specifically to cathepsin B-overexpressing cancer cells with 6.6 μM of IC50 value while its IC50 against normal cells is 230.7 μM. The MSC-guided administration of ApoLNPs efficiently inhibits tumor growth wherein the size of the tumor is 4.7- and 2.2-fold smaller than those treated with saline and intratumoral ApoLNP without MSC, respectively. Moreover, the ApoLNPs remarkably reduce the inhibitor of apoptosis proteins (IAPs) level in tumor tissues confirming their efficacy even in cancers with high drug resistance. The MSC-guided administration of LNPs greatly enhances the therapeutic efficiency of anticancer drugs via the slow diffusion mechanism through micro-syringe to tumor tissues for 6 h, whereas they bypass most hurdles of systemic delivery including hepatic metabolism, rapid renal clearance, and interaction with blood components or other normal tissues, resulting in the minimum toxicity to normal tissues. The negatively charged ApoLNPs with cancer cell-specific pro-apoptotic prodrug (SMAC-P-FRRG-DOX) show the highest tumor-targeting efficacy when they are treated with the MSC guidance, compared to their intravenous or intratumoral administration in 4T1 tumor-bearing mice. The MSC-guided administration of anticancer drug-encapsulated LNPs is expected to be a potent platform system that facilitates overcoming the limitations of systemic drug administration with low delivery efficiency and serious side effects.


Incidence of additional primary malignancies in patients with pancreatic and gastrointestinal neuroendocrine tumors.
The incidence of secondary malignancies is increased in patients with malignant and premalignant conditions. Although neuroendocrine tumors (NET) are uncommon, their incidence is increasing. We evaluated the rate of additional malignancies in patients with NET. Using the Surveillance, Epidemiology, and End Results (SEER) database, we identified a cohort of patients with pancreatic NET (PNET) or gastrointestinal NET (GINET). We determined the incidence of additional cancers diagnosed either before or after the diagnosis of PNET or GINET, by comparing these rates with the general population. Using multivariable regression, we evaluated factors that increased the risk of an additional malignancy. A cohort of 9,727 NET patients was identified. A total of 3,086 additional cancers occurred in 2,508 patients (25.8 %). The most common sites of additional malignancies included colorectal (21.1 %), prostate (14.5 %), breast (13.3 %), and lung (11.6 %). Among patients with PNET, the incidence of breast, lung, uterine, lymph, and pancreatic cancers was less than expected in the general population, whereas in patients with GINET, the observed incidence of nearly all malignancies exceeded that expected. Increasing age, marital status, and localized NET were associated with increased risk. Our study shows that the incidence of additional malignancies in patients with PNET and GINET is 25.8 %. Patients with GINET are at increased risk of additional malignancies, whereas patients with PNET have a decreased risk compared with the general population. More vigilant surveillance for secondary malignancies should be performed in patients with GINET. Studies investigating potential etiologic oncogenic pathways are warranted.


Neuronal and axonal loss are selectively linked to fibrillar amyloid-{beta} within plaques of the aged primate cerebral cortex.
The amyloid-beta peptide (Abeta) deposited in plaques in Alzheimer's disease has been shown to cause degeneration of neurons in experimental paradigms in vivo and in vitro. However, it has been difficult to convincingly demonstrate toxicity of native amyloid deposits in the aged and Alzheimer brains. Here we provide evidence that the fibrillar conformation of Abeta (fAbeta) deposited in compact plaques is associated with the pathologies observed in Alzheimer brains. fAbeta containing compact but not diffuse plaques in the aged rhesus cortex contained activated microglia and clusters of phosphorylated tau-positive swollen neurites. Scholl's quantitative analysis revealed that the area adjacent to fAbeta, containing compact but not diffuse plaques in aged rhesus, aged human, and Alzheimer's disease cortex, displays significant loss of neurons and small but statistically significant reduction in the density of cholinergic axons. These observations suggest that fAbeta toxicity may not be restricted to cultured cells and animal injection models. Rather, fAbeta deposited in native compact plaques in aged and AD brains may exert selective toxic effects on its surrounding neural environment.


