SPECT in the Kleine-Levin Syndrome, a Possible Diagnostic and Prognostic Aid?
Kleine-Levin syndrome (KLS) is a rare syndrome of periodic hypersomnia and behavioral and cognitive symptoms based on clinical criteria. In the setting of differential diagnosis of hypersomnia disorders, an objective diagnostic aid is desirable. A promising modality is single photon emission computed tomography (SPECT). As intraepisodal investigations are difficult to perform, an interepisodal investigation would be very helpful. Another aim of the study was to correlate SPECT findings to prognosis. Twenty-four KLS-patients were categorized as severe or non-severe based on clinical characteristics. The clinical characteristics were analyzed in relation to SPECT-examinations performed between hypersomnia periods (interepisodal) or after remission, as a clinical routine investigation. Forty-eight percent of the KLS-patients have hypoperfusion in the temporal or fronto-temporal regions. In patients that have undergone remission, 56% show that pattern. There were no specific findings related to prognosis. SPECT might be a diagnostic aid, in a setting of hypersomnia experience. With a sensitivity of 48%, interepisodal SPECT alone cannot be used for diagnosing KLS.


Primary tumor staging for oral cancer and a proposed modification incorporating depth of invasion: an international multicenter retrospective study.
The current American Joint Committee on Cancer (AJCC) staging system for oral cancer demonstrates wide prognostic variability within each primary tumor stage and provides suboptimal staging and prognostic information for some patients. To determine if a modified staging system for oral cancer that integrates depth of invasion (DOI) into the T categories improves prognostic performance compared with the current AJCC T staging. Retrospective analysis of 3149 patients with oral squamous cell carcinoma treated with curative intent at 11 comprehensive cancer centers worldwide between 1990 and 2011 with surgery ± adjuvant therapy, with a median follow-up of 40 months. We assessed the impact of DOI on disease-specific and overall survival in multivariable Cox proportional hazard models and investigated for institutional heterogeneity using 2-stage random effects meta-analyses. Candidate staging systems were developed after identification of optimal DOI cutpoints within each AJCC T category using the Akaike information criterion (AIC) and likelihood ratio tests. Staging systems were evaluated using the Harrel concordance index (C-index), AIC, and visual inspection for stratification into distinct prognostic categories, with internal validation using bootstrapping techniques. The mean and median DOI were 12.9 mm and 10.0 mm, respectively. On multivariable analysis, DOI was a significantly associated with disease-specific survival (P < .001), demonstrated no institutional prognostic heterogeneity (I² = 6.3%; P = .38), and resulted in improved model fit compared with T category alone (lower AIC, P < .001). Optimal cutpoints of 5 mm in T1 and 10 mm in T2-4 category disease were used to develop a modified T staging system that was preferred to the AJCC system on the basis of lower AIC, visual inspection of Kaplan-Meier curves, and significant improvement in bootstrapped C-index. We propose an improved oral cancer T staging system based on incorporation of DOI that should be considered in future versions of the AJCC staging system after external validation.


Time spent above optimal cerebral perfusion pressure is not associated with failure to improve in outcome in traumatic brain injury.
Optimal cerebral perfusion pressure (CPPopt) has emerged as a promising personalized medicine approach to the management of moderate-to-severe traumatic brain injury (TBI). Though literature demonstrating its association with poor outcomes exists, there is yet to be work done on its association with outcome transition due to a lack of serial outcome data analysis. In this study we investigate the association between various metrics of CPPopt and failure to improve in outcome over time. CPPopt was derived using three different cerebrovascular reactivity indices; the pressure reactivity index (PRx), the pulse amplitude index (PAx), and the RAC index. For each index, % times spent with cerebral perfusion pressure (CPP) above and below its CPPopt and upper and lower limits of reactivity were calculated. Patients were dichotomized based on improvement in Glasgow Outcome Scale-Extended (GOSE) scores into Improved vs. Not Improved between 1 and 3 months, 3 and 6 months, and 1- and 6-month post-TBI. Logistic regression analyses were then conducted, adjusting for the International Mission for Prognosis and Analysis of Clinical Trials (IMPACT) variables. This study included a total of 103 patients from the Winnipeg Acute TBI Database. Through Mann-Whitney U testing and logistic regression analysis, it was found that % time spent with CPP below CPPopt was associated with failure to improve in outcome, while % time spent with CPP above CPPopt was generally associated with improvement in outcome. Our study supports the existing narrative that time spent with CPP below CPPopt results in poorer outcomes. However, it also suggests that time spent above CPPopt may not be associated with worse outcomes and is possibly even associated with improvement in outcome.


The release of microparticles by apoptotic cells and their effects on macrophages.
Microparticles are small membrane vesicles released from the cell membrane by exogenous budding. To elucidate the interactions of microparticles with macrophages, the effect of microparticles released from Jurkat T cells on RAW 264.7 cells was determined. Microparticles were isolated by differential centrifugation, using FACS analysis with annexin V and cell surface markers for identification. Various inducers of apoptosis increased the release of microparticles from Jurkat cells up to 5-fold. The released microparticles were then cultured with RAW 264.7 cells. As shown by confocal microscopy and FACS analysis, RAW 264.7 macrophages cleared microparticles by phagocytosis. In addition, microparticles induced apoptosis in RAW 264.7 cells in a dose-dependent manner with up to a 5-fold increase of annexin V positive cells and 9-fold increase in caspase 3 activity. Cell proliferation as determined by the MTT test was also reduced. Furthermore, microparticles stimulated the release of microparticles from macrophages. These effects were specific for macrophages, since no apoptosis was observed in NIH 3T3 and L929 cells. These findings indicate that microparticles can induce macrophages to undergo apoptosis, in turn resulting in a further increase of microparticles. The release of microparticles from apoptotic cells may therefore represent a novel amplification loop of cell death.


The spinach AtpC and AtpD genes contain elements for light-regulated, plastid-dependent and organ-specific expression in the vicinity of the transcription start sites.
Run-on assays with isolated nuclei demonstrate that the transcription rates of AtpC and AtpD (gene products: the CF1 subunits gamma and delta of the chloroplast ATP synthase) are comparable in spinach seedlings. However, chimeric GUS gene fusions with 5'-flanking regions of the AtpC gene direct an approximately 10-fold lower GUS level in transgenic tobacco compared with equivalent fragments from the AtpD gene. Both promoters contain sequences in the vicinity of the respective TATA boxes, which are sufficient to direct light-regulated, plastid-dependent and organ-specific expression of the GUS gene. In contrast, the upstream regions of both promoters differ the higher GUS level directed by the AtpD promoter is caused by enhancer-like elements located upstream of the region involved in the regulated expression, while nucleotides upstream of -73 in the AtpC promoter contribute relatively little to the promoter activity. 5'-Deletion analyses and site-directed mutagenesis studies indicated that the -73/-48 bp AtpC region contains cis-elements crucial for this regulated expression. If five nucleotides within this region (-59/-55) are exchanged, the GUS gene is constitutively expressed and the activity in etiolated seedlings, in seedlings with photobleached plastids and in roots increases to the level detectable in green cotyledons. It is concluded that signal transduction pathways from different regulators converge prior to gene regulation and that these five nucleotides are part of a cis-element which functions as a repressor in darkness, in tissues with impaired plastids and in roots.


Long-term coenzyme Q10 therapy: a major advance in the management of resistant myocardial failure.
Coenzyme Q10 (CoQ10) treatment, orally administered as 100 mg daily dose, was initiated in a series of patients with advanced heart failure in an open, controlled design. They were all showing an insufficient response to classical therapy with diuretics and digitalis. Twelve patients with various causes of heart failure, classified clinically by echocardiography (ECHO), (12/12), and heart catheterization with endomyocardial biopsy, (10/12), were followed prospectively for a mean period of seven months. Serial assessments: Clinical examination (with questionnaire), ECG, chest X-ray, ECHO, systolic time intervals (STI) and blood levels of CoQ10 were performed. With a mean latency period of 30 days, eight out of 12 patients (67%) showed definite clinical improvement. Subjectively, the patients felt less tired, their general activity tolerance increased and dyspnoea at rest disappeared. There were obvious signs of decreased right-sided stasis (hepatic congestion). The heart rate fell significantly, and the heart volume (chest X-ray) decreased in the eight responders (although n.s.). A significant reduction in the left atrial size (ECHO) was registered, suggesting a reduced preload of the left ventricle, Furthermore, a significant decline in the PEP/LVET ratio (STI) was indicative of an improved myocardial performance. Preliminary CoQ10 withdrawal results showed severe clinical relapse with subsequent improvement on CoQ10 reinstatement, supporting the interpretation that treatment of these patients corrected a myocardial deficiency of CoQ10 and increased contractility. Hence CoQ10 appears to be an effective therapeutic agent in advanced cases of heart failure. This is an attractive circumvention of the traditional principles of therapy: supporting the myocardium directly by ameliorating a supposed underlying mitochondrial dysfunction (exhausted bioenergetics).


Complications After Body Contouring Surgery in Postbariatric Patients.
Over recent years, body contouring procedures in postbariatric patients have been in exponentially growing demand resulting in high complication rates rendering a variety of ciphers in the literature. The purpose of this study is to determine the complication rate in patients who have undergone body contouring surgery after bariatric surgery between June 2012 and March 2015 at Hospital de San José. A cohort study including 153 individuals who underwent a total of 198 body contouring procedures after massive weight loss following bariatric surgery was conducted. Data on variables, such as complication rate according to the type of body contouring surgery, major and minor complications, weight of resected tissue, or intraoperation time, among other variables, were analyzed. A total of 198 procedures were performed in 153 patients. The mean (SD) age of the patients was 43.93 years (9.4 years). Of 198 procedures, 110 (55.5%) had complications. The rate for major complications was 13%, and for minor complications, 87%. Complication rates according to the type of operation were as follows: circumferential abdominal lipectomy, 55.7%; extended abdominal lipectomy, 53.7%; cruroplasty, 69%; breast surgery, 57%; and brachioplasty, 40%. Patients who presented with bleeding enough to require transfusion (P = 0.000) and with weight of the resected tissue greater than 2700 g in abdominoplasty (odds ratio, 3.26; 95% confidence interval, 1.48-7.1) had a higher complication rate. There were no thromboembolic events among this population. The overall complication rate was 55.5%. The great majority were minor complications. The thromboprophylaxis regimen used was 100% effective.


