Collaboration of MYC and RUNX2 in lymphoma simulates T-cell receptor signaling and attenuates p53 pathway activity.
MYC and RUNX oncogenes each trigger p53-mediated failsafe responses when overexpressed in vitro and collaborate with p53 deficiency in vivo. However, together they drive rapid onset lymphoma without mutational loss of p53. This phenomenon was investigated further by transcriptomic analysis of premalignant thymus from RUNX2/MYC transgenic mice. The distinctive contributions of MYC and RUNX to transcriptional control were illustrated by differential enrichment of canonical binding sites and gene ontology analyses. Pathway analysis revealed signatures of MYC, CD3, and CD28 regulation indicative of activation and proliferation, but also strong inhibition of cell death pathways. In silico analysis of discordantly expressed genes revealed Tnfsrf8/CD30, Cish, and Il13 among relevant targets for sustained proliferation and survival. Although TP53 mRNA and protein levels were upregulated, its downstream targets in growth suppression and apoptosis were largely unperturbed. Analysis of genes encoding p53 posttranslational modifiers showed significant upregulation of three genes, Smyd2, Set, and Prmt5. Overexpression of SMYD2 was validated in vivo but the functional analysis was constrained by in vitro loss of p53 in RUNX2/MYC lymphoma cell lines. However, an early role is suggested by the ability of SMYD2 to block senescence-like growth arrest induced by RUNX overexpression in primary fibroblasts.


Splicing variation of Long-IRBIT determines the target selectivity of IRBIT family proteins.
IRBIT [inositol 1,4,5-trisphosphate receptor (IP3R) binding protein released with inositol 1,4,5-trisphosphate (IP3)] is a multifunctional protein that regulates several target molecules such as ion channels, transporters, polyadenylation complex, and kinases. Through its interaction with multiple targets, IRBIT contributes to calcium signaling, electrolyte transport, mRNA processing, cell cycle, and neuronal function. However, the regulatory mechanism of IRBIT binding to particular targets is poorly understood. Long-IRBIT is an IRBIT homolog with high homology to IRBIT, except for a unique N-terminal appendage. Long-IRBIT splice variants have different N-terminal sequences and a common C-terminal region, which is involved in multimerization of IRBIT and Long-IRBIT. In this study, we characterized IRBIT and Long-IRBIT splice variants (IRBIT family). We determined that the IRBIT family exhibits different mRNA expression patterns in various tissues. The IRBIT family formed homo- and heteromultimers. In addition, N-terminal splicing of Long-IRBIT changed the protein stability and selectivity to target molecules. These results suggest that N-terminal diversity of the IRBIT family and various combinations of multimer formation contribute to the functional diversity of the IRBIT family.


Transcriptomic analysis identifies a role of PI3K-Akt signalling in the responses of skeletal muscle to acute hypoxia in vivo.
Changes in gene expression that occur within hours of exposure to hypoxia in in vivo skeletal muscles remain unexplored. Two hours of hypoxia caused significant down-regulation of extracellular matrix genes followed by a shift at 6 h to altered expression of genes associated with the nuclear lumen while respiratory and blood gases were stabilized. Enrichment analysis of mRNAs classified by stability rates suggests an attenuation of post-transcriptional regulation within hours of hypoxic exposure, where PI3K-Akt signalling was suggested to have a nodal role by pathway analysis. Experimental measurements and bioinformatic analyses suggested that the dephosphorylation of Akt after 2 h of hypoxic exposure might deactivate RNA-binding protein BRF1, hence resulting in the selective degradation of mRNAs. The effects of acute hypoxia have been widely studied, but there are few studies of transcriptional responses to hours of hypoxia in vivo, especially in hypoxia-tolerant tissues like skeletal muscles. We used RNA-seq to analyse gene expression in plantaris muscles while monitoring respiration, arterial blood gases, and blood glucose in mice exposed to 8% O2 for 2 or 6 h. Rapid decreases in blood gases and a slower reduction in blood glucose suggest stress, which was accompanied by widespread changes in gene expression. Early down-regulation of genes associated with the extracellular matrix was followed by a shift to genes associated with the nuclear lumen. Most of the early down-regulated genes had mRNA half-lives longer than 2 h, suggesting a role for post-transcriptional regulation. These transcriptional changes were enriched in signalling pathways in which the PI3K-Akt signalling pathway was identified as a hub. Our analyses indicated that gene targets of PI3K-Akt but not HIF were enriched in early transcriptional responses to hypoxia. Among the PI3K-Akt targets, 75% could be explained by a deactivation of adenylate-uridylate-rich element (ARE)-binding protein BRF1, a target of PI3K-Akt. Consistent decreases in the phosphorylation of Akt and BRF1 were experimentally confirmed following 2 h of hypoxia. These results suggest that the PI3K-Akt signalling pathway might play a role in responses induced by acute hypoxia in skeletal muscles, partially through the dephosphorylation of ARE-binding protein BRF1.


Attention and its coupling to action.
We discuss two commentaries that we have received on our target article (Humphreys et al., 2010). We elaborate on the evidence for action effects on extinction and discuss whether these effects occur pre or post the selection of a response. In addition, we discuss the neural basis of the effects of action relations on extinction and on the generalization of results on action relations to real-world examples.


Developments in ghrelin biology and potential clinical relevance.
The spiropiperidine, MK0677, has been exploited to characterize and expression clone the growth hormone secretagogue receptor (GHS-R). Cloning of this receptor led to identification of its natural ligands, ghrelin and adenosine. Targeted disruption of the Ghsr gene demonstrated unambiguously that the GH-releasing and orexigenic properties of ghrelin are dependent on Ghsr expression and that the orexigenic signal is mediated through neuropeptide Y and agouti-related peptide neurons. This review summarizes new developments in our understanding of the physiological roles of ghrelin and its receptor (GHS-R). Recent discoveries of the effects of ghrelin on the thymus and proinflammatory and chemotactic cytokine pathways stimulate renewed interest in potential clinical applications, which include age-associated disorders, such as metabolic disease, sarcopenia, congestive heart failure, atherosclerosis and anorexia.


Cytokine treatment of endothelial cells increases glycoprotein Ib alpha-dependent adhesion to von Willebrand factor.
Endothelial cells (EC) possess at least two membrane receptors for von Willebrand factor (vWF), the vitronectin receptor (VNR, alpha(v)beta3), which recognizes an Arg-Gly-Asp (RGD) sequence in the C-terminus of vWF, and glycoprotein Ib alpha (GP Ib alpha), which interacts with a region in the N-terminal A1 domain of vWF. In the absence of added cytokines, EC attachment to a vWF substratum is mediated largely through the alpha(v)beta3, with a smaller contribution by GP Ib alpha. In the present study, we have examined the effect of cytokines on the receptor specificity of EC attachment to wild-type vWF (WT-vWF) and to vWF, which had been mutated in the C-terminal RGDS sequence (RADS-vWF). Exposure of human umbilical vein EC (HUVEC) to tumor necrosis factor-alpha (TNF-alpha) or to TNF-alpha in combination with interferon-gamma (IFN-gamma), but not to interleukin-1beta (IL-1), increased attachment to RADS-vWF by about twofold. The TNF-alpha-induced increase in EC attachment was accompanied by an increase in cell surface GP Ib alpha expression; GP Ib alpha surface expression was not increased by IL-1. Attachment of untreated HUVEC to WT-vWF could be inhibited 60% to 70% by a monoclonal antibody (MoAb) (LM609) to the VNR and 30% to 40% by the A1 fragment of vWF (containing the GP Ib alpha binding domain). The pattern of inhibition of attachment to WT-vWF was largely unchanged after TNF-alpha treatment of HUVEC. In contrast, the attachment to WT-vWF of HUVEC, treated with TNF-alpha +IFN-gamma was completely inhibited by vWF-A1 and inhibited only 35% by the anti-VNR antibody LM609. Two MoAbs to GP Ib alpha produced similar, but incomplete, inhibition. Pretreatment of HUVEC with the combination of TNF-alpha + IFN-gamma produced a dramatic decrease in VNR expression, confirming previous findings of Defilippi et al. These results suggest that in the presence of the inflammatory cytokines TNF-alpha + IFN-gamma, the endothelial GP Ib complex is a major determinant of HUVEC adhesion to surface-bound vWF.


Detection of G-protein heterotrimers on large dense core and small synaptic vesicles of neuroendocrine and neuronal cells.
Heterotrimeric G proteins, initially believed to be exclusively present in the plasma membrane, have also been found to be associated with intracellular membrane compartments. There they are involved in various membrane trafficking processes including regulated secretion (reviewed in Bomsel, M., K. Mostov, Mol. Biol. Cell 3, 1317-1328 (1992)). Vesicles of two distinct types enter the regulated secretory pathway, i.e. large dense core vesicles and small synaptic vesicles, which differ in their membrane composition and content. Little is known about an association of heterotrimeric G proteins with regulated secretory vesicles, that would explain some aspects of the role heterotrimeric G proteins have during secretion. By immunofluorescence microscopy and immunoreplica analysis, we provide the first demonstration of the presence of complete sets of heterotrimeric G proteins, consisting of alpha-, beta-, and gamma-subunits, on large dense core vesicles from bovine adrenal medulla (chromaffin granules) and small synaptic vesicles from rodent and bovine brain. Each of the two types of secretory vesicles contains beta-subunits (at least beta 1 and beta 2), as well as gamma-subunits (at least gamma 2 or gamma 3). Interestingly, they differ in their composition of alpha-subunits. On small synaptic vesicles, we found two G(o) alpha-subunits (alpha o1 and alpha o2) and two Gi alpha-subunits (alpha i1 and alpha i2). In contrast, on chromaffin granules so far only one alpha o-subunit but no alpha i-subunits could be detected. Functional properties such as transmitter storage and/or exocytotic membrane fusion may be modulated by the various G-protein subunits associated with chromaffin granules and small synaptic vesicles.


[Pregnancy and breast cancer: 2 entities with complex interferences. Critical study of the dossier].
The author reviews the epidemiological, biological and clinical data concerning the association of carcinoma of the breast and pregnancy. He discusses, in particular, the therapeutic problems raised by this association. The author attracts attention to the fact that in certain circumstances, pregnancy may aggravate the disease, whilst in others, it seems to protect with regard to the cancer. As far as the therapeutic problems are concerned, he emphasizes in particular, the following conclusions: 1) in cases which require surgery, the prognosis is less poor than generally believed; 2) the total inefficacy of therapeutic abortion is now certain; 3) radiotherapy and chemotherapy should be prohibited during pregnancy; 4) a pregnancy occurring during the years following mastectomy for carcinoma, does not seem to have the unfavourable prognosis which was originally believed.


