Comparative effectiveness research on patients with acute ischemic stroke using Markov decision processes.
Several methodological issues with non-randomized comparative clinical studies have been raised, one of which is whether the methods used can adequately identify uncertainties that evolve dynamically with time in real-world systems. The objective of this study is to compare the effectiveness of different combinations of Traditional Chinese Medicine (TCM) treatments and combinations of TCM and Western medicine interventions in patients with acute ischemic stroke (AIS) by using Markov decision process (MDP) theory. MDP theory appears to be a promising new method for use in comparative effectiveness research. The electronic health records (EHR) of patients with AIS hospitalized at the 2nd Affiliated Hospital of Guangzhou University of Chinese Medicine between May 2005 and July 2008 were collected. Each record was portioned into two "state-action-reward" stages divided by three time points: the first, third, and last day of hospital stay. We used the well-developed optimality technique in MDP theory with the finite horizon criterion to make the dynamic comparison of different treatment combinations. A total of 1504 records with a primary diagnosis of AIS were identified. Only states with more than 10 (including 10) patients' information were included, which gave 960 records to be enrolled in the MDP model. Optimal combinations were obtained for 30 types of patient condition. MDP theory makes it possible to dynamically compare the effectiveness of different combinations of treatments. However, the optimal interventions obtained by the MDP theory here require further validation in clinical practice. Further exploratory studies with MDP theory in other areas in which complex interventions are common would be worthwhile.


New sonographically-guided test for anterior knee instability - preliminary report.
Ultrasound examination is widely used in orthopedic diagnostics, however sonographic evaluation of traumatic anterior cruciate ligament insufficiency is still inadequate. Aim of this study is to evaluate diagnostic capability of a new sonographically-guided test for diagnosing complete anterior cruciate ligament insufficiency. In 47 patients, with suspicion of unilateral anterior cruciate ligament injury (based on magnetic resonance imaging), the sonographically-guided test for anterior instability was performed. The translation of the intercondylar eminence against the patellar tendon was measured in both knees. Afterwards all patients underwent arthroscopy. In 37 patients, with arthroscopically confirmed complete anterior cruciate ligament insufficiency, the mean anterior knee translation was 8.3 mm (SD = 2.8) in affected knee vs. 3 mm (SD = 1.1) in uninjured knee (p < 0.001). In 10 patients with no anterior cruciate ligament insufficiency the difference between body sides was not significant (2.6 mm, SD = 1.4 in injured knee vs. 2.5 mm, SD = 1.1 in uninjured joint; p < 0.7753). The proposed test supports the clinician with fast and non-invasive examination that can facilitate evaluation of anterior knee instability.


Abrogation of H-2-restricted CTL responses and efficient recognition of HLA-A3 molecules in DBA/2 HLA/A24 responder mice.
DBA/2 (H-2d) HLA-B7 x human beta 2-microglobulin transgenic and DBA/2 nontransgenic mice stimulated by DBA/2 HLA-A3 x human beta 2-microglobulin transgenic mouse spleen cells developed potent H-2Kd restricted cytolytic responses with recognition of a peptide from the second domain of the HLA-A3 H chain. These H-2Kd-restricted responses obliterated, as a rule, cytolytic responses with direct recognition of the HLA-A3 molecules, even in HLA-B7 transgenic mice. These immunodominant H-2Kd-restricted responses could be abrogated in DBA/2 HLA-A24 mice because of cross-tolerance, the HLA-A3 derived-H-2Kd presented peptide being shared by several (including A24) HLA class I H chain allelic variants. Under such experimental circumstances, strong CTL responses with exclusive direct recognition of HLA-A3 molecules constantly developed. Further analysis of these responses in six DBA/2 HLA-A24 responder mice indicated that a large fraction of the mouse V beta and V alpha genes could be used to mount such CTL responses. Thus, by combining classical HLA class I transgenesis and selective destruction of H-2K and H-2D genes, it should be possible to derive useful strains of mice for the study of HLA class I-restricted CTL responses.


Investigation into inconsistent lateralisation of language functions as a potential risk factor for language impairment.
Disruption to language lateralisation has been proposed as a cause of developmental language impairments. In this study, we tested the idea that consistency of lateralisation across different language functions is associated with language ability. A large sample of adults with variable language abilities (N = 67 with a developmental disorder affecting language and N = 37 controls) were recruited. Lateralisation was measured using functional transcranial Doppler sonography (fTCD) for three language tasks that engage different language subprocesses (phonological decision, semantic decision and sentence generation). The whole sample was divided into those with consistent versus inconsistent lateralisation across the three tasks. Language ability (using a battery of standardised tests) was compared between the consistent and inconsistent groups. The results did not show a significant effect of lateralisation consistency on language skills. However, of the 31 individuals showing inconsistent lateralisation, the vast majority (84%) were in the disorder group with only five controls showing such a pattern, a difference that was higher than would be expected by chance. The developmental disorder group also demonstrated weaker correlations between laterality indices across pairs of tasks. In summary, although the data did not support the hypothesis that inconsistent language lateralisation is a major cause of poor language skills, the results suggested that some subtypes of language disorder are associated with inefficient distribution of language functions between hemispheres. Inconsistent lateralisation could be a causal factor in the aetiology of language disorder or may arise in some cases as the consequence of developmental disorder, possibly reflective of compensatory reorganisation.


Electronic medical records and genomics (eMERGE) network exploration in cataract: several new potential susceptibility loci.
Cataract is the leading cause of blindness in the world, and in the United States accounts for approximately 60% of Medicare costs related to vision. The purpose of this study was to identify genetic markers for age-related cataract through a genome-wide association study (GWAS). In the electronic medical records and genomics (eMERGE) network, we ran an electronic phenotyping algorithm on individuals in each of five sites with electronic medical records linked to DNA biobanks. We performed a GWAS using 530,101 SNPs from the Illumina 660W-Quad in a total of 7,397 individuals (5,503 cases and 1,894 controls). We also performed an age-at-diagnosis case-only analysis. We identified several statistically significant associations with age-related cataract (45 SNPs) as well as age at diagnosis (44 SNPs). The 45 SNPs associated with cataract at p<1×10(-5) are in several interesting genes, including ALDOB, MAP3K1, and MEF2C. All have potential biologic relationships with cataracts. This is the first genome-wide association study of age-related cataract, and several regions of interest have been identified. The eMERGE network has pioneered the exploration of genomic associations in biobanks linked to electronic health records, and this study is another example of the utility of such resources. Explorations of age-related cataract including validation and replication of the association results identified herein are needed in future studies.


Long-term functional outcome of pediatric stroke survivors.
To examine the long-term functional, psychosocial, and medical outcome of pediatric stroke survivors. This was a descriptive survey performed on patients with childhood stroke who participated in an earlier study. Measures included the Vineland Adaptive Behavior Scales (VABS) and the Diener Satisfaction with Life Scale. Current information on living situation, school placement, employment, and medical outcome were obtained. Twenty-nine (58%) patients participated. The mean age was 19.3 years (SD = 6.6), mean age of onset of stroke was 7.0 years (SD = 5.4), and mean follow-up time was 11.9 years (SD = 3.9). Diagnoses included hemorrhagic (31%) and ischemic (69%) stroke. All but one adult had finished high school, and the majority of participants had gone to college. 60% of patients over age 16 were employed. The average VABS levels for communication, daily living skills, socialization, and adaptive behavior fell into the moderately low range. Use of seizure medications and ADL dependence were the predictors for lower VABS levels (p <.05). Younger age, ischemic stroke, and previous dependence in mobility were risk factors for lower scores for the self-care domain, but not for lower life satisfaction. Patients who scored below adequate on VABS tended toward lower life satisfaction. Pediatric stroke survivors had good educational and mobility outcomes, but communication, ADL, and socialization fell into the low-moderate range. The different predictors of functional and subjective quality of life outcomes suggest that functional outcomes may mediate the relations between medical factors and satisfaction with life.


Notch1 and TGFbeta1 cooperatively regulate Foxp3 expression and the maintenance of peripheral regulatory T cells.
Notch and its ligands have been implicated in the regulation and differentiation of various CD4(+) T-helper cells. Regulatory T cells (T(regs)), which express the transcription factor Foxp3, suppress aberrant immune responses that are typically associated with autoimmunity or excessive inflammation. Previous studies have shown that transforming growth factor beta (TGFbeta1) induces Foxp3 expression and a regulatory phenotype in peripheral T cells. Here, we show that pharmacologic inhibition of Notch signaling using gamma-secretase inhibitor (GSI) treatment blocks (1) TGFbeta1-induced Foxp3 expression, (2) the up-regulation of Foxp3-target genes, and (3) the ability to suppress naive T-cell proliferation. In addition, the binding of Notch1, CSL, and Smad to conserved binding sites in the foxp3 promoter can be inhibited by treatment with GSI. Finally, in vivo administration of GSI results in reduced Foxp3 expression and development of symptoms consistent with autoimmune hepatitis, a disease previously found to result from dysregulation of TGFbeta signaling and regulatory T cells. Together, these findings indicate that the Notch and TGFbeta signaling pathways cooperatively regulate Foxp3 expression and regulatory T-cell maintenance both in vitro and in vivo.


Dosimetric evaluation of target coverage as a predictor of local failure following stereotactic body radiation therapy for spinal tumors.
To perform a dosimetric analysis of target coverage and determine parameters predictive for local failure (LF) in patients undergoing spinal stereotactic body radiation therapy (sSBRT).Materials and Methods: Sixty-seven spinal tumors in 59 patients were treated with image-guided linac-based sSBRT from 2008-2012. Median prescription dose was 18Gy (8-35) delivered in 1-5 fractions (87% single-fraction). Prescription dose was targeted to cover ≥ 80% of PTV within spinal cord (SC) dose constraints (9/11Gy to 0.1cc SC/SC+2mm). Twelve tumors had local failure (LF, median time-to-failure 3.7 months) and were compared to 14 tumors with >1-year follow-up and local control (LC). Univariate and multivariate analyses were performed to determine parameters predictive of LF. Median follow-up was 7.4 months and 24.7 months for LF and LC, respectively. Post-SBRT, 42% of LF patients had neurological symptoms due to tumor progression. No patients developed post-SBRT myelopathy. Pre-treatment PTV volumes were not statistically different (median/mean/range 61.8/74.5/19.9-206.4cc for LF vs 39.4/47.1/10.3-119.7cc for LC; p=0.13). LF tumors had larger volumes receiving <80% of prescription dose (5.2cc vs 1.9cc, p=0.02) and larger overlap volume between GTV/SC within 2 and 3mm (p=0.01/p=0.007). LF tumors had lower GTV minimum dose (5.6 vs 8.5Gy, p=0.001) and smaller GTV to SC distance (0.06 vs 0.19mm, p=0.049). Maximum SC doses were not statistically different (6.4Gy LC vs 9.2Gy LF, p=0.33). GTV minimum dose was predictive of LF, with a trend for overlapping GTV/SC volume within 2mm. Minimum GTV dose, PTV volume receiving <80% prescription dose, smaller GTV-SC distance, and large overlapping volume of PTV/SC are predictive of LF after SBRT. Given the absence of SC toxicity but neurological progression upon LF, less conservative SC constraints should be considered.