Pathologic consequences of increased angiotensin II activity.
Advances in molecular medicine and pharmacology have allowed clinicians to critically reassess the renin-angiotensin system. Angiotensin II (AII) participates in the control of cardiovascular function and electrolyte balance, and plays a part in the regulation of cellular oncogenes and the expression of growth factors. The expression of the proteins of the renin-angiotensin system in organs other than the kidneys suggests that these diverse actions are associated with the peptide in the local environment. Tissue renin-angiotensin activity has prompted the investigation of alternate pathways for the production of AII and characterization of novel forms of angiotensin peptides that counteract the vasoconstrictor and proliferative actions of AII. The heptapeptide angiotensin-(1-7) appears to be critically involved in regulating the angiotensinogen activity of AII through stimulation of vasodilator prostaglandins and release of nitric oxide. Study in this area has been accelerated by the identification of receptors that convey the actions of angiotensin peptides at the cellular level and the pharmacologic characterization of agents that inhibit the ability of AII to bind to target receptors. The introduction of a new class of orally active AII-receptor blockers has provided a specific test of the role of AII in the development of essential hypertension and the potential for improved therapy for hypertension and cardiac and vascular sequelae.


Biological variables in non-small cell lung cancer: comparison between immunocytochemical determination on fine needle aspirates from surgical specimens and immunohistochemical determination on tissue sections.
A number of biological and predictive markers of non-small cell lung cancer (NSCLC) have been sought, but these have so far been mainly evaluated on surgically resected specimens. Given that fine needle aspiration biopsy (FNAB) is being increasingly used in the diagnosis of NSCLC, its application could be extended to the immunocytochemical detection of biological parameters at the time of diagnosis before surgery. In order to assess the reliability of estimating biological markers on fine needle aspirates (FNAs) from NSCLC, the aim of this study was to compare Ki67 growth fraction, p53 and bcl-2 protein expression as revealed by the immuncytochemical assessment of FNAs obtained from surgical samples with the immunohistochemical results obtained from the corresponding histological sections. FNAs were performed on surgical specimens obtained from 29 NSCLC patients. Ki67, p53 and bcl-2 were cytologically and histologically evaluable in respectively 25, 27 and 19 cases. Concordance between FNAs and corresponding paraffin sections was 84% for Ki67, 93% for p53 and 95% for bcl-2. All of the specimens whose biological parameters were studied by immunocytohistochemistry also underwent flow cytometric DNA analysis of FNAs taken from fresh surgical specimens. Of the 29 cases, 22 were aneuploid and seven diploid. The S-phase fraction (SPF) was evaluable in 62% of cases. Comparison of SPF results on FNAs with Ki67 values evaluated on the corresponding histologic and cytologic specimens, revealed a significant correlation only with histology. Good reproducibility was also found in relation to the immunocytochemical results obtained on FNAs from different areas of the same tumour, showing that tumour heterogeneity does not affect the method. The concordance between the immunocytochemical and immunohistochemical results suggests that FNAB may be a reliable procedure for the biological characterization of NSCLC. Given its limited invasiveness, FNAB could be used in vivo for the preoperative assessment of biological parameters in patients with operable or metastatic NSCLC.


Effects of Early Auditory Deprivation on Working Memory and Reasoning Abilities in Verbal and Visuospatial Domains for Pediatric Cochlear Implant Recipients.
The overall goal of this study was to compare verbal and visuospatial working memory in children with normal hearing (NH) and with cochlear implants (CI). The main questions addressed by this study were (1) Does auditory deprivation result in global or domain-specific deficits in working memory in children with CIs compared with their NH age mates? (2) Does the potential for verbal recoding affect performance on measures of reasoning ability in children with CIs relative to their NH age mates? and (3) Is performance on verbal and visuospatial working memory tasks related to spoken receptive language level achieved by children with CIs? A total of 54 children ranging in age from 5 to 9 years participated; 25 children with CIs and 29 children with NH. Participants were tested on both simple and complex measures of verbal and visuospatial working memory. Vocabulary was assessed with the Peabody Picture Vocabulary Test (PPVT) and reasoning abilities with two subtests of the WISC-IV (Wechsler Intelligence Scale for Children, 4th edition): Picture Concepts (verbally mediated) and Matrix Reasoning (visuospatial task). Groups were compared on all measures using analysis of variance after controlling for age and maternal education. Children with CIs scored significantly lower than children with NH on measures of working memory, after accounting for age and maternal education. Differences between the groups were more apparent for verbal working memory compared with visuospatial working memory. For reasoning and vocabulary, the CI group scored significantly lower than the NH group for PPVT and WISC Picture Concepts but similar to NH age mates on WISC Matrix Reasoning. Results from this study suggest that children with CIs have deficits in working memory related to storing and processing verbal information in working memory. These deficits extend to receptive vocabulary and verbal reasoning and remain even after controlling for the higher maternal education level of the NH group. Their ability to store and process visuospatial information in working memory and complete reasoning tasks that minimize verbal labeling of stimuli more closely approaches performance of NH age mates.


A comparison of nine scales to detect depression in Parkinson disease: which scale to use?
The Methods of Optimal Depression Detection in Parkinson's Disease (MOOD-PD) study compared the psychometric properties of 9 depression scales to provide guidance on scale selection in Parkinson disease (PD). Patients with PD (n = 229) from community-based neurology practices completed 6 self-report scales (Beck Depression Inventory [BDI]-II, Center for Epidemiologic Studies Depression Rating Scale-Revised [CESD-R], 30-item Geriatric Depression Scale [GDS-30], Inventory of Depressive Symptoms-Patient [IDS-SR], Patient Health Questionnaire-9 [PHQ-9], and Unified Parkinson's Disease Rating Scale [UPDRS]-Part I) and were administered 3 clinician-rated scales (17-item Hamilton Depression Rating Scale [HAM-D-17], Inventory of Depressive Symptoms-Clinician [IDS-C], and Montgomery-Åsberg Depression Rating Scale [MADRS] and a psychiatric interview. DSM-IV-TR diagnoses were established by an expert panel blinded to the self-reported rating scale data. Receiver operating characteristic curves were used to estimate the area under the curve (AUC) of each scale. All scales performed better than chance (AUC 0.75-0.85). Sensitivity ranged from 0.66 to 0.85 and specificity ranged from 0.60 to 0.88. The UPDRS Depression item had a smaller AUC than the BDI-II, HAM-D-17, IDS-C, and MADRS. The CESD-R also had a smaller AUC than the MADRS. The remaining AUCs were statistically similar. The GDS-30 may be the most efficient depression screening scale to use in PD because of its brevity, favorable psychometric properties, and lack of copyright protection. However, all scales studied, except for the UPDRS Depression, are valid screening tools when PD-specific cutoff scores are used.


Light effects in yeast: inhibition by visible light of growth and transport in Saccharomyces cerevisiae grown at low temperatures.
Growth rate, sugar transport, and amino acid transport of yeast cells grown at 12 degrees C were inhibited by cool-white fluorescent light. At light intensities below 1,250 lx, growth and membrane transport were only slightly inhibited. Above 1,250 lx, there was increasing inhibition of both processes. Transport of histidine was completely inhibited after 3 to 5 days in cultures grown at 12 degrees C under 3,500-lx illumination. Cells grown at 20 degrees C were not inhibited by light intensities that caused complete loss of viability and membrane transport activity in cells grown at 12 degrees C.


Ethanol exposure affects cell movement during gastrulation and induces split axes in zebrafish embryos.
To explore the toxic effects of ethanol on axis formation during embryogenesis, zebrafish embryos at different developmental stages were treated with 3% ethanol for 3h. The effects of ethanol exposure appeared to be stage-dependent. The dome stage embryo was most sensible to form posterior split axes upon ethanol exposure. Morphological and histological observations and whole-mount in situ hybridization results showed that ethanol exposure at this stage caused a general gastrulation delay, and induced double notochords, double neural tubes and two sets of somites in the posterior trunk. Mechanistically, no ectopic organizer was found by examining the expression patterns of dorsoventral markers including goosecoid, chordin and eve1 at the onset of gastrulation. However, radial intercalation, epiboly and convergence extension were inhibited by ethanol exposure as revealed by cell labeling, phenotypic observation and the expression patterns of axial or paraxial markers. Further investigation showed that the cell aggregation might be affected by ethanol exposure, as indicated by the much more scattered expression pattern of chordin, eve1 and wnt11 at the early gastrula stage, and the discontinuous gsc positive cells during migration. These results imply that ethanol might affect cell movement before and during gastrulation and as a consequence, induces a split axes phenotype.


PIM kinase inhibition: co-targeted therapeutic approaches in prostate cancer.
PIM kinases have been shown to play a role in prostate cancer development and progression, as well as in some of the hallmarks of cancer, especially proliferation and apoptosis. Their upregulation in prostate cancer has been correlated with decreased patient overall survival and therapy resistance. Initial efforts to inhibit PIM with monotherapies have been hampered by compensatory upregulation of other pathways and drug toxicity, and as such, it has been suggested that co-targeting PIM with other treatment approaches may permit lower doses and be a more viable option in the clinic. Here, we present the rationale and basis for co-targeting PIM with inhibitors of PI3K/mTOR/AKT, JAK/STAT, MYC, stemness, and RNA Polymerase I transcription, along with other therapies, including androgen deprivation, radiotherapy, chemotherapy, and immunotherapy. Such combined approaches could potentially be used as neoadjuvant therapies, limiting the development of resistance to treatments or sensitizing cells to other therapeutics. To determine which drugs should be combined with PIM inhibitors for each patient, it will be key to develop companion diagnostics that predict response to each co-targeted option, hopefully providing a personalized medicine pathway for subsets of prostate cancer patients in the future.


Endogenous prostaglandins modulate autoregulation of renal blood flow in young rats.
After uninephrectomy in the young rat, renal blood flow (RBF) increases at normal renal perfusion pressure (RPP) but not at reduced RPP. To evaluate the role of endogenous prostaglandins on these effects, RBF was measured during reduction in RPP in rats that were uninephrectomized within 1 wk of age and studied at 33-41 days of age. After acute cyclooxygenase inhibition, autoregulation improved; RBF was unchanged at normal RPP but increased at lower pressures with concomitant decrease in renal venous plasma renin activity (PRA). Prostaglandin inhibition also improved autoregulatory efficiency in acutely uninephrectomized and sham-operated young rats. Infusion of a thromboxane synthesis inhibitor had no effect on the pressure-flow relationship or on PRA, whereas angiotensin converting enzyme inhibition increased RBF at all measured RPP without affecting autoregulation. We conclude that increased RBF at normal RPP in the uninephrectomized young rat is not maintained by increased prostaglandin synthesis or decreased thromboxane- or angiotensin-dependent vasoconstriction. However, by maintaining increased vasoconstriction at reduced RPP, prostaglandin-dependent renin release reduces autoregulatory efficiency in the young rat.