Development of Optimized Inhibitor RNAs Allowing Multisite-Targeting of the HCV Genome.
Engineered multivalent drugs are promising candidates for fighting infection by highly variable viruses, such as HCV. The combination into a single molecule of more than one inhibitory domain, each with its own target specificity and even a different mechanism of action, results in drugs with potentially enhanced therapeutic properties. In the present work, the anti-HCV chimeric inhibitor RNA HH363-10, which has a hammerhead catalytic domain and an aptamer RNA domain, was subjected to an in vitro selection strategy to isolate ten different optimised chimeric inhibitor RNAs. The catalytic domain was preserved while the aptamer RNA domain was evolved to contain two binding sites, one mapping to the highly conserved IIIf domain of the HCV genome's internal ribosome entry site (IRES), and the other either to IRES domain IV (which contains the translation start codon) or the essential linker region between domains I and II. These chimeric molecules efficiently and specifically interfered with HCV IRES-dependent translation in vitro (with IC50 values in the low µM range). They also inhibited both viral translation and replication in cell culture. These findings highlight the feasibility of using in vitro selection strategies for obtaining improved RNA molecules with potential clinical applications.


Imaging of congenital Zika virus infection: the route to identification of prognostic factors.
Zika virus (ZIKV) has recently emerged as a novel teratogenic agent associated with severe neurological complications. The risk associated with maternal infection remains to be exactly defined but appears to be significant. Like other TORCH agents (toxoplasmosis, other agents, rubella, cytomegalovirus and herpes simplex), it is unlikely that all affected fetuses will be symptomatic at birth. It is therefore urgent to better define the spectrum of anomalies observed in infected fetuses to provide adequate parental counseling. In this review, we provide a comprehensive analysis of major cases described to date and highlight specific prenatal and postnatal radiological findings of congenital ZIKV infection. A total of 19 reports were included in our analysis. ZIKV seemed to harbor a specific tropism for the central nervous system, and anomalies were mostly limited to the brain. Major radiological findings were ventriculomegaly, diffuse calcifications and signs of abnormal gyration as well as cortical development. In addition, a significant number of fetuses suffered from intra uterine growth restriction. Based on these findings, we provide recommendations for adequate radiological monitoring of at-risk pregnancies. © 2016 John Wiley & Sons, Ltd.


Temozolomide-induced aplastic anaemia and incidental low-grade B-cell non-Hodgkin lymphoma in a geriatric patient with glioblastoma multiforme.
Glioblastoma multiforme is an astrocyte-derived tumour representing the most aggressive primary brain malignancy. The median overall survival is 10-12 months, but it drops to 3-8.5 months for the cohort with more than 65 years old, which account to half of all patients. Initial management in this patient population aims to balance overall patient survival and quality of life with the inherent risks of treatment intervention, which include maximal safe tumour resection, radiation and temozolomide (TMZ) chemotherapy. This is accomplished through risk stratification as a function of patient age, functional status, comorbidities, tumour location and methylguanine methyltransferase promoter methylation status. We describe the care of a patient with prolonged febrile neutropaenia, with a rare but fatal complication from TMZ-induced idiosyncratic reaction, leading to aplastic anaemia and a provoking diagnosis of low-grade B-cell non-Hodgkin's lymphoma.


An Analysis of Tumor Margin Shrinkage in the Surgical Resection of Squamous Cell Carcinoma of the Oral Cavity.
Background Surgical resection of the oral cavity squamous cell carcinoma with clear surgical margins is the key to preventing local recurrence and avoiding the need for adjuvant treatment or margin re-resection. There is often a discrepancy observed between the clinically determined margins of the tumor when it is being resected and the histopathological result after the specimen has been processed. Methods A total of six patients who underwent primary surgical resection of oral squamous cell carcinoma between June and October 2020 were included. Surgical margins of the tumor were measured and recorded at three stages of tumor resection: pre-incision, post-resection, and post-formalin fixation. The 1 cm pre-incision anterior margin was compared to both the anterior post-resection and post-formalin fixation margins to document any shrinkage between the different stages of tumor resection. Results The overall mean surgical margin shrinkage was 26% (95% confidence interval {CI} 9.34-42.66, p=0.012). The greatest amount of margin shrinkage occurred between pre-incision and post-resection measurements, which is statistically significant at 19.7% (95% CI 7.49-31.83, p=0.009). To a lesser extent, tumor surgical margins also decreased by 12.7% (95% CI -2.66 to 28.09, p=0.083) between post-resection and post-formalin fixation. Conclusion Dimensions of tumor surgical margins in oral cavity squamous cell carcinoma specimens decrease from surgical resection to histopathological processing. Most of this shrinkage occurs between the clinically determined pre-incision and immediately after tumor resection in the post-resection measurement. These findings suggest that it might be prudent to consider surgical margin shrinkage when outlining initial margins to ensure adequate and complete resection of the tumor.


New Nuclear Magnetic Moment of ^{209}Bi: Resolving the Bismuth Hyperfine Puzzle.
A recent measurement of the hyperfine splitting in the ground state of Li-like ^{208}Bi^{80+} has established a "hyperfine puzzle"-the experimental result exhibits a 7σ deviation from the theoretical prediction [J. Ullmann et al., Nat. Commun. 8, 15484 (2017)NCAOBW2041-172310.1038/ncomms15484; J. P. Karr, Nat. Phys. 13, 533 (2017)NPAHAX1745-247310.1038/nphys4159]. We provide evidence that the discrepancy is caused by an inaccurate value of the tabulated nuclear magnetic moment (μ_{I}) of ^{209}Bi. We perform relativistic density functional theory and relativistic coupled cluster calculations of the shielding constant that should be used to extract the value of μ_{I}(^{209}Bi) and combine it with nuclear magnetic resonance measurements of Bi(NO_{3})_{3} in nitric acid solutions and of the hexafluoridobismuthate(V) BiF_{6}^{-} ion in acetonitrile. The result clearly reveals that μ_{I}(^{209}Bi) is much smaller than the tabulated value used previously. Applying the new magnetic moment shifts the theoretical prediction into agreement with experiment and resolves the hyperfine puzzle.


Manganese Enhanced MRI for Use in Studying Neurodegenerative Diseases.
MRI has been extensively used in neurodegenerative disorders, such as Alzheimer's disease (AD), frontal-temporal dementia (FTD), mild cognitive impairment (MCI), Parkinson's disease (PD), Huntington's disease (HD) and amyotrophic lateral sclerosis (ALS). MRI is important for monitoring the neurodegenerative components in other diseases such as epilepsy, stroke and multiple sclerosis (MS). Manganese enhanced MRI (MEMRI) has been used in many preclinical studies to image anatomy and cytoarchitecture, to obtain functional information in areas of the brain and to study neuronal connections. This is due to Mn2+ ability to enter excitable cells through voltage gated calcium channels and be actively transported in an anterograde manner along axons and across synapses. The broad range of information obtained from MEMRI has led to the use of Mn2+ in many animal models of neurodegeneration which has supplied important insight into brain degeneration in preclinical studies. Here we provide a brief review of MEMRI use in neurodegenerative diseases and in diseases with neurodegenerative components in animal studies and discuss the potential translation of MEMRI to clinical use in the future.


Promoting Professional Socialization Within the Experiential Curriculum: Implementation of a High-stakes Professionalism Rubric.
Objectives. To describe the implementation of a high-stakes rubric to assess student professionalism in introductory and advanced pharmacy practice experiences (IPPEs and APPEs) to promote the professional socialization of students in the doctor of pharmacy (PharmD) program at Western New England University (WNE). Findings. A professionalism rubric was adapted from the literature to assess the professional behavior of students enrolled in experiential courses based on evaluation of the following criteria: appropriate communication skills with patients and providers, appearance and dress code, timeliness, and initiative. The rubric was implemented in the fall semester of 2013 as a high-stakes component of the assessment within all experiential courses. Students were required to meet expectations for each of the four criteria in order to pass the practice experience, independent of their performance in other course components. Students were assessed by their preceptors at the midpoint and end of each practice experience using the appropriate evaluation tool. Each of the IPPE and APPE evaluation tools included the professionalism rubric as a requirement for assessment. Use of the Professionalism Rubric as a high-stakes assessment tool highlighted professionalism as an important component of the program, making expectations explicit to students and providing leverage to preceptors for holding students accountable. Summary. The Office of Experiential Affairs at WNE has raised awareness of the importance of professionalism and promoted the professional socialization of PharmD students with the use of a high-stakes professionalism rubric.


Influence of risk factors on coronary flow reserve in patients with 1-vessel coronary artery disease.
Coronary artery disease (CAD) risk factors influence the hyperemic response in patients without coronary artery stenosis. The aim of this study was to evaluate the influence of risk factors on coronary flow reserve (CFR) estimated by (99m)Tc-sestamibi cardiac imaging in patients with 1-vessel CAD. Forty-eight patients with 1-vessel CAD were enrolled in the study. Systemic hypertension, hypercholesterolemia, diabetes mellitus, and cigarette smoking were considered CAD risk factors. Within 48 h, all patients underwent coronary angiography and regional CFR assessment by (99m)Tc-sestamibi imaging with dipyridamole (0.74 mg/kg of body weight). Eight patients had no risk factors, 14 had 1 risk factor, 18 had 2 risk factors, and 8 had 3 risk factors. The mean +/- SD CFR of the narrowed coronary vessel was 1.28 +/- 0.43 in the overall study population (1.52 +/- 0.30 in patients with 50%-70% stenosis and 0.94 +/- 0.75 in patients with >70% stenosis; P < 0.001). Two-factor ANOVA showed that the number of risk factors significantly affected CFR (P < 0.001) in patients with 50%-70% stenosis, whereas significance was only borderline in patients with >70% stenosis. Finally, a significant interaction among the degree of coronary artery stenosis, the number of risk factors, and the estimated CFR was found (F(3) = 14.0; P < 0.001). CFR is inversely related to the number of conventional risk factors in patients with 1-vessel CAD and intermediate coronary artery stenosis, whereas this influence is less evident in patients with more severe stenosis.