Systematic review of reviews including animal studies addressing therapeutic interventions for sepsis.
Certain methodologic features of animal experiments such as random assignment have been found to reduce the risk of bias. Because animal research sometimes informs clinical practice, explicit acknowledgment of the risk of bias and clinical relevance cultivates realistic expectations on the part of clinicians reading preclinical studies. We assessed literature reviews of therapeutic interventions for sepsis that include animal experiments for explicit appraisals of the risk of bias and clinical relevance. MEDLINE and EMBASE. Systematic reviews or meta-analyses of animal experiments focusing on therapeutic interventions for sepsis. In teams of two, reviewers independently screened citations and abstracted data. We determined whether the reviews systematically incorporated critical appraisals for the risk of bias and clinical relevance of the underlying studies as well as explicit extrapolations from preclinical research to human patients. From 164 citations, we retained 45 reviews. Chance-corrected agreement for inclusion was moderate (κ 0.57). Three (7%) met our criteria for a systematic review and one (2%) systematically appraised the risk of bias and the clinical relevance of the primary animal experiments. Thirty-six (80%) were narrative reviews addressing issues related to diverse topics such as pathophysiology and diagnosis as well as multiple therapies and 40 of 45 (89%) included both clinical and animal studies. Twelve (27%) explicitly assumed that data from preclinical studies could apply to human patients. Although a significant proportion of reviews extrapolated preclinical study results to human patients, most did not systematically appraise the risk of bias or the clinical relevance of preclinical research. Because animal experiments may influence clinical practice, we propose a framework to enhance these features in future reviews of preclinical research.


Swedish national register for retinopathy of prematurity (SWEDROP) and the evaluation of screening in Sweden.
To evaluate screening for retinopathy of prematurity (ROP) in Sweden and to investigate possible modifications of the present screening guidelines. Infants in Sweden with a gestational age (GA) of 31 weeks + 6 days or less are screened for ROP. Data from the Swedish national register for ROP (SWEDROP) during 2008 and 2009 were extracted and compared with a national perinatal quality register. In SWEDROP, there were 1791 infants born before a GA of 32 weeks from January 1, 2008, through December 31, 2009. Another 70 infants were registered in the perinatal quality register but not in SWEDROP (dropout rate, 3.8% [70 of 1861 infants]). Seven infants died before termination of screening. In the final study cohort (1784 infants), 15.6% had mild ROP and 8.5% had severe ROP. Treatment was performed in 4.4% of the infants, none of whom had a GA at birth of more than 28 weeks. Nine infants with a GA of more than 28 weeks at birth developed stage 3 ROP, which regressed spontaneously. The total number of examinations was 9286 (964 in infants with a GA of 31 weeks), and the mean (range) number of examinations of each infant was 5.2 (1-30). The SWEDROP, a quality register for ROP, has a national coverage (ie, participation) of 96%. Data from 2008 to 2009 show that it seems possible to reduce the upper limit for screening in Sweden by 1 week, including only infants with a GA of 30 weeks + 6 days or less. However, such a change should be combined with a strong recommendation to neonatologists to refer also severely ill and more "mature" infants.


Intra-individual variation in serum C-reactive protein over 4 years: an implication for epidemiologic studies.
Data on long-term intra-individual variability in high-sensitivity C-reactive protein (hsCRP) are needed to determine whether one measurement adequately reflects usual levels in prospective studies of on the etiology of cancer and other chronic diseases; when not reflective, the ability to statistically detect modest to moderate associations is reduced. The authors estimated the size of this source of variability and consequent attenuation of the relative risk (RR). High-sensitivity C-reactive protein (hsCRP) concentration was measured using a high-sensitivity immunoturbidometric assay in sera collected at years 2, 4, and 6 from 50 men in the placebo arm of the Prostate Cancer Prevention Trial (PCPT). After natural logarithm-transformation of hsCRP, analysis of variance was used to estimate the within- and between-individual variances from which the intra-class correlation coefficient (ICC) was calculated. The observed RR due to an ICC < 1 was calculated by e((ln true RR*ICC)) for a range of true RRs. The 4-year ICC was 0.66. Measuring hsCRP once and assuming no other error, if the true RRs were 1.50, 2.00, and 3.00 when comparing high with low concentration, then the observed RRs would be 1.31, 1.58, and 2.06, respectively. Investigators planning to measure hsCRP only once should design adequately sized studies to preserve inferences for hypothesized modest to moderate RRs.


From Positivity to Negativity Bias: Ambiguity Affects the Neurophysiological Signatures of Feedback Processing.
Previous studies on the neurophysiological underpinnings of feedback processing almost exclusively used low-ambiguity feedback, which does not fully address the diversity of situations in everyday life. We therefore used a pseudo trial-and-error learning task to investigate ERPs of low- versus high-ambiguity feedback. Twenty-eight participants tried to deduce the rule governing visual feedback to their button presses in response to visual stimuli. In the blocked condition, the same two feedback words were presented across several consecutive trials, whereas in the random condition feedback was randomly drawn on each trial from sets of five positive and five negative words. The feedback-related negativity (FRN-D), a frontocentral ERP difference between negative and positive feedback, was significantly larger in the blocked condition, whereas the centroparietal late positive complex indicating controlled attention was enhanced for negative feedback irrespective of condition. Moreover, FRN-D in the blocked condition was due to increased reward positivity (Rew-P) for positive feedback, rather than increased (raw) FRN for negative feedback. Our findings strongly support recent lines of evidence that the FRN-D, one of the most widely studied signatures of reinforcement learning in the human brain, critically depends on feedback discriminability and is primarily driven by the Rew-P. A novel finding concerned larger frontocentral P2 for negative feedback in the random but not the blocked condition. Although Rew-P points to a positivity bias in feedback processing under conditions of low feedback ambiguity, P2 suggests a specific adaptation of information processing in case of highly ambiguous feedback, involving an early negativity bias. Generalizability of the P2 findings was demonstrated in a second experiment using explicit valence categorization of highly emotional positive and negative adjectives.


Does income-related health inequality change as the population ages? Evidence from Swedish panel data.
This paper explains and empirically assesses the channels through which population aging may impact on income-related health inequality. Long panel data of Swedish individuals is used to estimate the observed trend in income-related health inequality, measured by the concentration index (CI). A decomposition procedure based on a fixed effects model is used to clarify the channels by which population aging affects health inequality. Based on current income rankings, we find that conventional unstandardized and age-gender-standardized CIs increase over time. This trend in CIs is, however, found to remain stable when people are instead ranked according to lifetime (mean) income. Decomposition analyses show that two channels are responsible for the upward trend in unstandardized CIs - retired people dropped in relative income ranking and the coefficient of variation of health increases as the population ages.


Nonunions of the humerus - Treatment concepts and results of the last five years.
Fractures of the humerus account for 5%-8% of all fractures. Nonunion is found with an incidence of up to 15%, depending on the location of the fracture. In case of a manifest nonunion the surgeon faces a challenging problem and has to conceive a therapy based on the underlying pathology. The aim of this study was to describe our treatment concepts for this entity and present our results of the last five years. Twenty-six patients were treated for nonunion of the humerus between January 2013 and December 2017. Their charts were reviewed retrospectively and demographic data, pathology, surgical treatment and outcome were assessed. The most frequent location for a nonunion was the humeral shaft, with the most common trauma mechanism being multiple falls. Most often atrophic nonunion (n = 14), followed by hypertrophic and infection-caused nonunion (each n = 4), were found. Our treatment concept could be applied in 19 patients, of which in 90% of those who were available for follow-up consolidation could be achieved. Humeral nonunion is a heterogeneous entity that has to be analyzed precisely and be treated correspondingly. We therefore present a treatment concept based on the underlying pathology.


Bundled Bispectral Index Monitoring and Sedation During Paralysis in Acute Respiratory Distress Syndrome.
Clinical assessments of depth of sedation are insufficient for patients undergoing neuromuscular blockade during treatment of acute respiratory distress syndrome (ARDS). This quality initiative was aimed to augment objective assessment and improve sedation during therapeutic paralysis using the bispectral index (BIS). This quality improvement intervention provided education and subsequent implementation of a BIS monitoring and sedation/analgesia bundle in a large, urban, safety-net intensive care unit. After the intervention, a retrospective review of the first 70 admissions with ARDS assessed use and documented sedation changes in response to BIS. Therapeutic neuromuscular blockade was initiated for 58 of 70 patients (82.8%) with ARDS, of whom 43 (74%) had BIS monitoring and 29.3% had bundled BIS sedation-titration orders. Explicit documentation of sedation titration in response to BIS values occurred in 27 (62.8%) of those with BIS recordings. BIS sedation/analgesia bundled order sets are underused, but education and access to BIS monitoring led to high use of monitoring alone and subsequent sedation changes.