Terminating substrates of DNA polymerases: synthesis and functional study.
In order to study the mechanisms of DNA biosynthesis a number of modified nucleoside - substrates of DNA polymerases was synthesized. The absence of hydroxyl at 3'-position of ribose results in terminating properties of DNA biosynthesis of these analogues. A single step synthesis of triphosphates and alpha-thiotriphosphates of natural and 3'-modified 2'-deoxynucleosides is described.


Establishment of patient-derived three-dimensional organoid culture in renal cell carcinoma.
Renal cell carcinoma is a heterogeneous kidney cancer, and over 403,000 cases were reported worldwide in 2018. Current methods for studying renal cell carcinoma are limited to two-dimensional (2D) culture of primary cell lines and patient-derived xenograft models. Numerous studies have suggested that 2D culture poorly represents the diversity, heterogeneity, and drug-resistance of primary tumors. The time and cost associated with patient-derived xenograft models poses a realistic barrier to their clinical utility. As a biomimetic model, patient-derived three-dimensional (3D) organoid culture can overcome these disadvantages and bridge the gap between in vitro cell culture and in vivo patient-derived xenograft models. Here, we establish a patient-derived 3D organoid culture system for clear cell renal cell carcinoma and demonstrate the biomimetic characteristics of our model with respect to both primary kidney cancer and conventional 2D culture. Normal renal tissues and tumor tissues were collected from patients with clear cell renal cell carcinoma. The dissociated cells were cultured as conventional 2D culture and 3D organoid culture. The biomimetic characteristic of the two cultures were compared. Compared with 2D culture, the 3D organoid cultures retained the characteristic lipid-rich, clear cell morphology of clear cell renal cell carcinoma. Carbonic anhydrase 9 and vimentin were validated as biomarkers of renal cell carcinoma. Expression of the two validated biomarkers was more enhanced in 3D organoid culture. Patient-derived 3D organoid culture retains the characteristics of renal cell carcinoma with respect to morphology and biomarker expression.


Origins and Functional Significance of Eukaryotic Protein Folds.
Folds are the architecture and topology of a protein domain. Categories of folds are very few compared to the astronomical number of sequences. Eukaryotes have more protein folds than Archaea and Bacteria. These folds are of two types: shared with Archaea and/or Bacteria on one hand and specific to eukaryotic clades on the other hand. The first kind of folds is inherited from the first endosymbiosis and confirms the mixed origin of eukaryotes. In a dataset of 1073 folds whose presence or absence has been evidenced among 210 species equally distributed in the three super-kingdoms, we have identified 28 eukaryotic folds unambiguously inherited from Bacteria and 40 eukaryotic folds unambiguously inherited from Archaea. Compared to previous studies, the repartition of informational function is higher than expected for folds originated from Bacteria and as high as expected for folds inherited from Archaea. The second type of folds is specifically eukaryotic and associated with an increase of new folds within eukaryotes distributed in particular clades. Reconstructed ancestral states coupled with dating of each node on the tree of life provided fold appearance rates. The rate is on average twice higher within Eukaryota than within Bacteria or Archaea. The highest rates are found in the origins of eukaryotes, holozoans, metazoans, metazoans stricto sensu, and vertebrates: the roots of these clades correspond to bursts of fold evolution. We could correlate the functions of some of the fold synapomorphies within eukaryotes with significant evolutionary events. Among them, we find evidence for the rise of multicellularity, adaptive immune system, or virus folds which could be linked to an ecological shift made by tetrapods.


Fc gamma RII on human B cells can mediate enhanced antigen presentation.
The ability of Fc gamma R on monocytes and macrophages to enhance presentation of Ab-bound Ag has been well established. In contrast, studies suggest that Fc gamma RII on B cells functions primarily to suppress B cell responses. We found that presentation of tetanus toxoid (TT) by human EBV-transformed B cells was enhanced by 100- to 1000-fold, when TT was targeted to Fc gamma RII on B cells by using a conjugate consisting of TT covalently linked to anti-human Fc gamma RII mAb 41H16. This enhanced presentation could be blocked by mAb 41H16, heat-aggregated human IgG, or anti-MHC Class II mAb. Similarly, multimeric immune complexes composed to TT and anti-TT mAb, SA13, and 9F12 also enhanced presentation of TT. Furthermore, Fc gamma RII on purified human peripheral blood B cells could also enhance Ag presentation. These studies provide the first clear evidence that Fc gamma RII can participate in enhanced Ag presentation by human B cells and suggest that Fc gamma RII has the potential to play a stimulatory role in human B cell responses to Ag-Ab complexes.


Angiogenesis and antiangiogenic therapy for malignant gliomas.
Angiogenesis is crucial to the growth of malignant gliomas. Therefore, antiangiogenic therapy represents a new, promising therapeutic modality for malignant gliomas. This study was designed to define the malignant glioma cases most suitable for antiangiogenic therapy in humans and to demonstrate the efficacy of antiangiogenic therapy in animals. Protein expression of the most potent angiogenic factor, vascular endothelial growth factor (VEGF), and its specific natural inhibitor, soluble Flt-1, as well as vessel architecture, including vessel density, area, and diameter, was evaluated in human malignant glioma samples (24 glioblastomas, 13 anaplastic astrocytomas). Among these, VEGF >1000ng/ml, VEGF/soluble Fltl ratio >1, vessel density >30, and vessel area >7% were prognostic factors for malignant gliomas. Based on these results, we performed three different antiangiogenic experiments targeted to inhibit VEGF expression in a human malignant glioma (U87) mouse model: anti-VEGF neutralized antibody intraperitoneal injection; interferon-beta intramusclar injection; and transfection of an endogenous nonspecific angiogenesis inhibitor, thrombospondin-1, into glioma cells caused inhibition of VEGF secretion and/or mRNA expression and resulted in glioma growth inhibition of 70%, 84%, and 50%, respectively, compared with control. We conclude that malignant gliomas with high degrees of VEGF expression and vessel areas are good candidates for antiangiogenic therapy, especially that designed to inhibit VEGF expression.


Human Adipose-Derived Mesenchymal Stem Cells-Incorporated Silk Fibroin as a Potential Bio-Scaffold in Guiding Bone Regeneration.
Recently, stem cell-based bone tissue engineering (BTE) has been recognized as a preferable and clinically significant strategy for bone repair. In this study, a pure 3D silk fibroin (SF) scaffold was fabricated as a BTE material using a lyophilization method. We aimed to investigate the efficacy of the SF scaffold with and without seeded human adipose-derived mesenchymal stem cells (hASCs) in facilitating bone regeneration. The effectiveness of the SF-hASCs scaffold was evaluated based on physical characterization, biocompatibility, osteogenic differentiation in vitro, and bone regeneration in critical rat calvarial defects in vivo. The SF scaffold demonstrated superior biocompatibility and significantly promoted osteogenic differentiation of hASCs in vitro. At six and twelve weeks postimplantation, micro-CT showed no statistical difference in new bone formation amongst all groups. However, histological staining results revealed that the SF-hASCs scaffold exhibited a better bone extracellular matrix deposition in the defect regions compared to other groups. Immunohistochemical staining confirmed this result; expression of osteoblast-related genes (BMP-2, COL1a1, and OCN) with the SF-hASCs scaffold treatment was remarkably positive, indicating their ability to achieve effective bone remodeling. Thus, these findings demonstrate that SF can serve as a potential carrier for stem cells, to be used as an osteoconductive bioscaffold for BTE applications.


A theoretical and experimental study of light absorption and scattering by in vivo skin.
A theoretical treatment has been developed for the optical properties of a layered structure which absorbs and scatters light. This theory predicts that the logarithm of the inverse of reflectance (LIR) of the surface should be a useful parameter for the examination of that structure. This approach has been applied to a study of skin in vivo. An instrument was constructed for use in clinical situations to measure the LIR spectrum of skin over the visible region of the spectrum (450-760 nm). The contributions to the observed spectra made by pigments and the skin structure were deduced by reference to the theoretical model. Numerical indices were used to quantify the changes in skin haemoglobin content following the application of vasoconstricting preparations. The indices also provided a means of measuring erythema and melanin pigmentation induced in the skin by exposure to ultraviolet radiation. The assessments made using this instrument were more reproducible and sensitive than judgments made by eye.


MUTYH-null mice are susceptible to spontaneous and oxidative stress induced intestinal tumorigenesis.
MUTYH is a mammalian DNA glycosylase that initiates base excision repair by excising adenine opposite 8-oxoguanine and 2-hydroxyadenine opposite guanine, thereby preventing G:C to T:A transversion caused by oxidative stress. Recently, biallelic germ-line mutations of MUTYH have been found in patients predisposed to a recessive form of hereditary multiple colorectal adenoma and carcinoma with an increased incidence of G:C to T:A somatic mutations in the APC gene. In the present study, a systematic histologic examination revealed that more spontaneous tumors had developed in MUTYH-null mice (72 of 121; 59.5%) than in the wild type (38 of 109; 34.9%). The increased incidence of intestinal tumors in MUTYH-null mice (11 tumors in 10 of 121 mice) was statistically significant compared with the wild type (no intestinal tumors in 109 mice). Two adenomas and seven adenocarcinomas were observed in the small intestines, and two adenomas but no carcinomas were found in the colons. In MUTYH-null mice treated with KBrO(3), the occurrence of small intestinal tumors dramatically increased. The mean number of polyps induced in the small intestines of these mice was 61.88 (males, 72.75; females, 51.00), whereas it was 0.85 (males, 0.50; females, 1.00) in wild-type mice. The tumors developed predominantly in the duodenum and in the upper region of the (jejunum) small intestines. We conclude that MUTYH suppresses spontaneous tumorigenesis in mammals, thus providing experimental evidence for the association between biallelic germ-line MUTYH mutations and a recessive form of human hereditary colorectal adenoma and carcinoma.


Multiplane HiLo microscopy with speckle illumination and non-local means denoising.
HiLo microscopy synthesizes an optically sectioned image from two images, one obtained with uniform and another with patterned illumination, such as laser speckle. Speckle-based HiLo has the advantage of being robust to aberrations but is susceptible to residual speckle noise that is difficult to control. We present a computational method to reduce this residual noise without undermining resolution. In addition, we improve the versatility of HiLo microscopy by enabling simultaneous multiplane imaging (here nine planes). Our goal is to perform fast, high-contrast, multiplane imaging with a conventional camera-based fluorescence microscope. Multiplane HiLo imaging is achieved with the use of a single camera and z-splitter prism. Speckle noise reduction is based on the application of a non-local means (NLM) denoising method to perform ensemble averaging of speckle grains. We demonstrate the capabilities of multiplane HiLo with NLM denoising both with synthesized data and by imaging cardiac and brain activity in zebrafish larvae at 40 Hz frame rates. Multiplane HiLo microscopy aided by NLM denoising provides a simple tool for fast optically sectioned volumetric imaging that can be of general utility for fluorescence imaging applications.