CT of colon cancer metastases to the liver using modified RECIST criteria: determining the ideal number of target lesions to measure.
We sought to define the ideal number of target lesions to be measured to assess disease response in patients undergoing chemotherapy for colon cancer metastases to the liver. Thirty consecutive patients were recruited for this study. Patients were part of a multisite, randomized, double-arm, phase 3 clinical trial involving chemotherapy with an investigational drug for metastatic colon cancer. Patients were recruited from U.S. and international sites. Institutional review board approval was obtained, and informed consent was obtained from all patients. Our study included CT measurements of hepatic metastases. All patients (n = 30) had a minimum of five target lesions in the liver. Target-lesion size was defined by Response Evaluation Criteria in Solid Tumors (RECIST) criteria. We calculated the patient response at 2 months and at 6 months (complete response, partial response, stable disease, and progressive disease) using RECIST. Patient response was calculated based on the percentage increase or decrease at 2 and 6 months in the greatest diameter of the single largest lesion, two large lesions, three large lesions, four lesions, and five lesions, respectively. The concordance between five-target-lesion measurement and lesser numbers of lesions was analyzed using kappa statistics (StatView, 5.0). In 93.33% of patients (n = 28/30), there was agreement on patient response irrespective of the number of measurements made on CT. Of these 30 patients, 47% had a partial response (n = 14/30), 43% had stable disease (n = 13/30), and 10% had progressive disease at 2 months (n = 3/30). At 6 months, 43% had a partial response (n = 13/30), 47% had stable disease (n = 14/30), and 10% had progressive disease (n = 3/30). Agreement in response evaluation between lesion groups for multiple measurements was high, with values of 1.0 for multiple-lesion measurements and 0.88 for single-lesion measurements at 2 months. The concordance values were the same at 6 months. In the majority of patients with hepatic metastases of colorectal cancer, measuring the maximal diameter of the single largest lesion yielded the same treatment-response classification as measuring up to five target lesions. This result suggests that it may be possible to reduce the number of lesions measured in clinical trials.


Amidoxime-Functionalized Macroporous Carbon Self-Refreshed Electrode Materials for Rapid and High-Capacity Removal of Heavy Metal from Water.
Heavy metal pollution continues to be one of the most serious environmental problems which has attracted major global concern. Here, a rapid, high-capacity, yet economical strategy for deep cleaning of heavy metals ions in water is reported based on amidoxime-functionalized macroporous carbon electrode materials. The active sites of our material can be self-refreshed during the electrochemical removal process, which is different from traditional methods. The novel filter device in this work can purify contaminated water very rapidly (3000 L h-1 m-2), and can decrease heavy metal ion concentrations to below 5 ppb with a very short contact time (only 3 s). The original treatment efficiency of the device can be retained even after 1 week of continuous device operation. An extremely high removal capacity of over 2300 mg g-1 can be achieved with 2-3 orders of magnitude higher efficiency than that of surface adsorption-based commercial filters without any decay. Additionally, the cost of energy consumed in our method is lower than $6.67 × 10-3 per ton of wastewater. We envision that this approach can be routinely applied for the rapid, efficient, and thorough removal of heavy metals from both point-of-use water and industrial wastewater.


Exergames for subsyndromal depression in older adults: a pilot study of a novel intervention.
Subsyndromal depression (SSD) is several times more common than major depression in older adults and is associated with significant negative health outcomes. Physical activity can improve depression, but adherence is often poor. The authors assessed the feasibility, acceptability, and short-term efficacy and safety of a novel intervention using exergames (entertaining video games that combine game play with exercise) for SSD in older adults. Community-dwelling older adults (N = 19, aged 63-94 years) with SSD participated in a 12-week pilot study (with follow-up at 20-24 weeks) of Nintendo's Wii sports, with three 35-minute sessions a week. Eight-six percent of enrolled participants completed the 12-week intervention. There was a significant improvement in depressive symptoms, mental health-related quality of life (QoL), and cognitive performance but not physical health-related QoL. There were no major adverse events, and improvement in depression was maintained at follow-up. The findings provide preliminary indication of the benefits of exergames in seniors with SSD. Randomized controlled trials of exergames for late-life SSD are warranted.