Association of Neighborhood Gun Violence With Mental Health-Related Pediatric Emergency Department Utilization.
Many children and adolescents in the United States are exposed to neighborhood gun violence. Associations between violence exposure and children's short-term mental health are not well understood. To examine the association between neighborhood gun violence and subsequent mental health-related pediatric emergency department (ED) utilization. This location-based cross-sectional study included 128 683 ED encounters for children aged 0 to 19 years living in 12 zip codes in Philadelphia, Pennsylvania, who presented to an urban academic pediatric ED from January 1, 2014, to December 31, 2018. Children were included if they (1) had 1 or more ED visits in the 60 days before or after a neighborhood shooting and (2) lived within a quarter-mile radius of the location where this shooting occurred. Analysis began August 2020 and ended May 2021. Neighborhood violence exposure, as measured by whether a patient resided near 1 or more episodes of police-reported gun violence. ED encounters for a mental health-related chief complaint or primary diagnosis. A total of 2629 people were shot in the study area between 2014 and 2018, and 54 341 children living nearby had 1 or more ED visits within 60 days of a shooting. The majority of these children were Black (45 946 [84.5%]) and were insured by Medicaid (42 480 [78.1%]). After adjusting for age, sex, race and ethnicity, median household income by zip code, and insurance, children residing within one-eighth of a mile (2-3 blocks) of a shooting had greater odds of mental health-related ED presentations in the subsequent 14 days (adjusted odds ratio, 1.86 [95% CI, 1.20-2.88]), 30 days (adjusted odds ratio, 1.49 [95% CI, 1.11-2.03]), and 60 days (adjusted odds ratio, 1.35 [95% CI, 1.06-1.72]). Exposure to neighborhood gun violence is associated with an increase in children's acute mental health symptoms. City health departments and pediatric health care systems should work together to provide community-based support for children and families exposed to violence and trauma-informed care for the subset of these children who subsequently present to the ED. Policies aimed at reducing children's exposure to neighborhood gun violence and mitigating the mental symptoms associated with gun violence exposure must be a public health priority.


Fisetin Attenuates Lipopolysaccharide-Induced Inflammatory Responses in Macrophage.
Several studies have reported the efficacy and safety of polyphenols in human health; however, the verification of their efficacy remains insufficient. The aim of this study was to examine whether fisetin, one of flavonoids prevalently present in fruits and vegetables, could suppress lipopolysaccharide- (LPS-) induced inflammatory responses in macrophages. LPS increased proinflammatory mRNA abundance (MCP 1, IL-1β, and iNOS) but were suppressed by fisetin. The increment of nitric oxide by LPS, an oxidative stress factor, was attenuated by fisetin. In addition, LPS-enhanced phosphorylation of mitogen-activated protein kinase (ERK and JNK) was reduced. Finally, fisetin attenuated the expression or activity of uPA, uPAR, MMP-2, and MMP-9, which are known as associated factors of macrophage recruitment or infiltration. In conclusion, fisetin is a promising therapeutic agent for macrophage-related inflammation diseases, like sepsis and atherosclerosis.


Volume and neuron number of the lateral geniculate nucleus in schizophrenia and mood disorders.
Subjects with schizophrenia show deficits in visual perception that suggest changes predominantly in the magnocellular pathway and/or the dorsal visual stream important for visiospatial perception. We previously found a substantial 25% reduction in neuron number of the primary visual cortex (Brodmann's area 17, BA17) in postmortem tissue from subjects with schizophrenia. Also, many studies have found reduced volume and neuron number of the pulvinar--the large thalamic association nucleus involved in higher-order visual processing. Here, we investigate if the lateral geniculate nucleus (LGN), the visual relay nucleus of the thalamus, has structural changes in schizophrenia. We used stereological methods based on unbiased principles of sampling (Cavalieri's principle and the optical fractionator) to estimate the total volume and neuron number of the magno- and parovocellular parts of the left LGN in postmortem brains from nine subjects with schizophrenia, seven matched normal comparison subjects and 13 subjects with mood disorders. No significant schizophrenia-related structural differences in volume or neuron number of the left LGN or its major subregions were found, but we did observe a significantly increased total volume of the LGN, and of the parvocellular lamina and interlaminar regions, in the mood group. These findings do not support the hypothesis that subjects with schizophrenia have structural changes in the LGN. Therefore, our previous observation of a schizophrenia-related reduction of the primary visual cortex is probably not secondary to a reduction in the LGN.


Nucleolar release of Hand1 acts as a molecular switch to determine cell fate.
The bHLH transcription factor Hand1 is essential for placentation and cardiac morphogenesis in the developing embryo. Here we implicate Hand1 as a molecular switch that determines whether a trophoblast stem cell continues to proliferate or commits to differentiation. We identify a novel interaction of Hand1 with a protein that contains an I-mfa (inhibitor of myogenic factor) domain that anchors Hand1 in the nucleolus where it negatively regulates Hand1 activity. In the trophoblast stem-cell line Rcho-1, nucleolar sequestration of Hand1 accompanies sustained cell proliferation and renewal, whereas release of Hand1 into the nucleus leads to its activation, thus committing cells to a differentiated giant-cell fate. Site-specific phosphorylation is required for nucleolar release of Hand1, for its dimerization and biological function, and this is mediated by the non-canonical polo-like kinase Plk4 (Sak). Sak is co-expressed in Rcho-1 cells, localizes to the nucleolus during G2 and phosphorylates Hand1 as a requirement for trophoblast stem-cell commitment to a giant-cell fate. This study defines a novel cellular mechanism for regulating Hand1 that is a crucial step in the stem-cell differentiation pathway.


Visor-display design based on planar holographic optics.
A method for designing and recording visor displays based on planar holographic optics is presented. This method can deal with the problem of recording-readout wavelength shift. The display system is composed of two holographic optical elements that are recorded on the same substrate. One element collimates the waves from each data point in the display into a plane wave that is trapped inside the substrate by total internal reflection. The other diffracts the plane waves into the eye of an observer. Because the chromatic dispersion of the first element can be corrected by the dispersion of the second, this configuration is relatively insensitive to source wavelength shifts. The method is illustrated by the design, recording, and testing of a compact holographic doublet visor display. The recording was at a wavelength of 458 nm, and readout was at 633 nm. The results indicate that diffraction-limited performance and relatively low chromatic dispersion over a wide field of view can be obtained.


Loss of the HVEM Tumor Suppressor in Lymphoma and Restoration by Modified CAR-T Cells.
The HVEM (TNFRSF14) receptor gene is among the most frequently mutated genes in germinal center lymphomas. We report that loss of HVEM leads to cell-autonomous activation of B cell proliferation and drives the development of GC lymphomas in vivo. HVEM-deficient lymphoma B cells also induce a tumor-supportive microenvironment marked by exacerbated lymphoid stroma activation and increased recruitment of T follicular helper (TFH) cells. These changes result from the disruption of inhibitory cell-cell interactions between the HVEM and BTLA (B and T lymphocyte attenuator) receptors. Accordingly, administration of the HVEM ectodomain protein (solHVEM(P37-V202)) binds BTLA and restores tumor suppression. To deliver solHVEM to lymphomas in vivo, we engineered CD19-targeted chimeric antigen receptor (CAR) T cells that produce solHVEM locally and continuously. These modified CAR-T cells show enhanced therapeutic activity against xenografted lymphomas. Hence, the HVEM-BTLA axis opposes lymphoma development, and our study illustrates the use of CAR-T cells as "micro-pharmacies" able to deliver an anti-cancer protein.


Mechanism mediating basolateral transport of 2,4-dichlorophenoxyacetic acid in rat kidney.
The organic anions, p-aminohippurate (PAH) and fluorescein, are transported across the basolateral membrane of the renal proximal tubule in exchange for intracellular alpha-ketoglutarate (alpha KG), a mechanism indirectly coupled to sodium via Na+/alpha KG cotransport. To determine whether this mechanism mediates the basolateral transport of other organic anions, transport of the herbicide, 2,4-dichlorophenoxyacetic acid (2,4-D), was examined in rat renal cortical slices and basolateral membrane vesicles. In slices, uptake of 2,4-D increased steadily over time, approaching steady-state tissue/medium ratios of approximately 8 after 60 min. Probenecid, PAH and chlorophenol red inhibited steady-state uptake of 2,4-D. Accumulation of 10 microM 2,4-D was stimulated 2-fold by 60 microM glutarate; other dicarboxylic acids failed to stimulate uptake. In the presence of sodium, the addition of 5 mM LiCl or 2 mM ouabain to the bathing medium abolished glutarate stimulation. Removal of sodium from the bathing medium reversibly inhibited uptake as much as 75%. Furthermore, PAH inhibited 2,4-D uptake by slices in a dose-dependent manner, and increasing the external 2,4-D concentration decreased the inhibitory potency of PAH. In basolateral membrane vesicles, unlabeled 2,4-D inhibited sodium glutarate-coupled uptake of 3H-labeled PAH and 2,4-D in a concentration-dependent manner. Moreover, concentrative uptake of 2,4-D into vesicles could be driven by an outwardly directed gradient of glutarate or alpha KG that was generated by lithium-sensitive Na+/dicarboxylate cotransport or imposed experimentally. An outwardly directed gradient of unlabeled 2,4-D or PAH also stimulated uptake of 2,4-D. Based on these data, basolateral accumulation of 2,4-D by the renal proximal tubule is mediated by 2,4-D/alpha KG exchange, a mechanism energetically coupled to Na+/alpha KG cotransport and shared with PAH.


Targeting the p53 pathway of apoptosis.
The tumor suppressor protein, p53 is regarded as a key player in tumor suppression, as it promotes growth arrest, apoptosis and cellular senescence, while also blocking angiogenesis. The plethora of mechanisms underlying the p53 efficient death response involves transcriptional activation or repression of target genes, as well as the recently identified microRNAs, and transcription-independent functions. Pathological conditions such as cancer, neurodegeneration, ischemia, cholestasis or atherosclerosis are all strongly associated with deregulated levels of apoptosis in which p53 dysfunction has a prominent role. The effect of targeting cell death signaling proteins has been established in preclinical models of human diseases. In this regard, therapeutic strategies aimed at reactivation of p53 in tumors emerge as a promising approach for the treatment of cancer patients, as well as chemical inhibitors of p53 that may prove effective in suppressing disorders associated with widespread p53 activation. This review highlights recent developments of p53-induced apoptosis in human diseases. In addition, we will discuss controversies arising from the double-edge sword of targeting p53 in disease. Finally, ursodeoxycholic acid (UDCA), an endogenous bile acid used to treat cholestatic liver diseases, was recently described as a fine modulator of the complex control of p53 by Mdm-2. We will also review recent therapeutic strategies and clinical applications of targeted agents, and their progress in drug lead discovery, with particular emphasis on the potential use of UDCA.