Children's perspectives and experiences of health, diet, physical activity and weight in an urban, multi-ethnic UK population: A qualitative study.
Children from Black and South Asian ethnic groups are at risk for childhood obesity in the United Kingdom. To inform local action for childhood obesity prevention, it is crucial to explore the basis of ethnic disparities and consider the perspectives of children. This study aimed to understand cultural and contextual factors influencing childhood obesity in an ethnically diverse population using child-centred methodology. 'Draw, write and tell' interviews were held with children aged 9-10 years in Coventry, an urban, multi-ethnic city in the United Kingdom. Data were analysed thematically using framework analysis. Twenty-six children participated (85% from Black or minority ethnic groups). Children's perspectives revealed universal themes around health, diet, physical activity and weight and highlighted issues specific to ethnic groups and those living in deprived areas. An underlying feature was weight-based stigmatization and group stereotyping, and an emphasis on internal factors as the cause of obesity. Children described some experiences of social disadvantage but did not regard these as a barrier to being physically active. Children identified cultural or religious practices or experiences of migration that influenced diet and physical activity. These findings allow a broad range of children's perspectives to inform future intervention design. In addition, the study was able to identify the many similarities and small amount of diversity in children's perspectives across ethnic groups.


Renal dysplasia in Bardet-Biedl syndrome.
Bardet-Biedl syndrome (BBS) is a multisystem genetic disorder characterized with central obesity, pigmentary retinopathy, polydactyly, mental retardation, and hypogenitalism. Renal abnormalities have been recognized as a cardinal feature of the disease with serious prognostic implication. The aim of this study was to analyze the renal status in children with BBS and to implement appropriate interventions in those with progressive course Patients and methods: The diagnosis of BBS was established on the basis of criteria proposed by Beales et al. (J Med Genet 1999). Imaging of the kidneys and urinary tract was performed with ultrasound study, Tc99(m)DMSA scan and a cystographic study. Twenty four hour urine collections were obtained for estimation of proteinuria and creatinine clearance. Blood pressure was monitored at clinical visits or as 24-hour ambulatory monitoring. There were 4 children (2 males, 2 females). All four children displayed abnormal kidney ultrasound and Tc99(m)DMSA scan resembling dysplastic kidney(s). Two of them had overt proteinuria (glomerulo-tubular pattern). Three children had normal blood pressure and glomerular filtration rate (GFR): 107, 145 and 95 ml/min/1.73m(2), and the fourth had hypertension and progressive worsening of the GFR at 65 ml/min/1.73m(2). Children with BBS should undergo imaging studies of the kidneys and urinary tract at initial work up; in those with renal dysplasia proteinuria, GFR and blood pressure should be regularly monitored to slow down progression to terminal renal failure.


Inhibition of endogenous nitric oxide activity impairs the colonic sparing effect of rofecoxib, the cyclooxygenase-2 inhibitor and resveratrol, the preferential cyclooxygenase-1 inhibitor in the course of experimental colitis. Role of oxidative stress biomarkers and proinflammatory cytokines.
The gut mucosal barrier plays a key role in the physiology of gastrointestinal (GI) tract, preventing under homeostatic conditions, the epithelial cells of the gastric mucosa from hydrochloric acid and intestinal mucosa from alkaline secretion, food toxins and pathogenic microbiota. Previous studies have documented that blockade of both isoforms of cyclooxygenase (COX): constitutive (COX-1) and inducible (COX-2), as well NO synthase in the stomach exacerbated the gastric damage induced by various ulcerogens, however, such as effects of non-selective and selective inhibition of COX-1, COX-2 and NOS enzymes on colonic damage have been little studied. The supplementation of NO by intragastric (i.g.) treatment with NO-releasing compound NO-aspirin (NO-ASA) or substrate for NO synthase L-arginine ameliorated the damage of upper GI-tract, but whether similar effect can be observed in colonic mucosa associated with the experimental colitis, and if above mentioned compounds can be effective in aggravation or protection of experimental colitis remains less recognized. In this study rats with experimental colitis induced by intrarectal administration of 2,4,6-trinitrobenzosulphonic acid (TNBS) were daily treated for 7 days with: 1) vehicle (i.g.), 2) ASA 40 mg/kg (i.g.), 3) rofecoxib 10 mg/kg (i.g.), 4) resveratrol 10 mg/kg (i.g.), 5) NO-ASA 40 mg/kg (i.g.), 6) L-arginine 200 mg/kg (i.g.) with or without of L-NNA 20 mg/kg (i.p.). The macroscopic and microscopic area of colonic damage was determined planimetrically, the colonic blood flow (CBF) was assessed by Laser flowmetry, and the oxidative stress biomarkers malondialdehyde and 4-hydroxynonenal (MDA+4-HNE), the antioxidative factors superoxide dismutase (SOD) and glutathione (GSH), as well as proinflammatory cytokines in the colonic mucosa (tumor necrosis factor alpha (TNF-α) and interleukin-1beta (IL-1β)) were measured. We have documented that administration of TNBS produced gross and microscopic colonic damage and significantly decreased CBF (p<0.05). Treatment with ASA significantly increased the area of colonic damage (p<0.05), an effect accompanied by a significant decrease in the CBF, the significant increment of MDA+4-HNE, and the attenuation of the antioxidative properties in colonic mucosa, documented by a significant decrease of SOD activity and GSH concentration, and elevation of the colonic tissue levels of TNF-α and IL-1β comparing to control Veh-treated TNBS rats. Administration of rofecoxib or resveratrol also significantly increased the colonic damage and significantly decreased the CBF, causing an increase in MDA+4-HNE and mucosal content of TNF-α and IL-1α and a significant decrease of the SOD activity and GSH content (p<0.05), however, these changes were significantly less pronounced as compared with ASA. On the contrary, the treatment with NO-ASA, or L-arginine, significantly diminished the area of colonic lesions, the MDA+4-HNE concentration, attenuated the TNF-α and IL-1β levels, while increasing the CBF, SOD activity and GSH content (p<0.05). The concomitant treatment of L-NNA with rofecoxib or resveratrol reversed an increase in area of colonic damage and accompanying changes in CBF, colonic mucosa TNF-α and IL-1β levels, the MDA+4-HNE concentration, and SOD activity and GSH content comparing to those observed in TNBS rats treated with these COX-inhibitors alone (p<0.05). In contrast, co-treatment with L-NNA and NO-ASA or L-arginine failed to significantly affect the decrease of colonic lesions accompanied by the rise in CBF, the attenuation of MDA+4-HNE concentration, TNF-α and IL-1β levels, SOD activity and GSH content exerted by NO-ASA- or L-arginine treatment of the respective control TNBS-rats without L-NNA administration. These observations suggest that 1) the increase of NO availability either from NO-releasing donors such as NO-ASA or NO precursors such as L-arginine, can inhibit the inflammatory and microvasculature alterations, as well as increase in lipid peroxidation due to the enhanced efficacy of these compounds to increase the antioxidative properties of colonic mucosa, 2) unlike ASA which exacerbated the severity of colitis, the treatment with rofecoxib, the specific 'safer' COX-2 inhibitor or resveratrol, the polyphenolic compound known to act as the dual COX-1 and COX-2 inhibitor, can attenuate the colonic damage during course of TNBS colitis possibly via anti-inflammatory and antioxidative properties, and 3) the blockade of endogenous NO activity by L-NNA which also exacerbated the severity of mucosal damage in colitis, can abolish the sparing effect of rofecoxib and resveratrol indicating the NO bioavailability plays an important role in enhanced efficacy of both specific and dual COX inhibitors to ameliorate the experimental colitis.


Detection of Mycoplasma synoviae in poultry environment samples by culture and polymerase chain reaction.
Successful detection of Mycoplasma synoviae (MS) by culture and PCR from samples collected in the environment of experimentally infected chickens and turkeys, or under field conditions, is described. Results showed that in the experimental infection, 10/96 and 46/96 samples of food, drinking water, feathers, droppings or dust were positive by culture and Mycoplasma-PCR. In field conditions, the number of positive results for environmental samples were respectively 7/28 and 17/28. These observations highlight the high disseminating capacities of this mycoplasma and show the usefulness of the PCR method for epidemiological studies.


COVID-19 in children: what did we learn from the first wave?
A pandemic caused by the novel coronavirus, severe acute respiratory syndrome - coronavirus 2 (SARS-CoV-2), has caused high rates of mortality, predominantly in adults. Children are significantly less affected by SARS-CoV-2 with far lower rates of recorded infections in children compared to adults, milder symptoms in the majority of children and very low mortality rates. A suspected late manifestation of the disease, paediatric inflammatory multisystem syndrome - temporally associated with SARS-CoV-2 (PIMS-TS), has been seen in small numbers of children and has a more severe disease course than acute SARS-CoV-2. The pandemic has meant that children around the world have been kept off school, isolated from their extended family and friends and asked to stay inside. The UK has been declared as being in an economic recession and unemployment rates are increasing. These indirect effects of SARS-CoV-2 are likely to have a significant impact on many children for years to come. Consolidating the knowledge that has accumulated during the first wave of this pandemic is essential for recognising the clinical signs, symptoms and effective treatment strategies for children; identifying children who may be at increased risk of severe SARS-CoV-2 infection; planning the safe delivery of healthcare and non-health related services that are important for childrens' wellbeing; and engaging in, and developing, research to address the things that are not yet known. This article summarises the evidence that has emerged from the early phase of the pandemic and offers an overview for those looking after children or planning services.