Permeation and block of the skeletal muscle chloride channel, ClC-1, by foreign anions.
A distinctive feature of the voltage-dependent chloride channels ClC-0 (the Torpedo electroplaque chloride channel) and ClC-1 (the major skeletal muscle chloride channel) is that chloride acts as a ligand to its own channel, regulating channel opening and so controlling the permeation of its own species. We have now studied the permeation of a number of foreign anions through ClC-1 using voltage-clamp techniques on Xenopus oocytes and Sf9 cells expressing human (hClC-1) or rat (rClC-1) isoforms, respectively. From their effect on channel gating, the anions presented in this paper can be divided into three groups: impermeant or poorly permeant anions that can not replace Cl- as a channel opener and do not block the channel appreciably (glutamate, gluconate, HCO3-, BrO3-); impermeant anions that can open the channel and show significant block (methanesulfonate, cyclamate); and permeant anions that replace Cl- at the regulatory binding site but impair Cl- passage through the channel pore (Br-, NO3-, ClO3-, I-, ClO4-, SCN-). The permeability sequence for rClC-1, SCN- approximately ClO4- > Cl- > Br- > NO3- approximately ClO3- > I- >> BrO3- > HCO3- >> methanesulfonate approximately cyclamate approximately glutamate, was different from the sequence determined for blocking potency and ability to shift the Popen curve, SCN- approximately ClO4- > I- > NO3- approximately ClO3- approximately methanesulfonate > Br- > cyclamate > BrO3- > HCO3- > glutamate, implying that the regulatory binding site that opens the channel is different from the selectivity center and situated closer to the external side. Channel block by foreign anions is voltage dependent and can be entirely accounted for by reduction in single channel conductance. Minimum pore diameter was estimated to be approximately 4.5 A. Anomalous mole-fraction effects found for permeability ratios and conductance in mixtures of Cl- and SCN- or ClO4- suggest a multi-ion pore. Hydrophobic interactions with the wall of the channel pore may explain discrepancies between the measured permeabilities of some anions and their size.


Repair of persistent perineal sinuses by means of a pedicle flap of musculus gracilis. Case report.
Chronic suppurating perineal sinuses are common following proctocolectomy for ulcerative colitis and Crohn's disease. Repeated surgery along conservative lines is often likely to result in failure. If a tissue with normal potentiality for healing is brought to the area of delayed healing, a reasonable chance of success will be achieved. This hypothesis was put forward by Graham (1965) who supported his theory by presenting outstanding results by using a pedicle flap of gracilis in the treatment of rectovaginal fistulas following irradiation. In the present study, Graham's technique was adapted in the treatment of chronic persistent perineal sinuses. The results are promising and the procedure is worth trying in the treatment of longstanding sinuses which fail to heal by conservative methods.


Alpha-Band Frequency and Temporal Windows in Perception: A Review and Living Meta-analysis of 27 Experiments (and Counting).
Temporal windows in perception refer to windows of time within which distinct stimuli interact to influence perception. A simple example is two temporally proximal stimuli fusing into a single percept. It has long been hypothesized that the human alpha rhythm (an 8- to 13-Hz neural oscillation maximal over posterior cortex) is linked to temporal windows, with higher frequencies corresponding to shorter windows and finer-grained temporal resolution. This hypothesis has garnered support from studies demonstrating a correlation between individual differences in alpha-band frequency (IAF) and behavioral measures of temporal processing. However, nonsignificant effects have also been reported. Here, we review and meta-analyze 27 experiments correlating IAF with measures of visual and audiovisual temporal processing. Our results estimate the true correlation in the population to be between .39 and .53, a medium-to-large effect. The effect held when considering visual or audiovisual experiments separately, when examining different IAF estimation protocols (i.e., eyes open and eyes closed), and when using analysis choices that favor a null result. Our review shows that (1) effects have been internally and independently replicated, (2) several positive effects are based on larger sample sizes than the null effects, and (3) many reported null effects are actually in the direction predicted by the hypothesis. A free interactive web app was developed to allow users to replicate our meta-analysis and change or update the study selection at will, making this a "living" meta-analysis (randfxmeta.streamlit.app). We discuss possible factors underlying null reports, design recommendations, and open questions for future research.


In situ Metabolite Mass Spectrometry Imaging: New Insights into the Adrenal Gland.
The adrenal gland integrates catecholamine-producing neuroendocrine cells and steroid-producing cells with mesenchymal origin in a structured manner under one capsule and is a key regulator for vital bioactivity. In addition to adrenal-specific disease, dysregulation of adrenal hormones is associated with systemic effects, leading to undesirable metabolic and cardiovascular consequences. Mass spectrometry imaging (MSI) technique can simultaneously measure a broad range of biomolecules, including metabolites and hormones, which has enabled the study of tissue metabolic and hormone alterations in adrenal and adrenal-related diseases. Furthermore, this technique coupled with labeled immunohistochemistry staining has enabled the study of the pathophysiological adaptation of the adrenal gland under normal and abnormal conditions at different molecular levels. This review discusses the recent applications of in situ MSI in the adrenal gland. For example, the combination of formalin-fixed paraffin-embedded tissue microarray and MSI to tissues from patient cohorts has facilitated the discovery of clinically relevant prognostic biomolecules and generated promising hypotheses for new sights into physiology and pathophysiology of adrenal gland. MSI also has enabled the discovery of clinically significant tissue molecular (i. e., biomarker) and pathway changes in adrenal disease, particularly in adrenal tumors. In addition, MSI has advanced the ability to optimally identify and detect adrenal gland specific molecules. Thus, as a novel analytical methodology, MSI has provided unprecedented capabilities for in situ tissue study.


Novel alleles, hemizygosity and deletions at an Alu-repeat within the neurofibromatosis type 1 (NF1) gene.
Neurofibromatosis type 1 (NF1) (von Recklinghausen) is a common autosomal dominant disorder, characterised by the presence of peripheral neurofibromas, café-au-lait spots and Lisch nodules of the iris. Due to the high mutation rate at the NF1 locus, most patients are expected to have different mutations, limiting molecular analysis and genetic counseling to the identification of the mutation in each patient or family, or to the use of DNA polymorphisms. We have analysed an Alu-repeat polymorphic sequence (AAAT), located in intron 27 of the NF1 gene, in 70 NF1 and 40 CEPH families and we have detected several genetic and molecular abnormalities. In two families the NF1 individuals were hemizygous at the AAAT-repeat and/or at the CA-repeat of intron 27 of NF1, due to interstitial deletions, which include intron 27 to exon 37 of the NF1 gene. A 71-bp deletion at the Alu sequence was detected in non-NF1 chromosomes of members of three NF1 families. New alleles at the AAAT-repeat were found in one NF1 family and in three CEPH families giving a mutation rate for this AAAT-repeat of 0.36% per allele, which is one of the highest detected for a microsatellite locus.


Intravenous feeding of the rat with short chain fatty acid esters. I. Glycerol monobutyrate.
Intravenous nutrition was investigated using butyric acid because it is oxidized independent of carnitine transport into the cell mitochondria. This carnitine independent fatty acid, in the form of water soluble monobutyrin, was continuously infused into rats for seven days at 27 g monobutyrin per kilogram of body weight per day and provided half the daily energy requirement. All experimental animals survived the alimentation in good health and were free of detectable physiological and behavioral abnormalities. The intravenous infusion depressed the test animals spontaneous food intake to half their preinfusion level. These experimental rats still demonstrated continuous weight pain in contrast to weight loss by pair-fed controls. At decapitation, the monobutyrin infused rats had hepatic glycogen levels three times that of the controls, along with lower soluble hepatic protein, and normal lipid and water content. The plasma acetoacetate was also elevated in experimental rats. It was inferred from these results that monobutyrin was hydrolyzed, and the metabolites were oxidized by the rat. It is concluded from these observations that monobutyrin produces no obvious toxic affects during short infusion periods and provides calories for the rat when given intravenously.


A comparative study of cone-beam computed tomography and periapical radiographs in decision-making after endodontic instrument fractures.
This study aimed to compare the diagnostic efficacies of periapical radiographs (PRs) and cone-beam computed tomography (CBCT) scans on clinicians' decision-making in diagnosing and providing treatment of fractured instruments in root canals during glide path preparation. 40 maxillary molar teeth were selected and randomly divided into 4 groups. In each experimental group (n = 10), Hedstroem stainless steel instruments were fractured in mesiobuccal (MB) or distobuccal (DB) root canals. In group 1 (#0.06), group 2 (#0.08), group 3 (#0.10), and group 4 (#0.15), instruments were forced to fracture. PRs and CBCT scans were evaluated and compared. Observers were asked to use the following scoring system to decide whether to: 1. remove the fragment; 2. leave the fragment in situ; 3. bypass the fragment; or 4. use a surgical approach for removing the fragment. CBCT observers mostly decided on removing and bypassing the fractured fragment, while radiograph observers mostly decided to leave fragments in situ. However, there was no significant difference between these parameters when CBCT scans were compared to PRs for decision-making of the fractured instruments (p≥0.05) regarding different sizes of root canal instruments. In decision-making regarding fractured instruments during glide path preparation, clinicians should strategize treatment options based on variables using high-resolution images that reflect accurate scenarios through PRs or CBCT scans that also take into account low radiation dosage in situ PRs have precedence for a diagnostic approach.


Phorbol esters and cyclic AMP activate AMP deaminase in adult rat cardiac myocytes.
Using rapid deenergization as a probe for adenylate deaminase activity in intact adult rat cardiac myocytes, we have previously established that IMP formation is enhanced by alpha-adrenergic agonists. In the present study, the effect of adrenergic agents on adenylate deaminase was further characterized. Phenylephrine (PE)3 increased IMP production in a dose-dependent fashion with an EC50 of 8 x 10(-7) M. The response to PE was reversed within 10 min by the alpha 1-antagonist, prazosin. Likewise, adenylate deaminase was also activated in ventricular myocytes challenged with phorbol 12-myristate 13-acetate (PMA, EC50 = 5 nM); cardiac cells presented with 100 nM PMA increased IMP production from 4.4 +/- 0.5 (control) to 15.7 +/- 0.9 nmol/mg protein when subsequently deenergized. The effects of PMA and PE were attenuated 85 +/- 5% and 96 +/- 4%, respectively, by pretreatment of cells with 150 nM staurosporine, an inhibitor of protein kinase C. Furthermore, incubation of cardiac cells with 1 microM PMA for 24 h blunted the response to both PMA and phenylephrine 85-90%. Elevating cyclic AMP (cAMP) content to greater than 15 pmol/mg by treatment with forskolin or isoproterenol plus isobutylmethylxanthine also resulted in enhanced adenylate deaminase activity, but this stimulatory effect was not abolished by 24 h incubation with 5 microM PMA. Forskolin and PMA-induced increases in IMP production appeared to be additive. However, 0.5 microM isoproterenol inhibited the cellular response to phenylephrine by about 30% but did not affect PMA-stimulated adenylate deaminase activity. We conclude that both cAMP and protein kinase C stimulate adenylate deaminase, perhaps through selective activation of different isoforms. However, cAMP also exerts partial inhibition on alpha-adrenoreceptor-mediated increases in IMP production.