Chronotype: Implications for Epidemiologic Studies on Chrono-Nutrition and Cardiometabolic Health.
Chrono-nutrition is an emerging research field in nutritional epidemiology that encompasses 3 dimensions of eating behavior: timing, frequency, and regularity. To date, few studies have investigated how an individual's circadian typology, i.e., one's chronotype, affects the association between chrono-nutrition and cardiometabolic health. This review sets the directions for future research by providing a narrative overview of recent epidemiologic research on chronotype, its determinants, and its association with dietary intake and cardiometabolic health. Limited research was found on the association between chronotype and dietary intake in infants, children, and older adults. Moreover, most of the evidence in adolescents and adults was restricted to cross-sectional surveys with few longitudinal cohorts simultaneously collecting data on chronotype and dietary intake. There was a gap in the research concerning the association between chronotype and the 3 dimensions of chrono-nutrition. Whether chronotype modifies the association between diet and cardiometabolic health outcomes remains to be elucidated. In conclusion, further research is required to understand the interplay between chronotype, chrono-nutrition, and cardiometabolic health outcomes.


Nucleosomal poly(ADP-ribose) polymerase: properties and relaxation of the chromatin structure.
The properties of poly(ADP-ribose) polymerase associated to nucleosomes and nuclei were analyzed by measuring enzymatic activity, isolating poly(ADP-ribosylated) histones and by electron microscopy. The enzyme was found to be most active in tri- and tetranucleosomes. The presence of NaC1 (0 to 200 mM) in the incubation medium showed a very small inhibitory effect on the enzymatic activity of isolated nucleosomes; however, the enzyme activity of incubated nuclei showed a gradual reduction with increasing salt concentrations leading to an almost complete inhibition at 200 mM NaC1. In parallel, the histones H1 extracted from isolated nucleosomes incubated at 200 mM NaC1 were hyper(ADP-ribosylated), whereas the histones H1 extracted from nuclei chromatin showed no highly modified forms at 200 mM NaC1. The effect of ethidium bromide (EB) was also analyzed and was found to stimulate enzyme activity in tetranucleosomes and octanucleosomes but not in dinucleosomes and nucleosomes of 20 repeat units. Electron microscopic analysis of poly(ADP-ribosylated) nucleosomes does not show the presence of aggregates at low ionic strength. However at higher ionic strengths the presence of localized relaxed domains of chromatin is evidenced in condensed nucleosomes.


Mechanisms of Antiviral Activity of Iminosugars Against Dengue Virus.
The antiviral mechanism of action of iminosugars against many enveloped viruses, including dengue virus (DENV), HIV, influenza and hepatitis C virus, is believed to be mediated by inducing misfolding of viral N-linked glycoproteins through inhibition of host endoplasmic reticulum-resident α-glucosidase enzymes. This leads to reduced secretion and/or infectivity of virions and hence lower viral titres, both in vitro and in vivo. Free oligosaccharide analysis from iminosugar-treated cells shows that antiviral activity correlates with production of mono- and tri-glucosylated sugars, indicative of inhibition of ER α-glucosidases. We demonstrate that glucose-mimicking iminosugars inhibit isolated glycoprotein and glycolipid processing enzymes and that this inhibition also occurs in primary cells treated with these drugs. Galactose-mimicking iminosugars that have been tested do not inhibit glycoprotein processing but do inhibit glycolipid processing, and are not antiviral against DENV. By comparison, the antiviral activity of glucose-mimetic iminosugars that inhibit endoplasmic reticulum-resident α-glucosidases, but not glycolipid processing, demonstrates that inhibition of α-glucosidases is responsible for iminosugar antiviral activity against DENV. This monograph will review the investigations of many researchers into the mechanisms of action of iminosugars and the contribution of our current understanding of these mechanisms for optimising clinical delivery of iminosugars. The effects of iminosugars on enzymes other than glucosidases, the induction of ER stress and viral receptors will be also put into context. Data suggest that inhibition of α-glucosidases results in inhibited release of virus and is the primary antiviral mechanism of action of iminosugars against DENV.


The polysaccharide capsule of the pathogenic fungus Cryptococcus neoformans enlarges by distal growth and is rearranged during budding.
The capsule of Cryptococcus neoformans can undergo dramatic enlargement, a phenomenon associated with virulence. A prior study that used Ab to the capsule as a marker for older capsular material concluded that capsule growth involved the intermixing of new and old capsular material with displacement of older capsular polysaccharide towards the surface. Here we have revisited that question using complement (C), which binds to capsular polysaccharide covalently, and cannot redistribute by dissociation and binding at different sites. The experimental approach involved binding of C to cells with small capsules, inducing capsule growth, and following the location of C relative to the cell wall as the capsule enlarged. C remained close to the cell wall during capsule growth, indicating that capsule enlargement occurred by addition of new polysaccharide near the capsule edge. This conclusion was confirmed by an independent method that employed radioactive metabolic labelling of newly synthesized capsule with 3H-mannose followed by gradual capsular stripping with gamma-radiation. Capsule growth proceeded to a certain size, which was a function of cell size, and was not degraded when the cells were transferred to a non-inducing medium. During budding, an opening appeared in the capsule of the mother cell that permitted the nascent bud to separate. Scanning EM suggested that a physical separation formed between the capsules of the mother and daughter cells during budding, which may avoid mixture between both capsules. Our results indicate that C. neoformans capsular enlargement also occurs by apical growth and that budding results in capsular rearrangements.


Fondaparinux compared with enoxaparin for the prevention of venous thromboembolism after hip-fracture surgery.
Surgery for hip fracture carries a high risk of venous thromboembolism, despite the use of current thromboprophylactic treatments. Fondaparinux, a synthetic pentasaccharide, is a new antithrombotic agent that may reduce this risk. In a double-blind study, were randomly assigned 1711 consecutive patients undergoing surgery for fracture of the upper third of the femur to receive subcutaneous doses of either 2.5 mg of fondaparinux once daily, initiated postoperatively, or 40 mg of enoxaparin once daily, initiated preoperatively, for at least five days. The primary efficacy outcome was venous thromboembolism up to postoperative day 11. Venous thromboembolism was defined as deep-vein thrombosis detected by mandatory bilateral venography, documented symptomatic deep-vein thrombosis, or documented symptomatic pulmonary embolism. The main safety outcomes were major bleeding and mortality from all causes. The duration of follow-up was six weeks. The incidence of venous thromboembolism by day 11 was 8.3 percent (52 of 626 patients) in the fondaparinux group and 19.1 percent (119 of 624 patients) in the enoxaparin group (P<0.001). The reduction in risk with fondaparinux was 56.4 percent (95 percent confidence interval, 39.0 to 70.3 percent). There were no significant differences between the two groups in the incidence of death or clinically relevant bleeding. In patients undergoing surgery for hip fracture, fondaparinux was more effective than enoxaparin in preventing venous thromboembolism and equally safe.


Multilayer descriptors for medical image classification.
In this paper, we propose a new method for improving the performance of 2D descriptors by building an n-layer image using different preprocessing approaches from which multilayer descriptors are extracted and used as feature vectors for training a Support Vector Machine. The different preprocessing approaches are used to build different n-layer images (n=3, n=5, etc.). We test both color and gray-level images, two well-known texture descriptors (Local Phase Quantization and Local Binary Pattern), and three of their variants suited for n-layer images (Volume Local Phase Quantization, Local Phase Quantization Three-Orthogonal-Planes, and Volume Local Binary Patterns). Our results show that multilayers and texture descriptors can be combined to outperform the standard single-layer approaches. Experiments on 10 datasets demonstrate the generalizability of the proposed descriptors. Most of these datasets are medical, but in each case the images are very different. Two datasets are completely unrelated to medicine and are included to demonstrate the discriminative power of the proposed descriptors across very different image recognition tasks. A MATLAB version of the complete system developed in this paper will be made available at https://www.dei.unipd.it/node/2357.


Antimycin-insensitive mutants of Candida utilis II. The effects of antimycin on Cytochrome b.
1. Cytochrome b-562 is more reduced in submitochondrial particles of mutant 28 during the aerobic steady-state respiration with succinate than in particles of the wild type. When anaerobiosis is reached, the reduction of cytochrome b is preceded by a rapid reoxidation in the mutnat. A similar reoxidation is observed in the wild type in the present of low concentrations of antimycin. 2. In contrast to the wild type, inhibition of electron transport in the mutant has a much higher antimycin titre than effects on cytochromes b (viz., aerobic steady-state reduction; reduction in the presence of substrate, cyanide and oxygen; the 'red shift' and lowering of E'-o of cytochrome b-562). Moreover, the titration curve of electron transport is hyperbolic whereas the curves for the reduction are sigmoidal. The conclusion is, that in both mutant and wild type, the actions of antimycin on electron transport and cytochromes b are separable. 3. The red shift in the mutant is more extensive than in the wild type. 4. Cytochrome b-558 and cytochrome b-566 (that absorbs in mutant and wild type at 564.5 nm) do not respond simultaneously to addition of antimycin, indicating that they are two separate cytochromes. 5. The difference between the effect of antimycin on electron transport and cytochromes b reduction is also found in intact cells of the mutant. 6. A model is suggested for the wild-type respiratory chain in which (i) the cytochromes b lie, in an uncoupled system, out of the main electron-transfer chain, (ii) antimycin induces a conformation change in QH-2-cytochrome c reductase resulting in effects on cytochrome b and inhibition of electron transport, (iii) a second antimycin-binding site with low affinity to the antibiotic is present, capable of inhibiting electron transport.


Fasting hepatic de novo lipogenesis is not reliably assessed using circulating fatty acid markers.
Observational studies often infer hepatic de novo lipogenesis (DNL) by measuring circulating fatty acid (FA) markers; however, it remains to be elucidated whether these markers accurately reflect hepatic DNL. We investigated associations between fasting hepatic DNL and proposed FA markers of DNL in subjects consuming their habitual diet. Fasting hepatic DNL was assessed using 2H2O (deuterated water) in 149 nondiabetic men and women and measuring the synthesis of very low-density lipoprotein triglyceride (VLDL-TG) palmitate. FA markers of blood lipid fractions were determined by gas chromatography. Neither the lipogenic index (16:0/18:2n-6) nor the SCD index (16:1n-7/16:0) in VLDL-TG was associated with isotopically assessed DNL (r = 0.13, P = 0.1 and r = -0.08, P = 0.35, respectively). The relative abundances (mol%) of 14:0, 16:0, and 18:0 in VLDL-TG were weakly (r ≤ 0.35) associated with DNL, whereas the abundances of 16:1n-7, 18:1n-7, and 18:1n-9 were not associated. When the cohort was split by median DNL, only the abundances of 14:0 and 18:0 in VLDL-TG could discriminate between subjects having high (11.5%) and low (3.8%) fasting hepatic DNL. Based on a subgroup, FA markers in total plasma TG, plasma cholesteryl esters, plasma phospholipids, and red blood cell phospholipids were generally not associated with DNL. The usefulness of circulating FAs as markers of hepatic DNL in healthy individuals consuming their habitual diet is limited due to their inability to discriminate clearly between individuals with low and high fasting hepatic DNL.