Optimal siting of heat flux transducers for the assessment of body heat loss when immersed in water.
Regional heat flux (intensity of heat flow rate, W m-2) was measured using heat flux transducers affixed to the skin surface at 14 sites on the left arm and 15 sites on both the left leg and the trunk (total of 44 locations) of human subjects during head-out immersion in water at temperatures between 11 and 28 degrees C. The heat flux data and skin temperature data were collected continuously over a 30-min period taking approximately 2.5 min to scan through all transducers in sequence before repeating the process. There were 3 immersions per individual, monitoring the arm or the leg or the trunk. The local measurements of heat flux from each of the body segments were summated and meaned to obtain a value of overall flux for each area of the body for each 2- to 3-min interval. This mean value was taken as the optimal value (Hopt) for any particular body segment. Every permutation of the positions of any 1, 2, 3, 4, or 5 heat flux transducers from the possible 14 or 15 sites was used to obtain a mean value which could be compared to the Hopt over the total duration of each immersion. Comparisons could be made between the best individual sites (Iopt) and Hopt as well as between the overall best site for the group of 10 people (Gopt) and Hopt. All possible permutations could be ranked in ascending order of preference based on a decreasing sum of the squares of the differences between Hopt and either Iopt or Gopt. Confirmation of the validity of this simple ranking system was based on an analysis of the percentage differences between Iopt or Gopt and Hopt. Such an analysis indicated that the better choice of discs when using a small number of transducers was between 3 and 5 sites, affording both reasonable accuracy and practicality. It is recommended that between 3 and 5 discs be used per body segment at specific locations and the calculated heat loss from each segment be weighted according to its area of representation to give a value for the mean rate of total body heat loss. A reasonable level of accuracy was demonstrated using Gopt permutations for measuring both the rate of heat flow as well as the integral of the data to obtain a value of the quantity of heat (Joules) lost from the body.


Proteins of the bovine interphotoreceptor matrix: retinoid binding and other functions.
Bovine interphotoreceptor matrix (IPM) is rich in protein, containing a limited and specific set of these macromolecules. The matrix proteins are synthesized by adjacent tissues (neural retina and pigment epithelium), but are not, themselves, major constituents of these tissues. Of possible functional importance are the presence of some lysosomal hydrolases and of glycoprotein components that foster the aggregation of RPE cells. The IPM contains also a unique retinoid-binding protein - IRBP, a large glycoprotein located only in the IPM. IRBP carries more all-trans-retinol in light- than in dark-adapted eyes; this finding, coupled with IRBP's strategic position, indicates that IRBP may be a shuttle for vitamin A in the visual cycle. Furthermore, IRBP may mediate exchange of retinal among opsin molecules in the dark. The molecular weight of bovine IRBP is 133,000; the molecular radius is 56A. These measurements show that IRBP is probably an elongated molecule. Retinol binds relatively loosely to IRBP (with dissociation constant 1.3 X 10(-6) M), implying that IRBP should be able to transfer its vitamin A to cellular retinoid carriers in the retina and RPE. The absorbance, fluorescence, and circular dichroic characteristics of IRBP are compared to those of other retinol-binding proteins.


Synthesis, physico-chemical and biological characterization of a paclitaxel macromolecular prodrug.
Paclitaxel was attached to poly(hydroxyethylaspartamide) via a succinic spacer arm by a two-step protocol: (1) synthesis of 2'-O-succinyl-paclitaxel; (2) synthesis of PHEA-2'-O-succinyl-paclitaxel. The 2'-O-succinyl-paclitaxel derivative and the macromolecular conjugate were characterized by UV, IR, NMR and mass spectrometry analysis. The reaction yields were over 95% and the purity of products over 98%. Paclitaxel release and degradation from 2'-O-succinyl-paclitaxel occurred at a faster rate at pH 5.5 than 7.4. After 30 h of incubation at pH 5.5 and 7.4 the released free paclitaxel was about 40 and 20%, respectively. In plasma both drug release and degradation were found to occur at a higher rate than in buffer at pH 7.4 suggesting that an enzymatic mechanism could be involved. The paclitaxel release and degradation from PHEA-2'-O-succinyl-paclitaxel were negligible at pH 5.5 and 7.4 and very slow in plasma. Investigation carried out using murine myeloid cell line showed that the polymeric prodrug maintains partial pharmacological activity of paclitaxel. The DL50 of the conjugate (over 40 ng/ml) as compared to free paclitaxel (about 1 ng/ml) was correlated to the slow drug release. Finally a pharmacokinetic study carried out by intravenous inoculation of the macromolecular prodrug to mice demonstrated that the polymer conjugation modify dramatically the in vivo fate of the drug. The conjugate disappeared from the bloodstream much more quickly as compared to both free drug and naked polymer. Massive accumulation of bioconjugate in the liver (80% of the dose) was found to persist throughout 1 week.


Mallard ducks - a waterfowl species with high risk of distributing Escherichia coli pathogenic for humans.
Mallard ducks may pose a so far underestimated risk to human and animal health by transmitting pathogenic Escherichia coli via their faecal deposits to various environmental sources. We processed Mallard duck faecal samples for E. coli and unique clones, as defined by pulsed-field gel electrophoresis (PFGE), were subsequently investigated for their virulence genotype and phylogenetic background. Multilocus sequence typing and in vivo experiments were performed for selected clones. Of 60 clones identified among 142 E. coli isolated from 175 samples, 15 (25%) were recovered from multiple individuals (up to 23 per clone). None of the clones harboured stx1 and stx2 genes and other intestinal pathogenic E. coli virulence-associated genes were only occasionally identified in the collection. In contrast, the clones possessed considerable numbers of VAGs (up to 30) linked with extraintestinal pathogenic E. coli (ExPEC). Their pathogenic potential was confirmed in chicken infection experiments. Moreover, selected clones were assigned to sequence types (STs) being most prominent for human ExPEC strains, including ST95 and ST73. One clone exhibited a multi-resistant phenotype against several antibiotics including beta-lactams, tetracyclines and sulfonamides. Mallard ducks have therefore to be considered as an important reservoir for zoonotic E. coli strains, thus serving as a substantial non-point source especially of strains capable of causing extraintestinal diseases.


Below Safety Limits, Every Unit of Glomerular Filtration Rate Counts: Assessing the Relationship Between Renal Function and Cancer-specific Mortality in Renal Cell Carcinoma.
The hypothesis that renal function could influence oncological outcomes is supported by anecdotal literature. To determine whether estimated glomerular filtration rate (eGFR) is related to cancer-specific mortality (CSM) in patients who had undergone surgery for renal cell carcinoma (RCC). A retrospective analysis of 3457 patients who underwent radical (39%) or partial nephrectomy (61%) for cT1-2 RCC between 1990 and 2015. The eGFR was calculated by the Chronic Kidney Disease Epidemiology Collaboration equation. CSM was analyzed in a multivariable competing-risk framework, estimating the subdistribution hazard ratio (SHR) accounting for deaths from other causes. The relationship between eGFR and CSM was investigated from multiple statistical approaches-extended Cox regression with eGFR incorporated as a time-dependent covariate, landmark analysis, and joint modeling. Other predictors were selected by competing-risk random forest method and backward elimination. The relationship between eGFR and CSM was graphically described by a linear spline, i.e. a continuous piecewise linear function with two lines joined by a knot. For eGFR treated as a time-dependent covariate, the knot was located at 65ml/min; at landmark analysis with eGFR at the baseline, 12 mo, and last functional follow-up, the knots were 85, 60, and 65ml/min, respectively. In multivariable competing-risk analysis, CSM was associated with eGFR only for values of eGFR below these cutoffs, with SHRs for every 10ml/min of reduction in eGFR of 1.25 (p=0.003), 1.16 (p=0.028), 1.44 (p=0.02), and 1.16 (p=0.042), corresponding to time-dependent eGFR, and eGFR at baseline, 12 mo, and last functional follow-up, respectively. Joint modeling confirmed these results. A retrospective design with inherent biases in data collection represents a limitation. In patients undergoing surgery for RCC, renal function should be preserved in order to improve cancer-related survival. The relationship between renal function and probability of dying due to renal cancer is complex. The present study found a correlation between glomerular filtration rate and cancer specific mortality that could reconsider the oncological role of renal function in patients undergoing surgery for renal cancer.


Rapid synchronized fabrication of vascularized thermosets and composites.
Bioinspired vascular networks transport heat and mass in hydrogels, microfluidic devices, self-healing and self-cooling structures, filters, and flow batteries. Lengthy, multistep fabrication processes involving solvents, external heat, and vacuum hinder large-scale application of vascular networks in structural materials. Here, we report the rapid (seconds to minutes), scalable, and synchronized fabrication of vascular thermosets and fiber-reinforced composites under ambient conditions. The exothermic frontal polymerization (FP) of a liquid or gelled resin facilitates coordinated depolymerization of an embedded sacrificial template to create host structures with high-fidelity interconnected microchannels. The chemical energy released during matrix polymerization eliminates the need for a sustained external heat source and greatly reduces external energy consumption for processing. Programming the rate of depolymerization of the sacrificial thermoplastic to match the kinetics of FP has the potential to significantly expedite the fabrication of vascular structures with extended lifetimes, microreactors, and imaging phantoms for understanding capillary flow in biological systems.


Evaluation of the Efficacy of Clinical Breast Examination Gloves in the Diagnosis of Breast Lumps.
Recent studies have questioned the efficacy of mammography in reducing breast cancer-related mortality. Additionally, the efficacies of commercially available gloves marketed as aiding the detection of breast lumps have not been independently verified. To evaluate the efficacy of clinical breast examination gloves in the detection of breast lumps. During the period from October 2011 to June 2012, patients were submitted to clinical examination with and without gloves. This prospective study involved 202 patients who underwent conventional clinical breast examination (test 1) or clinical breast examination with Sensifemme® gloves (test 2). All patients underwent subsequent bilateral ultrasonography (test 3) to confirm the findings of the physical examinations. The Chi-square test was used to compare values, while the kappa concordance index was used to determine the concordance between the diagnostic tests. The mean age of the patients was 43 years; 298 breast lumps were detected. In the clinical examination group (test 1), sensitivity was 54%, specificity was 78%, and accuracy was 57%. These rates for clinical breast examinations with gloves (test 2) were 68%, 58%, and 66%, respectively. The glove increased the diagnosis of breast nodules by 14%; the rate of false-positives was also higher (42% for test 2 compared to 22% for test 1). The accuracy of the glove was found to be superior to clinical examination after 100 patients had been examined. The kappa indices for test 1 vs. test 3 and for test 2 vs. test 3 were 0.15 and 0.16, respectively. Clinical examination using the glove was more effective than clinical examination with bare hands for the diagnosis of breast lumps, as it increased the sensitivity and accuracy of lump detection. However, this was at the expense of a higher false-positive rate, which can lead to further tests, unnecessary biopsies, and patient anxiety. The concordance of clinical examination results (whether performed with or without the glove) with those of ultrasonography is weak. Moreover, the glove has a steep learning curve that may discourage its use in certain circumstances.