Preventing depression after stroke: Results from a randomized placebo-controlled trial.
We designed this study to determine whether the daily treatment of nondepressed acute stroke patients with sertraline reduced the incidence of depression at follow-up. 111 patients with recent stroke (< 2 weeks; International Classification of Diseases, Tenth Revision criteria) were randomly assigned to treatment with placebo (N = 56) and sertraline (N = 55, 50 mg once daily) in this double-blind, placebo-controlled 24-week clinical trial. Subjects were recruited from the 2 largest teaching hospitals of Western Australia between June 2002 and June 2004. The primary endpoint of interest was development of clinically significant depressive symptoms as assessed by a Hospital Anxiety and Depression Scale-depression subscale score of 8 or above, or as diagnosed by the treating physician during 24 weeks. There was no significant difference in the incidence of depressive symptoms during 24 weeks of treatment (16.7% [8/48] sertraline vs. 21.6% [11/51] placebo, rate ratio = 0.8, 95% CI = 0.3 to 2.1, p = .590). The trial medication was discontinued by 51.8% (29/56) of patients assigned placebo and 47.3% (26/55) assigned sertraline (p = .634), most often because of perceived side effects or because the treating physician introduced an antidepressant medication. Twenty-four-week treatment with 50 mg of sertraline once daily initiated within 2 weeks of onset of acute stroke is not a significantly more effective strategy to prevent 6-month depression than usual care plus placebo among nondepressed stroke patients. New pharmacologic and nonpharmacologic strategies need to be developed to reduce the health and financial burden associated with depression after stroke.


[Morphometric characteristics of thrombocytes in renal amyloidosis].
Forty-three patients with primary, secondary and hereditary amyloidosis at different stages of renal lesions (proteinuric, nephrotic, chronic renal failure) were examined for the count and size of peripheral blood platelets. All the patients manifested a diminution of the mean diameter of platelets caused by predominance in the circulation of their minor forms. In half of the cases these alterations combined with thrombocytosis. A direct non-linear relationship was revealed between the size of platelets and their count in patients with primary and secondary amyloidosis. The possibility of primordial functional deficiency of platelets in patients with amyloidosis is discussed.


Effect of neural stimulation on contractile system (myoepithelial cells) in isolated salivary gland segments of rat.
The effect of cholinergic neural excitation by field stimulation on the tension of the contractile system (myoepithelial cells) was investigated in superfused segments of rat salivary glands (sublingual, submaxillary, and parotid). Electric field stimulation evoked gland tissue contraction of a few mg, which was blocked by addition of TTX or atropine. The order of the magnitude of the contraction per wet tissue weight was sublingual greater than submaxillary greater than parotid. This correlates with the order of the distribution of myoepithelial cells described in morphological investigations. Cholinergic stimulants induced contraction in all salivary glands, but adrenergic stimulants hardly evoked contraction in the sublingual gland, whereas those drugs evoked a considerable amount of contraction in the submaxillary gland. These findings seem to suggest that the contraction may originate in the myoepithelial cells. The omission of Na from the superfusing solution quickly abolished the contraction evoked by field stimulation but not the contraction induced by ACh. The omission of Ca from the solution reduced the contraction evoked not only by field stimulation but also by ACh and by high K. The results suggest that extracellular Ca is important for the initiation of myoepithelial contraction.


Symptoms of common mental disorder and cognitive associations with seropositivity among a cohort of people coming for testing for HIV/AIDS in Goa, India: a cross-sectional survey.
The majority of research on HIV/AIDS and mental health has been carried out among clinical populations: the time of onset of comorbid depression and the mechanisms for this are therefore unclear. Although there is evidence to suggest that asymptomatic people living with HIV/AIDS exhibit some cognitive deficits, the prevalence of poor cognitive functioning among people in low income settings at an early, pre-clinical stage has not yet been investigated. We used a cross-sectional survey design to test the hypotheses that symptoms of Common Mental Disorder (CMD) and low scores on cognitive tests would be associated with seropositivity among participants coming for testing for HIV/AIDS. Participants were recruited at the time of coming for testing for HIV/AIDS; voluntary informed consent was sought for participation in research interviews and data linkage with HIV test results. Baseline questionnaires including sociodemographic variables and measures of mental health (PHQ-9, GAD-7, panic disorder questions, AUDIT and delayed word list learning and recall and animal naming test of verbal fluency) were administered by trained interviews. HIV status data was extracted from clinical records. CMD and scoring below the educational norm on the test of verbal fluency were associated with testing positive for HIV/AIDS in bivariate analysis (OR = 2.26, 1.31-3.93; OR = 1.77, 1.26-2.48, respectively). After controlling for the effects of confounders, the association between CMD and seropositivity was no longer statistically significant (AOR = 1.56, 0.86-2.85). After adjusting for the effects of confounders, the association between low scores on the test of verbal fluency and seropositivity was retained (AOR = 1.77, 1.27-2.48). Our findings provide tentative evidence to suggest that low cognitive test scores (and possibly depressive symptoms) may be associated with HIV status among people who have yet to receive their HIV test results. Impaired cognitive functioning and depression-like symptoms may be the result of the same underlying neurological damage. CMD and cognitive impairment may overlap to a greater extent than previously assumed. If replicated, this may have implications for the way in which we measure and treat CMD and cognitive functioning among people living with HIV/AIDS.