Physician associates in England's hospitals: a survey of medical directors exploring current usage and factors affecting recruitment.
In the UK secondary care setting, the case for physician associates is based on the cover and stability they might offer to medical teams. We assessed the extent of their adoption and deployment - that is, their current usage and the factors supporting or inhibiting their inclusion in medical teams - using an electronic, self-report survey of medical directors of acute and mental health NHS trusts in England. Physician associates - employed in small numbers, in a range of specialties, in 20 of the responding trusts - were reported to have been employed to fill gaps in medical staffing and support medical specialty trainees. Inhibiting factors were commonly a shortage of physician associates to recruit and lack of authority to prescribe, as well as a lack of evidence and colleague resistance. Our data suggest there is an appetite for employment of physician associates while practical and attitudinal barriers are yet to be fully overcome.


Protein interactions central to stabilizing the K+ channel selectivity filter in a four-sited configuration for selective K+ permeation.
The structural and functional conversion of the nonselective NaK channel to a K(+) selective channel (NaK2K) allows us to identify two key residues, Tyr and Asp in the filter sequence of TVGYGD, that participate in interactions central to stabilizing the K(+) channel selectivity filter. By using protein crystallography and channel electrophysiology, we demonstrate that the K(+) channel filter exists as an energetically strained structure and requires these key protein interactions working in concert to hold the filter in the precisely defined four-sited configuration that is essential for selective K(+) permeation. Disruption of either interaction, as tested on both the NaK2K and eukaryotic K(v)1.6 channels, can reduce or completely abolish K(+) selectivity and in some cases may also lead to channel inactivation due to conformational changes at the filter. Additionally, on the scaffold of NaK we recapitulate the protein interactions found in the filter of the Kir channel family, which uses a distinct interaction network to achieve similar stabilization of the filter.


Fully automated quantification method (FQM) of coronary calcium in an anthropomorphic phantom.
Coronary artery calcium (CAC) score is a strong predictor for future adverse cardiovascular events. Anthropomorphic phantoms are often used for CAC studies on computed tomography (CT) to allow for evaluation or variation of scanning or reconstruction parameters within or across scanners against a reference standard. This often results in large number of datasets. Manual assessment of these large datasets is time consuming and cumbersome. Therefore, this study aimed to develop and validate a fully automated, open-source quantification method (FQM) for coronary calcium in a standardized phantom. A standard, commercially available anthropomorphic thorax phantom was used with an insert containing nine calcifications with different sizes and densities. To simulate two different patient sizes, an extension ring was used. Image data were acquired with four state-of-the-art CT systems using routine CAC scoring acquisition protocols. For interscan variability, each acquisition was repeated five times with small translations and/or rotations. Vendor-specific CAC scores (Agatston, volume, and mass) were calculated as reference scores using vendor-specific software. Both the international standard CAC quantification methods as well as vendor-specific adjustments were implemented in FQM. Reference and FQM scores were compared using Bland-Altman analysis, intraclass correlation coefficients, risk reclassifications, and Cohen's kappa. Also, robustness of FQM was assessed using varied acquisitions and reconstruction settings and validation on a dynamic phantom. Further, image quality metrics were implemented: noise power spectrum, task transfer function, and contrast- and signal-to-noise ratio among others. Results were validated using imQuest software. Three parameters in CAC scoring methods varied among the different vendor-specific software packages: the Hounsfield unit (HU) threshold, the minimum area used to designate a group of voxels as calcium, and the usage of isotropic voxels for the volume score. The FQM was in high agreement with vendor-specific scores and ICC's (median [95% CI]) were excellent (1.000 [0.999-1.000] to 1.000 [1.000-1.000]). An excellent interplatform reliability of κ = 0.969 and κ = 0.973 was found. TTF results gave a maximum deviation of 3.8% and NPS results were comparable to imQuest. We developed a fully automated, open-source, robust method to quantify CAC on CT scans in a commercially available phantom. Also, the automated algorithm contains image quality assessment for fast comparison of differences in acquisition and reconstruction parameters.


Having Outpatient Major Elective (HOME) Robotic Colon Resection Protocol: A Safe Approach to Ambulatory Colon Resection.
Within the past decade, colorectal surgery length of stay (LOS) has decreased from an average of 5-6 days to 2-3 days. However, elective colon resections have yet to become a common procedure with the potential for same-day discharge. During the COVID pandemic, hospital capacity was exceptionally strained and colon resections were delayed due to the lack of inpatient beds available. We sought to create a protocolized ERAS (enhanced recovery after surgery) pathway that would allow for safe and feasible ambulatory colon resections as well as decreasing overall hospital inpatient burden. Between November 2020 and March 2022, 15 patients were offered same-day discharges under the HOME protocol. Of the 15 patients, 11 patients agreed to be discharged home the day of surgery and followed prospectively for 30 days. All procedures were performed robotically. Patients were selected based on level of preoperative health (ASA class 1 and 2), low-risk for loss to follow-up, ability for close family supervision for 3 days postoperatively, and type of procedure (partial colectomy). Close follow-up was achieved with daily telephonic or televideo visits for 3 days post-operatively, as well as a 2-week outpatient clinic follow-up. A total of 11 patient underwent same-day surgery utilizing the protocol, 5 females and 6 males, between the ages of 34 and 62. All patients were ASA class 2. Indications for colon resection were cecal volvulus (1), recurrent sigmmoid diverticulitis (9), and Crohn's disease (1). Primary outcome was readmission rates within the 30-days. There were no readmissions or complications during the perioperative 30-day period. There was one emergency department return for pain who was not admitted. Average operative time was 132.1 minutes. Using a novel enhanced recovery protocol, we demonstrated the feasibility and safety of ambulatory partial colectomy in a highly select small subset of patients.


Variability in the prescribing of intravenous fluids: A cross sectional multicentre analysis of clinical practice.
Intravenous (IV) fluid administration continues to be a mainstay of care in General Surgery. Yet if they are prescribed incorrectly significant morbidity including electrolyte abnormalities, renal impairment and cardiac failure can develop. Despite this, it is frequently the responsibility of the most junior staff to prescribe IV fluids. We aim to analyse the understanding of IV fluid prescribing amongst junior doctors and to describe variability in clinical practice. We undertook a multicentre questionnaire study. Foundation doctors and specialty trainees were invited to undertake a two part paper-based questionnaire. Part one analysed baseline knowledge of the concentration of commonly prescribed fluids. Part two consisted of four clinical vignettes requiring a IV fluid prescribing decision by the surveyed doctor. A total of 143 Doctors working in 8 hospitals were recruited. 65 (45.5%) doctors correctly stated the daily maintenance fluid requirements of water for an adult (25-30 mls/kg/day), while only 54 (37.8%) knew the sodium concentration of 0.9% NaCl. Lack of postgraduate experience (p = 0.011), qualifying from a medical school outside the United Kingdom (p < 0.0001) and working in one of the eight hospitals in this study (p < 0.0001) were associated with a lower knowledge level. There was limited consensus in prescribing in the responses to the 4 clinical scenarios, with 69 unique combinations of fluid choice, rate and volume prescribed. Knowledge of the constituents of common IV fluids and routine requirement for fluid and common electrolytes is poor across junior doctors of all grades, driving large variation in clinical practice.


Sperm competition in bats.
Sperm competition is a widespread phenomenon influencing the evolution of male anatomy, physiology and behaviour. Bats are an ideal group for studying sperm competition. Females store fertile sperm for up to 200 days and the size of social groups varies from single animals to groups of hundreds of thousands. This study examines the relationship between social group size and investment in spermatogenesis across 31 species of microchiropteran bat using new and published data on testis mass and sperm length. In addition to male competition, I examined the effects of female reproductive biology on characteristics of spermatogenesis. Comparative studies indicate that relative testis mass is positively related to sperm competition risk in a wide range of taxa. Social group size may also influence the level of sperm competition, and one of the costs of living in groups may be decreased confidence of paternity. I used comparative analysis of independent contrast (CAIC) to control for phylogeny. Using two possible phylogenies and two measures of social group size, I found a significant positive relationship between social group size and testis mass. There was no relationship between testis mass and the dimension of the female reproductive tract or oestrus duration. Sperm length was not significantly related to body mass or group size, nor was it related to oestrus duration.


Hydrodistillation extraction time effect on essential oil yield, composition, and bioactivity of coriander oil.
Coriander (Coriandrum sativum L.) is a major essential oil crop grown throughout the world. Coriander essential oil is extracted from coriander fruits via hydrodistillation, with the industry using 180-240 min of distillation time (DT), but the optimum DT for maximizing essential oil yield, composition of constituents, and antioxidant activities are not known. This research was conducted to determine the effect of DT on coriander oil yield, composition, and bioactivity. The results show that essential oil yield at the shorter DT was low and generally increased with increasing DT with the maximum yields achieved at DT between 40 and 160 min. The concentrations of the low-boiling point essential oil constituents: α-pinene, camphene, β-pinene, myrcene, para-cymene, limonene, and γ-terpinene were higher at shorter DT (< 2.5 min) and decreased with increasing DT; but the trend reversed for the high-boiling point constituents: geraniol and geranyl-acetate. The concentration of the major essential oil constituent, linalool, was 51% at DT 1.15 min, and increased steadily to 68% with increasing DT. In conclusion, 40 min DT is sufficient to maximize yield of essential oil; and different DT can be used to obtain essential oil with differential composition. Its antioxidant capacity was affected by the DT, with 20 and 240 min DT showing higher antioxidant activity. Comparisons of coriander essential oil composition must consider the length of the DT.