Management of Extraocular Infections.
Extraocular infection is a frequently encountered problem in children including neonates and infants. Prompt recognition of the extraocular disease and management at first contact reduces the ocular morbidity and visual impairment. Delay in administering appropriate therapy and referral to an eye care personnel may result in blindness; sometimes bilateral, due to severe ophthalmia neonatorum. This review is mainly aimed at offering basic, brief information about common extraocular infections prevalent among infants and children living in developing countries as a ready reference to practising pediatricians and neonatologists. Through this review, the pediatricians, as the first contact may feel confident and comfortable in managing ocular infection in pediatric population particularly living in remote rural and tribal region. Literature search was made by retrieving related publications through peer reviewed journals, PUBMED and Cochrane meta-analysis.


Evaluating the feasibility of the KDIGO CKD referral recommendations.
In 2012, the international nephrology organization Kidney Disease Improving Global Outcomes (KDIGO) released recommendations for nephrology referral for chronic kidney disease (CKD) patients. The feasibility of adhering to these recommendations is unknown. We conducted a retrospective analysis of the primary care population at Brigham and Women's Hospital (BWH). We translated referral recommendations based upon serum creatinine, estimated glomerular filtration rate (eGFR), and albuminuria into a set of computable criteria in order to project referral volume if the KDIGO referral recommendations were to be implemented. Using electronic health record data, we evaluated each patient using the computable criteria at the times that the patient made clinic visits in 2013. We then compared the projected referral volume with baseline nephrology clinic volume. Out of 56,461 primary care patients at BWH, we identified 5593 (9.9%) who had CKD based on albuminuria or estimated GFR. Referring patients identified by the computable criteria would have resulted in 2240 additional referrals to nephrology. In 2013, this would represent a 38.0% (2240/5892) increase in total nephrology patient volume and 67.3% (2240/3326) increase in new referral volume. This is the first study to examine the projected impact of implementing the 2012 KDIGO referral recommendations. Given the large increase in the number of referrals, this study is suggestive that implementing the KDIGO referral guidelines may not be feasible under current practice models due to a supply-demand mismatch. We need to consider new strategies on how to deliver optimal care to CKD patients using the available workforce in the U.S. health care system.


Treatment of Stress Urinary Incontinence in Neurological Patients With an Injectable Elastomer Prosthesis: Preliminary Results.
Many treatment options for stress urinary incontinence are difficult to apply to neurological patients. Urolastic is a new agent that is primarily indicated for women with mild stress urinary incontinence or men after prostate surgery. In this report, we present a series of 5 cases describing the first use of Urolastic to treat neurological patients. All patients were evaluated with a voiding diary and the use of auxiliary devices as the main indicators of continence. The median operative time was 30.8 minutes, and no complications were observed. Of the 5 patients, 4 reported improved incontinence: 2 switched from diapers to small pads, while the other 2 patients were able to discontinue urinary condom use. The only instance of treatment failure occurred in a patient with a low-compliance bladder. The advantages of this procedure appear to include a soft-cuff effect, reversibility, and minimal invasiveness. However, a future randomized study would be necessary to validate this treatment option.


Peripheral neuropathy associated with malignant neoplasms in dogs.
Common peroneal and ulnar nerves of 21 dogs between 2 and 15 years of age with malignant tumors were examined for possible paraneoplastic effects. Percent of abnormalities in single-teased fiber studies exceeded the confidence limits (P = 0.99) established for age-matched control dogs in 16 of 21 dogs (76%) with malignancies, none of which manifested clinical signs of polyneuropathy. The incidence of abnormalities was higher in common peroneal nerve (73%) than in ulnar nerve (57%). Lesions were characterized by a mixture of demyelination/remyelination and axonal degeneration; however, the former lesions predominated in most affected dogs. The severity of the neuropathy varied with different tumors, with the most severe lesions observed in a bronchogenic carcinoma (59%), a mammary adenocarcinoma (59%), a malignant melanoma (48%), and an insulinoma (47%).


A Qualitative Evaluation of Clinical Audit in UK Dental Foundation Training.
Clinical Audit (CA) has been recognized as a useful tool for tool for improving service delivery, clinical governance, and the education and performance of the dental team. This study develops the discussion by investigating its use as an educational tool within UK Dental Foundation Training (DFT). The aim was to investigate the views of Foundation Dentists (FDs) and Training Programme Directors (TPDs) on the CA module in their FD training schemes, to provide insight and recommendations for those supervising and undertaking CA. A literature review was conducted followed by a qualitative research methodology, using group interviews. The interviews were transcribed and thematically analyzed using NVIVO, a Computer-Assisted Qualitative Data Analysis tool. CA was found to be a useful tool for teaching management and professionalism and can bring some improvement to clinical practice, but TPDs have doubts about the long-term effects on service delivery. The role of the Educational Supervisor (ES) is discussed and recommendations are given for those supervising and conducting CA.


Optical edge detection with adjustable resolution based on cascaded Pancharatnam-Berry lenses.
We designed a versatile optical edge detection setup with two cascaded Pancharatnam-Berry lenses (PBLs) placed at the Fourier plane of a 4f system. When the two PBLs are parallel and close to each other, owing to the moiré-like effect, one-dimensional edge detection with adjustable resolution is achieved by introducing a transverse displacement of one PBL. Furthermore, two-dimensional edge detection with adjustable resolution can also be realized by tuning the longitudinal distance between the PBLs, and the transverse displacement is exploited to adjust the edge resolution in specified directions. The proposed scheme is verified by a proof-of-principle experiment in which the resolution-adjustable edges of different targets and cells were clearly observed, showing its flexibility and potential application in image processing and high-contrast microscopy.


Autocatalytic association of proteins by covalent bond formation: a Bio Molecular Welding toolbox derived from a bacterial adhesin.
Unusual intramolecular cross-links present in adhesins from Gram-positive bacteria have been used to develop a generic process amenable to biotechnology applications. Based on the crystal structure of RrgA, the Streptococcus pneumoniae pilus adhesin, we provide evidence that two engineered protein fragments retain their ability to associate covalently with high specificity, in vivo and in vitro, once isolated from the parent protein. We determined the optimal conditions for the assembly of the complex and we solved its crystal structure at 2 Å. Furthermore, we demonstrate biotechnological applications related to antibody production, nanoassembly and cell-surface labeling based on this process we named Bio Molecular Welding.


Forkhead box O3 (FoxO3) regulates kidney tubular autophagy following urinary tract obstruction.
Autophagy has been shown to be important for normal homeostasis and adaptation to stress in the kidney. Yet, the molecular mechanisms regulating renal epithelial autophagy are not fully understood. Here, we explored the role of the stress-responsive transcription factor forkhead box O3 (FoxO3) in mediating injury-induced proximal tubular autophagy in mice with unilateral ureteral obstruction (UUO). We show that following UUO, FoxO3 is activated and displays nuclear expression in the hypoxic proximal tubules exhibiting high levels of autophagy. Activation of FoxO3 by mutating phosphorylation sites to enhance its nuclear expression induces profound autophagy in cultured renal epithelial cells. Conversely, deleting FoxO3 in mice results in fewer numbers of autophagic cells in the proximal tubules and reduced ratio of the autophagy-related protein LC3-II/I in the kidney post-UUO. Interestingly, autophagic cells deficient in FoxO3 contain lower numbers of autophagic vesicles per cell. Analyses of individual cells treated with autophagic inhibitors to sequentially block the autophagic flux suggest that FoxO3 stimulates the formation of autophagosomes to increase autophagic capacity but has no significant effect on autophagosome-lysosome fusion or autolysosomal clearance. Furthermore, in kidneys with persistent UUO for 7 days, FoxO3 activation increases the abundance of mRNA and protein levels of the core autophagy-related (Atg) proteins including Ulk1, Beclin-1, Atg9A, Atg4B, and Bnip3, suggesting that FoxO3 may function to maintain components of the autophagic machinery that would otherwise be consumed during prolonged autophagy. Taken together, our findings indicate that FoxO3 activation can both induce and maintain autophagic activities in renal epithelial cells in response to injury from urinary tract obstruction.


Patient-Reported Outcomes of Medical Tattooing for Capillary Malformations.
Patients with capillary malformations (CMs) may undergo medical tattooing (MT) as an alternative to laser therapy. But little is known about treatment results and impact from the patients' perspective. In this cross-sectional digital survey study, we evaluated the patient-reported outcomes of MT for CMs. MT practices were identified via the Dutch Association of Skin Therapists and Google. These practices invited all their CM patients who had undergone MT between January 2011 and September 2021 to participate. Baseline and treatment characteristics, tattooing effectiveness, patient satisfaction with treatment outcomes, and complications were evaluated using a custom-made online survey. Quality of life was assessed with the Dermatology Life Quality Index (DLQI) questionnaire. Factors associated with treatment effectiveness and patient satisfaction were identified via bivariate analysis and ordinal logistic regression analysis. Most of the 89 respondents were female (69%). Almost all CMs were located on the face (90%) and mainly (dark) red (74%). Nearly all patients had undergone laser therapy (91%). Median number of tattooing sessions was 5 (IQR: 4.0-8.0). Thirty-seven percent of the patients perceived >75% color reduction. Younger patients were more likely to obtain lower treatment effectiveness (OR 0.44, 95% CI: 0.20-0.97). Most patients (83%) were satisfied with treatment results. Patients with lighter (OR 0.30, 95% CI: 0.13-0.72), non-facial (OR 0.15, 95% CI: 0.03-0.89), and hypertrophic CMs (OR 0.30, 95% CI: 0.11-0.82) were less likely to be satisfied with treatment outcomes. Patients with lighter skin types were more satisfied (OR 2.89, 95% CI: 1.23-6.80). Complications included transient pain (23%), bleeding (3.4%), hypertrophic scarring (1.1%), hypopigmentation (1.1%), and a halo around the tattoo (1.1%). MT seems a valid alternative treatment in addition to laser therapy for CMs, with mild complications. Most patients are (very) satisfied with treatment results, while color reduction is incomplete. Hence, it seems appropriate to decide together with patients whether or not to use MT as primary treatment or secondary to laser therapy.