Physician executives straddle the digital divide.
e-Health is here to stay and experts predict that the Internet will become the hub of health care. Rapid advancements in biotechnology and medical research, increasingly curious patients who surf the Internet for medical information, and pressures from managed care companies to contain costs and speed treatments are the central components driving e-health. Despite physician reluctance to embrace the e-revolution, many hospitals and medical groups are employing the Internet and information technology to improve their customer interface, as well as to reduce business costs. This article offers seven e-strategies for health care performance improvement: (1) Supply chain management; (2) e-transactions; (3) care management; (4) improving quality; (5) boosting revenues; (6) outsourcing; and (7) provider networks (Intranets). By helping to incorporate these key e-solutions, physician executives can position their organizations for success in the new millennium.


Enhancing methodological clarity: principle-based concept analysis.
The aim of this paper is to operationalize the principle-based method of concept analysis. While nursing has embraced the use of concept analysis as a valid and significant entree into an area of research, methodological development has created strategies of inquiry that vary in purpose and in the nature of their findings. We propose that, as the primary utility of concept analysis is to determine the existing state of the science so that further work may be strategically and appropriately planned, the method described as principle-based concept analysis is superior in providing evidence to support subsequent inquiry into the concept of interest. Three problematic issues are discussed in an effort to clarify and procedurally explicate the strategies employed in this approach: selecting disciplinary literatures for inclusion in the analysis; conceptually-driven sampling issues; and within- and across-discipline analytic techniques. In this form of concept analysis, each principle contributes to an understanding of the strengths and limitations of the present state of the concept in the scientific literature. We believe that this perspective will enable nursing to begin to harness the power of concept analysis for advancing science rather than simply imagining what a concept could be or constructing what we believe it should be.


Ythdf2-mediated STK11 mRNA decay supports myogenesis by inhibiting the AMPK/mTOR pathway.
An emerging research focus is the role of m6A modifications in mediating the post-transcriptional regulation of mRNA during mammalian development. Recent evidence suggests that m6A methyltransferases and demethylases play critical roles in skeletal muscle development. Ythdf2 is a m6A "reader" protein that mediates mRNA degradation in an m6A-dependent manner. However, the specific function of Ythdf2 in skeletal muscle development and the underlying mechanisms remain unclear. Here, we observed that Ythdf2 expression was significantly upregulated during myogenic differentiation, whereas Ythdf2 knockdown markedly inhibited myoblast proliferation and differentiation. Combined analysis of high-throughput sequencing, Co-IP, and RIP assay revealed that Ythdf2 could bind to m6A sites in STK11 mRNA and form an Ago2 silencing complex to promote its degradation, thereby regulating its expression and consequently, the AMPK/mTOR pathway. Furthermore, STK11 downregulation partially rescued Ythdf2 knockdown-induced impairment of proliferation and myogenic differentiation by inhibiting the AMPK/mTOR pathway. Collectively, our results indicate that Ythdf2 mediates the decay of STK11 mRNA, an AMPK activator, in an Ago2 system-dependent manner, thereby driving skeletal myogenesis by suppressing the AMPK/mTOR pathway. These findings further enhance our understanding of the molecular mechanisms underlying RNA methylation in the regulation of myogenesis and provide valuable insights for conducting in-depth studies on myogenesis.