Efficient recovery of dysferlin deficiency by dual adeno-associated vector-mediated gene transfer.
Deficiency of the dysferlin protein presents as two major clinical phenotypes: limb-girdle muscular dystrophy type 2B and Miyoshi myopathy. Dysferlin is known to participate in membrane repair, providing a potential hypothesis to the underlying pathophysiology of these diseases. The size of the dysferlin cDNA prevents its direct incorporation into an adeno-associated virus (AAV) vector for therapeutic gene transfer into muscle. To bypass this limitation, we split the dysferlin cDNA at the exon 28/29 junction and cloned it into two independent AAV vectors carrying the appropriate splicing sequences. Intramuscular injection of the corresponding vectors into a dysferlin-deficient mouse model led to the expression of full-length dysferlin for at least 1 year. Importantly, systemic injection in the tail vein of the two vectors led to a widespread although weak expression of the full-length protein. Injections were associated with an improvement of the histological aspect of the muscle, a reduction in the number of necrotic fibers, restoration of membrane repair capacity and a global improvement in locomotor activity. Altogether, these data support the use of such a strategy for the treatment of dysferlin deficiency.


Comparison of the osteoblastic activity conferred on Si-doped hydroxyapatite scaffolds by different osteostatin coatings.
Parathyroid hormone-related protein (107-111) (osteostatin) induces osteogenic effects in osteoblasts in vitro and in regenerating bone in mice and rabbits. In this study we used osteoblastic MC3T3-E1 cell cultures to evaluate and compare the bioactivity of this peptide either adsorbed or covalently bound (by its C-terminus) to Si-doped hydroxyapatite (Si-HA) scaffolds after organic (-NH(2)) functionalization. By these means osteostatin can be locally released or kept anchored to the scaffold surface. This was confirmed by chemical analysis and by testing the efficiency of osteostatin-loaded Si-HA scaffolds (placed in Transwell chambers) in healing a scratch wound in mouse pluripotent mesenchymal C3H10T1/2 cells. Our results show that exposure of MC3T3-E1 cell monolayers to Si-HA scaffolds with both types of osteostatin coating (deliverable or immobilized), in contrast to those without peptide, similarly stimulated cell growth and matrix mineralization. These findings demonstrate that osteostatin release from Si-HA scaffolds is not essential to promote osteoblastic growth and function in vitro, and lend credence to considering osteostatin a bone regenerating factor.


Purification and characterization of antithrombotics from Syzygium aromaticum (L.) MErr. & PERRY.
Two antithrombotic polysaccharides with relatively high molecular weight (HMW) and low molecular weight (LMW) were isolated from the flower buds of Syzygium aromaticum (L.) MERR. & PERRY (clove) by anion-exchange chromatography, hydrophobic interaction column chromatography and size exclusion chromatography (LMW: EC-2B-IIIa-2, M.W. ca. 34000; HMW: EC-2C-Ia-2, M.W. ca. 103000). The LMW polysaccharide was mainly composed of Rha, Gal, GalA and Ara (molar %: 24.1, 18.9, 18.0 and 17.9, respectively) with 10.8% of sulfate and 18.2% of protein. The HMW fraction consisted of Ara, Gal, Glc and Rha (molar %: 26.0, 23.7, 17.5 and 12.4, respectively) with 15.4% of sulfate and 8.0% of protein. Both polysaccharides had the backbone of type I rhamnogalacturonan and the side chain of arabinan. Also, most of the sulfates were attached at the position 6 of 3-linked galactosyl residues. Compared to the antithrombotic activity of the HMW fraction (plasma clotting time of 145 s in APTT assay), the LMW fraction displayed a slightly low activity (90 s). However, animal studies indicated that crude LMW polysaccharide did not show acute toxicity, while the acute LD50 of the HMW fraction was approximately 2-fold lower than that of heparin.


Driving With Hemianopia VII: Predicting Hazard Detection With Gaze and Head Scan Magnitude.
One rehabilitation strategy taught to individuals with hemianopic field loss (HFL) is to make a large blind side scan to quickly identify hazards. However, it is not clear what the minimum threshold is for how large the scan should be. Using driving simulation, we evaluated thresholds (criteria) for gaze and head scan magnitudes that best predict detection safety. Seventeen participants with complete HFL and 15 with normal vision (NV) drove through 4 routes in a virtual city while their eyes and head were tracked. Participants pressed the horn as soon as they detected a motorcycle (10 per drive) that appeared 54 degrees eccentricity on cross-streets and approached toward the driver. Those with HFL detected fewer motorcycles than those with NV and had worse detection on the blind side than the seeing side. On the blind side, both safe detections and early detections (detections before the hazard entered the intersection) could be predicted with both gaze (safe 18.5 degrees and early 33.8 degrees) and head (safe 19.3 degrees and early 27 degrees) scans. However, on the seeing side, only early detections could be classified with gaze (25.3 degrees) and head (9.0 degrees). Both head and gaze scan magnitude were significant predictors of detection on the blind side, but less predictive on the seeing side, which was likely driven by the ability to use peripheral vision. Interestingly, head scans were as predictive as gaze scans. The minimum scan magnitude could be a useful criterion for scanning training or for developing assistive technologies to improve scanning.


Molecular Epidemiology, prenatal exposure and prevention of cancer.
Molecular Epidemiology was originally conceived as a preventive approach, providing a valuable tool for investigating risk factors for cancer in vulnerable populations. Biomarkers can be used as early indicators of risk for preventative purposes and risk assessment. The present contribution mainly refers to in utero exposures to carcinogens, since humans are especially vulnerable during fetal development. Environmental exposures in utero can increase risks for both childhood and adult cancers; their interactions with genetic and nutritional susceptibility factors may further increase risk. Thus, the early developmental period represents an important window for cancer prevention.


Cooperation of EBV DNA polymerase and EA-D(BMRF1) in vitro and colocalization in nuclei of infected cells.
Expression of the Epstein-Barr virus (EBV) DNA polymerase (EBVpol) open reading frame (BALF5) by in vitro transcription-translation yielded a 116-kDa primary translation product. Enzymatic DNA polymerase activity of the in vitro translated polypeptide required the presence of the 47-kDa BMRF1 (EA-D) gene product. Antiserum raised to the BALF5 gene product expressed in Escherichia coli specifically precipitated a 116-kDa polypeptide in extracts of latently infected lymphoblastoid cells induced for EBV replication. Immunofluorescence microscopy revealed colocalization of the EBVpol and EA-D(BMRF1) to discrete foci within the nuclei of induced cells; however, the blockade of viral DNA synthesis resulted in diffuse nuclear staining patterns for both antigens. Bromodeoxyuridine staining of these discrete foci colocalizing with EBVpol suggests that they are sites of early viral DNA synthesis. These observations suggest that EA-D(BMRF1) may be an accessory protein of the EBV DNA polymerase which colocalizes in vivo with EBVpol to sites of viral DNA replication and cooperates in vitro to form an active EBVpol holoenzyme.


Interventions using new digital media to improve adolescent sexual health: a systematic review.
New digital media (e.g., the Internet, text messaging, and social networking sites [SNS]) have dramatically altered the communication landscape, especially for youth. These communication platforms present new tools for engaging youth in sexual health promotion and risk reduction. We searched eight public databases across multiple disciplines for all peer-reviewed studies published between January 2000 and May 2011 that empirically evaluated the impact of digital media-based interventions on the sexual health knowledge, attitudes, and/or behaviors of adolescents aged 13-24 years. Of 942 abstracts, 10 met inclusion criteria. Seven studies were conducted in the United States. Eight described Web-based interventions, one used mobile phones, and one was conducted on an SNS. Two studies significantly delayed initiation of sex, and one was successful in encouraging users of an SNS to remove sex references from their public profile. Seven interventions significantly influenced psychosocial outcomes such as condom self-efficacy and abstinence attitudes, but at times the results were in directions unexpected by the study authors. Six studies increased knowledge of HIV, sexually transmitted infections, or pregnancy. This area of research is emerging and rapidly changing. More data from controlled studies with longer (>1 year) follow-up and measurement of behavioral outcomes will provide a more robust evidence base from which to judge the effectiveness of new digital media in changing adolescent sexual behavior.


Fertility preservation in patients with BRCA mutation.
Evidence suggests a likely negative impact of deleterious BRCA mutations on female fertility. Hence, different studies have aimed to address the reproductive potential and performance of fertility preservation strategies in BRCA-mutated breast cancer patients with a prime focus on their safety and efficacy. However, several uncertainties exist in many domains of this field. The aim of the current paper is to overview the reproductive potential and fertility preservation options in breast and ovarian cancer patients harbouring a BRCA mutation. We also discuss pre-implantation genetic testing in an attempt to help physicians during oncofertility counselling of these patients.


Quality control for quantitative PCR based on amplification compatibility test.
Quantitative qPCR is a routinely used method for the accurate quantification of nucleic acids. Yet it may generate erroneous results if the amplification process is obscured by inhibition or generation of aberrant side-products such as primer dimers. Several methods have been established to control for pre-processing performance that rely on the introduction of a co-amplified reference sequence, however there is currently no method to allow for reliable control of the amplification process without directly modifying the sample mix. Herein we present a statistical approach based on multivariate analysis of the amplification response data generated in real-time. The amplification trajectory in its most resolved and dynamic phase is fitted with a suitable model. Two parameters of this model, related to amplification efficiency, are then used for calculation of the Z-score statistics. Each studied sample is compared to a predefined reference set of reactions, typically calibration reactions. A probabilistic decision for each individual Z-score is then used to identify the majority of inhibited reactions in our experiments. We compare this approach to univariate methods using only the sample specific amplification efficiency as reporter of the compatibility. We demonstrate improved identification performance using the multivariate approach compared to the univariate approach. Finally we stress that the performance of the amplification compatibility test as a quality control procedure depends on the quality of the reference set.