Oral Care and Quality Evaluation: A Multicentric Study on Periodontal Treatment.
No information is available on the perception of the quality of care in patients treated for periodontitis. The purpose of this article was to assess how periodontitis-affected patients perceive the quality of periodontal treatment (PT) and to measure the factors which may influence it. 306 subjects who completed PT were invited to participate. Questionnaires and visual analogic scales (VAS) evaluating perception of quality of care, symptoms, and oral health related quality of life (OHRQoL) were handed out. Oral and periodontal indicators were collected before and after treatment. The impact of different factors on perception of quality was assessed with a regression model. Quality evaluation was high yet unrelated for both patients and clinicians (p = 0.983). Quality was negatively influenced by the number of residual oral infections (p < 0.001), patient's age (p = 0.07) and presence of residual pain at completion of PT (p = 0.02). Professionalism, kindness of the staff and communication skills were the characteristics mostly appreciated. The OHRQoL was influenced by the number of residual teeth (p < 0.001), increasing age of patients (p = 0.08), number of residual infections (p < 0.01) and pain (p = 0.04). Patients' quality perception appeared to be influenced by clinical and emotional aspects. Oral care providers should be aware of the impact of non-clinical factors in patients' appreciation of quality of treatment.


A double-blind, controlled comparison of piperacillin and cefoxitin in the prevention of postoperative infection in patients undergoing cesarean section.
Data are presented from a randomized, double-blind multicenter trial (six study sites, involving 346 patients) in which piperacillin was compared with cefoxitin as a prophylactic agent for patients undergoing cesarean section. One hundred and sixty-nine patients received piperacillin and 177 received cefoxitin; in each instance, the total dosage was 6 grams. Strict criteria were used to evaluate safety and efficacy. The courses of all of the patients were evaluable for safety and 183 courses were evaluable for efficacy (136 efficacy-evaluable courses were from patients treated with piperacillin and 147, from those treated with cefoxitin). Postoperative infection was prevented in 89 per cent of the patients treated with piperacillin and in 93 per cent of the those treated with cefoxitin. The difference was not statistically significant. The data from each of the individual study sites, as well as the pooled data, indicated that the short term perioperative administration of piperacillin in patients undergoing a cesarean section was as safe and effective as cefoxitin with regard to prophylactic response, duration of hospitalization and the usage of other systemic antibiotics.


Absolute thermodynamic properties of molten salts using the two-phase thermodynamic (2PT) superpositioning method.
We show that the absolute thermodynamic properties of molten salts (mixtures of KCl and LiCl) can be accurately determined from the two-phase thermodynamic (2PT) method that is based on superpositioning of solid-like and gas-like (hard-sphere) vibrational density of states (DoS). The 2PT predictions are in excellent accordance with those from the thermodynamic integration method; the melting point of KCl evaluated from the free energy and the absolute entropy shows close conformity with the experimental/NIST data. The DoS partitioning shows that the Li(+) ions in the eutectic LiCl-KCl molten mixture are largely solid-like, unlike the K(+) and Cl(-) ions, which have a significant gas-like contribution, for temperatures ranging from 773 K to 1300 K. The solid-like states of the Li(+) ions may have practical implications when employed for chemical and nuclear reprocessing applications.


Competition between crystallization and gelation: a local description.
We develop a model to describe the competition between gelation and crystallization in colloidal suspensions where particle interactions are represented by square well attractions. The competition is discussed locally in terms of the tendencies of individual particles to attain amorphous or crystalline configurations on cluster surfaces. These tendencies are dictated by three independent processes, the aggregation of particles onto, the dissociation of particles from, and the rearrangement of particles on the cluster surfaces. Models are developed to determine the rates of each of these processes. The relative magnitudes of these rates determine the probability that a particle arriving onto a cluster surface reaches a crystalline configuration, remains arrested in an amorphous configuration, or dissociates back into the suspension. These probabilities are employed to determine whether stable crystalline or amorphous clusters nucleate, resulting in predictions of the occurrence of crystallization or gelation as a function of solution conditions. Comparisons of model predictions with recent experiments on globular protein suspensions show excellent agreement, suggesting that the model captures much of the underlying physics of the competition between gelation and crystallization in attractive colloidal suspensions.


Sodium phenylbutyrate antagonizes prostate cancer through the induction of apoptosis and attenuation of cell viability and migration.
Prostate cancer (PCa) is a leading cause of cancer-related death in men. Sodium phenylbutyrate (SPB) has shown its potential as an anticancer therapy in numerous cancer types. In the present study, we attempted to assess the effect of SPB against PCa and whether this treatment was associated with the regulation of survivin. Two human PCa cancer cell lines, DU145 and PC3, were used in the present study. Cell Counting Kit-8 (CCK-8) assay was conducted to measure the proliferation of PCa cells incubated with SPB. The effect of SPB on the cell apoptosis, cell colony formation ability, and cell morphological change was also assessed. Transwell experiment and Western blotting assay were performed to determine the effect of SPB on the migration and invasion ability of both cell types. Moreover, the expression pattern of survivin and MAPK members in both cell types after the treatment of SPB was also detected. Additionally, an in vivo tumor formation assay was performed to evaluate the treatment potential of SPB against PCa. We found that the viability of PCa cells was significantly inhibited by SPB treatment. As illustrated by flow cytometry, for DU145 cell line the average apoptotic rate of SPB-treated cells was significantly lower than that of the control group (P<0.05); similar results were also seen for PC3 (P<0.05). SPB administration also attenuated the colony formation and migration abilities in both cell lines. The expression level of survivin in SPB-treated cells was significantly downregulated, while the phosphorylation of p-38 and ERK was enhanced. Furthermore, in vivo tumor formation of both cell lines was suppressed by SPB as well. The above results confirmed the potential of SPB as an effective therapeutic agent for the prevention or treatment of PCa. This amelioration might be due to the blockade of the survivin pathway.


Mechanisms of ischemic brain damage.
This article provides a brief review of recent developments regarding the pathophysiology of ischemic brain damage, and offers hypotheses explaining the pathogenesis of selective neuronal vulnerability and of tissue infarction, respectively. It is suggested that selective neuronal vulnerability, observed after brief periods of ischemia and after hypoglycemic coma, qualifies as an excitotoxic lesion, which causes postsynaptic damage to neurons innervated by excitatory amino acids by enhancing calcium influx. However, ischemic damage often involves glial and vascular cells as well, and causes infarction. It is hypothesized that this type of brain damage is related to acidosis and that enhanced acidosis is detrimental because it accelerates delocalization of protein-bound iron, with an ensuing free-radical damage to membrane lipids and proteins.


Genotypic analysis of invasive Streptococcus pneumoniae from Mali, Africa, by semiautomated repetitive-element PCR and pulsed-field gel electrophoresis.
As part of a large, ongoing study of invasive infections in pediatric patients in Bamako, Mali, 106 cases of invasive pneumococcal disease were identified from June 2002 to July 2003 (J. D. Campbell et al., Pediatr. Infect. Dis. J. 23:642-649, 2004). Of the 12 serotypes present, the majority of isolates were not contained in PCV7 (the 7-valent pneumococcal conjugate vaccine), including 1 isolate that was serotype 1, 12 isolates that were serotype 2, 58 isolates that were serotype 5, 7 isolates that were serotype 7F, and 1 isolate that was serotype 12F. To determine whether clonal dissemination of the predominant serotypes had taken place, genotyping was performed on 100 S. pneumoniae isolates by using two methods: pulsed-field gel electrophoresis (PFGE) of SmaI-digested genomic DNA, and the Bacterial Barcodes repetitive-element PCR (rep-PCR) method. Criteria for delineating rep-PCR genotypes were established such that isolates of different serotypes were generally not grouped together. The two methods were equally discriminatory within a given pneumococcal serotype. PFGE separated the isolates into 15 genotypes and 7 subtypes; rep-PCR separated isolates into 15 genotypes and 6 subtypes. Using either method, isolates within serotypes 2, 5, and 7 formed three large, separate clusters containing 1 genotype each. Both methods further distinguished related subtypes within serotypes 2 and 5. Interestingly, one of the PFGE subtypes of serotype 5 is indistinguishable from the Columbia(5)-19 clone circulating in Latin America since 1994. The data support that serotypes 2 and 5 were likely to be the result of dissemination of particular clones, some of which are responsible for invasive disease over a broad population range.


PICNIC Technology.
A key objective of the Professionals and Citizen Network for Integrated Care (PICNIC) project was to provide products for a European and potentially worldwide software market. The approach followed was through the delivery of a number of Open Source (OS) components, to be integrated into applications that deliver similar services across the participating regions, aiming at their exploitation by other regions and the industry. This chapter describes the technology developed during the lifecycle of the PICNIC project, focusing on the three core services of Clinical Messaging, Access to Patient Data, and Collaboration. For each service, the entire process of how to turn its functional specifications into reusable components and common data sets in order to support Information Technology (IT) services for the next generation of secure, user-friendly healthcare networks is presented by means of common documentation tools. Security and privacy issues are also addressed.


Serum hyaluronic acid and procollagen III, N-terminal propeptide levels are highly associated with disease severity and predict the progression of COVID-19.
The coronavirus disease 2019 (COVID-19) pandemic is a rapidly evolving global emergency and continuously poses a serious threat to public health, highlighting the urgent need of identifying biomarkers for disease severity and progression. In order to early identify severe and critical patients, we retrospectively analyze the clinical characteristics and risk indicators of severe disease in patients with corona virus disease 2019 (COVID-19). A total of 420 confirmed COVID-19 patients were included in the study. According to the "Diagnosis and Treatment of novel coronavirus Pneumonia (10th Edition)", the cases were divided into mild group (n = 243) and severe group (n =177). Laboratory parameters were analyzed in combination with clinical data. Male patients over 46 years who have smoking habits were more likely to suffer from severe COVID-19. Critically ill patients had lower lymphocyte counts and red blood cell counts, and higher white blood cell counts (P<0.05). Expectedly, serum inflammatory factors (NLR, PLR, LMR, CLR, PCT, CRP), coagulation markers (APTT, PT, TT, FIB, D-Dimer), Myocardial damage markers (hs-TNT, LDH) were significantly increased (P<0.05) in severe COVID-19 patients. Surprisedly, those patients showed obviously elevated levels of common tumor markers (ProGRP, CYFRA21-1, SCC, NSE) (P<0.05). In this case, the levels of tumor marker reflected more the condition of inflammation than the growth of tumor. More importantly, HA and PIIIN-P were highly associated with COVID-19 severity. The AUC of the ROC curve for the diagnosis of severe COVID-19 by HA and PIIIN-P was 0.826. Meanwhile, HA was positively correlated with myocardial damage markers (hs-TNT, LDH). PIIIN-P was positively correlated with myocardial damage markers (hs-TNT, LDH) and inflammatory factors (NLR, PLR, LMR, CLR, ProGRP, SCC, PCT, CRP). On the contrary, PIIIN-P was negatively correlated with pulmonary function indexes (oxygenation index and oxygen saturation of hemoglobin). HA and PIIIN-P are highly associated with disease severity and progression of COVID-19 and can be used as new markers for the prediction of severe COVID-19.