Fate of chlorinated fatty acids in migrating sockeye salmon and their transfer to arctic grayling.
To investigate whether biotransport constitutes an entry route into pristine ecosystems for nonpersistent, nonvolatile xenobiotic compounds, extractable organically bound halogen in sockeye salmon (Oncorhynchus nerka) from Alaska was determined before and after spawning migration. The major organohalogen compounds in the salmon were halogenated fatty acids, predominantly chlorinated species that accounted for up to 35% of the extractable, organically bound chlorine (EOCl) in the fish tissues. The amount of chlorinated fatty acids in the salmon muscle decreased as a result of spawning migration. The decrease was correlated with that of triacylglycerols in the salmon muscle, indicating the chlorinated fatty acids to be mobilized and metabolized to approximately the same extent as the other fatty acids. Chlorinated fatty acids were also transferred to the maturing roe in a manner similar to that of the unchlorinated fatty acids. Lipids of the Arctic grayling (Thymallus arcticus), a fish resident to the spawning lake of the salmon, contained higher concentrations of chlorinated fatty acids than grayling in a lake without migratory salmon. This may reflect a food-chain transfer of the chlorinated fatty acids originating from the salmon, demonstrating a long-range transport route for this type of pollutants to pristine areas.


From the Eukaryotic Molybdenum Cofactor Biosynthesis to the Moonlighting Enzyme mARC.
All eukaryotic molybdenum (Mo) enzymes contain in their active site a Mo Cofactor (Moco), which is formed by a tricyclic pyranopterin with a dithiolene chelating the Mo atom. Here, the eukaryotic Moco biosynthetic pathway and the eukaryotic Moco enzymes are overviewed, including nitrate reductase (NR), sulfite oxidase, xanthine oxidoreductase, aldehyde oxidase, and the last one discovered, the moonlighting enzyme mitochondrial Amidoxime Reducing Component (mARC). The mARC enzymes catalyze the reduction of hydroxylated compounds, mostly N-hydroxylated (NHC), but as well of nitrite to nitric oxide, a second messenger. mARC shows a broad spectrum of NHC as substrates, some are prodrugs containing an amidoxime structure, some are mutagens, such as 6-hydroxylaminepurine and some others, which most probably will be discovered soon. Interestingly, all known mARC need the reducing power supplied by different partners. For the NHC reduction, mARC uses cytochrome b5 and cytochrome b5 reductase, however for the nitrite reduction, plant mARC uses NR. Despite the functional importance of mARC enzymatic reactions, the structural mechanism of its Moco-mediated catalysis is starting to be revealed. We propose and compare the mARC catalytic mechanism of nitrite versus NHC reduction. By using the recently resolved structure of a prokaryotic MOSC enzyme, from the mARC protein family, we have modeled an in silico three-dimensional structure of a eukaryotic homologue.


Update of diagnostic and prognostic markers in cutaneous malignant melanoma.
The biomolecules described in this article generally have been studied as possible diagnostic or clinically prognostic markers in the context of melanoma disease progression as measured by the gold standards of tumor thickness and development of metastasis. Most of the markers showed variations in expression phenotype only during the deeply invasive or metastatic stage of tumor progression and were thus predictive of clinical outcome only for these subgroups of patients. Some of the markers may have utility in identifying patients with deeply invasive primary tumors who are likely to develop metastasis and thus should receive earlier, more aggressive treatments. In addition, some of the markers may identify patients likely to respond better to a new type of therapy (e.g., anti-angiogenic therapy in a patient whose tumor is overexpressing VEGF or immunotherapy for a patient whose tumor is expressing high levels of MART-1). In the future, it will probably be possible to employ new techniques, such as laser-guided microdissection of tissues, to isolate individual melanocytes in order to identify the earliest stage-specific defects that contribute to an aggressive biological behavior. Identifying the subset of patients with superficially invasive melanomas who will develop metastatic disease will continue to provide a challenge.


3D CT cinematic rendering of mycotic aneurysms.
Mycotic aneurysms are uncommon but emergent conditions in which infection of a vessel leads to a contained rupture. Progression to frank rupture, thrombosis, distal embolization, and death can occur. The widespread availability of computed tomography (CT) and its ability to obtain high-resolution, contrast-enhanced, volumetric images rapidly has made it the modality of choice for evaluating mycotic aneurysms. Three-dimensional CT visualizations can provide important information to surgeons and interventionalists prior to attempting repair of these lesions. In this case series, we demonstrate the appearance of mycotic aneurysms with the novel 3D CT visualization methodology known as cinematic rendering (CR). CR makes use of a more complex lighting model than has previously been utilized with other 3D CT techniques, allowing for enhanced surface detail and realistic shadowing effects. These features of CR may have utility in evaluating mycotic aneurysms and in pre-procedural/pre-operative planning, although a prospective study definitively evaluating this has not yet been performed.


Analysis of genome-wide association study data using the protein knowledge base.
Genome-wide association studies (GWAS) aim to identify causal variants and genes for complex disease by independently testing a large number of SNP markers for disease association. Although genes have been implicated in these studies, few utilise the multiple-hit model of complex disease to identify causal candidates. A major benefit of multi-locus comparison is that it compensates for some shortcomings of current statistical analyses that test the frequency of each SNP in isolation for the phenotype population versus control. Here we developed and benchmarked several protocols for GWAS data analysis using different in-silico gene prediction and prioritisation methodologies. We adopted a high sensitivity approach to the data, using less conservative statistical SNP associations. Multiple gene search spaces, either of fixed-widths or proximity-based, were generated around each SNP marker. We used the candidate disease gene prediction system Gentrepid to identify candidates based on shared biomolecular pathways or domain-based protein homology. Predictions were made either with phenotype-specific known disease genes as input; or without a priori knowledge, by exhaustive comparison of genes in distinct loci. Because Gentrepid uses biomolecular data to find interactions and common features between genes in distinct loci of the search spaces, it takes advantage of the multi-locus aspect of the data. Results suggest testing multiple SNP-to-gene search spaces compensates for differences in phenotypes, populations and SNP platforms. Surprisingly, domain-based homology information was more informative when benchmarked against gene candidates reported by GWA studies compared to previously determined disease genes, possibly suggesting a larger contribution of gene homologs to complex diseases than Mendelian diseases.


Highly Ordered Mesoporous Co3 O4 Electrocatalyst for Efficient, Selective, and Stable Oxidation of 5-Hydroxymethylfurfural to 2,5-Furandicarboxylic Acid.
Electrochemical oxidation of biomass substrates to valuable bio-chemicals is highly attractive. However, the design of efficient, selective, stable, and inexpensive electrocatalysts remains challenging. Here it is reported how a 3D highly ordered mesoporous Co3 O4 /nickel foam (om-Co3 O4 /NF) electrode fulfils those criteria in the electrochemical oxidation of 5-hydroxymethylfurfural (HMF) to value-added 2,5-furandicarboxylic acid (FDCA). Full conversion of HMF and an FDCA yield of >99.8 % are achieved with a faradaic efficiency close to 100 % at a potential of 1.457 V vs. reversible hydrogen electrode. Such activity and selectivity to FDCA are attributed to the fast electron transfer, high electrochemical surface area, and reduced charge transfer resistance. More impressively, remarkable catalyst stability under long-term testing is obtained with 17 catalytic cycles. This work highlights the rational design of metal oxides with ordered meso-structures for electrochemical biomass conversion.


Drug-drug interaction between paxlovid and tacrolimus in a patient with myasthenia gravis and SARS-CoV-2 infection.
Patients with both myasthenia gravis (MG) and SARS-CoV-2 infection face treatment challenges due to potential drug interactions. One common immunosuppressant for MG, Tacrolimus, is primarily metabolized by the cytochrome P450. However, Paxlovid, an antiviral medication, inhibits cytochrome P450 activity, which can lead to increased Tacrolimus levels and potential toxicity when the two drugs are combined. In this case report, we present the case of a 39-year-old woman with early-onset MG who was initially treated with Tacrolimus. Later, she received Paxlovid for SARS-CoV-2 infection, which resulted in a sudden spike in Tacrolimus levels due to the drug interaction. This case emphasizes the importance of personalized treatment plans and close monitoring of drug interactions in patients with multiple health conditions. Clinicians should exercise vigilance regarding potential Tacrolimus interactions and regularly monitor blood levels to prevent adverse effects. Caution and close monitoring of Tacrolimus levels are essential when administering Paxlovid to patients on Tacrolimus therapy.


Crystalloid inclusions in endothelial cells of cellular and capillary hemangiomas. A possible sign of cellular immaturity.
The fine structure of crystalloid inclusions were found in the endothelial cells of capillary (strawberry) hemangiomas and cellular hemangiomas in nine infants. The inclusions measured from 0.5 to 2.0 microns in diameter and showed parallel lamellar bands with a regular periodicity of about 180 to 300 A. The presence of crystalloid inclusions and the absence of typical rod-shaped tubulated Weibel-Palade bodies suggest the immaturity and embryonic character of the endothelium present in the blood vessels of these kinds of hemangiomas.


Role of long-chain fatty acyl-CoA esters in the regulation of metabolism and in cell signalling.
The intracellular concentration of free unbound acyl-CoA esters is tightly controlled by feedback inhibition of the acyl-CoA synthetase and is buffered by specific acyl-CoA binding proteins. Excessive increases in the concentration are expected to be prevented by conversion into acylcarnitines or by hydrolysis by acyl-CoA hydrolases. Under normal physiological conditions the free cytosolic concentration of acyl-CoA esters will be in the low nanomolar range, and it is unlikely to exceed 200 nM under the most extreme conditions. The fact that acetyl-CoA carboxylase is active during fatty acid synthesis (Ki for acyl-CoA is 5 nM) indicates strongly that the free cytosolic acyl-CoA concentration is below 5 nM under these conditions. Only a limited number of the reported experiments on the effects of acyl-CoA on cellular functions and enzymes have been carried out at low physiological concentrations in the presence of the appropriate acyl-CoA-buffering binding proteins. Re-evaluation of many of the reported effects is therefore urgently required. However, the observations that the ryanodine-senstitive Ca2+-release channel is regulated by long-chain acyl-CoA esters in the presence of a molar excess of acyl-CoA binding protein and that acetyl-CoA carboxylase, the AMP kinase kinase and the Escherichia coli transcription factor FadR are affected by low nanomolar concentrations of acyl-CoA indicate that long-chain acyl-CoA esters can act as regulatory molecules in vivo. This view is further supported by the observation that fatty acids do not repress expression of acetyl-CoA carboxylase or Delta9-desaturase in yeast deficient in acyl-CoA synthetase.