A Working Hypothesis for the Role of the Cerebellum in Impulsivity and Compulsivity.
Growing evidence associates cerebellar abnormalities with several neuropsychiatric disorders in which compulsive symptomatology and impulsivity are part of the disease pattern. Symptomatology of autism, addiction, obsessive-compulsive (OCD), and attention deficit/hyperactivity (ADHD) disorders transcends the sphere of motor dysfunction and essentially entails integrative processes under control of prefrontal-thalamic-cerebellar loops. Patients with brain lesions affecting the cortico-striatum thalamic circuitry and the cerebellum indeed exhibit compulsive symptoms. Specifically, lesions of the posterior cerebellar vermis cause affective dysregulation and deficits in executive function. These deficits may be due to impairment of one of the main functions of the cerebellum, implementation of forward internal models of the environment. Actions that are independent of internal models may not be guided by predictive relationships or a mental representation of the goal. In this review article, we explain how this deficit might affect executive functions. Additionally, regionalized cerebellar lesions have been demonstrated to impair other brain functions such as the emergence of habits and behavioral inhibition, which are also altered in compulsive disorders. Similar to the infralimbic cortex, clinical studies and research in animal models suggest that the cerebellum is not required for learning goal-directed behaviors, but it is critical for habit formation. Despite this accumulating data, the role of the cerebellum in compulsive symptomatology and impulsivity is still a matter of discussion. Overall, findings point to a modulatory function of the cerebellum in terminating or initiating actions through regulation of the prefrontal cortices. Specifically, the cerebellum may be crucial for restraining ongoing actions when environmental conditions change by adjusting prefrontal activity in response to the new external and internal stimuli, thereby promoting flexible behavioral control. We elaborate on this explanatory framework and propose a working hypothesis for the involvement of the cerebellum in compulsive and impulsive endophenotypes.


Stent thrombosis: role of compliance and nonresponsiveness to antiplatelet therapy.
Percutaneous coronary intervention with drug-eluting stents has revolutionized the management of patients with symptomatic coronary artery disease. Although this strategy significantly reduces the incidence of restenosis and repeat revascularization, concern has been raised about an increased frequency of late stent thrombosis with drug-eluting stents compared with bare-metal stents. The mechanism of stent thrombosis remains unclear, and various hypotheses have been described. Platelets are believed to play a pivotal role in the development of stent thrombosis, with pathological studies demonstrating an abundance of platelets within the occlusive thrombi. Premature discontinuation and nonadherence to antiplatelet therapy are considered important risk factors for late stent thrombosis. Early identification of vulnerable patients and definition of the role of antiplatelet nonresponsiveness in the development of stent thrombosis should be the focus of future diagnostic and therapeutic strategies.


The Consequences of Overlapping G-Quadruplexes and i-Motifs in the Platelet-Derived Growth Factor Receptor β Core Promoter Nuclease Hypersensitive Element Can Explain the Unexpected Effects of Mutations and Provide Opportunities for Selective Targeting of Both Structures by Small Molecules To Downregulate Gene Expression.
The platelet-derived growth factor receptor β (PDGFR-β) signaling pathway is a validated and important target for the treatment of certain malignant and nonmalignant pathologies. We previously identified a G-quadruplex-forming nuclease hypersensitive element (NHE) in the human PDGFR-β promoter that putatively forms four overlapping G-quadruplexes. Therefore, we further investigated the structures and biological roles of the G-quadruplexes and i-motifs in the PDGFR-β NHE with the ultimate goal of demonstrating an alternate and effective strategy for molecularly targeting the PDGFR-β pathway. Significantly, we show that the primary G-quadruplex receptor for repression of PDGFR-β is the 3'-end G-quadruplex, which has a GGA sequence at the 3'-end. Mutation studies using luciferase reporter plasmids highlight a novel set of G-quadruplex point mutations, some of which seem to provide conflicting results on effects on gene expression, prompting further investigation into the effect of these mutations on the i-motif-forming strand. Herein we characterize the formation of an equilibrium between at least two different i-motifs from the cytosine-rich (C-rich) sequence of the PDGFR-β NHE. The apparently conflicting mutation results can be rationalized if we take into account the single base point mutation made in a critical cytosine run in the PDGFR-β NHE that dramatically affects the equilibrium of i-motifs formed from this sequence. We identified a group of ellipticines that targets the G-quadruplexes in the PDGFR-β promoter, and from this series of compounds, we selected the ellipticine analog GSA1129, which selectively targets the 3'-end G-quadruplex, to shift the dynamic equilibrium in the full-length sequence to favor this structure. We also identified a benzothiophene-2-carboxamide (NSC309874) as a PDGFR-β i-motif-interactive compound. In vitro, GSA1129 and NSC309874 downregulate PDGFR-β promoter activity and transcript in the neuroblastoma cell line SK-N-SH at subcytotoxic cell concentrations. GSA1129 also inhibits PDGFR-β-driven cell proliferation and migration. With an established preclinical murine model of acute lung injury, we demonstrate that GSA1129 attenuates endotoxin-mediated acute lung inflammation. Our studies underscore the importance of considering the effects of point mutations on structure formation from the G- and C-rich sequences and provide further evidence for the involvement of both strands and associated structures in the control of gene expression.


Protective effect of crocin on BPA-induced liver toxicity in rats through inhibition of oxidative stress and downregulation of MAPK and MAPKAP signaling pathway and miRNA-122 expression.
Bisphenol A (BPA) is an artificial environmental endocrine disrupting chemical and commonly used as a monomer of polycarbonate plastics and epoxy resins. The aim of the present study is to investigate the hepatoprotective effects of crocin, a constituent of saffron, against BPA-induced liver toxicity. We showed that treatment of male Wistar rats with 0.5 mg/kg BPA for 30 days increased the level of 8-isoprostane, decreased the level of reduced glutathione, elevated serum levels of aspartate aminotransferase, lactate dehydrogenase, triglyceride, and glucose, and induced periportal inflammation. Western blot results revealed that BPA increased the phosphorylation of c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK1/2), and mitogen-activated protein kinase-activated protein kinase (MAPKAPK), but not p38. BPA also reduced the Akt signaling activation and upregulated microRNA (miR-122) expression. Moreover, we showed here that crocin 20 mg/kg administration ameliorated liver damage and improved elevated levels of TG and liver enzymes of BPA-treated rats possibly though antioxidant activity, downregulation of miR-122 transcript level and lowering the phosphorylation of JNK, ERK1/2, and MAPKAPK and subsequently their activities. Overall, the findings suggest that crocin possesses hepatoprotective effects against BPA-induced liver toxicity by enhancing the antioxidative defense system and regulation of important signaling pathway activities and miR-122 expression.


Reduced dietary intake of simple sugars alters perceived sweet taste intensity but not perceived pleasantness.
Individuals who adhere to reduced-sodium diets come to prefer less salt over time, but it is unclear whether sweet taste perception is modulated by reduced sugar intake. The objective was to determine how a substantial reduction in dietary intake of simple sugars affects sweetness intensity and pleasantness of sweet foods and beverages. Healthy men and women aged 21-54 y participated for 5 mo. After the baseline month, 2 subject groups were matched for demographic characteristics, body mass index, and intake of simple sugars. One group (n = 16; 13 of whom completed key experimental manipulations) was randomly assigned to receive a low-sugar diet during the subsequent 3 mo, with instructions to replace 40% of calories from simple sugars with fats, proteins, and complex carbohydrates. The other (control) group (n = 17; 16 of whom completed the study) did not change their sugar intake. During the final month, both groups chose any diet they wished. Each month subjects rated the sweetness intensity and pleasantness of vanilla puddings and raspberry beverages that varied in sucrose concentration. ANOVA showed no systematic differences between groups in rated sweetness during the baseline or first diet month. During the second diet month, the low-sugar group rated low-sucrose pudding samples as more intense than did the control group (significant group-by-concentration interaction, P = 0.002). During the third diet month, the low-sugar subjects rated both low and high concentrations in puddings as ∼40% sweeter than did the control group (significant effect of group, P = 0.01). A weaker effect on rated sweetness was obtained for the beverages. Rated pleasantness was not affected for either of the stimuli. This experiment provides empirical evidence that changes in consumption of simple sugars influence perceived sweet taste intensity. More work is needed to determine whether sugar intake ultimately shifts preferences for sweet foods and beverages. This trial was registered at clinicaltrials.gov as NCT02090478.


NOTCH3 signaling pathway plays crucial roles in the proliferation of ErbB2-negative human breast cancer cells.
ErbB2-negative breast tumors represent a significant therapeutic hurdle because of a lack of effective molecular targets. Although NOTCH proteins are known to be involved in mammary tumorigenesis, the functional significance of these proteins in ErbB2-negative breast tumors is not clear. In the present study, we examined the expression of activated NOTCH receptors in human breast cancer cell lines, including ErbB2-negative and ErbB2-positive cell lines. Activated NOTCH1 and NOTCH3 proteins generated by gamma-secretase were detected in most of the cell lines tested, and both proteins activated CSL-mediated transcription. Down-regulation of NOTCH1 by RNA interference had little or no suppressive effect on the proliferation of either ErbB2-positive or ErbB2-negative cell lines. In contrast, down-regulation of NOTCH3 significantly suppressed proliferation and promoted apoptosis of the ErbB2-negative tumor cell lines. Down-regulation of NOTCH3 did not have a significant effect on the ErbB2-positive tumor cell lines. Down-regulation of CSL also suppressed the proliferation of ErbB2-negative breast tumor cell lines, indicating that the NOTCH-CSL signaling axis is involved in cell proliferation. Finally, NOTCH3 gene amplification was detected in a breast tumor cell line and one breast cancer tissue specimen even though the frequency of NOTCH3 gene amplification was low (<1%). Taken together, these findings indicate that NOTCH3-mediated signaling rather than NOTCH1-mediated signaling plays an important role in the proliferation of ErbB2-negative breast tumor cells and that targeted suppression of this signaling pathway may be a promising strategy for the treatment of ErbB2-negative breast cancers.


Systematic review of interventions for reducing stigma experienced by children with disabilities and their families in low- and middle-income countries: state of the evidence.
To identify and assess the evidence for interventions to reduce stigma experienced by children with disabilities and their families in low- and middle-income settings. Systematic review of seven databases (MEDLINE, EMBASE, Global Health, PsycINFO, Social Policy and Practice, CINAHL, IBSS) for studies of interventions that aimed to reduce stigma for children with disabilities published from January 2000 to April 2018. Data were extracted on study population, study design, intervention level(s) and target group, and type(s) of stigma addressed. A narrative approach was used to synthesise the results. Twenty studies were included. The majority (65%) of interventions targeted enacted stigma (negative attitudes) and the most common intervention approach was education/training (63%). Over half (54%) of interventions were delivered at the organisational/institutional level, and only four studies targeted more than one social level. The most common disability targeted was epilepsy (50%) followed by intellectual impairment (20%). The majority of studies (n = 18/20, 90%) found a reduction in a component of stigma; however, most (90%) studies had a high risk of bias. This review highlights the lack of quality evidence on effective stigma-reduction strategies for children with disabilities. Validation and consistent use of contextually relevant scales to measure stigma may advance this field of research. Studies that involve people with disabilities in the design and implementation of these strategies are needed.