Focal Salmonella enteritidis infection in a patient with HIV infection and other multiple causes of immunodeficiency.
An HIV-infected man receiving antiretroviral therapy-who also had lupus-like vasculitis and membranous glomerulonephritis (treated with prednisolone and azathioprine), beta-thalassaemia minor trait and post-radiotherapy functional asplenia (mimicking sickle cell disease-induced hyposplenism)-developed focal soft issue and bone infection caused by Salmonella enteritidis at the site of previous mycobacterial infection.


Sporadic meningioangiomatosis-associated atypical meningioma mimicking parenchymal invasion of brain: a case report and review of the literature.
Meningioangiomatosis is a rare hamartomatous lesion or meningiovascular malformation in brain. In extremely rare condition, meningioma may occur together with meningioangiomatosis, and only 19 cases have been described in English literature until now. We now report a case of meningioangiomatosis-associated meningioma with atypical and clear cell variant. A 34-year-old man presented a 3-month history of progressive numbness and weakness of his left lower extremity. He had no stigmata of neurofibromatosis type 2. Magnetic resonance imaging (MRI) revealed multifocal lesions in the right frontoparietal lobe. The lesions were totally removed. Microscopically, parts of lesions were atypical and clear cell meningioma corresponding to WHO grade II. The adjacent brain parenchyma showed the histological features of meningioangiomatosis. Neoplastic cells in atypical meningioma area were immunoreactive to epithelial membrane antigen (EMA) with high MIB-1 index of up to 20%. However, the spindle cells in meningioangiomatosis area were negative for EMA with low MIB-1 index of up to 1%. The diagnosis of atypical meningioma associated with sporadic meningioangiomatosis was made. To our knowledge, this is the first case of a meningioangiomatosis-associated meningioma with atypical and clear cell variant component to be described. The patient had been followed-up for 11 months without adjuvant radiotherapy or chemotherapy. No tumor recurrence was found during this period. Meningioangiomatosis-associated meningioma is more likely to occur in younger patients and histologically to mimic parenchymal invasion of brain. We suggest that postoperative radiotherapy or chemotherapy should be given careful consideration to avoid over-treatment due to erroneously interpret as malignant meningioma.


Men, masculinity, and the contexts of help seeking.
Research on men's help seeking yields strategies for enhancing men's use of mental and physical health resources. Analysis of the assumptions underlying existing theory and research also provides a context for evaluating the psychology of men and masculinity as an evolving area of social scientific inquiry. The authors identify several theoretical and methodological obstacles that limit understanding of the variable ways that men do or do not seek help from mental and physical health care professionals. A contextual framework is developed by exploring how the socialization and social construction of masculinities transact with social psychological processes common to a variety of potential help-seeking contexts. This approach begins to integrate the psychology of men and masculinity with theory and methodology from other disciplines and suggests innovative ways to facilitate adaptive help seeking.


Loss of integrin alpha1beta1 ameliorates Kras-induced lung cancer.
The collagen IV binding receptor integrin alpha1beta1 has been shown to regulate lung cancer due to its proangiogenic properties; however, it is unclear whether this receptor also plays a direct role in promoting primary lung tumors. To investigate this possibility, integrin alpha1-null mice were crossed with KrasLA2 mice that carry an oncogenic mutation of the Kras gene (G12D) and develop spontaneous primary tumors with features of non-small cell lung cancer. We provide evidence that KrasLA2/alpha1-null mice have a decreased incidence of primary lung tumors and longer survival compared with KrasLA2/alpha1 wild-type controls. Tumors from KrasLA2/alpha1-null mice were also smaller, less vascularized, and exhibited reduced cell proliferation and increased apoptosis, as determined by proliferating cell nuclear antigen and terminal deoxynucleotidyl-transferase-mediated dUTP nick-end staining, respectively. Moreover, tumors from the KrasLA2/alpha1-null mice showed diminished extracellular signal-regulated kinase (ERK) but enhanced p38 mitogen-activated protein kinase activation. Primary lung tumor epithelial cells isolated from KrasLA2/alpha1-null mice showed a significant decrease in anchorage-independent colony formation, collagen-mediated cell proliferation, ERK activation, and, most importantly, tumorigenicity when injected into nude mice compared with KrasLA2/alpha1 wild-type tumor cells. These results indicate that loss of the integrin alpha1 subunit decreases the incidence and growth of lung epithelial tumors initiated by oncogenic Kras, suggesting that both Kras and integrin alpha1beta1 cooperate to drive the growth of non-small cell lung cancer in vivo.