Direct Liquid-Culture Screening for Evaluating the Production of Heterologous Proteins Using an Auxotrophic Mutant of Aspergillus Oryzae.
Aspergillus oryzae, a filamentous fungus, is one of the most widely used hosts for industrial applications including large-scale production of proteins. A polyethylene glycol (PEG)-mediated protoplast transformation method is generally used for the introduction of heterologous genes into A. oryzae. The conventional method typically requires three weeks for the screening of favorable transformants. Here, a new technique, the direct liquid-culture (DLC) screening method, is introduced which reduces the screening time to six days in a 200 mL flask format or to 10 days in a 24 well microplate format. The DLC screening method ensures the acquisition of positive transformants and evaluation of the secretory production of heterologous proteins in a single step, unlike the conventional screening method where two separate steps are required for the same. The protocol for PEG-mediated protoplast transformation of A. oryzae is described, which consists of five steps: preparation of fresh spore suspension, preculture, preparation of protoplasts, introduction of DNA, and DLC screening. For successful results in DLC screening, it is critical to use a nutrient-rich medium with optimized osmotic pressure. The protocol should further popularize the use of A. oryzae as a host of choice in the industrial production of proteins.


FDA-Approved Drug Screening for Compounds That Facilitate Hematopoietic Stem and Progenitor Cells (HSPCs) Expansion in Zebrafish.
Hematopoietic stem cells (HSCs) are a specialized subset of cells with self-renewal and multilineage differentiation potency, which are essential for their function in bone marrow or umbilical cord blood transplantation to treat blood disorders. Expanding the hematopoietic stem and progenitor cells (HSPCs) ex vivo is essential to understand the HSPCs-based therapies potency. Here, we established a screening system in zebrafish by adopting an FDA-approved drug library to identify candidates that could facilitate HSPC expansion. To date, we have screened 171 drugs of 7 categories, including antibacterial, antineoplastic, glucocorticoid, NSAIDS, vitamins, antidepressant, and antipsychotic drugs. We found 21 drugs that contributed to HSPCs expansion, 32 drugs' administration caused HSPCs diminishment and 118 drugs' treatment elicited no effect on HSPCs amplification. Among these drugs, we further investigated the vitamin drugs ergocalciferol and panthenol, taking advantage of their acceptability, limited side-effects, and easy delivery. These two drugs, in particular, efficiently expanded the HSPCs pool in a dose-dependent manner. Their application even mitigated the compromised hematopoiesis in an ikzf1-/- mutant. Taken together, our study implied that the larval zebrafish is a suitable model for drug repurposing of effective molecules (especially those already approved for clinical use) that can facilitate HSPCs expansion.


[Study of the carbohydrate metabolic enzyme activity of Act. noursei].
Activity of aldolase and threosophosphate dehydrogenase, transketolase and phosphogluconate dehydrogenase in Act. noursei, strain 153 and its inactive mutant 149 was studied comparatively. The enzyme activity of the inactive mutant was investigated in the absence of the antibiotic production and under conditions of reduced biosynthesis of nystatin in this strain after addition of the fermentation broth filtrate of the inactive mutant 369 to the medium. The activity of the enzymes of the hexosomonophosphate metabolic pathway in the active strain 153 of Act. noursei was 2-4 times higher than that of the inactive mutant 149. The activity of the enzymes of the hexosomonophosphate metabolic pathways increased and reached the level of the enzyme of the active mutant. The high level of the enzyme activity of the hexosomonophosphate glycolysis pathway is probably one of the necessary conditions for nystatin production.


Trail marking by caterpillars of the silverspot butterfly Dione juno huascuma.
A pheromone is implicated in the trail marking behavior of caterpillars of the nymphalid silverspot butterfly, Dione juno huascuma (Reakirt) (Lepidoptera: Heliconiinae) that feed gregariously on Passiflora (Malpighiales: Passifloraceae) vines in Mexico. Although they mark pathways leading from one feeding site to another with silk, this study shows that the silk was neither adequate nor necessary to elicit trail following behavior. Caterpillars marked trails with a long-lived pheromone that was deposited when they brushed the ventral surfaces of the tips of their abdomens along branch pathways. The caterpillars distinguished between pathways deposited by different numbers of siblings and between trails of different ages. Caterpillars also preferentially followed the trails of conspecifics over those of another nymphalid, Nymphalis antiopa L., the mourning cloak butterfly.


[Effect of electronic moxibustion on memory function in patients with amnestic mild cognitive impairment].
To observe the effect of electronic moxibustion on memory function in the patients with amnestic mild cognitive impairment (aMCI). A total of 59 aMCI patients were randomized into an electronic moxibustion group (30 cases) and a placebo moxibustion group (29 cases). In the electronic moxibustion group, the electronic moxibustion was applied to Baihui (GV 20), Dazhui (GV 14), Mingmen (GV 4) and Taixi (KI 3), 45 ℃ in temperature, 20 min each time. The treatment was given once a day, 5 times a week. The treatment for 4 weeks was as one course and 2 courses were required totally. In the placebo moxibustion group, the moxa-free patch was used, 38 ℃ in temperature. The acupoint selection and the treatment frequency were same as the electronic moxibustion group. Before and after treatment, Rivermead behavior memory test (RBMT) was adopted to evaluate the global memory function of the patients in the two groups and the N-back task test was adopted to evaluate working memory function separately. Additionally, the mini-mental state examination (MMSE) and its immediate memory, Montreal cognitive assessment (MoCA) and its delay recall were adopted to evaluate the global cognitive function and memory function. In the electronic moxibustion group, after treatment, RBMT score, N-back accuracy rates, MMSE and MoCA scores and the scores of immediate memory and delay recall were improved significantly as compared with those before treatment (P<0.01). In the placebo moxibustion group, the accuracy rates of 1-back and 2-back task and the scores of immediate memory and delay recall were improved obviously as compared with those before treatment (P<0.05, P<0.01). After treatment, the improvements of RBMT score, the accuracy rates of N-back task and MMSE and MoCA scores in the electronic moxibustion group were higher than those in the placebo moxibustion group (P<0.05). Electronic moxibustion improves memory function in the patients with amnestic mild cognitive impairment.


ADHD: Is There an App for That? A Suitability Assessment of Apps for the Parents of Children and Young People With ADHD.
Attention-deficit hyperactivity disorder (ADHD) is a highly comorbid disorder that can impact significantly on the individual and their family. ADHD is managed via pharmacological and nonpharmacological interventions. Parents also gain support from parent support groups, which may include chat rooms, as well as face-to-face meetings. With the growth of technology use over recent years, parents have access to more resources that ever before. A number of mobile apps have been developed to help parents manage ADHD in their children and young people. Unfortunately many of these apps are not evidence-based, and little is known of their suitability for the parents or whether they are helpful in ADHD management. The aim of this study was to explore the (1) parents' views of the suitability of the top ten listed apps for parents of children and young people with ADHD and (2) the views of clinicians that work with them on the suitability and value of the apps. The top 10 listed apps specifically targeted toward the parents of children and young people with ADHD were identified via the Google Play (n=5) and iTunes store (n=5). Interviews were then undertaken with 7 parents of children or young people with ADHD and 6 clinicians who specialize in working with this population to explore their opinions of the 10 apps identified and what they believe the key components are for apps to be suitable and valuable for this population. Four themes emerged from clinician and parent interviews: (1) the importance of relating to the app, (2) apps that address ADHD-related difficulties, (3) how the apps can affect family relationships, and (4) apps as an educational tool. Two additional themes emerged from the clinician interviews alone: monitoring ADHD symptoms and that apps should be practical. Parents also identified an additional theme: the importance of the technology. Overall, the characteristics of the current top 10 listed apps did not appear to match well to the views of our sample. Findings suggest that these apps may not fully meet the complex needs of this parent population. Further research is required to explore the value of apps with this population and how they can be tailored to their very specific needs.


The induction of cytoplasmic petite mutants of Saccharomyces cerevisiae by hydrostatic pressure.
This study demonstrates that hydrostatic pressure is a potent inductive agent of the petite mutation in cultures of Saccharomyces cerevisiae. The inductive capacity of this mutagen is dependent on the magnitude and the duration of the pressure treatment. Furthermore, the extent of petite induction varies with the growth stage of the culture. Induction occurs in pressure-treated (1-4 X 1-(4) lbf in.-2 or 9-66 X 10(4) kN m-2 for 4 h) log growth cultures but not in stationary or lag phase cultures. Petite induction and cell survival are also dependent on the particular strain of yeast which is pressure-treated. Tetrad analysis and complementation assays demonstrate that pressure-induced petite cells are cytoplasmic in nature. Moreover, induced petite cells show a wide range of suppressivity (2--99%) with a large proportion of the petite cells being highly suppressive.


Mood changes after watching pornography on the Internet are linked to tendencies towards Internet-pornography-viewing disorder.
Internet-pornography-viewing disorder (IPD) is considered one type of Internet-use disorder. For IPD's development, it was assumed theoretically that a dysfunctional use of Internet pornography to cope with depressive mood or stress might be considered to be a risk factor. To address the effect of Internet pornography use on mood, an online study with three measuring points with a sample of male participants was conducted. Participants were investigated regarding their tendencies towards IPD, personal use of Internet pornography, general mood, perceived stress, and their Internet pornography use motivation. Moreover, participants were asked regarding their current mood, sexual arousal, and need to masturbate before and after they watched Internet pornography self-determinedly in a private environment. Data showed that tendencies towards IPD were associated negatively with feeling generally good, awake, and calm and were correlated positively with perceived stress in daily life and using Internet pornography for excitation seeking and emotional avoidance. Self-determined use of Internet pornography in their private environment was accompanied by changes in mood and indicators of sexual arousal. Moreover, tendencies towards IPD were negatively related to mood before and after Internet-pornography use as well as an actual increase of good and calm mood. The results showed effects of watching Internet pornography on mood and sexual arousal which can be considered having reinforcing effects for the user. Thus, the results are in line with theoretical assumptions on IPD's development, in which the positive (and negative) reinforcement received by Internet-pornography use is related to cue-reactivity and craving reactions.


Knotted vs. unknotted proteins: evidence of knot-promoting loops.
Knotted proteins, because of their ability to fold reversibly in the same topologically entangled conformation, are the object of an increasing number of experimental and theoretical studies. The aim of the present investigation is to assess, on the basis of presently available structural data, the extent to which knotted proteins are isolated instances in sequence or structure space, and to use comparative schemes to understand whether specific protein segments can be associated to the occurrence of a knot in the native state. A significant sequence homology is found among a sizeable group of knotted and unknotted proteins. In this family, knotted members occupy a primary sub-branch of the phylogenetic tree and differ from unknotted ones only by additional loop segments. These "knot-promoting" loops, whose virtual bridging eliminates the knot, are found in various types of knotted proteins. Valuable insight into how knots form, or are encoded, in proteins could be obtained by targeting these regions in future computational studies or excision experiments.