Ward managers' perceptions of the role of healthcare assistants.
While unqualified nurses form an integral part of the UK's healthcare system, accounting for a large percentage of the workforce, there is limited evidence to identify the actual role of the healthcare assistant. This study explored hospital ward managers' views on the current and potential expansion of the healthcare assistant's role. A confidential survey was sent to all ward managers (n = 35) in one large acute hospital trust. A response rate of 94% (n = 33) was obtained. Data were analysed using descriptive statistics and open-ended questions were analysed for emerging themes. The results indicated that healthcare assistants are undertaking a wide diversity of procedures raging from general tidying to carrying out complex clinical procedures. Additionally, 82% (n = 27) of the managers reported that the healthcare assistant's role could be expanded, although concerns were raised regarding additional training and supervision.


Comparative Study of Laparoscopic versus Conventional Open Surgical Staging Procedure for Endometrial Cancer: Our Institutional Experience.
The aim of this study was to compare laparoscopic surgical staging of endometrial cancer with conventional open methods. The retrospective data were collected from 90 endometrial cancer patients who were operated between 2013 and 2018 in our government institution. The safety and morbidity of the two groups were compared based on mean operative time, blood loss, intraoperative complications, postoperative complication, etc., and the results were analyzed. Statistical analysis was performed using IBM SPSS statistics version 20. Clinical and pathologic factors were compared between two groups with Fisher's exact test and Student's t-test for data analysis. The survival data were analyzed using Kaplan-Meier survival curves. P < 0.05 was considered statistically significant. Thirty-two patients underwent laparoscopic staging, 45 underwent laparotomy, and 13 cases lap converted to open surgery. The mean operative time for the laparoscopic procedure was lower than the open procedure (P = 0.001). The mean hospital stay of patients who underwent laparoscopic staging was around 7.1 days significantly less compared to the laparotomy group. Intraoperative blood loss of patients undergoing laparoscopic surgery was significantly less than that of the laparotomy group (P = 0.015). There was no statistically significant difference in nodal retrieval between laparotomy and laparoscopy group (P = 0.172). The mean duration of hospitalization was statistically significantly greater in the laparotomy group than the laparoscopic group (13 and 7 days, P < 0.001). Based on the Kaplan-Meier survival curve, there was no difference in survival rates between the groups. Median follow-up duration was 32 months. Laparoscopic surgical staging is oncologically safe for the management of endometrial cancer with acceptable morbidity compared to the open approach with far less blood loss and shorter postoperative stay.


Organ donation after cardiac determination of death (DCD): a swine model.
Donors after Cardiac Death (DCD) may reduce the organ scarcity; however, their use is limited because of warm ischemia time. Fortunately, this is less important in a subclass of DCD called expected (e-DCD), those with irreversible but incomplete brain injury. This study analyzed hemodynamic/pulmonary data to establish a clinically relevant model of cardiac death that would simulate an e-DCD setting. Hemodynamics, pulmonary artery flows, arterial blood gasses, and left atrial pressure were recorded q 5 minutes in anesthetized swine. After baseline data collection, the ventilator was discontinued and heparin was administered. Cardiac death was defined: as asystole, or mean arterial presusure < or = 25 mm Hg with a pulse pressure < or = 20 mm Hg. The time to death was approximately 14.8 minutes. Within 5 minutes of removal of the ventilator, there was a hyperdynamic period. Blood gases throughout the apneic time showed a rapid hypercapnia and acidosis. The hyperdynamic reflex response was followed by hypotension, bradycardia, and finally asystole or ventricular fibrillation. The protocol of withdrawal of ventilation, systemic anticoagulation, determination of death was developed to closely resemble the clinical e-DCD scenario. The physiologic changes that happen before death in DCD were described. An e-DCD model that can be used in studies related to organ transplantation was established.


Oxidant production and SOD1 protein expression in single skeletal myofibers from Down syndrome mice.
Down syndrome (DS) is a genetic condition caused by the triplication of chromosome 21. Persons with DS exhibit pronounced muscle weakness, which also occurs in the Ts65Dn mouse model of DS. Oxidative stress is thought to be an underlying factor in the development of DS-related pathologies including muscle dysfunction. High-levels of oxidative stress have been attributed to triplication and elevated expression of superoxide dismutase 1 (SOD1); a gene located on chromosome 21. The elevated expression of SOD1 is postulated to increase production of hydrogen peroxide and cause oxidative injury and cell death. However, it is unknown whether SOD1 protein expression is associated with greater oxidant production in skeletal muscle from Ts65Dn mice. Thus, our objective was to assess levels of SOD1 expression and oxidant production in skeletal myofibers from the flexor digitorum brevis obtained from Ts65Dn and control mice. Measurements of oxidant production were obtained from myofibers loaded with 2',7'-dichlorodihydrofluorescein diacetate (DCFH2-DA) in the basal state and following 15min of stimulated unloaded contraction. Ts65Dn myofibers exhibited a significant decrease in basal DCF emissions (p < 0.05) that was associated with an approximate 3-fold increase in SOD1 (p < 0.05). DCF emissions were not affected by stimulating contraction of Ts65Dn or wild-type myofibers (p > 0.05). Myofibers from Ts65Dn mice tended to be smaller and myonuclear domain was lower (p < 0.05). In summary, myofibers from Ts65Dn mice exhibited decreased basal DCF emissions that were coupled with elevated protein expression of SOD1. Stimulated contraction in isolated myofibers did not affect DCF emissions in either group. These findings suggest the skeletal muscle dysfunction in the adult Ts65Dn mouse is not associated with skeletal muscle oxidative stress.


Chart review of the impact of attention-deficit/hyperactivity disorder comorbidity on response to lithium or divalproex sodium in adolescent mania.
Although adolescent onset of bipolar disorder is common, the optimal treatment approach for mania in this age group remains understudied. Comorbid attention-deficit/hyperactivity disorder (ADHD) has been reported to predict lithium resistance in adolescents with bipolar disorder. Little is known about response to dival-proex sodium in adolescents with bipolar disorder comorbid with ADHD. This study was conducted to evaluate comparative response rates to lithium and divalproex sodium in adolescent mania with and without this comorbidity. Medical records were reviewed for 42 patients (ages 12-19 years) who were hospitalized for acute mania and discharged with a diagnosis of DSM-III-R or DSM-IV bipolar disorder on either lithium (N = 29) or divalproex sodium (N = 13) treatment. A clinician blinded to treatment status rated improvement on the basis of abstracted notes in each case utilizing the Clinical Global Impressions Scale modified for use in bipolar illness (CGI-BP). Response was defined as a discharge CGI-BP overall change score of 1 or 2 (much or very much improved). Data were collected from January 1992 through May 1999. 36/42 (85.7%) patients presented with mixed mania, and 14/41 (34.1%) patients had a history of ADHD. The overall response rate was 80.9% (34/42). 92.6% (25/27) of patients without ADHD were responders versus 57.1% (8/14) of subjects with comorbid ADHD (p =.007). There were no significant differences in response rates for lithium versus divalproex sodium in subjects with and without ADHD. These retrospective data suggest overall equivalent response rates for lithium and divalproex sodium in predominantly mixed adolescent mania. However, a history of ADHD was associated with a significantly diminished acute response to both divalproex sodium and lithium as a primary treatment for the manic phase of bipolar disorder.


Interstitial fibrosis evolution on early sequential screening renal allograft biopsies using quantitative image analysis.
Screening renal biopsies (RB) may assess early changes of interstitial fibrosis (IF) after transplantation. The aim of this study was to quantify IF by automatic color image analysis on sequential RB. We analyzed RB performed at day (D) 0, month (M) 3 and M12 from 140 renal transplant recipients with a program of color segmentation imaging. The mean IF score was 19 ± 9% at D0, 27 ± 11% at M3 and 32 ± 11% at M12 with a 8% progression during the first 3 months and 5% between M3 and M12. IF at M3 was correlated with estimated glomerular rate (eGFR) at M3, 12 and 24 (p < 0.02) and IF at M12 with eGFR at M12 and 48 (p < 0.05). Furthermore, IF evolution between D0 and M3 (ΔIFM3-D0) was correlated with eGFR at M24, 36 and 48 (p < 0.03). IF at M12 was significantly associated with male donor gender and tacrolimus dose (p = 0.03). ΔIFM3-D0 was significantly associated with male donor gender, acute rejection episodes (p = 0.04) and diabetes mellitus (p = 0.02). Thus, significant IF is already present before transplantation. IF evolution is more important during the first 3 months and has some predictive ability for change in GFR. Intervention to decrease IF should be applied early, i.e. before 3 months, after transplantation.


Cadaveric skin allograft-associated cytomegalovirus transmission in a mouse model of thermal injury.
As a routine procedure to provide temporary coverage for burn wounds, cadaveric skin allografts have been used in patients with massive thermal injuries. In this study, CMV infection associated with skin grafting was investigated. Graft-associated CMV transmission was shown in a mouse model of thermal injury. Skins from mice 100 days after a nonlethal dose of murine CMV (MCMV) infection contained MCMV DNA and mRNA, although the virus was not isolated from these murine skins. When these skins were grafted to burned mice, the marked growth of MCMV was demonstrated in salivary glands. No viral growth was shown in the salivary glands of unburned mice or CMV sero(+) mice after grafting with these skins. When severe combined immunodeficient beige (SCID-beige) mice were used as recipients for CMV sero(+) skins, all mice died within 30 days after the grafting. Only 1 PFU/mouse of MCMV was shown to be 1 LD(50) in SCID-beige mice, while a 50% mortality rate was shown in normal unburned mice infected with 5 x 10(5) PFU/mouse of MCMV. This indicates that a very small amount of CMV contained in skins is sufficient to induce CMV infection in immunocompromised hosts. On the other hand, human CMV (HCMV) DNA and mRNA were detected by PCR analysis in 55% (DNA) and 33% (mRNA) of cadaveric skins, although the isolation of HCMV from cadaveric skin homogenates was not achieved in tissue cultures. CMV sero(-) patients with severe burn injuries may have a high risk for CMV infection associated with allografts of cadaveric skins.