Editorial: Child psychology and psychiatry - using science to make a difference.
The Journal of Child Psychology and Psychiatry has, I think it is fair to say, a special place in the hearts of scientists and scientist-practitioners working broadly in the field of developmental psychopathology. How would you put into words what it is we all love about the journal? Answers on a postcard please! For me, in addition to the high quality of the science, there is something unique about JCPP's open-minded, eclectic yet rigorous and methodologically pluralistic style that makes it stand out from the rest.


Time-resolved voltage measurements of Z-pinch radiation sources with a vacuum voltmeter.
A vacuum-voltmeter (VVM) was fielded on the Saturn pulsed power generator during a series of argon gas-puff Z-pinch shots. Time-resolved voltage and separately measured load current are used to determine several dynamic properties as the load implodes, namely, the inductance, L(t), net energy coupled to the load, E(coupled)(t), and the load radius, r(t). The VVM is a two-stage voltage divider, designed to operate at voltages up to 2 MV. The VVM is presently being modified to operate at voltages up to 6 MV for eventual use on the Z generator.


Nanoextraction from a flow of a highly diluted solution for much-improved sensitivity in offline chemical detection and quantification.
Preconcentration of the target compound is a critical step that ensures the accuracy of the subsequent chemical analysis. In this work, we present a straightforward yet effective liquid-liquid extraction approach based on surface nanodroplets (i.e., nanoextraction) for offline analysis of highly diluted sample solutions. The extraction and sample collection were streamlined in a 3-m microcapillary tube. The concentration of the target analyte in surface nanodroplets was significantly increased compared to the concentration in the sample solution, reaching several orders of magnitude. A limit of detection (LOD) was decreased by a factor of ∼103 for an organic model compound in Fourier-transform infrared spectroscopy (FTIR) measurements and ∼105 for a model fluorescent dye in fluorescence detection. The quantitative analysis of the organic compound was also achieved in a wide concentration region from 10-3 M to 10-4 M. The total volume of surface nanodroplets can be manipulated to further enhance extraction efficiency, according to the principle that governs droplet formation by solvent exchange. Additionally, our method exhibited significantly improved sensitivity compared to traditional dispersive liquid-liquid microextraction (DLLME). The LOD of the fluorescent dye and the organic model compound obtained with DLLME was 3 orders of magnitude and 20 times higher than the LOD achieved through nanoextraction approach. The nanoextraction developed in this work can be applied to preconcentrate multi-compounds from river water samples, without clear interference from each other. This can further extend its applicability for the detection and quantification of target analytes in complex aqueous samples by common analytical instruments.


A Novel Joint Power and Feedback Bit Allocation Interference Alignment Scheme for Wireless Sensor Networks.
It is necessary to improve the energy efficiency of batteries in wireless sensor networks (WSNs). The multiple-input multiple-output (MIMO) technique has become an important means to ameliorate WSNs, and interference management is the core of improving energy efficiency. A promising approach is interference alignment (IA), which effectively reduces the interference and improves the throughput of a system in the MIMO interference channels. However, the IA scheme requires perfect channel state information (CSI) at all transceivers in practice, which results in considerable feedback overhead. Thus, limited IA feedback has attracted much attention. In this paper, we analyze the throughput loss of the K-user MIMO interference channels when each transmitter delivers multiple streams in one slot, and derives the upper-bound of the system interference leakage and throughput loss. Then, to reduce the interference leakage and throughput loss for the MIMO interference alignment with limited feedback, a joint power and feedback bit allocation optimization scheme is proposed. The simulation results show that, compared with the conventional schemes, the presented optimal scheme achieves less residual interference and better performance in the system throughput.


RhoJ integrates attractive and repulsive cues in directional migration of endothelial cells.
During angiogenesis, VEGF acts as an attractive cue for endothelial cells (ECs), while Sema3E mediates repulsive cues. Here, we show that the small GTPase RhoJ integrates these opposing signals in directional EC migration. In the GTP-bound state, RhoJ interacts with the cytoplasmic domain of PlexinD1. Upon Sema3E stimulation, RhoJ released from PlexinD1 induces cell contraction. PlexinD1-bound RhoJ further facilitates Sema3E-induced PlexinD1-VEGFR2 association, VEGFR2 transphosphorylation at Y1214, and p38 MAPK activation, leading to reverse EC migration. Upon VEGF stimulation, RhoJ is required for the formation of the holoreceptor complex comprising VEGFR2, PlexinD1, and neuropilin-1, thereby preventing degradation of internalized VEGFR2, prolonging downstream signal transductions via PLCγ, Erk, and Akt, and promoting forward EC migration. After conversion to the GDP-bound state, RhoJ shifts from PlexinD1 to VEGFR2, which then terminates the VEGFR2 signals. RhoJ deficiency in ECs efficiently suppressed aberrant angiogenesis in ischemic retina. These findings suggest that distinct Rho GTPases may act as context-dependent integrators of chemotactic cues in directional cell migration and may serve as candidate therapeutic targets to manipulate cell motility in disease or tissue regeneration.


Targeting a cryptic allosteric site for selective inhibition of the oncogenic protein tyrosine phosphatase Shp2.
Protein tyrosine phosphatases (PTPs) have been the subject of considerable pharmaceutical-design efforts because of the ubiquitous connections between misregulation of PTP activity and human disease. PTP-inhibitor discovery has been hampered, however, by the difficulty in identifying cell-permeable compounds that can selectively target PTP active sites, and no PTP inhibitors have progressed to the clinic. The identification of allosteric sites on target PTPs therefore represents a potentially attractive solution to the druggability problem of PTPs. Here we report that the oncogenic PTP Shp2 contains an allosteric-inhibition site that renders the enzyme sensitive to potent and selective inhibition by cell-permeable biarsenical compounds. Because Shp2 contains no canonical tetracysteine biarsenical-binding motif, the enzyme's inhibitor-binding site is not readily predictable from its primary or three-dimensional structure. Intriguingly, however, Shp2's PTP domain does contain a cysteine residue (C333) at a position that is removed from the active site and is occupied by proline in other classical PTPs. We show that Shp2's unusual cysteine residue constitutes part of a Shp2-specific allosteric-inhibition site, and that Shp2's sensitivity to biarsenicals is dependent on the presence of the naturally occurring C333. The determinative role of this residue in conferring inhibitor sensitivity is surprising because C333's side chain is inaccessible to solvent in Shp2 crystal structures. The discovery of this cryptic Shp2 allosteric site may provide a means for targeting Shp2 activity with high specificity and suggests that buried-yet-targetable allosteric sites could be similarly uncovered in other protein families.


Nonbacterial thrombotic endocarditis and nondisseminated malignancy associated with osteopetrosis.
Nonbacterial thrombotic endocarditis (NBTE) is a presentation of an occult malignancy. The clinical, laboratory and postmortem findings of a 44-year-old man with osteopetrosis and NBTE are reported. A small apical bronchogenic carcinoma was found on postmortem examination. A thorough search for malignancy should be made in patients with unexplained cerebral emboli, especially if hematologic abnormalities are present. There is presently no evidence that anticoagulation or other treatment is beneficial.


Extending the coordination capabilities of tertiary phosphines and arsines: preparation, molecular structure, and reactivity of dinuclear rhodium complexes with PR3 and AsR3 in a doubly bridging coordination mode.
The reactions of [Rh2(kappa2-acac)2(mu-CPh2)2(mu-PR3)] (PR3= PMe34, PMe2Ph 7, PEt38) with an equimolar amount of Me3SiX (X = Cl, Br, I) afforded the unsymmetrical complexes [Rh2X(kappa2-acac)(mu-CPh2)2(mu-PR3)]5, 9-12, which contain the phosphine in a semi-bridging coordination mode. From 4 and excess Me3SiCl, the tetranuclear complex [[Rh2Cl(mu-Cl)(mu-CPh2)2(mu-PMe3)]2]6 was obtained. In contrast, the reaction of 4 with an excess of Me3SiX (X = Br, I) yielded the dinuclear complexes [Rh2X2(mu-CPh2)2(mu-PMe3)]13, 14 in which, as shown by the X-ray crystal structure analysis of 14, the bridging phosphine is coordinated in a truly symmetrical bonding mode. While related compounds with PEt3 and PMe2Ph as bridging ligands were prepared on a similar route, the complex [Rh2Cl2(mu-CPh2)2(mu-PiPr3)]19 was obtained from the mixed-valence species [(PiPr3)Rh(mu-CPh2)2Rh(kappa2-acac)2]17 and HCl. The reaction of [Rh2(kappa2-acac)2(mu-CPh2)2(mu-SbiPr3)]3 with AsMe3 gave the related Rh(mu-AsMe3)Rh compound 21. With Me3SiCl, the acac ligands of 21 can be replaced stepwise by chloride to give [Rh2Cl(kappa2-acac)(mu-CPh2)2(mu-AsMe3)]23 and [[Rh2Cl(mu-Cl)(mu-CPh2)2(mu-AsMe3)]2]24, the latter being isomorphous to the phosphine-bridged dimer 6.


Stretching and solvency of charged cellulose chains.
The transition between good and poor solvency conditions was studied for cellulose interacting in an aqueous environment as a function of the pH and ionic strength as well as for cellulose with weakly ionizable grafted groups. For uncharged cellulose, poor solvent conditions were observed for a pH range of 4-10 and an ionic strength between 0.1 and 10 mM, as demonstrated by the characteristic constant force plateau in the single-molecule force spectroscopy data as a function of the polymer extension. The magnitude of the force plateaus was quantized, indicating that on occasion more than a single cellulose chain was stretched into solution. Charged cellulose was also studied with a transition from poor to good solvency observed by an increase in the pH of the solution well above the pK(a) of the carboxyl groups. This transition was characterized by a fundamental change in the form of the force data as a function of extension from the constant force plateau (poor solvent conditions) to the nonlinear increase in adhesion due to elastic stretching (good solvent conditions). Using this single-molecule technique, the Kuhn length of cellulose was determined to be 3.8 A from the fit to a freely jointed chain model. Furthermore, the loop distribution under the good solvent conditions was studied as a function of the pH and ionic strength. The distribution became more compact as the influence of the charged groups was reduced through an increase in the screening or a reduction in the charge density, with chain association dominating because of strong hydrogen bonding.