Islet Transplantation to the Anterior Chamber of the Eye-A Future Treatment Option for Insulin-Deficient Type-2 Diabetics? A Case Report from a Nonhuman Type-2 Diabetic Primate.
Replacement of the insulin-secreting beta cells through transplantation of pancreatic islets to the liver is a promising treatment for type-1 diabetes. However, low oxygen tension, shear stress, and the induction of inflammation lead to significant islet dysfunction and loss. The anterior chamber of the eye (ACE) has gained considerable interest and represents an alternative therapeutic islet transplantation site because of its accessibility, high oxygen tension, and immune-privileged milieu. We have previously demonstrated the feasibility of intraocular islet transplant in mouse and nonhuman primate models of type-1 diabetes and are now assessing its efficacy on glucose homeostasis in a nonhuman primate model of type-2 diabetes. We transplanted allogeneic donor islets (1,500 islet equivalents/kg) into the anterior chamber of one eye in a cynomolgus monkey with high-fat-diet-induced type-2 diabetes. Repeated examinations of the anterior and posterior segments of both eyes were done to monitor the engrafted islets and assess the overall ocular health. Fasting blood glucose level, blood biochemistry, and other metabolic parameters were routinely evaluated to determine the function of the islet graft and diabetes status. The transplanted islets were rapidly engrafted onto the iris and became vascularized 1 month after transplantation. We did not detect changes in intraocular pressure, cataract formation, ophthalmitis, or retinal vessel deformation. A significant lower fasting blood glucose level was observed while the graft was in place, and the transplantation reverts the progression of diabetes. The metabolic markers, hemoglobin A1C and fructosamine, demonstrated improvement following islet transplantation. As a conclusion, intraocular islet transplantation in one eye of a cynomolgus monkey with type-2 diabetes improved its overall plasma glucose homeostasis, as evidenced by short-term measures and long-term metabolic markers. These results further support the future application of the ACE as an alternative site for clinical islet transplants in the context of type-2 diabetes.


New perspectives on the role of Escherichia coli O157:H7 and other enterohaemorrhagic E. coli serotypes in human disease.
This review compares the rates of detection of non-O157:H7 enterohaemorrhagic Escherichia coli (EHEC) with EHEC O157:H7 in outbreaks and sporadic cases of human disease by analysing Australian data and the world literature. Numerous outbreaks of disease have been attributed to EHEC O157:H7. In many studies, isolation rates of this organism have been low and attempts to seek other EHEC have not been made. Ease of isolation and identification of the O157:H7 serotype may have given the impression that this serotype was the sole organism responsible for the outbreaks. Careful review and analysis shows that serotypes other than O157:H7 also play an important role in human disease. Evidence is presented from several overseas outbreaks described in the literature, as well as from investigations of the Adelaide O111:H- outbreak, that suggests an association between severity of disease and multiple infecting serotypes. While not diminishing the role of the O157:H7/H- clone, this review indicates that other serotypes can be responsible for outbreaks as well as cases of sporadic human disease. The current focus on O157:H7 has major implications in terms of diagnosis, the food industry and human health.