MICB typing by PCR amplification with sequence specific primers.
MICB is a member of the MIC (MHC class I chain-related gene) family. Sixteen MICB alleles have been described; however, the functional relevance and population distribution of MICB alleles or their potential association to disease has not yet been evaluated. In this study, we have developed a PCR system using sequence-specific primers (PCR-SSP) that allows unambiguous amplification of all MICB alleles. This approach has been applied to type 100 healthy unrelated individuals from the Spanish population. The extent of polymorphism in this population is lower than that initially expected, and only nine alleles were detected. The alleles MICB01021 (46%), MICB0103101 (13.5%), MICB0104 (13.5%) and MICB0106 (12.5%) were found to be the most frequent alleles. HLA-B and MICA transmembrane polymorphism typing were also performed in this population. Our data showed that MICB is in linkage disequilibrium with MICA and even with HLA-B. Thus, the linkage disequilibrium with MICA and HLA-B suggests that MICB is a potential candidate for those diseases classically associated with HLA class I alleles.


Characterization of mouse liver alpha-L-fucosidase. Demonstration of unusual basic isoelectric forms of the enzyme that appear to be developmentally regulated.
Mouse tissues contain unusual basic isoelectric forms of alpha-L-fucosidase (with approximate isoelectric points of 8.3 and 9.0) in addition to the usual acidic and neutral forms previously described in tissues of other species. These unusual forms are very prominent in placenta and foetal tissues and comprise approx, 50-80% of total activity up to 11 days of postnatal development. By 15 days of postnatal development, the basic forms are diminished in amount and comprise not more than 25% of total activity. Neuraminidase treatment of adult mouse liver alpha-L-fucosidase led to significantly decreased amounts of acidic forms and increased amounts of the basic forms, suggesting that these forms are chemically related at least in part by sialic acid residues. Comparative kinetic studies on mouse liver, human liver and mouse placental alpha-L-fucosidases indicated that they have the same Km (0.05-0.06 mM) for 4-methylumbelliferyl alpha-L-fucopyranoside but different pH optima and thermostability properties. Mouse liver alpha-L-fucosidase has one pH optimum (5.5) and an acidic shoulder (centred around pH 4.0) compared with two distinct optima (4.3 and 6.8) for the human liver enzyme. Mouse placental alpha-L-fucosidase has a pH-activity curve comparable with that of the mouse liver enzyme except that the acidic shoulder is absent. Mouse liver alpha-L-fucosidase is considerably more thermolabile after preincubation at 50 degrees C than are the human liver and mouse placental enzymes, which gave similar thermodenaturation curves. Immunochemical studies indicated that mouse and human alpha-L-fucosidases are dissimilar antigenically but exhibit some cross-reactivity. The IgG fraction of antibody prepared in goat against human liver alpha-L-fucosidase was ineffective by itself in immunoprecipitating mouse liver alpha-L-fucosidase, but 63% and 72% of the mouse liver and placental enzymes respectively could be immunoprecipitated in the double-antibody experiments under conditions that immunoprecipitated 92% of the human liver enzyme.


Hippocampal pyramidal cells: the reemergence of cortical lamination.
The increasing resolution of tract-tracing studies has led to the definition of segments along the transverse axis of the hippocampal pyramidal cell layer, which may represent functionally defined elements. This review will summarize evidence for a morphological and functional differentiation of pyramidal cells along the radial (deep to superficial) axis of the cell layer. In many species, deep and superficial sublayers can be identified histologically throughout large parts of the septotemporal extent of the hippocampus. Neurons in these sublayers are generated during different periods of development. During development, deep and superficial cells express genes (Sox5, SatB2) that also specify the phenotypes of superficial and deep cells in the neocortex. Deep and superficial cells differ neurochemically (e.g. calbindin and zinc) and in their adult gene expression patterns. These markers also distinguish sublayers in the septal hippocampus, where they are not readily apparent histologically in rat or mouse. Deep and superficial pyramidal cells differ in septal, striatal, and neocortical efferent connections. Distributions of deep and superficial pyramidal cell dendrites and studies in reeler or sparsely GFP-expressing mice indicate that this also applies to afferent pathways. Histological, neurochemical, and connective differences between deep and superficial neurons may correlate with (patho-) physiological phenomena specific to pyramidal cells at different radial locations. We feel that an appreciation of radial subdivisions in the pyramidal cell layer reminiscent of lamination in other cortical areas may be critical in the interpretation of studies of hippocampal anatomy and function.


Humoral immunity in malnutrition.
Malnutrition affects the humoral immune system in diverse fashions. B lymphocyte subpopulations, serum IgG, and IgA levels, and immunoglobulin synthesis and metabolism are usually normal or increased. Hypogammaglobulinemia may occur in very young, severely malnourished infant. Although usually normal, deficient antibody responses to injected antigens are occasionally present, depending on the severity of the malnutrition. Serum IgE levels are usually high in malnutrition, probably due both to the increased incidence of intestinal parasites and to the lack of T cell control of IgE. Despite these high IgE levels, allergy is unusual. Secretory IgA levels in the respiratory and gastrointestinal (GI) fluids are generally decreased, as are secretory IgA antibody responses. This local antibody deficiency may increase GI permeability, stimulate immune complex formation, and result in the formation of IgG food antibodies. Breast milk secretory IgA levels are equivalent in malnourished mothers but because of lessened milk volume, the total amount of secretory IgA delivered to their offspring is diminished. Complement levels are somewhat diminished in the presence of malnutrition with a resultant opsonic deficiency. Although these B cell system aberrations undoubtedly contribute to the enhanced susceptibility to infection of malnourished patients, they are usually less severe than are concomitant T cell deficiencies.


The Relevance of Sex Differences in Performance Fatigability.
Performance fatigability differs between men and women for a range of fatiguing tasks. Women are usually less fatigable than men, and this is most widely described for isometric fatiguing contractions and some dynamic tasks. The sex difference in fatigability is specific to the task demands so that one mechanism is not universal, including any sex differences in skeletal muscle physiology, muscle perfusion, and voluntary activation. However, there are substantial knowledge gaps about the task dependency of the sex differences in fatigability, the involved mechanisms, and the relevance to clinical populations and with advanced age. The knowledge gaps are in part due to the significant deficits in the number of women included in performance fatigability studies despite a gradual increase in the inclusion of women for the last 20 yr. Therefore, this review 1) provides a rationale for the limited knowledge about sex differences in performance fatigability, 2) summarizes the current knowledge on sex differences in fatigability and the potential mechanisms across a range of tasks, 3) highlights emerging areas of opportunity in clinical populations, and 4) suggests strategies to close the knowledge gap and understanding the relevance of sex differences in performance fatigability. The limited understanding about sex differences in fatigability in healthy and clinical populations presents as a field ripe with opportunity for high-impact studies. Such studies will inform on the limitations of men and women during athletic endeavors, ergonomic tasks, and daily activities. Because fatigability is required for effective neuromuscular adaptation, sex differences in fatigability studies will also inform on optimal strategies for training and rehabilitation in both men and women.


Transcription factor ZNF488 accelerates cervical cancer progression through regulating the MEK/ERK signaling pathway.
Cervical cancer (CC) is one of the most common gynecological malignancies worldwide. Zinc Finger Protein 488 (ZNF488) has been identified as an oncogene in nasopharyngeal carcinoma. However, its biological role and potential mechanism in CC remain to be elucidated. In the present study, upregulation of ZNF488 expression in human CC tissues was found in clinical samples and analyzed in The Cancer Genome Atlas (TCGA) dataset, which was associated with clinical staging and lymph node metastasis. Quantitative real time polymerase chain reaction (PCR) and western blot assays indicated that the expression of ZNF488 was up-regulated in CC cells. Cell colony formation and cell cycle analysis assays suggested that ZNF488 promoted CC cell proliferation and cycle progression. Knockdown of ZNF488 inhibited tumor growth of xenograft tumor mice in vivo, in agreement with the levels of ZNF488 and Ki-67. Moreover, transwell and western assays demonstrated that ZNF488 enhanced CC cell migration and invasion. Additionally, knockdown of ZNF488 also inhibited lung metastasis of CC cells in vivo. Further mechanism analysis implied that ZNF488 promoted the MEK/ERK signaling pathway. ERK inhibitor PD98059 significantly weakened the proliferation and epithelial-mesenchymal transformation (EMT) promotion effect of ZNF488. Collectively, ZNF488 exerts its oncogene function partially through modulating MEK/ERK signaling pathway in CC, indicating that ZNF488 may provide a promising therapeutic target for the treatment of CC.


Noncompetitive, reversible inhibition of aminoacylase-1 by a series of L-alpha-hydroxyl and L-alpha-fluoro fatty acids: ligand specificity of aspergillus oryzae and porcine kidney enzymes.
L-lactate and L-beta-phenyllactate have been identified in the culture broth of Streptomyces sp. KY-11 as reversible noncompetitive inhibitors of Aspergillus oryzae aminoacylase-1 and porcine kidney aminoacylase I. A series of alpha-hydroxyl acids (DL-R-CH(OH)-COOH, R = Et, n-pro, n-butyl, n-pentyl, n-hexyl) also inhibited the two enzymes in reversible noncompetitive kinetics, and the inhibition potency (-log K(i)) increased with the increased hydrophobicity of the R group. The two eukaryotic enzymes showed distinct preferences to the ligand alpha-alkyl group, and the fungus enzyme was inhibited by L-beta-phenyllactate (R = benzyl) 10(3)-fold more potently than the mammalian enzyme. L-alpha-Fluoro-beta-phenyl-propionate and its D-isomer were used to show that the L-configuration of the alpha-substituent was important for potent inhibition of both the enzymes. The fungus aminoacylase-1 steeply decreased the affinity to alpha-fluoro- and alpha-hydroxy-n-caproate as pH was raised from 7 to 11, whereas the mammalian enzyme retained the affinity to these ligands under alkaline conditions. These results suggest that A. oryzae aminoacylase-1 has an acidic residue that interacts with -OH or -F, while the mammalian enzyme would have a basic residue that recognizes the alpha-substituents.


Traumatic fronto-ethmoidal encephalocele: a rare case.
Encephalocele is defined as herniation of the brain tissue from a bone defect in the cranium. It may be congenital, traumatic, tumoural or develop spontaneously. Traumatic fronto-ethmoidal encephalocele is rare and should be kept in mind for patients who have suffered trauma. Early diagnosis is important in terms of preventing life-threatening complications such as meningitis. Encephalocele and cerebrospinal fluid (CSF) leakage can be easily determined with basic radiological imaging methods and clinical findings. The computed tomography (CT) and magnetic resonance imaging (MRI) findings are here presented of a case with traumatic encephalocele.