Microwave-mediated reduction of disulfide bridges with supported (tris(2-carboxyethyl)phosphine) as resin-bound reducing agent.
We report on the synthesis and use of a new supported reagent consisting in tris(2-carboxyethyl)phosphine (TCEP) immobilized on hydrophilic PEG based resin beads. Used in conjunction with a 5 min microwave (MW) irradiation, "supported TCEP" reduced disulfide bridges in free thiols in peptides having two or more cysteine residues. Separation of reaction products from reducing agent was easily performed by simple filtration.


Contrast enhancement versus vasculature of uterine tumors.
The relative contrast enhancement of normal myometrium and small or medium size uterine fibroids and clinical stage I endometrial carcinomas were correlated to their microangiographically demonstrable vascularity. In four cases small uterine fibroids showed contrast enhancement equal to normal myometrium, independent of their vascularity in microangiography. In five, endometrial carcinomas contrast enhancement was always below that of the surrounding myometrium in CT. Intratumour vasculature showed great variability in microangiography. No correlation between the topographic variations in the arrangement or density of tumour blood vessels and the degree of contrast enhancement was found. Contrast enhancement of small uterine fibroids was greater than that of small adenocarcinomas. More contrast material accumulates tissue of the fibroid than into the extravascular space of endometrial carcinoma.


Amodiaquine and Ciprofloxacin Combination in Plasmodiasis Therapy.
Objective. The study was designed to determine the efficacy of combined Amodiaquine and Ciprofloxacin in plasmodiasis therapy. Method. The in vivo antiplasmodial effect of different dosage levels of Amodiaquine, Ciprofloxacin, and their combinations against Plasmodium berghei berghei was evaluated using Swiss albino mice. Results. Amodiaquine (a known antiplasmodial agent) had a fairly significant antiplasmodial effect reducing the parasites for every 100 red blood cells (RBC) from 66 to 16 (75.75%) at the tolerable dosage level of 7.5 mg/kg body weight while Ciprofloxacin (an antibiotic known to have antimalarial effect) showed an insignificant antiplasmodial effect reducing the parasites for every 100 RBC from 65 to 64 (1.53%) at the tolerable dosage level of 10.7 mg/kg body weight. Conversely, the combination therapy of Amodiaquine and Ciprofloxacin had a significant antiplasmodial effect at all the doses administered. The combination of 7.5 mg/kg of Amodiaquine and 12.8 mg/kg of Ciprofloxacin, however, showed the most significant antiplasmodial effect of the doses used reducing the number of parasites per 100 RBC from 60 to 10 (83.33%). Conclusions. Appropriate Amodiaquine and Ciprofloxacin combination will be effective for the treatment of malaria and better than either Amodiaquine or Ciprofloxacin singly at their recommended dosage levels.


Using the child behaviour checklist in ordinary primary schools.
The study considers the application of the Child Behaviour Checklist (Achenbach & Edelbrock, 1986) to the identification of children with emotional and behavioural difficulties in ordinary primary schools. Teachers provided checklist data on a sample of 320 boys and 118 girls previously referred to the schools psychological services and a further 183 boys and 39 girls who had not been referred. There was a high level of agreement between referred and non-referred student status and subsequent classification using the CBCL criteria for psychiatric disturbance. An analysis of variance was carried out to determine how far teacher expectations may contribute to CBCL scores. The results indicated that teacher differences within schools are more important than the differences which exist between teachers working in different schools. The results support the validity of the CBCL. The future application of the CBCL in ordinary schools is discussed.


Short-term dietary choline supplementation alters the gut microbiota and liver metabolism of finishing pigs.
Choline is an essential nutrient for pig development and plays a role in the animal's growth performance, carcass characteristics, and reproduction aspects in weaned pigs and sows. However, the effect of choline on finishing pigs and its potential regulatory mechanism remains unclear. Here, we feed finishing pigs with 1% of the hydrochloride salt of choline, such as choline chloride (CHC), under a basic diet condition for a short period of time (14 days). A 14-day supplementation of CHC significantly increased final weight and carcass weight while having no effect on carcass length, average backfat, or eye muscle area compared with control pigs. Mechanically, CHC resulted in a significant alteration of gut microbiota composition in finishing pigs and a remarkably increased relative abundance of bacteria contributing to growth performance and health, including Prevotella, Ruminococcaceae, and Eubacterium. In addition, untargeted metabolomics analysis identified 84 differently abundant metabolites in the liver between CHC pigs and control pigs, of which most metabolites were mainly enriched in signaling pathways related to the improvement of growth, development, and health. Notably, there was no significant difference in the ability of oxidative stress resistance between the two groups, although increased bacteria and metabolites keeping balance in reactive oxygen species showed in finishing pigs after CHC supplementation. Taken together, our results suggest that a short-term supplementation of CHC contributes to increased body weight gain and carcass weight of finishing pigs, which may be involved in the regulation of gut microbiota and alterations of liver metabolism, providing new insights into the potential of choline-mediated gut microbiota/metabolites in improving growth performance, carcass characteristics, and health.


The adherence paradox: guideline deviations contribute to the increased 5-year survival of breast cancer patients.
In German breast cancer care, the S1-guidelines of the 1990s were substituted by national S3-guidelines in 2003. The application of guidelines became mandatory for certified breast cancer centers. The aim of the study was to assess guideline adherence according to time intervals and its impact on survival. Women with primary breast cancer treated in three rural hospitals of one German geographical district were included. A cohort study design encompassed women from 1996-97 (N = 389) and from 2003-04 (N = 488). Quality indicators were defined along inpatient therapy sequences for each time interval and distinguished as guideline-adherent and guideline-divergent medical decisions. Based on all of the quality indicators, a binary overall adherence index was defined and served as a group indicator in multivariate Cox-regression models. A corrected group analysis estimated adjusted 5-year survival curves. From a total of 877 patients, 743 (85 %) and 504 (58 %) were included to assess 104 developed quality indicators and the resuming binary overall adherence index. The latter significantly increased from 13-15 % (1996-97) up to 33-35 % (2003-04). Within each time interval, no significant survival differences of guideline-adherent and -divergent treated patients were detected. Across time intervals and within the group of guideline-adherent treated patients only, survival increased but did not significantly differ between time intervals. Across time intervals and within the group of guideline-divergent treated patients only, survival increased and significantly differed between time intervals. Infrastructural efforts contributed to the increase of process quality of the examined certified breast cancer center. Paradoxically, a systematic impact on 5-year survival has been observed for patients treated divergently from the guideline recommendations. This is an indicator for the appropriate application of guidelines. A maximization of guideline-based decisions instead of the ubiquitous demand of guideline adherence maximization is advocated.


Computer-guided implant insertion in a patient with impacted maxillary canines: case report.
The use of minimally invasive surgical techniques undoubtedly represents a huge advantage for both the clinician and the patient. This case report shows the possibility of making an implant-prosthetic rehabilitation in an upper jaw in a patient who presented dental elements 1.3 and 2.3 in bone inclusion. The use of computer-guided surgery offers us the possibility to carry out this type of implant rehabilitation without performing the extraction of the above-mentioned dental elements combined with the bone regeneration of the extraction sockets. Consequently, surgery invasiveness and post-surgery complications are reduced to a minimum. Additionally, the predictability of the methodology is evaluated with specific software that enables a comparison between what is virtually planned and what occurs in clinical practice.


A Simple Outline of Methods for Protein Isolation and Purification.
At the summer workshop of the Korean Endocrine Society held in 2016, some examples of protein experiments were discussed in the session entitled "All about working with proteins." In contrast to what the title suggested, it was unrealistic to comprehensively discuss all protein analytical methods. Therefore, the goal was to outline protein experimental techniques that are useful in research or in bench work. In conversations with clinicians, however, I have always felt that researchers who do not engage in bench science have different demands than those who do. Protein research tools that are useful in bench science may not be very useful or effective in the diagnostic field. In this paper, I provide a general summary of the protein analytical methods that are used in basic scientific research, and describe how they can be applied in the diagnostic field.


RNA m6A methylation regulates the ultraviolet-induced DNA damage response.
Cell proliferation and survival require the faithful maintenance and propagation of genetic information, which are threatened by the ubiquitous sources of DNA damage present intracellularly and in the external environment. A system of DNA repair, called the DNA damage response, detects and repairs damaged DNA and prevents cell division until the repair is complete. Here we report that methylation at the 6 position of adenosine (m6A) in RNA is rapidly (within 2 min) and transiently induced at DNA damage sites in response to ultraviolet irradiation. This modification occurs on numerous poly(A)+ transcripts and is regulated by the methyltransferase METTL3 (methyltransferase-like 3) and the demethylase FTO (fat mass and obesity-associated protein). In the absence of METTL3 catalytic activity, cells showed delayed repair of ultraviolet-induced cyclobutane pyrimidine adducts and elevated sensitivity to ultraviolet, demonstrating the importance of m6A in the ultraviolet-responsive DNA damage response. Multiple DNA polymerases are involved in the ultraviolet response, some of which resynthesize DNA after the lesion has been excised by the nucleotide excision repair pathway, while others participate in trans-lesion synthesis to allow replication past damaged lesions in S phase. DNA polymerase κ (Pol κ), which has been implicated in both nucleotide excision repair and trans-lesion synthesis, required the catalytic activity of METTL3 for immediate localization to ultraviolet-induced DNA damage sites. Importantly, Pol κ overexpression qualitatively suppressed the cyclobutane pyrimidine removal defect associated with METTL3 loss. Thus, we have uncovered a novel function for RNA m6A modification in the ultraviolet-induced DNA damage response, and our findings collectively support a model in which m6A RNA serves as a beacon for the selective, rapid recruitment of Pol κ to damage sites to facilitate repair and cell survival.


The treatment of refractory anaemia of chronic renal failure with cobalt chloride.
Twelve anephric patients on maintenance haemodialysis received treatment with oral enteric coated cobalt chloride 25 to 50 mg daily. The change in haemoglobin concentration, and packed cell volume, are recorded and discussed with special reference to possible toxicity and mechanism of action of cobalt. Six of eight patients who completed the first course with cobalt chloride 50 mg daily for 12 weeks showed a significant rise in haemoglobin concentration of 26 to 70 per cent and a fall to near pre-therapeutic levels when cobalt was withdrawn. Evidence of a response was present within two months of starting treatment. Four patients showed a diminution in their blood transfusion requirements and three patients experienced a definite sense of increased well-being during treatment. One patient suffered from side effects of the drug and failed to complete the study because of gastrointestinal disturbance. The improvment in haemoglobin concentration was reproducible in four patients who were given second, and in one case third courses courses with varying doses of cobalt over differing periods of time. Serum cobalt levels tended to stabilize after two months continuous treatment to the therapeutic range of 40-100 mumg per 100 ml. A rapid fall in serum cobalt was seen on cessation of treatment. It is suggested that therapy with enteric coated cobalt chloride at a dose of between 25 and 50 mg per day has a definite place in the treatment of the refractory anaemia of chronic renal failure.