IgE-mediated allergy to corn: a 50 kDa protein, belonging to the Reduced Soluble Proteins, is a major allergen.
Although corn is often cited as an allergenic food, very few studies have been devoted to the identification of corn allergens and corn allergy has been rarely confirmed by double-blind, placebo-controlled food challenge (DBPCFC). Recently, Pastorello et al. (1) identified some salt-soluble IgE-binding proteins of corn flour as potential allergens. One of these, corresponding to corn Lipid Transfer Protein (LTP), appeared to be the major one. The aim of this study was to verify the clinical significance of the skin prick test (SPT) and CAP-FEIA CAP-System IgE fluozoenzyme immunosorbent assay (Pharmacia Diagnostic, Uppsala, Sweden) positivities to corn and to identify the presence of IgE-binding proteins in the corn flour salt-insoluble protein fractions (comprising up to 96% of the total protein) using sera of patients with DBPCFC-documented food allergy to corn. In addition the effect of cooking and proteolytic digestion on the corn allergens was investigated. Sixteen subjects with SPT and CAP-FEIA positivities to corn flour were examined. Only six of them complained of suffering from urticaria and/or other symptoms after ingestion of corn-based foods. The patients were food challenged with cooked corn flour (polenta). IgE-binding proteins were detected by immunoblotting. The digestibility of the IgE-binding proteins was examined during a pepsin attack followed by a pancreatin digestion performed on a cooked corn flour sample. Oral challenge was positive only for six patients with symptoms after ingestion of corn. A 50 kDa protein, belonging to the corn Reduced Soluble Protein (RSP) fraction was recognized by the serum IgE of all the DBPCFC-positive subjects and resulted to be resistant to both heating and peptic/pancreatic digestion. SPT with the purified RSP fraction gave positive results for all of the DBPCFC-positive patients examined. SPT and CAP-FEIA positivities to corn flour had no clinical significance for most of the patients and food allergy to corn has to be proved by DBPCFC. A salt-unextractable protein of 50 kDa, belonging to the RSP fraction, represents a potential allergen in food hypersensitivity to corn because of its stability to cooking and digestion.


The behavioral phenotype of pituitary adenylate-cyclase activating polypeptide-deficient mice in anxiety and depression tests is accompanied by blunted c-Fos expression in the bed nucleus of the stria terminalis, central projecting Edinger-Westphal nucleus, ventral lateral septum, and dorsal raphe nucleus.
Pituitary adenylate-cyclase activating polypeptide (PACAP) has been implicated in the (patho)physiology of stress-adaptation. PACAP deficient (PACAP(-/-)) mice show altered anxiety levels and depression-like behavior, but little is known about the underlying mechanisms in stress-related brain areas. Therefore, we aimed at investigating PACAP(-/-) mice in light-dark box, marble burying, open field, and forced swim paradigms. We also analyzed whether the forced swim test-induced c-Fos expression would be affected by PACAP deficiency in the following stress-related brain areas: magno- and parvocellular paraventricular nucleus of the hypothalamus (PVN); basolateral (BLA), medial (MeA), and central (CeA) amygdaloid nuclei; ventral (BSTv), dorsolateral (BSTdl), dorsomedial (BSTdm), and oval (BSTov) nuclei of the bed nucleus of stria terminalis; dorsal (dLS) and ventral parts (vLS) of lateral septal nucleus, central projecting Edinger-Westphal nucleus (EWcp), dorsal (dPAG) and lateral (lPAG) periaqueductal gray matter, dorsal raphe nucleus (DR). Our results revealed that PACAP(-/-) mice showed greatly reduced anxiety and increased locomotor activity compared with wildtypes. In forced swim test PACAP(-/-) mice showed increased depression-like behavior. Forced swim exposure increased c-Fos expression in all examined brain areas in wildtypes, whereas this was markedly blunted in the DR, EWcp, BSTov, BSTdl, BSTv, PVN, vLS, dPAG, and in the lPAG of PACAP(-/-) mice vs. wildtypes, strongly suggesting their involvement in the behavioral phenotype of PACAP(-/-) mice. PACAP deficiency did not influence the c-Fos response in the CeA, MeA, BSTdm, and dLS. Therefore, we propose that PACAP exerts a brain area-specific effect on stress-induced neuronal activation and it might contribute to stress-related mood disorders.


The ferroportin Q248H mutation protects from anemia, but not malaria or bacteremia.
Iron acquisition is critical for life. Ferroportin (FPN) exports iron from mature erythrocytes, and deletion of the Fpn gene results in hemolytic anemia and increased fatality in malaria-infected mice. The FPN Q248H mutation (glutamine to histidine at position 248) renders FPN partially resistant to hepcidin-induced degradation and was associated with protection from malaria in human studies of limited size. Using data from cohorts including over 18,000 African children, we show that the Q248H mutation is associated with modest protection against anemia, hemolysis, and iron deficiency, but we found little evidence of protection against severe malaria or bacteremia. We additionally observed no excess Plasmodium growth in Q248H erythrocytes ex vivo, nor evidence of selection driven by malaria exposure, suggesting that the Q248H mutation does not protect from malaria and is unlikely to deprive malaria parasites of iron essential for their growth.