[Preservation of women fertility facing potential premature ovarian failure].
Numerous pathologies and their treatments may alter ovarian reserve in women and girls and may harm their future fertility. Oocytes cryopreservation techniques may be used in a limited number of cases, excluding young patients. We hereby report results obtained with our and other teams' ovarian tissue vitrification techniques. This new approach can be offered to young patients before puberty and should not delay both chemotherapy and radiotherapy when needed. Major drawbacks include potential alteration of ovarian reserve, as well as recurrence of the original malignancy. The later can be circumvented using in vitro maturation techniques. It is important to discuss fertility preservation opportunities with every woman or girls facing potential premature ovarian failure.


Effect of dutasteride on castration-resistant prostate cancer.
It has previously been demonstrated that the intratumoral generation of the potent androgen dihydrotestosterone (DHT), contributes critically to the progression of prostate cancer and its castration-resistant form, castration-resistant prostate cancer (CRPC). Circulating testosterone is converted into DHT by 5α-reductase (SRD5A). Dutasteride is a dual inhibitor of type I and II SRD5A. The present study assessed the effectiveness of dutasteride in the treatment of CRPC. Between 2010 and 2013, CRPC was diagnosed in 41 patients at the Tokyo Metropolitan Tama Medical Center. Following diagnosis, the patients received 0.5 mg dutasteride daily. The patients' median age was 77.3 years (range, 63-90). Bone metastases were recognized in 12 patients. All the patients received dexamethasone. Twenty-four (59%) patients had previously undergone chemotherapy, while 11 (27%) received docetaxel, and 24 (59%) estramustine. The prostatic-specific antigen (PSA) level declined in 17 (41%) patients from the baseline value, following dutasteride treatment. The median value for the PSA decrease was 23% (range, 4.3-89.8%), and the median duration of the response was 4 months (range, 1-10). The PSA response rate (defined as >50% decline in PSA from the baseline value) was recognized in 7 (17%) patients. The median duration of the response was 3 months (range, 2-10). Dutasteride was efficacious against CRPC in certain patients and may be a promising option in CRPC treatment. A prospective randomized trial is necessary to verify the efficacy of dutasteride.


CAV1 regulates primordial follicle formation via the Notch2 signalling pathway and is associated with premature ovarian insufficiency in humans.
What is the function of CAV1 in folliculogenesis and female reproduction? CAV1 regulates germline cyst breakdown and primordial follicle (PF) formation in mice, and CAV1 mutation may be related to premature ovarian insufficiency (POI). Pre-granulosa cells are essential for the establishment of the PF pool, which determines female fertility and reproductive lifespan. Cav1 participates in vascularization in fetal mouse ovaries. However, the role of CAV1 in early folliculogenesis and POI pathogenesis remains unclear. Cav1 function was investigated in mice and Human Embryonic Kidney 293 cells. Ovaries (six per group) were randomly assigned to Cav1-vivo-morpholino, control and control-morpholino groups, and all experiments were repeated at least three times. To investigate CAV1 mutations in women, 200 Chinese women with POI and 200 control individuals with regular menstrual cycles and normal endocrine profiles were recruited from the Center for Reproductive Medicine of Shandong University between September 2012 and December 2013. Wild-type CD1 mice, Lgr5-EGFP-ires-CreERT2 (Lgr5-KI) reporter mice and Human Embryonic Kidney 293 cells were used for these experiments. Protein expression was detected by Western blot, and quantitative RT-PCR was used to detect gene expression. The expression pattern of CAV1 in mouse ovaries and the phenotype of Cav1 deficiency in mice were detected by immunofluorescence. Pre-granulosa cell proliferation in ovaries was detected by bromodeoxyuridine (BrdU) assay and immunofluorescence. The coding region of the CAV1 gene was sequenced in 200 women with POI and 200 controls. The functional effect of the novel mutation c.142 G > C (p.Glu48Gln) was investigated by Cell Counting Kit-8 (CCK8) assays and Western blot. We confirmed that Cav1 deficiency in mouse ovary induced by CAV1-vivo-morpholino resulted in more multi-oocyte follicles than in the control and control-morpholino groups (P < 0.01). Suppression of Cav1 decreased Leucine rich repeat containing G protein coupled receptor 5 (Lgr5)-positive cell proliferation (P < 0.01) and reduced the number of Lgr5 and Forkhead box L2 (Foxl2) double-positive cells (P < 0.01). Furthermore, suppression of Cav1 inhibited ovarian epithelial Lgr5-positive cell proliferation and differentiation through the Notch2 signalling pathway. Two of the POI women carried novel CAV1 mutations (c.45 C > G synonymous and c.142 G > C [Glu48Gln]). The deleterious effect of p.Glu48Gln was corroborated by showing that it adversely affected the function of CAV1 in cell proliferation and NOTCH2 expression in HEK293FT cells. N/A. The novel Glu48Gln mutation was only detected in one of 200 POI patients and we were unable to investigate its effects in the ovary. The identification of CAV1 as a potentially causative gene for POI provides a theoretical basis to devise treatments for POI in women. This work was supported by the National Basic Research Program of China (973 Programs: 2012CB944700; 2013CB945501; 2013CB911400; 2014CB943202), the National Key Research and Development Program of China (2016YFC1000604, 2017YFC1001301), the State Key Program of National Natural Science Foundation of China (81430029), and the National Natural Science Foundation of China (31571540, 81522018, 81471509, 81601245, 81701406, 81571406). The authors declare no competing financial interests.


Amacrine cells in scotopic vision.
Signals from rod bipolar cells of cat retina are processed by a variety of rod amacrine cells before finally arriving at ganglion cells. Three of these rod amacrine cells (AII, A13, and A17 ) have been studied at the physiological and anatomical levels; the results suggest that each carries out a unique visual function: AII cells appear to quicken the response time of the rod system in the mid-scotopic range, while A17 cells may increase the light-gathering area of rod bipolars near visual threshold. Stimulation of A13 cells may disinhibit ganglion cells, thus heightening their responsiveness at low levels of illumination.


Low Dose Spironolactone Monotherapy in the Management of Stage I Essential Hypertension: A Pilot Randomized, Double-Blind, Placebo-Controlled Trial.
High blood pressure (BP) is a common chronic disease needs long life drug consumption to control. Spironolactone could be used as the fourth-line therapy in patients with resistant hypertension. However, there is no study to determine the effects of low dose spironolactone as a first line therapy in treatment of essential hypertension. The aim of this study is to investigate the effect of low dose spironolactone monotherapy in management of essential hypertension. In this double blind randomized clinical trial, 40 patients who had stage I essential hypertension were randomly divided into two groups: intervention group received spironolactone 25 milligram once daily for one month and control group received placebo once daily. At the baseline and after one month, 24-hour BP holter-monitoring and serum potassium assay were done. Systolic BP was reduced from 143.5 ± 8.2 mmHg to 137.10 ± 7.57 mmHg in the intervention group, while it did not change significantly in control (between group treatment difference = -4.5 mmHg, p = 0.004). There was no significant reduction of diastolic BP in the intervention group in comparison to placebo group (between group treatment difference = -1.3 mmHg, p = 0.099). Short course monotherapy with low dose spironolactone is effective in reducing systolic BP in patients with stage I essential hypertension.


Effect of recombinant human alpha interferon (INF) and antilymphocyte globulin (ALG) on normal and aplastic anaemia haematopoietic progenitor cell growth.
The effect of recombinant alpha interferon (INF) and of antilymphocyte globulin (ALG) to the colony stimulating factor (CSF) production was examined with in vitro culture of the bone marrow of healthy and of aplastic anaemia (AA) persons. In healthy persons the supernatant of lymphocytes preincubated with PHA and ALG was found to show a stimulating effect to clonogenic properties of marrow progenitors, the mentioned effect being not in proportion to the concentration value. Similar properties are shown by interferon in these persons. In patients with aplastic anaemia, a considerable stimulating ALG effect to the granulocytic formation of colonies and a lesser stimulating effect of interferon were shown.


[Long-term outcome of incontinence and prolapse surgery at the Heidelberg University Gynecologic Clinic 1980-1992].
Long-term results after different types of operations for urinary stress incontinence (minimum follow-up: 18 months) as well as multiple risk factors for the pelvic floor were analysed in a retrospective study. Between 1980 and 1992 1283 patients underwent surgery because of urinary stress incontinence at the University Women's Hospital in Heidelberg. The data of 478 patients, 430 of these after primary and 48 after recurrent surgery, were evaluated by questionnaires with regard to the long-term-results. 57% of patients after primary surgical therapy and 37% after recurrent surgery were cured for longer than 5 years or since the operation. A cure or improvement of the incontinence could be observed in 80% after primary and in 73% after recurrent surgery. Among the vaginal approaches for primary surgery the hysterectomy combined with colporrhaphy was most successful (60% cured or more than 5 years continent, 80.5% at least improved). The Burch colposuspension revealed even better results among the abdominal approaches (64% cured or longer than 5 years continent, 86% at least improved) compared to the Marshall-Marchetti-Krantz procedure with a cure rate of 33%. For therapy of the recurrent urinary incontinence the abdominal Burch colposuspension showed the best results with cure rates of 50% and cure or improvement in 75%. Therefore the abdominal approach seems to be superior to vaginal techniques such as sling operations (33% cure rate, 67% at least improved) or only re-colporrhaphy (27% cure rate, 78% at least improved). For primary incontinence the hysterectomy with vaginal repair or the Burch colposuspension have proved to be most successful. For recurrent urinary incontinence the abdominal colposuspension (Burch procedure) seems to be superior to other approaches.


Transfer factor II: results of therapy.
Transfer factor is a dialyzable extract of sensitized leukocytes, which transfers reactivity from skin test-positive donors to skin test-negative recipients. Transfer factor supplied by our laboratory has been used therapeutically to induce cellular immunity in 78 patients around the world. Many patients received multiple doses of transfer factor ranging from 1 unit given every 6 months for 3 years to 1 unit every week for 6 months to as much as 8 units per week for a brief period. A total of 299 units of transfer factor have been given. Diseases in which transfer factor appeared to cause improvement include the Wiskott-Aldrich syndrome, severe combined immunodeficiency disease, mucocutaneous candidiasis, chronic active hepatitis, coccidioidmycosis, dysgammaglobulinemia, Behcet disease, aphthous stomatitis, linear morphea, familial keratoacanthoma and malignancy.